VYSOKÉ UČENÍ TECHNICKÉ V BRNĚ - COREcore.ac.uk/download/pdf/30295600.pdf · vysokÉ uČenÍ technickÉ v brnĚ brno university of technology fakulta chemickÁ Ústav chemie potravin

VYSOKEacute UČENIacute TECHNICKEacute V BRNĚBRNO UNIVERSITY OF TECHNOLOGY

FAKULTA CHEMICKAacuteUacuteSTAV CHEMIE POTRAVIN A BIOTECHNOLOGIIacute

FACULTY OF CHEMISTRYINSTITUTE OF FOOD SCIENCE AND BIOTECHNOLOGY

ZMĚNY PROTEINOVEacuteHO PROFILU V PRŮBĚHU SLADOVAacuteNIacute

JEČMENE

DIPLOMOVAacute PRAacuteCEDIPLOMA THESIS

AUTOR PRAacuteCE MARTINA ŠOPIacuteKOVAacuteAUTHOR

BRNO 2008




ZMĚNY PROTEINOVEacuteHO PROFILU V PRŮBĚHUSLADOVAacuteNIacute JEČMENECHANGES OF PROTEIN PROFILE IN BARLEY DURING MALTING



VEDOUCIacute PRAacuteCE ING JANETTE BOBAacuteĽOVAacute CSCSUPERVISOR

BRNO 2008

3

Abstrakt

Tato diplomovaacute praacutece se zabyacutevaacute studiem změn proteinoveacuteho profilu ječmene během procesu sladovaacuteniacute Podstatnaacute čaacutest praacutece se věnuje proteomickeacute identifikaci ječnyacutech proteinů ktereacute se během sladovaacuteniacute měniacute staacutevajiacute se stabilnějšiacutemi a v konečneacutem důsledku ovlivňujiacute vlastnosti piva (např pěnu a zaacutekal) Jako vzorek byl použit ječmen odrůdy Jersey

V literaacuterniacute čaacutesti jsou uvedeny obecneacute informace o pivu o procesu jeho vyacuteroby s popisem důležityacutech surovin pro vyacuterobu a informace o sladovnickeacutem ječmeni a procesu sladovaacuteniacute Daacutele jsou popsaacuteny použiteacute metody separace proteinů (1D a 2D gelovaacute elektroforeacuteza) hmotnostniacute spektrometrie MALDI TOFTOF a jejiacute využitiacute k analyacuteze a identifikaci proteinů použitiacute matric a způsoby přiacutepravy vzorků

V experimentaacutelniacute čaacutesti byla provedena optimalizace daacutevkovaacuteniacute vzorku pro 1D gelovou elektroforeacutezu a optimalizace vizualizace Nejlepšiacute vyacutesledky byly dosaženy u 15 TRIS-HCl gelu kteryacute byl obarven barvivem Commassie Brilliant Blue G-250 Pro ilustraci změn byla provedena i 2D gelovaacute elektroforeacuteza

Pomociacute metody peptidoveacuteho mapovaacuteniacute a MSMS byly identifikovaacuteny ječneacute proteiny ndash protein Z α-amylasa subtilisin inhibitor β-amylasa a peroxidasa Byla provedena analyacuteza intaktniacutech proteinů kteraacute byla zaměřena na sledovaacuteniacute změn u vyacuteznamneacuteho proteinu ječmene LTP 1

Abstract

This diploma thesis is focused on studies of changing of protein profile during barley malting Substantial part of this work is devoted to the proteomics identification of barley proteins which change during malting and so become more stationary and they influence quality of beer (haze and foam in beer) For this experiment was used barley variety Jersey

In the theoretical part of this thesis there is information about beer manufacturing of beer with description of important commodities for manufacturing of beer and information about barley malting and information about malting process Next there is description of methods for separation of proteins (1D gel electrophoresis and 2D gel electrophoresis) MALDI TOFTOF mass spectrometry and this use for the analysis and identification of proteins the use of matrices and ways of the sample preparation

In the experimental part of this thesis there was carried out the optimisation of the dosage of sample for 1D gel electrophoresis and the optimisation of staining The 15 TRIS-HCl gel was the best this gel was stained by Commassie Brilliant Blue G-250 For illustration of changes was made 2D gel electrophoresis

With help of method peptide mass fingerprinting and MSMS proteins of barley ndash protein Z α-amylase subtilisin inhibitor β-amylase a peroxidase were identificated The analysis of barley extract intact proteins was carried out this analysis was focused on changes of important barley protein LTP 1

4

Kliacute čovaacute slova 1D gelovaacute elektroforeacuteza 2D gelovaacute elektroforeacuteza MALDI TOFTOF MS proteiny sladovaacuteniacute ječmen Keywords 1D gel electrophoresis 2D gel electrophoresis MALDI TOFTOF MS proteins malting barley

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ŠOPIacuteKOVAacute M Změny proteinoveacuteho profilu v průběhu sladovaacuteniacute ječmene Brno Vysokeacute učeniacute technickeacute v Brně Fakulta chemickaacute 2008 69 s Vedouciacute diplomoveacute praacutece Ing Janette Bobaacuteľovaacute CSc

PROHLAacuteŠENIacute

Prohlašuji že diplomovaacute praacutece byla vypracovaacutena samostatně a že všechny použiteacute literaacuterniacute zdroje jsou spraacutevně a uacuteplně citovaacuteny Tato praacutece je z hlediska obsahu majetkem Fakulty chemickeacute VUT v Brně a může byacutet využita ke komerčniacutem uacutečelům jen se souhlasem vedouciacuteho diplomoveacute praacutece a děkana FCH VUT

helliphelliphelliphelliphelliphelliphelliphelliphelliphelliphellip

podpis diplomanta

Poděkovaacuteniacute Děkuji za spolupraacuteci vedouciacutemu diplomoveacute praacutece Ing Janette Bobaacuteľoveacute CSc a celeacutemu kolektivu na Akademii věd ČR v Brně s jejiacutež pomociacute byla tato praacutece realizovaacutena Daacutele bych na tomto miacutestě chtěla poděkovat svojiacute rodině za podporu během studia Tato diplomovaacute praacutece byla podporovaacutena Vyacutezkumnyacutem centrem pro studium obsahovyacutech laacutetek ječmene a chmele ndash 1M0570 ndash MŠMT ČR

6

OBSAH 1 UacuteVOD 8 2 CIacuteL PRAacuteCE 9 3 TEORETICKAacute ČAacuteST 10

31 Pivo 10 311 Zaacutekladniacute suroviny pro vyacuterobu piva 10

312 Sladovaacuteniacute 11 313 Vyacuteroba piva 13

32 Ječmen 15

321 Zkoumaneacute odrůdy 17

322 Dusiacutekateacute laacutetky v ječmeni 19

33 Proteomika 20

34 Separačniacute techniky 21

341 Gelovaacute elektroforeacuteza proteinů 21

342 Elektoforeacuteza v polyakrylamidoveacutem gelu (PAGE) 21

343 SDS elektroforeacuteza 22 344 2D elektroforeacuteza 22 345 Detekce a vizualizace biacutelkovin v gelech 22

35 Hmotnostiacute spektrometrie 23

351 Hmotnostniacute spektrometrie s ionizaciacute laserem v přiacutetomnosti matrice 24

352 Hmotnostniacute analyzaacutetor bdquodoby letuldquo (Time of flight - TOF) 25

353 Tandemovyacute hmotnostniacute analyzaacutetor TOFTOF 26

354 Matrice 27 355 Identifikace proteinů metodou peptidoveacuteho mapovaacuteniacute 29 356 Sekvenovaacuteniacute proteinů a peptidů 29

4 EXPERIMENTAacuteLNIacute ČAacuteST 32

41 Seznam chemikaacuteliiacute 32

42 Materiaacutel 33

43 Extrakce biacutelkovin pro 1D elektroforeacutezu v polyakrylamidoveacutem gelu 33

44 1D elektroforeacuteza v polyakrylamidoveacutem gelu (1D PAGE) 33


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47 Přiacuteprava vzorku pro hmotnostně spektrometrickou analyacutezu 36

48 Hmotnostniacute spektrometrie 37

49 Přiacuteprava vzorků na měřeniacute intaktniacutech proteinů 37 5 VYacuteSLEDKY A DISKUSE 39

51 Separace proteinů 1D (SDS-PAGE) elektroforeacutezou 39

511 Optimalizace daacutevkovaacuteniacute vzorku 39 512 Vizualizace proteinů 41

513 Elektroforetickyacute profil proteinů ječmene 42

52 Separace proteinů 2D gelovou elektroforeacutezou 47

53 Identifikace proteinů metodou peptidoveacuteho mapovaacuteniacute 49 531 Identifikace standardů pomociacute hmotnostniacute spektrometrie 50 532 Identifikace proteinu Z pomociacute hmotnostniacute spektrometrie 52

533 MS spektra dalšiacutech identifikovanyacutech proteinů ječmene ziacuteskanyacutech z teacutehož gelu 55

534 Změny proteinu Z v průběhu celeacuteho procesu sladovaacuteniacute (MS a MSMS spektra) 57

54 Měřeniacute intaktniacutech proteinů ječmene 61 6 ZAacuteVĚR 64 7 LITERATURA 66 8 SEZNAM SYMBOL Ů 69

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1 UacuteVOD

Proteiny jsou podstatou všech živyacutech organismů jsou to z aminokyselin složeneacute vysokomolekulaacuterniacute přiacuterodniacute laacutetky s relativniacute molekulaacuterniacute hmotnostiacute 103 až 106 [1] Peptidy jsou anhydropolymery 20 α-L-aminokyselin ktereacute jsou spojeny peptidovyacutemi vazbami za vzniku lineaacuterniacutech polypeptidovyacutech řetězců Podle počtu aminokyselin v molekule rozlišujeme oligopeptidy (2ndash10 aminokyselin) polypeptidy (11ndash100) a proteiny (viacutece než 100 aminokyselin) Pořadiacute aminokyselin v řetězci proteinu označujeme jako primaacuterniacute strukturu nebo takeacute sekvenci Rozlišujeme primaacuterniacute sekundaacuterniacute terciaacuterniacute a u některyacutech složitějšiacutech molekul ještě kvarteacuterniacute strukturu peptidoveacuteho řetězce Peptidoveacute řetězce proteinů jsou syntetizovaacuteny na ribozomech procesem zvanyacutem translace Většina z nich podleacutehaacute kotranslačniacutem a posttranslačniacutem modifikaciacutem Pořadiacute aminokyselin v polypeptidoveacutem řetězci je pro každyacute protein jedinečneacute a geneticky daneacute Drobneacute změny primaacuterniacute struktury (vyvolaneacute mutacemi genu) mohou veacutest k podstatneacute změně vlastnostiacute daneacuteho proteinu V buňce se vyskytuje několik set až tisiacutec různyacutech proteinů ktereacute zajišťujiacute jejiacute zaacutekladniacute funkce a lišiacute se jak chemickou stavbou (předevšiacutem pořadiacutem aminokyselin v peptidoveacutem řetězci - tzv primaacuterniacute strukturou) tak prostorovyacutem uspořaacutedaacuteniacutem Univerzaacutelniacute systeacutem klasifikace proteinů neexistuje Lze je třiacutedit např na zaacutekladě rozpustnosti na zaacutekladě isoelektrickeacuteho bodu (kyseleacute neutraacutelniacute basickeacute) na zaacutekladě molekuloveacute hmotnosti (maleacute do 40 kDa velkeacute obvykle nad 200 kDa) nebo na zaacutekladě tvaru molekuly zvlaacuteštniacute skupinu tvořiacute proteiny zabudovaneacute do biologickyacutech membraacuten Podle složeniacute lze proteiny dělit na proteiny jednoducheacute a složeneacute důležitějšiacute je však děleniacute proteinů podle funkce [2] Mezi zaacutekladniacute funkce patřiacute funkce stavebniacute (kolagen elastin keratin) transportniacute a skladovaciacute (hemoglobin transferin) zajišťujiacuteciacute pohyb (aktin myosin) katalytickeacute řiacutediacuteciacute a regulačniacute (enzymy hormony) a ochranneacute a obranneacute (imunoglobulin fibrin fibrinogen)

Pro pochopeniacute funkce biologickeacuteho systeacutemu je nutneacute znaacutet jeho chemickeacute složeniacute Stav životniacuteho cyklu buňky v ktereacutemkoliv okamžiku je daacuten předevšiacutem složeniacutem jejich proteinů tj jejiacutem proteomem

Existuje několik způsobů identifikace proteinů ale většinou jsou to časově naacuteročneacute postupy Moderniacute a uacutečinnyacutem naacutestrojem je proteomika Ta je založena na srovnaacuteniacute znaacutemyacutech sekvenciacute DNA a primaacuterniacutech struktur proteinů ktereacute jsou dostupneacute v databaacuteziacutech s experimentaacutelně stanovenyacutemi molekulovyacutemi hmotnostmi peptidů ziacuteskanyacutech enzymatickyacutem rozkladem sledovaneacuteho proteinu Identifikace proteinů je nyniacute důležityacutem uacutekolem v mnoha oblastech včetně identifikace odrůd sladovnickeacuteho ječmene neboť proteiny jsou často označovaacuteny jako biochemickeacute markery určityacutech vlastnostiacute (např odrůdoveacute přiacuteslušnosti) sladovnickeacuteho ječmene [3]

9

2 CIacuteL PRAacuteCE

Diplomovaacute praacutece je věnovanaacute studiu změn proteinů v průběhu sladovaacuteniacute ječmene protože se ukazuje že sladovnickou kvalitu ovlivňujiacute i proteiny

Ciacutelem teoretickeacute čaacutesti diplomoveacute praacutece bylo proveacutest literaacuterniacute rešerši o proteomice ječmene principu metod použiacutevanyacutech pro studium proteinů ječmene a uveacutest přehled důležityacutech proteinů ječmene v technologickeacutem procesu vyacuteroby piva

Ciacutelem experimentaacutelniacute čaacutesti byla extrakce vodorozpustnyacutech proteinů z ječmene a sladu separace těchto extrahovanyacutech proteinů a naacuteslednaacute identifikace pomociacute hmotnostniacute spektrometrie - MALDI TOFTOF

10

3 TEORETICKAacute ČAacuteST 31 Pivo

Pivo je jeden z nejstaršiacutech osvěžujiacuteciacutech miacuterně alkoholickyacutech naacutepojů lidstva Pivo je disperzniacute soustavou různyacutech sloučenin Jde o koloidniacute roztok různyacutech makromolekul ndash proteinů nukleovyacutech kyselin sacharidů a lipidů Chemickeacute složeniacute piva se měniacute v širokyacutech meziacutech V zaacutevislosti na extraktu původniacute mladiny a stupni prokvašeniacute obsahuje pivo asi 2 ndash 6 procent extraktivniacutech laacutetek Hlavniacute součaacutestiacute extraktu jsou sacharidy (dextriny mono- a oligosacharidy maltoacuteza maltotrioacuteza pentoacuteza) Asi 6 až 9 procent extraktu tvořiacute dusiacutekateacute laacutetky V pivu jsou mimo jineacute daacutele polyfenoloveacute laacutetky (cca 100 až 180 mgl) hořkeacute laacutetky z chmele barviva heterocyklickeacute laacutetky a vitamiacuteny [4] Důležitou vlastnostiacute piva ze zdravotnickeacuteho hlediska je jeho antioxidačniacute schopnost Antioxidačniacute antimutagenniacute antikarcinogenniacute antimikrobiaacutelniacute a dalšiacute uacutečinky jsou v pivu přisuzovaacuteny polyfenolům Polyfenoly se do piva dostaacutevajiacute z ječmene resp ze sladu chmele a chmelovyacutech vyacuterobků jako přiacuterodniacute složky ktereacute ovlivňujiacute jeho senzorickeacute vlastnost i trvanlivost [56] Piva českeacuteho typu vykazujiacute o cca 30 procent vyššiacute obsah polyfenolů a vyššiacute pH což je to důsledkem technologickeacuteho postupu [7]

311 Zaacutekladniacute suroviny pro vyacuterobu piva

Zaacutekladniacute princip vyacuteroby piva se dodnes od dob Sumeřanů a Egypťanů nezměnil Pivo je

vyrobeno jednou z nejstaršiacutech technologiiacute ktereacute člověk zvlaacutedl Enzymy ječmene a kvasinek jsou využiacutevaacuteny k přeměně suroviny na pivo Klasickaacute vyacuteroba trvaacute 40 až 60 dniacute v zaacutevislosti na typu piva [8] Zaacutekladniacute suroviny jsou sladovnickyacute ječmen chmel pivovarskeacute kvasnice a voda a) Sladovnickyacute ječmen

Z ječmene se vyraacutebiacute slad kteryacute se velmi vyacuteznamně podiacuteliacute na charakteru pěny zlataveacute barvě a specifickeacute chuti Nejběžněji vyraacuteběnyacutemi druhy sladů v Českeacute republice jsou světlyacute slad a bavorskyacute slad kteryacute je charakteristickyacute vysokou barvou a vyacuteraznějšiacutem aromatem Pro vyacuterobu sladu a sladovyacutech vyacutetažků se na našem uacutezemiacute pěstujiacute vybraneacute odrůdy jarniacuteho dvouřadeacuteho niacuteciacuteho ječmene (Hordeum distichum var nutans) ktereacute patřiacute k nejkvalitnějšiacutem odrůdaacutem na světě Mnoheacute zahraničniacute odrůdy majiacute genetickyacute zaacuteklad pochaacutezejiacuteciacute z našich odrůd zejmeacutena z oblasti Haneacute b) Chmel a chmeloveacute vyacuterobky

Chmel jako jedna ze třiacute zaacutekladniacutech pivovarskyacutech surovin je představovaacuten usušenyacutemi chmelovyacutemi hlaacutevkami samičiacutech rostlin chmele evropskeacuteho (Humulus lupulus var europeus) Poskytuje pivu typickou hořkou chuť přispiacutevaacute k tvorbě charakteristickeacuteho aroma Chmele pěstovaneacute u naacutes v žateckeacute oblasti patřiacute mezi vysoce kvalitniacute jemneacute aromatickeacute odrůdy chmele evropskeacuteho otaacutečiveacuteho Chmeloveacute hlaacutevky ktereacute se skliacutezejiacute pro pivovarskeacute uacutečely se sklaacutedajiacute ze

11

stopky vřeteacutenka pravyacutech a kryciacutech listenů a při oplozeniacute obsahujiacute naviacutec semeno neboli pecku Na vnitřniacute straně listenů se při zraacuteniacute chmele vylučujiacute pryskyřičnaacute zrnka lupulinu obsahujiacuteciacute chmeloveacute pryskyřice a silice [9] c) Pivovarskeacute kvasnice

Kvasinky druhu Saccharomyces cerevisiae jsou fakultativně anaerobniacute mikroorganismy Přeměňujiacute cukr na alkohol a oxid uhličityacute d) Voda

Voda je ve sladařskeacutem a pivovarskeacutem průmyslu důležitou surovinou protože přiacutemo ovlivňuje kvalitu piva Na vyacuterobu 1 t sladu se spotřebuje 10 - 15 hl vody a na vyacuterobu 1 hl piva se spotřebuje 12 - 15 hl vody

312 Sladovaacuteniacute

Ciacutelem sladovaacuteniacute je přeměnit ječmen na slad bohatyacute na enzymy a extrakt [10] Proces

vyacuteroby sladu lze z hlediska jednotlivyacutech vyacuterobniacutech faacuteziacute rozdělit na několik čaacutestiacute (viz obrč1)

Přiacutejem čištěniacute třiacuteděniacute a skladovaacuteniacute ječmene

darrdarrdarrdarr

Maacutečeniacute ječmene

darrdarrdarrdarr

Kliacutečeniacute namočeneacuteho ječmene

darrdarrdarrdarr

Hvozděniacute zeleneacuteho sladu

darrdarrdarrdarr

Odkliacutečeniacute leštěniacute baleniacute a expedice hotoveacuteho sladu

Obrč1 Scheacutema vyacuteroby sladu [12]

Prvniacute z nich je maacutečeniacute Samotneacutemu maacutečeniacute předchaacuteziacute přiacutejem čištěniacute a skladovaacuteniacute

ječmene Skladovanyacute ječmen představuje živyacute organismus jehož životniacute pochody jsou utlumeny nikoliv však zastaveny Energii potřebnou pro životniacute projevy ziacuteskaacutevaacute zrno odbouraacutevaacuteniacutem rezervniacutech polysacharidů hlavně škrobu Podle okamžityacutech podmiacutenek ziacuteskaacutevaacute energii buď aerobniacutem dyacutechaacuteniacutem v přiacutetomnosti kysliacuteku nebo anaerobniacutem kvašeniacutem v nepřiacutetomnosti kysliacuteku Při skladovaacuteniacute se čerstvě sklizenyacute a vytřiacuteděnyacute ječmen nachaacuteziacute ve

12

stadiu zaacutekladniacuteho klidu tzv dormance a neniacute schopen kliacutečit Je to způsobeno přiacutetomnostiacute inhibitorů kliacutečeniacute tzv dormitů Teprve po jejich odbouraacuteniacute oxidaciacute dormance zanikaacute uvolňuje se činnost stimulaacutetorů (giberelinů) a zrno se staacutevaacute schopnyacutem kliacutečit Skladovaacuteniacute probiacutehaacute v silech [11] Vyčištěnyacute a odleženyacute ječmen se maacutečiacute v namaacutečeciacutech naacuteduvniacuteciacutech Ciacutelem maacutečeniacute je dodaacuteniacute potřebneacute vegetačniacute vody z 12-15 na 42-48 kteraacute je nutnaacute na zahaacutejeniacute enzymatickyacutech reakciacute a pro kliacutečeniacute zrna Maacutečeniacute se dnes považuje za nejdůležitějšiacute uacutesek vyacuteroby sladu Stupeň domočeniacute se lišiacute podle typu vyraacuteběneacuteho sladu Vyacuteznamnyacutem efektem je vypraacuteniacute ječmene protože se z ječmene vyloužiacute barevneacute a hořkeacute laacutetky kyselina křemičitaacute a biacutelkoviny z pluch Tyto laacutetky jsou nežaacutedouciacute neboť zhoršujiacute senzorickeacute vlastnosti piva a podporujiacute tvorbu zaacutekalu v pivu

Druhou faacuteziacute je kliacutečeniacute Kliacutečivost a kliacutečivaacute energie patřiacute mezi důležiteacute fyziologickeacute znaky sladovnickeacuteho ječmene Udaacutevajiacute procentuaacutelniacute podiacutel zrn schopnyacutech vykliacutečit za stanovenyacutech podmiacutenek během třiacute až pěti dnů Ciacutelem sladařskeacuteho kliacutečeniacute ječmene je aktivace a tvorba enzymů (největšiacute technologickyacute vyacuteznam majiacute amylasy fosfatasy cytasy proteasy) a dosaženiacute požadovaneacuteho stupně rozluštěniacute Dosahuje se toho umělyacutem modelovaacuteniacutem optimaacutelniacutech podmiacutenek přirozeneacuteho kliacutečeniacute V průběhu kliacutečeniacute rozlišujeme tvorbu enzymů a přeměnu laacutetek růstoveacute změny a projevy růstu S vyacutejimkou α-amylasy kteraacute neniacute v ječmeni obsažena jsou ostatniacute enzymy v maleacutem množstviacute již v ječmeni přiacutetomny Synteacuteza novyacutech enzymů je iniciovaacutena prostřednictviacutem činnosti fytohormonů Nejprve vznikaacute β-glukonasa poteacute α-amylasa a proteasy Amylasy jsou nejdůležitějšiacutemi enzymy ve sladu s jejich pomociacute jsou později odbouraacutevaacuteny škroby při rmutovaacuteniacute bull α-amylasa neniacute v ječmeni přiacutetomnaacute Největšiacute množstviacute tohoto enzymu se tvořiacute od

druheacuteho do čtvrteacuteho dne kliacutečeniacute bull β-amylasa je přiacutetomna v maleacutem množstviacute v ječmeni Po přechodneacute maleacute ztraacutetě v prvniacutem

dni kliacutečeniacute se jejiacute množstviacute od druheacuteho a třetiacuteho dne daacutele zvyšuje Tvorba β-amylasy je spojena s dyacutechaacuteniacutem v prvniacutem dni kliacutečeniacute Proto je pro jejiacute tvorbu důležiteacute dostatečneacute provětraacutevaacuteniacute už v prvniacute faacutezi kliacutečeniacute

Obsah amylas je odrůdovou zaacutevislostiacute a je rovněž ovlivněn klimatickyacutemi podmiacutenkami

ročniacuteku [11] Během kliacutečeniacute neprobiacutehaacute jen tvorba a zvyacutešeniacute množstviacute enymů Řiacutezeneacute kliacutečeniacute ječmene ve

sladovně se nazyacutevaacute vedeniacute hromad Jednaacute se o fyziologickyacute proces při ktereacutem se v zaacuterodečneacute čaacutesti zrna vyviacutejejiacute zaacuterodky kořiacutenků kliacutečků a listů za využitiacute zaacutesobniacutech laacutetek z endospermu [12] Současně se měniacute i vnitřniacute znaky zrna Působeniacutem enzymů se štěpiacute rezervniacute laacutetky a zvyšuje se rozpustnost a luštitelnost Vyacuteslednyacutem produktem kliacutečeniacute je tzv zelenyacute slad Zelenyacute slad maacute vysokyacute obsah vody a neniacute na rozdiacutel od hotoveacuteho sladu skladovatelnyacute Posledniacute faacuteziacute je hvozděniacute sladu Zelenyacute slad je dopraven na tzv hvozd V průběhu hvozděniacute je šetrně odstraněna většina vody dodanaacute zrnu při maacutečeniacute a slad je převeden do skladovatelneacuteho a stabilniacuteho stavu Dojde k zastaveniacute životniacutech projevů a luštiacuteciacutech pochodů v zrně Během hvozděniacute se vytvaacuteřiacute aromatickeacute a barevneacute laacutetky charakteristickeacute pro druhy sladu Tyto laacutetky vznikajiacute při vyššiacutech teplotaacutech interakcemi štěpnyacutech produktů polysacharidů a biacutelkovin zejmeacutena monosacharidů a aminokyselin Jednaacute se o řadu chemickyacutech změn označovanou jako Maillardovy reakce Mezi laacutetky vznikajiacuteciacute při hvozděniacute sladu během Maillardovyacutech reakciacute patřiacute hlavně melanoidiny a glykosylovaneacute proteiny ktereacute zvyšujiacute viskozitu [1314]

13

a podporujiacute pěnivost piva na zaacutekladě iontoveacute vazby s proteiny při tvorbě kostry pěny [15] Odhvozděnyacute slad se sklaacutepiacute do košů a dopravuje se do odkličovačce kde se zbavuje kořiacutenků zvanyacutech sladovyacute květ

313 Vyacuteroba piva

Vyacuteroba piva začiacutenaacute šrotovnou Ta dokaacuteže slad rozemliacutet na přesně požadovaneacute složeniacute

poměru mouky krupice a pluh Vyacuteroba v sobě zahrnuje čtyři procesy Vařeniacute kvašeniacute dokvašovaacuteniacute a staacutečeniacute (viz obrč2) 1) Vařeniacute

Probiacutehaacute ve varně ve čtyřech naacutedobaacutech ndash vystiacuteraciacute kaacuteď rmutovaciacute kotel scezovaciacute kaacuteď

a mladinovyacute kotel Sladovyacute šrot padaacute do vystiacuteraciacute kaacutedě kde se miacutesiacute s vodou Ze sladu se uvolňuje extrakt do roztoku probiacutehaacute rmutovaacuteniacute Ciacutelem rmutovaacuteniacute je rozštěpeniacute a převedeniacute optimaacutelniacuteho podiacutelu extraktu surovin do roztoku Zaacutekladniacutem požadavkem všech rmutovaciacutech postupů je převeacutest do roztoku veškeryacute škrob i vhodnyacute podiacutel biacutelkovin a dalšiacutech laacutetek (škrob je při rmutovaacuteniacute štěpen sladovyacutemi amylasamy) Naopak přiacutetomnost jinyacutech složek jako např polyfenolů a sladovyacutech pluch se snažiacuteme omezit [16] Při tomto procesu probiacutehajiacute mechanickeacute chemickeacute fyzikaacutelniacute a hlavně enzymoveacute děje (Důležiteacute teploty při rmutovaacuteniacute jsou teplota kyselinotvornaacute 35-38 degC peptonizačniacute 45-55 degC nižšiacute cukrotvornaacute 60-65 degC vyššiacute cukrotvornaacute 70-75 degC nejvyššiacute cukrotvornaacute 78 degC) Po rmutovaacuteniacute se celeacute diacutelo přečerpaacutevaacute na scezovaciacute kaacuteď kde se po 30 minutaacutech vytvořiacute filtračniacute vrstva pevnyacutech podiacutelů a probiacutehaacute vlastniacute filtrace tedy odděleniacute kapalneacuteho podiacutelu sladiny od pevneacuteho podiacutelu sladoveacuteho mlaacuteta (mlaacuteto se použiacutevaacute jako krmivo pro dobytek) Sladkyacute roztok neboli sladina se převede do mladinoveacuteho kotle kde se povařiacute s chmelem Chmel dodaacute hořkost Ziacuteskanaacute mladina se musiacute ochladit aby mohla začiacutet dalšiacute faacuteze vyacuteroby kterou je kvašeniacute 2) Kvašeniacute

Pro kvašeniacute mladiny se použiacutevajiacute buď svrchniacute pivovarskeacute kvasinky (Saccharomyces cerevisiae) kvasiacuteciacute při teplotaacutech až 24 degC ktereacute stoupajiacute ode dna na hladinu nebo spodniacute pivovarskeacute kvasinky (Saccharomyces cerevisiae var uvarum) kvasiacuteciacute při teplotaacutech 6-12 degC ktereacute klesajiacute od hladiny na dno Staršiacute způsob kvašeniacute se provaacuteděl ve spilce Spilkou rozumiacuteme otevřenou kvasnou kaacuteď Modernějšiacute způsob se provaacutediacute v uzavřenyacutech cylindrokonickyacutech tanciacutech Českaacute piva se vyraacutebějiacute spodniacutem kvašeniacutem Zchlazenaacute a provzdušněnaacute mladina se po zakvašeniacute spodniacutemi pivovarskyacutemi kvasnicemi přivaacutediacute do uzavřenyacutech nerezovyacutech cylindrokonickyacutech tanků o objemu 3 600 hl Nejdůležitějšiacutemi reakcemi hlavniacuteho kvašeniacute jsou přeměny zkvasitelnyacutech sacharidů glukosy maltosy a maltotriosy na ethanol a oxid uhličityacute anaerobniacutem kvašeniacutem

C6H12O6 rarr 2C2H5OH + 2CO2 (1)

Dřiacuteve bylo kvašeniacute teacuteměř nekontrolovatelnyacute proces ale dnes se použiacutevajiacute speciaacutelně vyšlechtěneacute kmeny kvasnic Vyacutesledkem kvašeniacute je mladeacute pivo

14

3) Dokvašovaacuteniacute

Dokvašovaacuteniacute se provaacutediacute aby pivo ziacuteskalo spraacutevnou chuť (syceniacute CO2) Po prokvašeniacute se mladeacute pivo chladiacute v tanciacutech v ležaacuteckyacutech sklepiacutech Ještě zde dobiacutehajiacute posledniacute zbytky kvašeniacute pivo zraje a zbavuje se mnoha negativniacutech laacutetek Svrchně kvašenaacute piva zrajiacute jeden tyacuteden až rok spodně kvašenaacute zrajiacute čtyři tyacutedny až rok Dokvašovaacuteniacute se ukončiacute filtraciacute a pasteraciacute [17] Pasterace se použiacutevaacute ke zvyacutešeniacute biologickeacute stability piva Hotoveacute pivo je připraveno ke staacutečeniacute do transportniacutech naacutedob 4) Staacutečeni a expedice

Staacutečeniacute piva do transportniacutech a spotřebitelskyacutech obalů je konečnou faacuteziacute vyacuteroby U naacutes se pivo staacutečiacute do cisteren pro dislokovaneacute staacutečiacuterny a pro export do sudů lahviacute a plechovek pro vnitřniacute obchodniacute siacuteť i pro export Při staacutečeniacute je nutneacute zamezit ztraacutetaacutem oxidu uhličiteacuteho aby neutrpěla kvalita piva proto jsou staacutečeciacute stroje konstruovaacuteny na izobarickeacutem principu [18]

Obrč2 Scheacutema vyacuterobniacuteho procesu

15

32 Ječmen

Zařazeniacute ječmene do systeacutemu

Podle způsobu růstu se ječmeny děliacute na divoce rostouciacute planeacute ječmeny z nichž je u naacutes nejrozšiacuteřenějšiacute ječmen myšiacute a ječmeny seteacute ktereacute se vyskytujiacute v kultuře a jsou jednoletou jarniacute nebo ozimou traacutevou Kulturniacute ječmeny se pak ještě děliacute na ječmeny dvouřadeacute a viacuteceřadeacute [16 20]

Stručnyacute morfologickyacute popis ječmene Kořenovaacute soustava svazčiteacute kořeny Steacuteblo odnože steacuteblo ječmene tvořiacute 4 - 8 člaacutenků (internodiiacute) oddělenyacutech koleacutenky (nody)

a dosahuje vyacutešky 80 až 130 cm Listy listy maacute ječmen pravotočiveacute a jsou umiacutestěny nad sebou ve dvou řadaacutech Květenstviacute a květ složenyacute nerozvětvenyacute klas (lichoklas) tvořenyacute smaacutečknutyacutem vřetenem

na stranaacutech obrvenyacutem ktereacute je rozděleno na jednotliveacute člaacutenky Obilka obilka (zrno) je složena ze třiacute čaacutestiacute obalu endospermu a zaacuterodku Endosperm

(biacutelek) vyplňuje hlavniacute podiacutel zrna Jeho vnějšiacute vrstva se nazyacutevaacute aleuronovaacute Buňky aleuronoveacute vrstvy obsahujiacute zaacutesobniacute proteiny tuk a menšiacute množstviacute škrobovyacutech zrn Na počaacutetku kliacutečeniacute se v nich aktivujiacute enzymy ktereacute degradujiacute obsah škroboveacuteho endospermu Obilky jsou pluchateacute nebo bezplucheacute (naheacute) [21] U pluchateacuteho ječmene je obilka na hřbetniacute straně kryta pluchou kteraacute svyacutemi okraji překryacutevaacute menšiacute plušku (viz obrč3) Plucha spolu s pluškou chraacuteniacute obilku před vnějšiacutemi vlivy Svou strukturou se přiacuteliš nelišiacute od listů a jejich zvraacutesněniacute určuje jemnost [20] Zrno bezpluchyacutech forem ječmene se na rozdiacutel od pluchateacuteho nemusiacute loupat maacute lepšiacute senzorickeacute vlastnosti zrna Nevyacutehodou je citlivost na poškozeniacute kliacutečku V současneacute době je bezpluchyacute ječmen asi nejviacutece ceněn jako vynikajiacuteciacute zdroj rozpustneacute vlaacutekniny Uvaacutediacute se že obsah celkoveacute potravinaacuteřskeacute vlaacutekniny u různyacutech odrůd koliacutesaacute od 15 do 24 z toho nerozpustnaacute vlaacuteknina tvořiacute 11 - 19 a rozpustnaacute 3 - 6 Pluchatyacute ječmen se tradičně použiacutevaacute jako krmivo pro zviacuteřata nebo pro vyacuterobu sladu a malaacute čaacutest sloužiacute pro lidskou vyacuteživu [22]

Řiacuteše Rostliny (Plantae)

Podřiacuteše Ceacutevnateacute rostliny (Tracheobionta)

Odděleniacute Krytosemenneacute (Magnoliophyta)

Třiacuteda Jednoděložneacute (Liliopsida)

Řaacuted Lipnicotvareacute (Poales)

Čeleď Lipnicoviteacute (Poaceae)

Podčeleď Vlastniacute lipnicoviteacute (Pooideae)

Rod Ječmen (Hordeum) [19]

16

Tab č 1 Chemickeacute složeniacute obilky ječmene (Mac Gregor a kolektiv 1993)

Obrč3 Přiacuteklad dvoukvěteacuteho klaacutesku trav 1- plevy 2- pluchy 3- osiny 4- plušky 5- tyčinky

17

Ke sladovnickyacutem uacutečelům se pěstujiacute převaacutežně jarniacute formy ječmene V roce 1999 bylo v Českeacute republice registrovaacuteno 40 odrůd ječmene jarniacuteho 6 odrůd ječmene ozimeacuteho dvouřadeacuteho a 9 odrůd ječmene ozimeacuteho viacuteceřadeacuteho [26] Ale odrůdovaacute skladba se neustaacutele měniacute Sladovnickyacute ječmen se staacutevaacute specifickou plodinou na kterou jsou kladeny mimořaacutedně vysokeacute požadavky ze strany sladovnickeacuteho a pivovarskeacuteho průmyslu Vyacuteběr vhodneacute odrůdy rozhodujiacuteciacutem způsobem ovlivňuje kvalitu sladu Požadavky na sladovnickou kvalitu jsou zcela specifickeacute podle jednotlivyacutech pivovarů a regionů nejen v Evropě ale i v zaacutemořskyacutech zemiacutech [23] Ječmen pro vyacuterobu sladu musiacute současně vykazovat optimaacutelniacute zpracovatelskeacute vlastnosti vlhkost nejvyacuteše 16 kliacutečivost nejmeacuteně 90 obsah biacutelkovin maximaacutelně 125 v sušině co nejvyššiacute hladinu škrobu a řadu dalšiacutech kriteacuteriiacute [24] Požadavky uvaacutediacute norma ČSN 46 1100-5 kteraacute byla novelizovaacutena v roce 2006 [25] Vysokeacute sladovnickeacute hodnoty je dosahovaacuteno jen za určityacutech půdně klimatickyacutech podmiacutenek V Čechaacutech je to předevšiacutem v Polabskeacute niacutežině a nižšiacutech polohaacutech Středočeskeacute pahorkatiny na Moravě celaacute středniacute Morava s jaacutedrem uacuterodneacute Haneacute [26]

Hlavniacute skupinu sladovnickyacutech ječmenů tvořiacute ječmeny dvouřadeacute Ječmen dvouřadyacute se vyskytuje v několika varietaacutech z nichž nejdůležitějšiacute jsou bull Varieta nutans (ječmen niacuteciacute haacutečkujiacuteciacute) tvořiacute klas dlouhyacute 50 ndash 130 mm maacute dlouheacute

souběžneacute přileacutehajiacuteciacute osiny v době zralosti se klas ohyacutebaacute (haacutečkuje) Patřiacute sem většina sladovnickyacutech odrůd

bull Varieta nudum (ječmen nahyacute) u něj obilka nesrůstaacute s pluchami při vyacutemlatu zůstaacutevaacute asi

20 obilek obaleno pluchami ktereacute však rovněž s obilkou nesrůstajiacute Obilky se vyznačujiacute niacutezkyacutem obsahem vlaacutekniny vysokou krmnou hodnotou v posledniacute době se uplatňuje jako potravina v cereaacutelniacute vyacuteživě [20]

321 Zkoumaneacute odrůdy

- Jersey byl vyšlechtěn v Holandsku firmou CEBECO SEEDS BV a v ČR byl registrovaacuten v roce 2000 Jednaacute se o vyacuteběrovou sladovnickou odrůdu s kraacutetkyacutem obdobiacutem posklizňoveacuteho dozraacutevaacuteniacute Jersey je polopozdniacute středně vysokyacute ječmen (80-85 cm) s dobrou odnoživostiacute Hodiacute se pro pěstovaacuteniacute ve všech oblastech pro vyacuterobu kvalitniacutech sladů Na zaacutekladě vynikajiacuteciacutech parametrů kvality a vyacutesledků pěstovaacuteniacute je Jersey v současnosti nejrozšiacuteřenějšiacute odrůdou v ČR Odrůda maacute vynikajiacuteciacute uacuteroveň extraktu v sušině sladu optimaacutelniacute enzymatickou aktivitu a optimaacutelniacute složeniacute sladiny [26] - Tolar byl vyšlechtěn firmou PLANT SELECT spol s ro v Hrubčiciacutech a zaregistrovaacuten v ČR a SR v roce 1997 Tolar je polopozdniacute niacutezkaacute až středně vysokaacute odrůda se středně velkyacutem až velkyacutem zrnem [26] Jednaacute se o sladovnickyacute ječmen vhodnyacute zejmeacutena pro slady na vyacuterobu typicky českyacutech piv (ležaacuteků) Je vhodnyacute pro pěstovaacuteniacute ve všech vyacuterobniacutech oblastech kde dosahuje standardně nadprůměrnyacutech vyacutenosů - KM V raacutemci řešeniacute vyacutezkumnyacutech projektů v Kroměřiacuteži se podařilo vytvořit několik novyacutech liniiacute bezplucheacuteho ječmene s odlišnyacutemi hospodaacuteřskyacutemi vlastnostmi a rozdiacutelnyacutem

18

chemickyacutem složeniacutem zrna Linie KM 1910 vyznačuje se dobrou energetickou hodnotou zrna danou niacutezkyacutem

obsahem nerozpustneacute vlaacutekniny vyššiacutem obsahem škrobů a sacharidů (viz tabulka č1) Linie KM 2283 perspektivniacute linie s vysokyacutem obsahem rozpustneacute vlaacutekniny a β-glukanů

Testy potenciaacutelu produktivity ukaacutezaly že je dlouhodobě nejvyacutekonnějšiacutem bezpluchyacutem ječmenem kteryacute se v meacuteně přiacuteznivyacutech pěstitelskyacutech podmiacutenkaacutech svyacutem vyacutenosem často přibližuje i některyacutem sladařskyacutem odrůdaacutem [27] Nevyacutehodou je vyššiacute naacutechylnost bezplucheacuteho zrna k mechanickeacutemu poškozeniacute

Obecně je nahyacute ječmen svyacutemi chemickyacutemi a sladařskyacutemi vlastnostmi srovnatelnyacute s ječmenem pluchatyacutem ale maacute o 3 vyššiacute obsah extraktu takže se z něj ziacuteskaacute viacutece sladiny a piva [28]

Tab č 2 Chemickeacute složeniacute zrna bezplucheacuteho ječmene [27]

Typ škrobu N-laacutetky ()

Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555

Tabč3 Porovnaacuteniacute složeniacute zrn mezi odrůdami

α-amylaza

(DU)

β-amylaza

(Ug) škrob () N-laacutetky ()

KM 1910

ječmen 0 283 598 232

zelenyacute slad 37 480 hellip hellip

slad 46 387 hellip hellip

Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

322 Dusiacutekateacute laacutetky v ječmeni

Tvorba dusiacutekatyacutech laacutetek v ječmeni je založena na přiacutejmu amoniaku a organickyacutech kyselin

ktereacute vznikly jako meziprodukty štěpeniacute sacharidů Synteacuteza proteinů a aminokyselin probiacutehaacute v systeacutemu enzymovyacutech reakciacute za uacutečasti ATP ribonukleovyacutech kyselin a ribozomů [16]

Dusiacutekateacute laacutetky lze rozdělit na - Dusiacutekateacute laacutetky typu proteinů a jejich štěpnyacutech produktů (aminokyseliny peptidy

peptony) - Dusiacutekateacute laacutetky nebiacutelkovinneacute povahy (dusiacutekateacute baacuteze složky fosfotidů maleacute množstviacute

amidů a amonnyacutech soliacute) ktereacute nejsou složkami proteinů

Aminokyseliny jsou nejjednoduššiacute dusiacutekateacute sloučeniny Pro vyacuteživu lidiacute jsou důležiteacute tzv esenciaacutelniacute aminokyseliny (valin leucin izoleucin threonin methionin lyzin fenylalanin tryptofan) ktereacute živočišnyacute organismus neniacute schopen syntetizovat

Proteiny patřiacute k faktorům pozitivně ovlivňujiacuteciacutem tvorbu a stabilitu pivniacute pěny avšak mohou byacutet přiacutečinou vzniku pivniacutech zaacutekalů Jsou to molekuly velkyacutech rozměrů a jsou tvořeny řetězci aminokyselin vaacutezanyacutech peptidovou vazbou Proteiny se dajiacute klasifikovat z několika hledisek Podle morfologickeacuteho původu se rozlišujiacute na proteiny endospermu proteiny aleuronoveacute vrstvy a proteiny zaacuterodku Podle biologickeacute funkce se rozlišujiacute na zaacutesobniacute a metabolicky aktivniacute daacutele se děliacute podle velikosti na niacutezko a vysokomolekulaacuterniacute Podle chemickeacuteho složeniacute se děliacute na jednoducheacute proteiny a složeneacute peptidy Na zaacutekladě fyzikaacutelně chemickyacutech vlastnostiacute a rozpustnosti v různyacutech rozpouštědlech se děliacute na [69] - Albuminy (leukosidy) rozpustneacute ve vodě představujiacute 4 všech ječnyacutech proteinů

Během sladovaacuteniacute se většina štěpiacute

- Globuliny (edestiny) rozpustneacute v roztociacutech elektrolytů (soliacute) představujiacute asi 18 celkovyacutech ječnyacutech biacutelkovin Velkyacute vyacuteznam se přisuzuje β-globulinu (je možnyacutem původcem chladovyacutech zaacutekalů piva)

- Prolaminy (hordeiny) rozpustneacute ve vodnyacutech roztociacutech alkoholů Tvořiacute asi 37 - 50

proteinů ječneacuteho zrna a nachaacutezejiacute se předevšiacutem v aleuronoveacute vrstvě jako zaacutesobniacute proteiny Během sladovaacuteniacute se silně štěpiacute

- Gluteliny (gluteminy) zčaacutesti rozpustneacute ve zředěnyacutech roztociacutech kyselin a zaacutesad tvořiacute

37 z veškeryacutech biacutelkovin a nachaacuteziacute se předevšiacutem v aleuronoveacute vrstvě odkud teacuteměř nezměněny přechaacutezejiacute do mlaacuteta Při jejich vyššiacutem obsahu dochaacuteziacute k horšiacutemu rozluštěniacute sladu Jsou znaacutemy jejich čtyři frakce [20]

Albuminy a globuliny představujiacute převaacutežně strukturniacute a metabolickeacute biacutelkoviny Prolaminy

jsou hlavniacute skupinou zaacutesobniacutech biacutelkovin s největšiacutem procentuaacutelniacutem zastoupeniacutem v zrnu [29] Obsah biacutelkovin ve zraleacutem zrnu ječmene byacutevaacute mezi 9 a 13 sucheacute vaacutehy zrna [30]

20

Obsah proteinů (a obdobně i sacharidů) na odrůdě a agroekologickyacutech podmiacutenkaacutech v roce pěstovaacuteniacute 33 Proteomika

Proteomika je vědniacute obor kteryacute se zabyacutevaacute systematickou analyacutezou proteinů z hlediska jejich identity množstviacute a funkciacute [31 62] Proteomickeacute studie zahrnujiacute zpravidla tři zaacutekladniacute kroky vhodnou separačniacute techniku hmotnostniacute spektrometrii a bioinformatiku kteraacute sloužiacute jako kliacuteč k identifikaci daneacuteho proteinu (viz obr č4) Hlavniacutem ciacutelem je identifikace proteinů včetně stanoveniacute molekuloveacute hmotnosti sekvence (pořadiacute) aminokyselin a určeniacute posttranslačniacutech modifikaciacute V dnešniacute době lze ziacuteskat vyacuteznamneacute informace o struktuře biacutelkoviny aniž by bylo nutneacute miacutet protein ve zcela čisteacute formě Přesto neexistuje žaacutednaacute universaacutelniacute metoda na přiacutepravu proteomickeacuteho vzorku ani na extrakci pro různeacute typy vyacutechoziacutech materiaacutelů Tradičniacute metodou analytickeacute proteomiky je dvourozměrnaacute gelovaacute elektroforeacuteza na polyakrylamidoveacutem gelu kteraacute se využiacutevaacute k separaci proteinů i k ziacuteskaacutevaacuteniacute dvourozměrnyacutech map proteinů ktereacute lze přiacutemo využiacutet k proteomickeacute charakterizaci organismů Proteinovaacute mapa představuje dvourozměrnyacute obraz znaacutezorňujiacuteciacute (v ideaacutelniacutem přiacutepadě všechny) buněčneacute proteiny jako skvrny (spoty) [61]

Proteomika se během posledniacute doby stala jedniacutem s nejdynamičtěji se rozviacutejejiacuteciacutech oborů v přiacuterodniacutech vědaacutech tento rozvoj byl umožněn zejmeacutena diacuteky pokrokům hmotnostniacute spektrometrie a to hlavně využitiacutem novyacutech ionizačniacutech technik MALDI a ionizace elektrosprejem ze ktereacute byla v roce 2002 udělena Nobelova cena K Tanakovi a J Fennovi Mezi hmotnostně spektrometrickeacute způsoby identifikace proteinů patřiacute stanoveniacute molekuloveacute hmotnosti intaktniacutech proteinů peptidoveacute mapovaacuteniacute fragmentačniacute analyacuteza peptidů a fermentačniacute analyacuteza intaktniacutech biacutelkovin [31]

Obr č 4 Scheacutema proteomickeacuteho studia

21

34 Separačniacute techniky Separačniacute techniky na ktereacute byla zaměřena pozornost teacuteto praacutece jsou jednorozměrnaacute

a dvourozměrnaacute gelovaacute elektroforeacuteza

341 Gelovaacute elektroforeacuteza proteinů

Gelovaacute elektroforeacuteza patřiacute mezi elektromigračniacute metody ktereacute jsou důležitou skupinou

metod pro analytickeacute děleniacute směsiacute biacutelkovin Diacuteky polaacuterniacutem aminokyselinovyacutem zbytkům biacutelkovinnyacutech řetězců ktereacute mohou byacutet ve vodneacutem prostřediacute různě disociovaacuteny v zaacutevislosti na pH prostřediacute se proteiny chovajiacute jako laacutetky amfoterniacute (amfolyty) V kyseleacutem prostřediacute přijiacutemajiacute protony a majiacute kladnyacute naacuteboj Při vysokeacutem pH dochaacuteziacute k odštěpovaacuteniacute protonů Hodnota pH při niacutež je počet kladnyacutech a zaacutepornyacutech naacutebojů v molekule proteinu stejnyacute tj molekula se na venek projevuje jako elektricky neutraacutelniacute se nazyacutevaacute izoelektrickyacute bod (označuje se pI)

Elektromigračniacute metody jsou tedy založeny na pohybu makromolekuly v elektrickeacutem poli Ten je způsoben nenulovou hodnotou celkoveacuteho naacuteboje molekuly v daneacutem prostřediacute Hnaciacute silou je působeniacute elektrickeacuteho pole proti pohybu molekul vyvolaneacutemu elektrickyacutem polem působiacute třeniacute ndash odpor prostřediacute Vyacutesledniciacute těchto sil je pohyb molekul v elektrickeacutem poli konstantniacute rychlostiacute Podstatou elektroforeacutezy je děleniacute nabityacutech čaacutestic na zaacutekladě jejich různyacutech elektroforetickyacutech pohyblivostiacute [32]

Gelovaacute elektroforeacuteza využiacutevaacute agarosoveacute nebo polyakrylamidoveacute gely Agarosa je polysacharid z mořskyacutech řas Jde o lineaacuterniacute polymer galaktosy a 36-anhydrogalaktosy Rozpouštiacute se v horkeacute vodě a po ochlazeniacute tuhne Elektroforeacuteza v agarosoveacutem gelu se využiacutevaacute předevšiacutem pro separaci nukleovyacutech kyselin

342 Elektoforeacuteza v polyakrylamidoveacutem gelu (PAGE)

Elektroforeacuteza v polyakrylamidoveacutem gelu patřiacute v současnosti k nejčastěji použiacutevanyacutem elektroforetickyacutem technikaacutem k analyacuteze proteinů PAGE se často využiacutevaacute v analytice proteinů a to ke zjištěniacute homogenity preparaacutetu a různyacutech stupniacutech isolačniacuteho postupu a k čaacutestečně fyzikaacutelně-chemickeacute charakterizaci proteinu

Uacutespěšnost elektroforetickeacute separace zaacutevisiacute na volbě vhodnyacutech podmiacutenek zejmeacutena na použiteacutem pH

Laacutetky ktereacute se nachaacutezejiacute v izoelektrickeacutem stavu nenesou žaacutednyacute vnějšiacute naacuteboj a nebudou se proto v elektrickeacutem poli pohybovat Naopak laacutetky nesouciacute naacuteboje se budou pohybovat směrem k elektrodě s opačnyacutem naacutebojem tedy kationty ke katodě a anionty k anodě Velice důležityacutem faktorem při elektroforeacuteze na nosičiacutech a tedy i při PAGE je koncentrace gelu resp stupeň zesiacutetěniacute gelu Gel s většiacutemi či menšiacutemi poacutery je mechanickou překaacutežkou pro molekuly určiteacute velikosti Maacute-li se tedy separovanaacute směs dělit pouze na zaacutekladě velikosti naacutebojů nesmiacute gel braacutenit molekulaacutem v jejich průchodu V přiacutepadě různeacute velikosti separovanyacutech molekul se pak vhodně zvolenaacute koncentrace gelu (porosita) staacutevaacute dalšiacutem faktorem ovlivňujiacuteciacute separaci

Gel se připravuje polymeraciacute akrylamidu je inertniacute je omezeno rozšiřovaacuteniacute zoacuten difuziacute maacute dobrou mechanickou pevnost je průhlednyacute Postupem přiacutepravy lze regulovat hustotu

22

siacuteťovaacuteniacute gelu [33] Gel se naleacutevaacute mezi dvě skleněneacute destičky po stranaacutech se umiacutestiacute spacery a na horniacute stranu hřebiacutenek Zvoliacute se hřebiacutenek s dostatečnyacutem členěniacutem čiacutem viacutece bude hřebiacutenek členityacute tiacutem viacutece vzorků můžeme analyzovat Jednotliveacute vzorky můžeme mezi sebou porovnaacutevat Separovaneacute laacutetky postupujiacute shora dolů (jednaacute se o vertikaacutelniacute a horizontaacutelniacute systeacutem u proteinů pak většinou o vertikaacutelniacute) Dobreacuteho vyacutesledku separace lze dosaacutehnout pouze tehdy jsou-li jednotliveacute zoacuteny laacutetek dostatečně vzdaacuteleny a vzaacutejemně se neproliacutenajiacute

343 SDS elektroforeacuteza

Elektroforeacuteza v polyakrylamidoveacutem gelu s dodecylsulfaacutetem sodnyacutem (SDS) je elektroforetickyacute způsob separace na zaacutekladě velikosti molekul separovanyacutech laacutetek jehož použitiacute je vymezeno specielně pro proteiny Dodecylsulfaacutet sodnyacute je ionogenniacute detergent kteryacute uděluje v podmiacutenkaacutech separace proteinům zaacutepornyacute naacuteboj jehož velikost je (s jistou aproximaciacute) uacuteměrnaacute velikosti molekul resp jejich relativniacute molekuloveacute hmotnosti (poměr naacutebojhmotnost je viacutecemeacuteně konstantniacute) [34] Dodecylsulfaacutet sodnyacute se vaacuteže na proteiny v poměru 14 g SDS na 1 g proteinu vznikajiacute tak asociaacutety ktereacute majiacute stejnyacute naacuteboj a přibližně tyčinkovyacute tvar [33]

344 2D elektroforeacuteza

K dokonalejšiacute separaci biacutelkovin se využiacutevaacute kombinace izoelektrickeacute fokusace (IEF)

s gelovou elektroforeacutezou ndash dvojrozměrnaacute elektroforeacuteza (2DE) Tato technika poskytuje efektivniacute separaci komplexniacute směsi proteinů [35] Směs proteinů je nejprve separovaacutena IEF v přiacutetomnosti močoviny na tzv stripu gelu (proteiny migrujiacute do bodu kde nemajiacute žaacutednyacute naacuteboj pH v tomto bodě odpoviacutedaacute jejich isoelektrickeacutemu bodu pI) Naacutesledně je IEF gel přenesen na gel pro SDS-PAGE (přiacutedavek rekukčniacuteho činidla a dodecylsulfaacutetu sodneacuteho analyzovaneacute proteiny denaturujiacute a propůjčiacute jim negativniacute naacuteboj potřebnyacute pro jejich elektroforetickou migraci v elektrickeacutem poli) a elektroforeacuteza proběhne ve směru 2 dimenze (90o vůči 1 dimenzi) [3670] Proteiny migrujiacute ve druheacutem rozměru v zaacutevislosti na sveacute velikosti Rutinně lze takto separovat 1000 ndash 2000 proteinů v zaacutevislosti na formaacutetu gelu

Poprveacute byla tato technika popsaacutena OrsquoFarrelem [63] a Klosem [64] už v roce 1975 ale opravdu širokeacuteho rozšiacuteřeniacute dosaacutehla až v posledniacutech letech Jde o instrumentaacutelně naacuteročnou techniku jejiacutemž vyacutesledkem je dvourozměrnaacute mapa skvrn proteinů jejichž polohu v jednom směru určuje izoelektrickyacute bod a ve druheacutem směru ji determinuje jejich elektroforetickaacute pohyblivost kteraacute je funkciacute jejich molekulovyacutech hmotnostiacute [70]

345 Detekce a vizualizace biacutelkovin v gelech

Pro vizualizaci separovanyacutech proteinů se použiacutevaacute několik technik barveniacute Mezi

nejpoužiacutevanějšiacute techniky barveniacute ktereacute se lišiacute citlivostiacute a naacuteročnostiacute provedeniacute patřiacute barveniacute modryacutem barvivem Commassie Brilliant Blue a barveniacute střiacutebrem Barveniacute musiacute byacutet citliveacute aby bylo dosaženo i vizualizace proteinů s niacutezkyacutemi molekulovyacutemi hmotnostmi ale zaacuteroveň musiacute byacutet vhodneacute i pro dalšiacute MS analyacutezu [37]

23

1) Commassie Brilliant Blue (CBB) Je nespecifickeacute kvantitativniacute barveniacute Vykazuje různou uacutečinnost na různeacute proteiny Barviacute

lysin arginin histidin tyrozin (tryptofan leucin) Jde o citlivou a jednoduchou metodu barveniacute proteinů Princip barveniacute je ten že protein musiacute byacutet zafixovaacuten v gelu [37] Gel se ponořiacute do roztoku barvy a nechaacute se několik hodin miacuterně třepat Po obarveniacute se omyje barva navaacutezanaacute na pozadiacute Na gelu zůstane barva jen tam kde se navaacutezala na fixovanyacute protein - Klasickaacute CBB (CBB-R250)

Barveniacute se provaacutediacute v kyseleacutem prostřediacute (7 kyselina octovaacute) v přiacutetomnosti alkoholu Citlivost je asi 50-100 ng Odbarvovaacuteniacute se provaacutediacute roztokem methanolu a kyseliny octoveacute Vyacutehodou tohoto typu barveniacute je niacutezkaacute cena - Koloidniacute CBB (CBB-G250)

Je citlivějšiacute než klasickaacute CBB jejiacute citlivost je 10-30 ng Vyacutehodou je dobraacute kvantifikace nevyacutehodou středně vysokaacute cena Koloidniacute barveniacute se provaacutediacute 008 CBB v prostřediacute 8 siacuteranu amonneacuteho 16 kyseliny fosforečneacute a 20 (vv) methanolu Odbarvuje se destilovanou vodou

2) Barveniacute střiacutebrem Nejednaacute se o kvantitativniacute barveniacute Ag+ ionty se v proteinech vaacutežou na -SH a -COOH

skupiny Barviacute lysin arginin histidin tyrozin tryptofan Každyacute protein se tedy střiacutebrem barviacute v zaacutevislosti na aminokyselinoveacutem složeniacute Uvaacutediacute se že je až 50 až 100 kraacutet citlivějšiacute než Commassie ale samotneacute barveniacute je mnohem obtiacutežnějšiacute Jednaacute se o viacutecekrokovyacute postup [38] Principem je redukce dusičnanu střiacutebrneacuteho na kovoveacute střiacutebro Proteinoveacute paacutesy se vizualizujiacute jako žluteacute oranžoveacute přes hnědou až k černyacutem Je to daacuteno rozptylem světla na vyredukovanyacutech čaacutestečkaacutech střiacutebra Existuje několik variant barveniacute ale pouze některeacute z nich jsou vhodneacute pro MALDI-MS [39] 35 Hmotnostiacute spektrometrie

Jednou ze zaacutekladniacutech charakteristickyacutech vlastnostiacute kteraacute definuje laacutetky niacutezkomolekulaacuterniacute i makromolekulaacuterniacute je jejich molekulovaacute hmotnost Hmotnostniacute spektrometrie je metoda jejiacutemž principem je rozděleniacute iontů podle jejich tzv efektivniacutech hmotnostiacute (mz kde m je hmotnost iontu a z je jeho naacuteboj) Zaacutekladniacute uspořaacutedaacuteniacute hmotnostniacuteho spektrometru obsahuje tři kliacutečoveacute součaacutesti zdroj iontů (kteryacute poskytuje ionty analyzovanyacutech sloučenin) jejich analyzaacutetor (kteryacute pomociacute elektromagnetickeacuteho pole třiacutediacute ionty na zaacutekladě rozdiacutelů v hodnotě mz) a detektor Hmotnostniacute spektrometrie je rychlaacute citlivaacute specifickaacute metoda s jednoduchou interpretaciacute dat Principem je ionizace laacutetek (analytů) v evakuovaneacute komoře jejich převedeniacute do plynneacute faacuteze a pak rozděleniacute buď v prostoru nebo v čase podle poměru jejich hmotnosti a naacuteboje

Kvantitativniacute analyacuteza spočiacutevaacute v měřeniacute počtu určityacutech iontů (plochy nebo vyacutešky jejich piacuteku) Zaacuteznam četnosti vyacuteskytu jednotlivyacutech iontů se pak nazyacutevaacute hmotnostniacute spektrum

Ionizačniacute techniky jsou voleny dle charakteru analyzovaneacuteho vzorku a analytickyacutech požadavků Pro velkeacute molekuly je zapotřebiacute speciaacutelniacuteho způsobu ionizace aby nedochaacutezelo k fragmentaciacutem a byla možnaacute jejich ionizace v pevneacute faacutezi Technika na kterou je zaměřena

24

pozornost teacuteto praacutece je desorpce a ionizace laserem v přiacutetomnosti matrice (matrix-assisted laser desorptionionization MALDI) řadiacuteciacute se mezi tzv měkkeacute či šetrneacute ionizace

351 Hmotnostniacute spektrometrie s ionizaciacute laserem v přiacutetomnosti matrice

(MALDI-MS)

Matrice se k desorpci a ionizaci velkyacutech biomolekul začaly využiacutevat koncem osmdesaacutetyacutech let 20 stoletiacute Prvniacute použitiacute matrice ukaacutezali F Hillenkamp a M Karas pro analyacutezu biacutelkovin jako matrici použili kyselinu nikotinovou a laser s vlnovou deacutelkou 266 nm Prvniacute proteiny analyzovaneacute touto metodou byly chymotrypsinogen o hmotnosti 25 717 Da karboxypeptidasa A (34 472 Da) a cytochrom c (12 384 Da) [65] Za rozvoj analyacutezy biomolekul metodou laseroveacute desorpce a ionizace byla K Tanakovi udělena v roce 2002 Nobelova cena za chemii [66]

Matrice jsou kliacutečovou součaacutestiacute teacuteto hmotnostně spektrometrickeacute metody Přiacutetomnost martice nejčastěji slabeacute kyseliny zajišťuje desorpci a ionizaci velkyacutech molekul analytu aniž by přitom došlo k jejich fragmentaci Pro desorpci a ionizaci velkyacutech makromolekul je nutnaacute spraacutevnaacute kombinace vlnoveacute deacutelky a energie laseru chemickyacutech a fyzikaacutelniacutech vlastnostiacute matrice jako absorbance přestup tepla rozpustnosti v daneacutem rozpouštědle a na chemickeacute struktuře molekul analytu a použiteacute matrice [50] Přiacuteprava vzorku spočiacutevaacute v naneseniacute analytu s matriciacute (v poměru koncentraciacute cca 1104) na speciaacutelniacute kovovou destičku kde se nechajiacute společně vykrystalizovat Matrice absorbuje energii laseroveacuteho pulzu (1-10 ns) a šetrně ji předaacute molekulaacutem analytu (viz obraacutezek č5) Takto přednostně vznikajiacute v plynneacute faacutezi protonovaneacute molekuloveacute ionty analytu s vyacuterazně nižšiacute energiiacute než při ionizaci laserem v nepřiacutetomnosti matrice [40]

Obr č 5 Scheacutema Matrix Assisted Laser DesorptionIonization (MALDI)

25

Vyacutehodou tvorby pouze molekulovyacutech iontů je možnost aplikace metody při analyacuteze směsiacute kteraacute tak neniacute komplikovanaacute přiacutetomnostiacute viacutecečetnyacutech signaacutelů [41 42] Mechanismus ionizace neniacute doposud znaacutem Na průběh ionizace existujiacute dva naacutezory ndash ionty se tvořiacute buď v pevneacute faacutezi nebo až v plynneacutem skupenstviacute Ve skutečnosti probiacutehajiacute oba procesy a to v různě velkeacute miacuteře podle toho o jakyacute se jednaacute vzorek a jakaacute matrice byla použita [43] MALDI vynikaacute niacutezkyacutemi naacuteroky na spotřebu vzorku přiacuteprava krystalů matrice-analyt vyžaduje piko - až mikro - moly stanovovaneacute laacutetky [44]

352 Hmotnostniacute analyzaacutetor bdquodoby letuldquo (Time of flight - TOF)

Hmotnostniacute spektrometr pro MALDI se sklaacutedaacute z iontoveacuteho zdroje analyzaacutetoru

a vyhodnocovaciacuteho zařiacutezeniacute Dnešniacute přiacutestroje jsou kompletně ovlaacutedaacuteny a řiacutezeny počiacutetačem MALDI je v současnosti nejčastěji použiacutevaacutena s hmotnostniacutem analyzaacutetorem TOF (Time of flight analyzaacutetor doby letu) Ionty analytu jsou urychleny silnyacutem elektrickyacutem polem (25-30 kV) a přes uzemněnou mřiacutežku vstupujiacute do evakuovaneacute letoveacute trubice V tomto prostoru bez přiacutetomnosti siloveacuteho pole se ionty pohybujiacute rychlosti charakteristickou pro jejich hmotnost Na konci trubice je detekovaacutena doba letu jednotlivyacutech iontů kteraacute je za danyacutech podmiacutenek uacuteměrnaacute hodnotě (mz) a je tedy miacuterou hmotnosti analytu [45] Vztah mezi dobou letu a hmotnostiacute iontu je popsaacuten rovniciacute

Vez

mLt

2= (2)

kde t je doba letu L je draacuteha mezi iontovyacutem zdrojem a detektorem m hmotnost iontu z naacuteboj iontu e elementaacuterniacute naacuteboj iontu (160210-19 C) a V urychlovaciacute potenciaacutel elektrickeacuteho pole Ze vztahu vyplyacutevaacute že skutečně měřenou veličinou neniacute hmotnost ale poměr hmotnostnaacuteboj pro každyacute ion Typickeacute doby letu jsou v řaacutedech několika mikrosekund až několika stovek mikrosekund

Analyzaacutetor lze užiacutet ve dvou moacutedech Pozitivniacute moacuted měřiacute kladneacute ionty negativniacute moacuted zaacuteporneacute ionty Analyzaacutetor pracuje v lineaacuterniacutem nebo reflektronoveacutem uspořaacutedaacuteniacute (viz obraacutezek č6) V prvniacutem přiacutepadě (lineaacuterniacute uspořaacutedaacuteniacute) jsou ionty detekovaacuteny na konci přiacutemeacuteho letu Lineaacuterniacute moacuted se použiacutevaacute pro měřeniacute proteinů maacute nižšiacute rozlišeniacute a je vhodnyacute pro měřeniacute velkyacutech molekul Analyzaacutetor doby letu byl vyvinut v padesaacutetyacutech letech 20 stoletiacute V hmotnostniacute spektrometrii velkyacutech molekul jej poprveacute použily MacFarlane a Torgerson v roce 1976 kteřiacute jako iontovyacute zdroj použily desorpci plazmou 252Cf

V druheacutem přiacutepadě (reflektronoveacute uspořaacutedaacuteniacute) je zapojeno iontoveacute zrcadlo ktereacute zlepšuje rozlišeniacute tiacutem že dopadajiacuteciacute ionty odraacutežiacute ke zpaacutetečniacutemu letu po miacuterně odkloněneacute draacuteze k excentricky umiacutestěneacutemu detektoru Sestaacutevaacute ze seacuterie mřiacutežek nebo kruhovyacutech elektrod Na ně je vloženo napětiacute ktereacute postupně roste až k hodnotě o něco většiacute než je urychlovaciacute napětiacute za iontovyacutem zdrojem Rozlišeniacute se zvyšuje jednak prodlouženiacutem draacutehy letu [46] jednak zaostřovaciacutem efektem neboť ionty se stejnyacutem poměrem hmotnostnaacuteboj ale vyššiacute kinetickou energiiacute proniknou hlouběji do reflektronu čiacutemž se prodloužiacute doba letu vůči iontům s nižšiacute kinetickou energiiacute [47] Rozšiacuteřeniacute piacuteků iontů způsobeneacute počaacutetečniacutem rozptylem kinetickeacute energie je nepřiacuteznivyacutem jevem kteryacute je pomociacute reflektronoveacuteho uspořaacutedaacuteniacute korigovaacuten To se

26

projeviacute předevšiacutem u velkyacutech iontů Přesnost stanoveniacute molekuloveacute hmotnosti zaacutevisiacute na velikosti analytu Reflektronovyacute moacuted použiacutevaacuteme na měřeniacute peptidů

U polypeptidů s molekulovou hmotnostiacute většiacute než 40 kDA lze dosaacutehnout přesnosti cca 001 u biopolymerů nad touto hmotnostiacute je běžnaacute přesnost 01 ndash 02 Největšiacute přednostiacute analyzaacutetoru TOF je velkyacute rozsah detekovanyacutech hmotnostiacute (do cca 1 MDa) Kalibrace hmotnostniacuteho spektrometru se provaacutediacute pomociacute definovanyacutech referenčniacutech sloučenin buď metodou vnitřniacuteho standardu (přidaacutevaacuteniacutem ke směsi analytu a matriciacute) většinou však oddělenou analyacutezou jako vnějšiacute standard

Obr č 6 Scheacutema hmotnostniacuteho spektrometru MALDI-TOF

V oblasti biologickyacutech věd je MALDI-TOF hmotnostniacute spektrometrie v současneacute době

využiacutevaacutena předevšiacutem k detekci a identifikaci proteinů [48] sekvenci DNA identifikaci bodovyacutech mutaciacute sekvenaci peptidů [49] a identifikaci bakteriaacutelniacutech kmenů

353 Tandemovyacute hmotnostniacute analyzaacutetor TOFTOF

Time-of-flightTime-of-flight analyzaacutetor neboli tandemovyacute TOF MS je tvořen dvěma TOF akceleraacutetory mezi ktereacute je umiacutestěna kolizniacute cela Tandemoveacute spojeniacute dvou nebo viacutece hmotnostniacutech analyzaacutetorů bylo původně určeno vyacutehradně pro studium fragmentačniacutech iontů organickyacutech molekul [32] Tato metoda je charakteristickaacute svou citlivostiacute a vyššiacute selektivitou

Celaacute procedura začiacutenaacute tiacutem že ze skupiny iontů různeacuteho druhu přiacutetomnyacutech po ionizaci v iontoveacutem zdroji vybereme iont kteryacute patřiacute molekule kterou chceme identifikovat Mluviacuteme tak o tzv rodičovskeacutem iontu (k selekci a izolaci rodičovskeacuteho iontu dochaacuteziacute v prvniacutem akceleraacutetoru) Vybranyacute iont daacutele postupuje do kolizniacute cely kde dochaacuteziacute ke sraacutežkaacutem s molekulami kolizniacuteho plynu (např helium argon) (Collision-Induced Decomposition CID)

27

Dochaacuteziacute k rozsaacutehlyacutem fragmentaciacutem a generovaneacute fragmentovanaacute ionty tzv dceřineacute ionty jsou ve druheacutem TOF akceleraacutetoru detekovaacuteny (viz obrč7)

Obrč7 Scheacutema tandemoveacute hmotnostniacute spektrometrie

354 Matrice

Pro měřeniacute daneacuteho analytu je důležiteacute najiacutet vhodnou matrici Matrice hrajiacute kliacutečovou roli u

mnohyacutech hmotnostně spektrometrickyacutech metod [50] Vhodnost matrice spočiacutevaacute hlavně ve schopnosti ziacuteskat co nejkvalitnějšiacute spektrum s velkou reprodukovatelnostiacute vyacutesledků a co největšiacutem rozlišeniacutem Velmi zjednodušeně platiacute že pro hydrofilniacute laacutetky jsou vhodneacute hydrofilniacute matrice a opačně ndash pro hydrofobniacute laacutetky hydrofobniacute matrice Často to vyplyacutevaacute z nutnosti naleacutezt vhodneacute rozpouštědlo ale existujiacute i dalšiacute faktory ktereacute nejsou dosud obecně vysvětleny [51] Matrice by měla byacutet rozpustnaacute ve stejneacutem rozpouštědle jako vzorek aby během krystalizace byly tyto laacutetky v kontaktu a takeacute aby nedochaacutezelo k inkorporaci molekul analytu do krystalů matrice Matrice musiacute absorbovat vlnovou deacutelku zaacuteřeniacute laseru Se vzorkem by neměla reagovat ndash v přiacutepadě peptidů nelze napřiacuteklad použiacutevat matrice ktereacute by mohly oxidovat sulfo-skupiny cysteinu a metioninu nebo aldehydy vytvaacuteřejiacuteciacute s aminokyselinami Schiffovy baacuteze V přiacutetomnosti laseru musiacute matrice vykazovat fotostabilitu Daacutele je u matric důležitaacute přiacutetomnost -OH skupin ktereacute zajišťujiacute ionizaci analytu [52] Vyacuteběr matrice je tedy poměrně složityacute a ve většině přiacutepadů je nutneacute optimaacutelniacute podmiacutenky hledat Rovněž se voliacute matrice dle typu analyzovanyacutech iontů Jinaacute matrice se použije pro měřeniacute iontů v pozitivniacutem moacutedu jinaacute pro měřeniacute iontů v negativniacute moacutedu nebo pro analyacutezu fragmentovyacutech iontů v reflektronoveacutem uspořaacutedaacuteniacute [43] Uacutespěšnost analyacutezy spočiacutevaacute takeacute v tom jakeacute krystaly tvořiacute matrice samotnaacute jakeacute krystaly tvořiacute ve směsi s analytem a za jakyacutech podmiacutenek krystalizace probiacutehaacute Optimaacutelniacute je homogenniacute krystalizace jejiacutež krystaly jsou rovnoměrně rozmiacutestěny po celeacute naneseneacute ploše Takovaacute matrice zajišťuje reprodukovatelneacute vyacutesledky a lze ji použiacutet i pro kvantitativniacute analyacutezy [53 54]

bull Matrice použiteacute pro analyacutezu proteinů a peptidů v teacuteto praacuteci a) Kyselina sinapovaacute (35-dihydroxy-4-hydroxyskořicovaacute kyselina SA)

Tato matrice se použiacutevaacute pro analyacutezu peptidů a biacutelkovin za použitiacute UV laseru S vyacutehodou se použiacutevaacute u proteinů s hmotnostiacute většiacute než 10 000 Da Literatura [45] uvaacutediacute meze použitiacute 400 ndash 100 000 Da Pro analyacutezu proteinů je připravovaacuten roztok matrice ve směsi acetonitrilu s vodou Při použitiacute kyseliny sinapoveacute nerušiacute analyacutezu ani relativně vysokeacute koncentrace anorganickyacutech a organickyacutech sloučenin (koncentrace v rozmeziacute 2 ndash 200 mmoll-1)

28

Obr č 8 Kyselina sinapovaacute M = 22422 Da b) Kyselina α-kyano-4-hydroxyskořicovaacute (HCCA)

Matrice se uplatňuje při identifikaci biacutelkovin metodou peptidoveacuteho mapovaacuteniacute (peptide mass fingerprinting) Jejiacute hlavniacute použitiacute se zaměřuje na analyacutezu niacutezkomolekulaacuterniacutech peptidů o hmotnosti menšiacute než 10 000 Da HCCA vytvaacuteřiacute několik mnohonaacutesobně nabityacutech molekulaacuterniacutech iontů [55] Reprodukovatelnost vyacutesledků při použitiacute teacuteto matrice velmi zaacutevisiacute na složeniacute roztoku a způsobu přiacutepravy vzorku [56]

Obr č 9 Kyselina α-kyano-4-hydroxyskořicovaacute M = 18917 Da bull Vyacuteběr substraacutetu

Role substraacutetu v procesu ještě neniacute dostatečně prostudovaacutena Zaacutekladniacute vlastnostiacute substraacutetu je vodivost Vodivost je důležitaacute pro zachovaacuteniacute homogenity elektrickeacuteho pole při vypuzovaacuteniacute iontů z iontoveacuteho zdroje směrem k analyzaacutetoru Destičky na MALDI jsou zpravidla vyraacuteběny z antikorozniacute oceli nebo hliniacuteku Tyto materiaacutely jsou charakteristickeacute inertnostiacute vůči použityacutem matriciacutem a rozpouštědlům Destičky musiacute byacutet snadno čistitelneacute do hladkeacuteho povrchu kteryacute je důležityacute pro dobreacute rozlišeniacute a vysokou přesnost určovaacuteniacute hmotnosti bull Metody nanaacutešeniacute vzorku na destičku

I způsob naneseniacute vzorku na destičku vyacuterazně ovlivňuje vyacutesledky měřeniacute Použiacutevajiacute se zpravidla dva způsoby Prvniacutem způsobem je tzv metoda Dried-Dropletldquo (DD) kteraacute byla zavedena autory Karasem a Hillenkampem [57] Roztok vzorku je smiacutechaacutem s matriciacute a pak je směs v množstviacute 05 až 2 microl nanesena na destičku a usušena při laboratorniacute teplotě

29

Pro urychleniacute můžeme použiacutet proud vzduchu nebo vakuum Vyacutehoda teacuteto metody nanaacutešeniacute je jejiacute jednoduchost Naopak nevyacutehoda je vylučovaacuteniacute krystalů matrice a vzorku na obvodu vysušeneacute kapky

Druhyacute způsob je označovaacuten jako Thin layerrdquo (TL) Na destičku nanaacutešiacuteme matrici a vzorek odděleně Nejprve je nanesena matrice a až po odpařeniacute rozpouštědla je nanesen roztok analytu Přednostiacute teacuteto metody je vytvořeniacute krystalickeacute vrstvy 10 až 100 kraacutet tenčiacute než u předchoziacute metody což vede ke zlepšeniacute rozlišeniacute a spraacutevnosti určeniacute molekuloveacute hmotnosti Přiacuteliš tenkaacute vrstva však může vest k rychleacute spotřebě materiaacutelu během laserovyacutech pulsů což je nežaacutedouciacute [58]

355 Identifikace proteinů metodou peptidoveacuteho mapovaacuteniacute

Čiacutem je molekulovaacute hmotnost proteinů vyššiacute tiacutem je analyacuteza obtiacutežnějšiacute Pro identifikaci proteinů se použiacutevaacute jejich specifickeacute chemickeacute či enzymatickeacute štěpeniacute a měřeniacute hmotnostniacutech spekter proteolytickyacutech štěpů Identifikace začiacutenaacute separaciacute proteinu z vyizolovaneacute směsi Velmi důležitaacute je uacutečinnaacute separace Nejčastěji jednorozměrnou nebo dvojrozměrnou elektroforeacutezou gel se obarvi přiacuteslušneacute zoacuteny se vyřiacuteznou provede se redukce (obvykle dithiotreitolem) alkylace (jodacetamidem) a naacutesledně specifickeacute štěpeniacute nejčastěji se využiacutevaacute trypsin (štěpiacute řetězec za bazickyacutemi aminokyselinami Lys a Arg) nebo chymotrypsin (štěpiacute za aromatickyacutemi aminokyselinami) [4867] Naacutesledně se provede hmotnostniacute analyacuteza vznikleacute směsi peptidů Ziacuteskaacute se tak řada hodnot ktereacute se zadajiacute databaacuteze (např Mascot Protein Prospector Proteomics) Databaacuteze pak hledajiacute peptidoveacute štěpy ktereacute majiacute stejnou hmotnost jako zadaneacute hodnoty Pokud je primaacuterniacute struktura studovaneacuteho proteinu v databaacutezi zadaacutena podařiacute se ji identifikovat

356 Sekvenovaacuteniacute proteinů a peptidů

Pomociacute současnyacutech hmotnostniacutech spektrometrů je možneacute stanovit hmotnost peptidu či

proteinů s velkou přesnostiacute ale ani takto velkaacute přesnost často nestačiacute k jejich jednoznačneacute identifikaci Tu lze zjistit sekvenovaacuteniacutem polypeptidů pomociacute tandemoveacute hmotnostniacute spektrometrie (MSMS) Peptidu jehož sekvenci chceme určit se dodaacute energie kteraacute vyvolaacute fragmentaci a naacutesledně se změřiacute hmotnostniacute spektrum fragmentů

Peptidy a proteiny jsou většinou lineaacuterniacute polymery přerušeniacutem jedineacute kovalentniacute vazby v řetězci vzniknou různeacute typy iontů v zaacutevislosti na miacutestě přerušeniacute peptidoveacuteho řetězce Přerušeniacutem vzniknou dvě čaacutestice z nichž jedna obsahuje N- a druhaacute C-koncovou čaacutest peptidu K tomu aby byla vzniklaacute čaacutestice detegovaacutena je potřeba aby nesla naacuteboj Jestliže je naacuteboj zadržen N-koncovou čaacutestiacute peptidu ion je klasifikovaacuten jako a b nebo c je-li zadržen C-koncovou čaacutestiacute je ion klasifikovaacuten jako x y nebo z a to podle vazby v niacutež došlo k fragmentaci (viz obrč10) [67]

30

Obrč10 Scheacutema fragmentace

Typy fragmentace

a) Fragmentace CID (Collision-induced decomposition) V současneacute době je nejčastěji použiacutevanou metodou fragmentace peptidu tzv koliziacute

vyvolanaacute disociace a to zejmeacutena ve spojeniacute s trojityacutem kvadrupoacutelem nebo TOF-TOF jako hmotnostniacutemi analyzaacutetory U TOF-TOF hmotnostniacuteho spektrometru prvniacute TOF analyzaacutetor vybiacuteraacute prekursoroveacute ionty ktereacute vstupujiacute do kolizniacute cely mezi TOF analyzaacutetory a spektrum fragmentů je měřeno druhyacutem TOF analyzaacutetorem Množstviacute a typ fragmentovyacutech iontů zaacutevisiacute na hodnotě kolizniacute energie (až 800 eV) Čiacutem je tato hodnota vyššiacute tiacutem viacutece fragmentovyacutech iontů lze ziacuteskat Při vyššiacutech hodnotaacutech se objevujiacute i fragmenty typu x a a ktereacute se při nižšiacutech hodnotaacutech nevygenerujiacute [68]

b) Fragmentace ve zdroji (In-source decay ISD)

K rozpadu molekul na fragmenty dochaacuteziacute přiacutemo v iontoveacutem zdroji Fragmentace je vyvolanaacute vysokou excitaciacute analyzovanyacutech molekul při ionizaci Jde o vznik tzv fokusovanyacutech iontů Tiacutemto způsobem lze analyzovat pouze čisteacute laacutetky protože interpretace spekter ve kteryacutech by se vyskytovaly fragmenty několika prekurzorů by byla velice obtiacutežnaacute U MALDI lze ISD dosaacutehnout zvyacutešenyacutem vyacutekonem laseru [67] Fragmenty ISD lze detekovat v lineaacuterniacutem uspořaacutedaacuteniacute

c) Fragmentace za zdrojem (Post-source decay)

Využiacutevaacute jevu rozpadu iontů za iontovyacutem zdrojem Ionty ktereacute opustiacute iontovyacute zdroj majiacute

stejnou kinetickou energii Kvůli rozdiacutelům v desorpčniacutech a ionizačniacutech procesech majiacute různou vnitřniacute energii Během jejich letu v oblasti bez elektrickeacuteho pole se mohou některeacute ionty s vysokou vnitřniacute energiiacute rozpadnout Vznikleacute fragmenty se pohybujiacute stejnou rychlostiacute jako původniacute iont jednaacute se o tzv metastabilniacute či nefokusovaneacute ionty ktereacute lze detekovat pouze v reflektronoveacutem uspořaacutedaacuteniacute Jejich množstviacute je přiacutemo uacuteměrneacute době po kterou se zdržujiacute

31

přiacutemo ve zdroji (pro ziacuteskaacuteniacute PSD spektra je tedy nutneacute minimalizovat ISD fragmentaci) V reflektronu dochaacuteziacute k jejich odraženiacute v různyacutech potenciaacutelech elektrickeacuteho pole Fragmentovaneacute ionty vystupujiacute z reflektronu se stejnou rychlostiacute jako ionty původniacute ale v různyacutech časech Při sekvenaci peptidu se postupuje naacutesledujiacuteciacutem způsobem trubiciacute detektoru se nechaacute prochaacutezet pouze iont kteryacute chceme fragmentovat Ostatniacute ionty se odchyacuteliacute z draacutehy pomociacute detektoru (iontoveacute braacutenyldquo) nevstupujiacute do detektoru Ve vyacutesledneacutem spektru lze pozorovat fragmenty vybraneacuteho iontu ndash peptidu a podle jejich hmotnostiacute lze určit jeho sekvenci [49] Techniku MALDI PSD lze použiacutevat ke strukturniacute analyacuteze sloučenin největšiacute oblast použitiacute metody představuje určovaacuteniacute a ověřovaacuteniacute sekvence peptidů

32

4 EXPERIMENTAacuteLNIacute ČAacuteST 41 Seznam chemikaacuteliiacute Aceton Lachema Neratovice ČR Acetonitril (ACN) Fluka Buchs Švyacutecarsko Akrylamid Lachema Neratovice ČR Biolyt 310 BIO-RAD Hercules CA USA Bromofenolovaacute modř Sigma - Aldrich Schnelldorf Německo Commassie Brilliant Blue R (CBB R-250) Sigma - Aldrich Schnelldorf Německo Deionizovanaacute voda (DV) Dithiothreitol (DTT) Sigma - Aldrich Schnelldorf Německo Dodecylsulfaacutet sodnyacute (SDS) Sigma - Aldrich Schnelldorf Německo Dusičnan střiacutebrnyacute (AgNO3) Fluka Buchs Švyacutecarsko Ethanol (EtOH) Merck Kvas Darmstadt Německo Formaldehyd Sigma - Aldrich Schnelldorf Německo Glycerol Lachema Neratovice ČR 3-[(3-cholamidopropyl)dimethylamonio]propan-1-sulfonaacutet (CHAPS) BIO-RAD Hercules CA USA Jodoacetamid Sigma - Aldrich Schnelldorf Německo Kyselina mravenčiacute Sigma - Aldrich Schnelldorf Německo Kyselina octovaacute Penta Chrudim ČR Kyselina trifluorooctovaacute (TFA) Fluka Buchs Švyacutecarsko Kyselina trichloroctovaacute (TCA) Fluka Buchs Švyacutecarsko 2-Merkaptoethanol Sigma - Aldrich Schnelldorf Německo Methanol Penta Chrudim ČR Močovina BIO-RAD Hercules CA USA N Nrsquo- Mythylenbisakrylamid (Bisakrylamid) Sigma - Aldrich Schnelldorf Německo N N Nrsquo Nrsquo- Tetramethylethyldiamin (TEMED) Sigma - Aldrich Schnelldorf Německo Persiacuteran amonnyacute Sigma - Aldrich Schnelldorf Německo Thiosiacuteran sodnyacute (Na2S2O3) Lachema Neratovice ČR Tris Sigma - Aldrich Schnelldorf Německo Uhličitan sodnyacute (Na2CO3) Lachema Neratovice ČR

33

42 Materiaacutel

Jako vyacutechoziacute materiaacutel pro extrakci proteinů byly použity pomleteacute obilky ječmene (zrno zelenyacute slad slad a vzorky odebraneacute v průběhu sladovaacuteniacute ndash prvniacute až paacutetyacute den) Pro uacutečel teacuteto praacutece byl použit ječmen odrůdy Jersey Pozornost byla soustředěna na biacutelkoviny rozpustneacute ve vodě 43 Extrakce biacutelkovin pro 1D elektroforeacutezu v polyakrylamidoveacutem gelu

Navaacutežka ječmene 50 mg byla extrahovaacutena deionizovanou vodou čiacutemž došlo k extrakci proteinů rozpustnyacutech ve vodě Extrakce byla provedena dvakraacutet s 05 ml deionizovaneacute vody na třepačce po dobu 30 minut při laboratorniacute teplotě Extrakty byly odstředěny při 13 000 g po dobu 10 minut Po odstředěniacute byly extrakty spojeny a vysušeny na vakuoveacute odparce

44 1D elektroforeacuteza v polyakrylamidoveacutem gelu (1D PAGE)

Vysušeneacute extrakty byly rozpuštěny ve 150 microl vzorkovaciacuteho pufru (Laemmli Sample Buffer BIO-RAD 625 mM Tris-HCl (pH 68) 25 glycerol 2 dodecylsulfaacutet sodnyacute 001 bromofenolovaacute modř) kteryacute byl smiacutechaacuten v poměru 191 s β-merkaptoethanolem Vzorek byl povařen 10 minut a množstviacute tohoto vzorku (množstviacute vzorku v microl se voliacute podle velikosti gelu) bylo naneseno na polyakrylamidovyacute gel Pro separaci biacutelkovin byly použity komerčniacute gely od firmy BIO-RAD a naleacutevaneacute gely 125 15 (Tris ndash HCl BIO-RAD) a gradientoveacute gely (4 - 20 Tris ndash HCl BIO-RAD) Rozměry naleacutevanyacutech gelů byly kompetentniacute s použitou aparaturou rozměry gelu pro aparaturu Mini PROTEAN 3 Cell od firmy BIO-RAD byla 70 mm x 100 mm x 1 mm a rozměry gelu pro aparaturu OWL Separation Systems byla 140 mm x 120 mm x 15 mm - Naleacutevaacuteniacute gelu

Gel připravenyacute na elektroforeacutezu se sklaacutedal ze dvou čaacutestiacute separačniacuteho gelu a gelu zaostřovaciacuteho

Separačniacute gel (125 ) se připravil smiacutechaacuteniacutem 25 ml roztoku A (A30 g akrylamidu a 08 g bisakrylamidu bylo doplněno do 100 ml destilovanou vodou) 30 ml roztoku C (C 91 g TRIS byl rozpuštěno v 50 ml destilovaneacute vody dotitrovaacuteno koncentrovanou HCl na pH 88 a doplněno do 100 ml destilovanou vodou) 06 ml roztoku B (B 10 g dodecylsulfaacutelu sodneacuteho bylo doplněno do 100 ml destilovanou vodou) 38 ml destilovaneacute vody 20 microl TEMEDU a 015 ml roztoku F (F 01 g persiacuteranu amonneacuteho bylo doplněno do 1 ml destilovanou vodou) Takto připravenyacute gelovyacute roztok se nalil mezi dvě skla a nechal se polymerovat Po ztuhnutiacute se na vrstvu separačniacuteho gelu opatrně nalil gel zaostřovaciacute (vizobr č11)

Zaostřovaciacute gel se připravil smiacutechaacuteniacutem 18 ml roztoku A 5 ml roztoku D (D3 g TRIS byly rozpuštěny v 50 ml destilovaneacute vody roztok byl dotitrovaacuten koncentrovanou HCl na pH 68 a doplněn do 100ml destilovanou vodou) 01 ml roztoku B 3 ml destilovaneacute vody 10 microl TEMEDU a 015 ml roztoku F Roztoky A B a F byly stejneacute jako u separačniacuteho gelu

34

Ihned po nalitiacute zaostřovaciacuteho gelu byl opatrně vložen mezi skla hřebiacutenek kteryacute vytvořil v gelu jamky na daacutevkovaacuteniacute vzorků

Obr 11A 1-jamka v gelu pro vzorek 2-skla mezi ktereacute naleacutevaacuteme gel 3-polyakrylamidovyacute gel 4-plastoveacute spacery11B 1-naleacutevaacuteniacute separačniacuteho gelu 2-naleacutevaacuteniacute zaostřovaciacuteho gelu

3-hřebiacutenek 4- manipulace je provaacuteděna opatrně

Po ztuhnutiacute zaostřovaciacuteho gelu byl hřebiacutenek opatrně vytažen aby nedošlo k poškozeniacute jamek naacutesledně byl na gel nalit elektrodovyacute pufr (přiacuteprava elektrodoveacuteho pufru 3 g TRIS 144 g glycinu a 1 g dodecylsulfaacutetu sodneacuteho byl rozpuštěn v 1000 ml destilovaneacute vody) a do jamek byly pomociacute mikropipet naneseny vzorky do krajniacutech jamek byly zpravidla naneseny standardy (SDS-PAGE Molecular Weight Standards Low Range BIO-RAD nebo SDS-PAGE Molecular Weight Standards Broad Range BIO-RAD) Elektroforeacuteza byla provedena na aparatuře Mini PROTEAN 3 Cell od firmy BIO-RAD a na aparatuře Owl Separation Systems za konstantniacuteho napětiacute 180 V Po proběhnutiacute analyacutezy byl gel zafixovaacuten ve fixačniacutem roztoku (fixačniacute roztok byl zvolen podle způsobu barveniacute gelu) - Barveniacute gelu

K barveniacute biacutelkovin v gelu bylo použito barvivo Commassie Brilliant Blue R-250 a barvivo Commassie Brilliant Blue G-250

a) Barveniacute barvivem Commassie Brilliant Blue R-250 Před samotnyacutem barveniacutem byly proteiny v gelu fixovaacuteny fixačniacutem roztokem (45

methanol 5 kyselina octovaacute a destilovanaacute voda) a gel byl dvakraacutet omyt destilovanou vodou Naacutesledně byl vložen do barviciacute směsi (0025 Commassie Brilliant Blue R-250 40 methanol 7 kyselina octovaacute a destilovanaacute voda) ve ktereacute se za miacuterneacuteho třepaacuteniacute nechal barvit dokud nebyly vidět na gelu bandy standardů i vzorků Po obarveniacute se gel nechal 30 minut odbarvovat v odbarvovaciacutem roztoku č1 (40 methanol 7 kyselina octovaacute a destilovanaacute voda) a potom dokud nebylo pozadiacute průsvitneacute v odbarvovaciacutem roztoku č2 (7 kyselina octovaacute 5 methanol a destilovanaacute voda)

35

b) Barveniacute barvivem Commassie Brilliant Blue G-250 Před barveniacutem byl gel 20 minut fixovaacuten 12 roztokem kyseliny trichloroctoveacute a omyt

destilovanou vodou (třikraacutet po 5 minutaacutech) Samotneacute barveniacute barviciacutem roztokem (008 Commassie Brilliant Blue G-250 8 siacuteran amonnyacute 16 kyselina fosforečnaacute a 20 methanol byla použita komerčniacute barviciacute směs od firmy Fermentas Vilnius Litva) probiacutehalo za miacuterneacuteho třepaacuteniacute přes noc Po obarveniacute se gel odbarvoval destilovanou vodou tak dlouho dokud nebylo pozadiacute gelu průsvitneacute

45 Extrakce biacutelkovin pro 2D elektroforeacutezu v polyakrylamidoveacutem gelu

Pro 2D gelovou elektroforeacutezu byly použity stejneacute vzorky ječmene jako pro 1D gelovou elektroforeacutezu Navaacutežka 100 mg ječmene byla extrahovaacutena 05 ml deionizovaneacute vody na třepačce po dobu 2 hodin při laboratorniacute teplotě Extrakt byl odstředěn při 14 000 g po dobu 15 minut Po odstředěniacute byl supernatant přenesen do čisteacute mikrozkumavky Supernatant byl opět odstředěn při 14 000 g po dobu 5 minut

- Sraacuteženiacute

Naacutesledovalo sraacuteženiacute supernatantu ktereacute se provaacutedělo roztokem 10 kyseliny trichloroctoveacute v acetonu a 12 (wv) dithiothreitolu v poměru 13 Sraacuteženiacute probiacutehalo při teplotě -20 degC přes noc Po sraacuteženiacute byla sraženina odstředěna při 10 000 g po dobu 5 minut Supernatant byl odstraněn a sediment byl dvakraacutet promyt acetonem (centrifugace při 10 000 g po dobu 1 minuty) a naacutesledně vysušen na vakuoveacute odparce - Rehydratace

Vysušenaacute sraženina byla rozpuštěna v rehydratačniacutem pufru (8 M močovina 50 mM dithiothreitol 2 chaps 02 biolyt bromfenolovaacute modř) a naacutesledně odstředěna při 14 000 g po dobu 3 minut 125 microl takto vznikleacuteho supernatantu bylo přeneseno na strip gelu (lineaacuterniacute ReadyStrip IPG STRIP 7 cm 3-10 pI BIO-RAD) strip gelu byl pokryt kvůli vysychaacuteniacute 1 ml mineraacutelniacuteho oleje Rehydratace proběhla přes noc 46 2D elektroforeacuteza v polyakrylamidoveacutem gelu (2D PAGE) - Isoelektrickaacute fokusace

Po rehydrataci se strip gelu přenesl do fokusaacutetoru Fokusace byla provedena na fokusaacutetoru PROTEAN IEF Cell od firmy BIO-RAD Fokusace proběhla v několika na sebe navazujiacuteciacutech krociacutech [35] po dobu několika hodin Po fokusaci byl strip gelu uchovaacuten přes noc v mraziciacutem boxu při teplotě -20 degC - Ekvilibrace Strip gelu byl rozmražen na laboratorniacute teplotu a ekvilibrovaacuten v naacutesledujiacuteciacutech roztociacutech Nejprve po dobu 10 minut v roztoku prvniacuteho ekvilibračniacuteho pufru (6 M močovina 2 dodecylsulfaacutet sodnyacute 0375 M Tris-HCl (pH 88) 20 glycerol 2 (vv) dithiothreitol) potom dalšiacutech 10 minut v roztoku druheacuteho ekvilibračniacuteho pufru (6 M močovina 2 dodecylsulfaacutet sodnyacute 0375 M Tris-HCl (pH 88) 20 glycerol 25 (vv) jodoacetamid) Po proběhnutiacute ekvilibrace byl strip gelu umiacutestěn na separačniacute gel kde proběhla separace ve druheacutem směru Přechod mezi stripem gelu a separačniacutem gelem byl zprostředkovaacuten agaroacutezou

36

(viz obrč12) Do krajniacute jamky určeneacute pro standard byl nanesen standard (SDS-PAGE Molecular Weight Standards Low Range BIO-RAD nebo SDS-PAGE Molecular Weight Standards Broad Range BIO-RAD) Elektroforeacuteza byla provedena na aparatuře Mini PROTEAN 3 Cell od firmy BIO-RAD za konstantniacuteho napětiacute 180 V Po proběhnutiacute elektroforeacutezy byly proteiny v gelu 20 minut fixovaacuteny 12 roztokem kyseliny trichloroctoveacute omyty destilovanou vodou (třikraacutet po dobu 5 minut) a obarveny barviciacutem roztokem Commassie Brilliant Blue G-250

Obr12 1- isoelektricky fokusovanyacute strip gelu 2-SDS gel

47 Přiacuteprava vzorku pro hmotnostně spektrometrickou analyacutezu

Vzorky pro hmotnostně spektrometrickou analyacutezu byly připraveny podle naacutevodu

Shevchenka a Fernandez-Patrona [59 60]

- Vyřiacuteznutiacute proteinů z gelu Po separaci pomociacute 1D PAGE byl gel omyt ve vodě Skalpelem byly vyřiacuteznuty spoty

proteinů Byl vyřiacuteznut rovněž kousek gelu z oblasti kteraacute neobsahuje proteiny (kontrolniacute pokus ndash blank) Vyřiacuteznuteacute kousky gelu byly nakraacutejeny na kostičky o velikosti asi 1 x 1 mm a přeneseny do mikrozkumavky

- Omyacutevaacuteniacute kousků gelu

Kousky gelu byly omyty ve vodě a směsi acetonitrilvoda 11 vv (dvakraacutet po 15 minutaacutech) Použiteacute množstviacute rozpouštědla pro omytiacute bylo asi 30 microl ndash přibližně dvojnaacutesobek objemu gelu Po omytiacute byla kapalina odstraněna a kousky gelu byly převrstveny acetonitrilem Až se kousky gelu srazily (zbělaly a slepily k sobě) byl acetonitril odstraněn a kousky gelu byly rehydratovaacuteny 01 M hydrogenuhličitanem amonnyacutem (NH4HCO3) Po 5 minutaacutech byl přidaacuten stejnyacute objem acetonitrilu aby byly kousky inkubovaacuteny v roztoku 01 M NH4HCO3acetonitril 11 vv Po 15 minutaacutech byla odstraněna veškeraacute kapalina a kousky gelu byly vysušeny na vakuoveacute odparce

- Redukce a alkylace

K vysušenyacutem kouskům gelu bylo přidaacuteno 30 microl roztoku dithiotreitolu v 01 M NH4HCO3 Proteiny byly redukovaacuteny 45 minut při 56 degC Mikrozkumavky byly ochlazeny na laboratorniacute teplotu Kapalina byla odstraněna a nahrazena stejnyacutem objemem 55 mM roztoku

37

jodoacetamidu v 01 M NH4HCO3 Mikrozkumavky byly ponechaacuteny 30 minut ve tmě při laboratorniacute teplotě Po odstraněniacute roztoku jodoacetamidu byly kousky promyacutevaacuteny směsiacute 01 M NH4HCO3acetonitril 11 vv (dvakraacutet po 15 minutaacutech) Toto omyacutevaacuteniacute bylo opakovaacuteno až do odbarveniacute kousků gelu

- Štěpeniacute v gelu

Kousky gelu byly vysušeny ve vakuoveacute odparce a při 4 degC převrstveny 30 microl roztoku obsahujiacuteciacuteho 50 mM NH4HCO3 5 mM CaCl2 a 12 ngml chymotrypsinu Roztok byl čaacutestečně absorbovaacuten gelem Po 45 minutaacutech byl supernatant odstraněn a kousky gelu převrstveny stejnyacutem roztokem avšak neobsahujiacuteciacutem chymotrypsin (tedy roztokem obsahujiacuteciacutem pouze 50 mM NH4HCO3 a 5 mM CaCl2) Enzymatickeacute štěpeniacute probiacutehalo při 37 degC přes noc

- Extrakce peptidů

Během nočniacuteho enzymatickeacuteho štěpeniacute došlo k uvolněniacute hlavniacuteho podiacutelu peptidů z gelu do roztoku Zbyleacute peptidy byly extrahovaacuteny z gelu přidaacuteniacutem dostatečneacuteho objemu 25 mM NH4HCO3 k převrstveniacute kousků gelů (asi 30 microl) Po 15 minutaacutech byl přidaacuten stejnyacute objem acetonitrilu a po dalšiacutech 15 minutaacutech byl veškeryacute roztok odstraněn (nevyhazuje se přidaacutevaacute se k hlavniacutemu peptidoveacutemu podiacutelu) Extrakce byla ještě dvakraacutet opakovaacutena směsiacute 5 kyselina mravenčiacuteacetonitril 11 vv supernatant byl smiacutechaacuten s hlavniacutem peptidovyacutem podiacutelem a vysušen na vakuoveacute odparce - Analyacuteza peptidů hmotnostniacute spektrometriiacute

Peptidy byly rozpuštěny ve 20 microl 01 kyseliny trifluorooctoveacute a naacutesledně přečištěny pomociacute pipetovyacutech špiček ZipTip C18 (Millipore USA) a metodou TL (thin layer postupneacute nanaacutešeniacute) naneseny na destičku Nanaacutešeneacute množstviacute bylo 03 microl vzorku a 03 microl matrice Jako matrice byla použita kyselina α-kyano-4-hydroxyskořicovaacute (koncentrace 6 mg ml rozpouštědlo acetonitril01 kyselina trifluorooctovaacute 11 vv) 48 Hmotnostniacute spektrometrie

Hmotnostniacute spektra byla měřena na hmotnostniacutem spektrometru MALDI TOFTOF (4700 Proteomics Analyzer Applied Biosystems Framingham USA) Přiacutestroj je vybaven NdYAG laserem emitujiacuteciacutem zaacuteřeniacute o vlnoveacute deacutelce 355 nm laseroveacute pulsy jsou vysiacutelaacuteny v časovyacutech periodaacutech 3-7 ns MSMS experimenty byly provaacuteděny na stejneacutem přiacutestroji a jako kolizniacute plyn byla použita atmosfeacutera 49 Přiacuteprava vzorků na měřeniacute intaktniacutech proteinů

Navaacutežka ječmene 60 mg byla extrahovaacutena deionizovanou vodou aby došlo k uvolněniacute proteinů rozpustnyacutech ve vodě Extrakce byla provedena 1 ml deionizovaneacute vody na třepačce po dobu 45 minut při laboratorniacute teplotě Extrakty byly odstředěny při 13 000 g po dobu 10 minut Po odstředěniacute byly extrakty koncentrovaacuteny na vakuoveacute odparce asi na polovinu původniacuteho množstviacute (cca 05 ml) Naacutesledně byly koncentrovaneacute extrakty přeneseny pomociacute mikropipet do nanosepů (Nanosep Centrifugal Devices 3000 Da MWCO- molecular weight cut off od firmy Pall Life Sciences Dreieich Německo) nanosepy byly odstředěny při

38

10 000 g po dobu 20 minut (Nanosep je odstřediveacute zařiacutezeniacute obsahujiacuteciacute membraacutenu kteraacute propouštiacute proteiny o niacutezkeacute molekuloveacute hmotnosti Použiacutevaacute se pro rychlou a pohodlnou koncentraci rozděleniacute a odsoleniacute vzorků) Po odstředěniacute byly vzorky odebraacuteny z horniacutech diacutelů nanosepů a metodou TL (postupneacuteho nanaacutešeniacute) naneseny na destičku Nanaacutešeneacute množstviacute bylo 03 microl vzorku a 03 microl matrice Jako matrice byla použita kyselina sinapovaacute (koncentrace 20 mgml rozpouštědlo acetonitril01 kyselina trifluorooctovaacute 32 vv) Hmotnostniacute spektra byla naměřena na hmotnostniacutem spektrometru MALDI TOFTOF v pozitivniacutem iontoveacutem moacutedu při lineaacuterniacutem uspořaacutedaacuteniacute

39

5 VYacuteSLEDKY A DISKUSE V proteomickeacute analyacuteze je nutneacute před vlastniacute analyacutezou pomociacute hmotnostniacute spektrometrie komplexniacute vzorky nejdřiacuteve separovat V teacuteto praacuteci byly využity separačniacute techniky jednorozměrnaacute (SDS-PAGE) a dvojrozměrnaacute (2D PAGE) gelovaacute elektroforeacuteza 51 Separace proteinů 1D (SDS-PAGE) elektroforeacutezou

Jednaacute se o elektroforetickyacute způsob separace na zaacutekladě velikosti molekul separovanyacutech laacutetek (viz kapitola 343) V prvniacutem kroku je nutneacute určit jakeacute množstviacute vzorku se bude daacutevkovat na gel a v naacutesledujiacuteciacutem kroku jakyacutem barvivem se separovaneacute proteiny zafixovaneacute v gelu budou barvit (způsob fixace je zaacutevislyacute na použiteacutem barvivu)

511 Optimalizace daacutevkovaacuteniacute vzorku

Pro optimalizaci daacutevkovaacuteniacute na 125 TRIS-HCl gel byly vybraacuteny dva vzorky ndash pomleteacute obilky zrna a pomleteacute obilky hotoveacuteho sladu ktereacute se na gel nadaacutevkovaly v různeacutem množstviacute Zrno a hotovyacute slad byl vybraacuten zaacuteměrně protože reprezentujiacute počaacutetečniacute a konečnyacute stav procesu sladovaacuteniacute

a) Pomleteacute obilky zrna ječmene odrůdy Jersey (obr č13)

Obrč13 Na odraacutezku je znaacutezorněno rostouciacute daacutevkovaacuteniacute vzorku na gel od 4 ndash 11 microl A-standard

Na gelu je vidět že rostouciacute množstviacute nadaacutevkovaneacuteho vzorku na gel způsobuje lepšiacute

viditelnost separovanyacutech proteinů

A A

40

b) Pomleteacute obilky sladu ječmene odrůdy Jersey (obr č14)

Obrč14 Rostouciacute daacutevkovaacuteniacute vzorku na gel od 5-12 microl A-standard Za danyacutech podmiacutenek bylo zvoleno (jak u zrna tak u sladu) pro uvedenyacute formaacutet gelu

daacutevkovaacuteniacute 10 microl vzorku na gel

A A

41

512 Vizualizace proteinů

Porovnaacuteniacute dvou nejčastěji použiacutevanyacutech technik barveniacute klasickeacute CBB (CBB-R250)

a koloidniacute CBB (CBB-G250) lišiacuteciacutech se předevšiacutem citlivostiacute a provedeniacutem (viz kapitola 44) a) Barveniacute barvivem Commassie Brilliant Blue R-250 (obr č15)

b) Barveniacute barvivem Commassie Brilliant Blue G-250 (obr č16)

Obr č 15 a 16 A ndash standard B ndash zrno C ndash odběr ze 3dne sladovaacuteniacute D ndash hotovyacute slad

42

Porovnaacuteniacutem barviciacutech směsiacute bylo zjištěno že pro uacutečely teacuteto praacutece je vyacutehodnějšiacute použiacutet barvivo Commassie Brilliant Blue G-250 ktereacute ukaacutezalo že barviacute proteiny zafixovaneacute v gelu citlivěji a barva je sytějšiacute Toto barvivo se použilo k barveniacute všech naacutesledujiacuteciacutech gelů

513 Elektroforetickyacute profil protein ů ječmene

Po zjištěniacute optimaacutelniacuteho daacutevkovaacuteniacute vzorku a vhodneacuteho způsobu barveniacute gelu byly na gel

naneseny vzorky představujiacuteciacute celyacute profil sladovaacuteniacute (zrno odběry z prvniacuteho až paacuteteacuteho dne sladovaacuteniacute zelenyacute slad a slad) K separaci byl použit 125 gel (viz obrč17) vhodnyacute pro systeacutem Mini PROTEAN 3 Cell od firmy BIO-RAD Vyacutehodou použitiacute aparatury Mini PROTEAN 3 Cell je rychlaacute separace (cca 50 minut) a spotřeba maleacuteho množstviacute vzorků a použityacutech chemikaacuteliiacute

Obr č 17 A H - standardy B - zrno C - odběr z 1dne sladovaacuteniacute D - odběr ze 2dne sladovaacuteniacute E - odběr ze 3dne sladovaacuteniacute F - odběr ze 4dne sladovaacuteniacute G - odběr z 5dne

sladovaacuteniacute I - zelenyacute slad J - hotovyacute slad

I když byly na gelu zachyceny rozdiacutely v proteinoveacutem profilu během procesu sladovaacuteniacute za

uvedenyacutech podmiacutenek gel neposkytl očekaacutevaneacute rozlišeniacute Byla snaha dosaacutehnout většiacuteho rozlišeniacute proto byly stejneacute vzorky v různeacutem daacutevkovaacuteniacute naneseny na 125 gel při použitiacute aparatury OWL Separation Systems (viz obrč18)

43

Obr č18 A - standardy B - zrno (v různeacutem množstviacute nadaacutevkovaneacuteho vzorku 12 microl16 microl

20 microl a 25 microl) C - odběr ze 3dne sladovaacuteniacute D - hotovyacute slad u C a D byl vzorek nadaacutevkovaacuten ve stejneacutem množstviacute jako v přiacutepadě A

Ziacuteskaneacute vyacutesledky ukazujiacute že daacutevkovaacuteniacute 20 microl na uvedenyacute gel je optimaacutelniacute Stejneacute daacutevkovaacuteniacute vzorku na gel bylo použito i u naacutesledujiacuteciacuteho gelu (obrč19) ndash profilu procesu sladovaacuteniacute (jednaacute se o 125 gel při použitiacute aparatury OWL Separation Systems)

44

Obr č19 A J ndash standardy B - zrno C - odběr z 1dne sladovaacuteniacute D - odběr ze 2dne

sladovaacuteniacute E - odběr ze 3dne sladovaacuteniacute F - odběr ze 4dne sladovaacuteniacute G - odběr z 5dne sladovaacuteniacute H - zelenyacute slad I - hotovyacute slad

Elektroforetickyacute postup separace ječnyacutech proteinů při použitiacute aparatury OWL Separation Systems poskytl lepšiacute rozděleniacute proteinů Na gelu jsme mohli pozorovat proteiny v rozmeziacute molekulovyacutech hmotnostiacute 21 - 100 kDa

45

Naše pozornost byla nadaacutele zaměřena na separaci proteinů ječmene při použitiacute komerčniacutech gelů firmy BIO-RAD a s nimi kompetentniacute aparaturou Mini PROTEAN 3 Cell K tomuto uacutečelu byl použit gradientovyacute gel 4 ndash 20 TRIS ndash HCl (obr č20) a 15 TRIS ndash HCl gel (obr č21)

a) Gradientovyacute gel 4 ndash 20 TRIS ndash HCl



Gradientovyacute gel 4 ndash 20 TRIS-HCl naacutem umožnil za danyacutech podmiacutenek separaci proteinů od 6 ndash 200 kDa Změny proteinoveacuteho profilu je možneacute na gelu pozorovat už od druheacuteho dne sladovaacuteniacute kdy dochaacuteziacute k naacuterustu intenzity některyacutech bandů K největšiacutem změnaacutem dochaacuteziacute v rozmeziacute proteinovyacutech hmotnostiacute 40 ndash 60 kDa

46

b) 15 TRIS ndash HCl gel

Obr č21 A J ndash standardy B - zrno C - odběr z 1dne sladovaacuteniacute D - odběr ze 2dne sladovaacuteniacute E - odběr ze 3dne sladovaacuteniacute F - odběr ze 4dne sladovaacuteniacute G - odběr z 5dne

sladovaacuteniacute H - zelenyacute slad I - hotovyacute slad

15 TRIS ndash HCl gel naacutem poskytl podobnyacute proteinovyacute profil jako gel gradientovyacute rovněž jsme mohli pozorovat proteiny v rozmeziacute molekulovyacutech hmotnostiacute 6 - 200 kDa

Gel byl daacutele použit k identifikaci proteinů pomociacute metody peptidoveacuteho mapovaacuteniacute (peptide mass fingerptinting) a MSMS Pozornost byla soustředěna na oblast proteinů mezi 22 ndash 60 kDa

47

52 Separace proteinů 2D gelovou elektroforeacutezou

Nespornou vyacutehodou 1D elektroforeacutezy je možnost nadaacutevkovaacuteniacute viacutece vzorků na jeden gel a naacutesledneacute porovnaacutevaacuteniacute vzorků mezi sebou ale protože se jednaacute o velmi komplexniacute pohled na proteiny byla provedena i 2D elektroforeacuteza jejiacutež vyacutehodou je separace proteinu podle hmotnosti i podle izoelektrickeacuteho bodu (viz kapitola 344) takže můžeme od sebe odseparovat i proteiny ktereacute majiacute stejnou molekulovou hmotnost ale různyacute naacuteboj

2D gely v teacuteto praacuteci sloužiacute pouze na ukaacutezku daacutele s nimi pracovaacuteno nebylo Pro ilustraci změn proteinoveacuteho profilu během procesu sladovaacuteniacute byly vybraacuteny gely pomletyacutech obilek

-gel zrna (obr č22) reprezentujiacuteciacute počaacutetek procesu -gel odběru ze třetiacuteho dne sladovaacuteniacute (obr č23) kdy dochaacuteziacute asi k nejvyacuteraznějšiacutem změnaacutem proteinoveacuteho profilu během celeacuteho procesu -gel hotoveacuteho sladu (obr č24) kteryacute demonstruje konec procesu sladovaacuteniacute

a) Zrno (obr č22)

48

b) Odběr ze třetiacuteho dne sladovaacuteniacute (obr č23)

c) Hotovyacute slad (obr č24)

49

Z uvedenyacutech 2D gelů je zřejmyacute naacuterůst spotů u sladu v rozmeziacute molekulovyacutech hmotnostiacute 31 ndash 70 kDa kteryacute koresponduje s možnyacutemi posttranslačniacutemi modifikacemi


Identifikace proteinů byla zaměřenaacute na proteiny ktereacute prochaacuteziacute celyacutem procesem sladovaacuteniacute během tohoto procesu se měniacute staacutevajiacute se stabilnějšiacutemi a v konečneacutem důsledku ovlivňujiacute kvalitu piva - jeho technologickeacute i senzorickeacute vlastnosti (např pěnu a zaacutekal) Na obr č25 jsou znaacutezorněny proužky gelu na kteryacutech byly změny nejmarkantnějšiacute Tyto proužky byly z gelu vyřezaacuteny a identifikovaacuteny



50

V naacutesledujiacuteciacute tabulce jsou uvedeny identifikovaneacute proteiny

čiacuteslo spotu identifikovanyacute protein molekulovaacute

hmotnost

1 ovalbumin 45000 kDa

2 karbonaacutet anhydrasa 31000 kDa

3 α-amylasa subtilisin

inhibitor 22378 kDa

4 protein Z 43363 kDa

5 peroxidasa BP1 32151 kDa

6 β-amylasa 59912 kDa

7 až 13 protein Z v průběhu

sladovaacuteniacute

531 Identifikace standardů pomociacute hmotnostniacute spektrometrie

Z gelu (obrč25) byly vyřiacuteznuty proužky č1 a 2 ktereacute byly podrobeny enzymatickeacutemu

štěpeniacute chymotrypsinem Po jeho proběhnutiacute bylo změřeno MS spektrum směsi peptidů v reflektrononeacutem uspořaacutedaacuteniacute v kladneacutem modu na přiacutestroji 4700 Proteomics Analyzer Hmotnosti peptidů byly zadaacuteny do databaacuteze Mascot vyacutesledkem vyhledaacutevaacuteniacute v prvniacutem přiacutepadě byl ovalbumin (obr č26) a ve druheacutem přiacutepadě karbonaacutet anhydrasa (obr č27)

a) ovalbumin (45 kDa)

800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Reflector Spec 1 MC[BP = 10025 11520]

1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51

a) karbonaacutet anhydrasa (31 kDa)

800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52

532 Identifikace proteinu Z pomociacute hmotnostniacute spektrometrie

Identifikace reaacutelnyacutech vzorků proběhla stejně jako u standardů Proužky z gelu byly vyřezaacuteny podrobeny enzymatickeacutemu štěpeniacute byla naměřena MS spektra (obrč28) a naacutesledně MSMS spektra (obrč29) Posledniacutem krokem bylo vyhledaacutevaacuteniacute v databaacutezoveacutem programu Mascot (obrč30)

a) Přiacuteklad MS spektra směsi peptidů vzniklyacutech enzymatickyacutem štěpeniacutem

(chymotrypsinem) proteinu Z ziacuteskaneacuteho z gelu (ze zrna) (obrč28)

800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269

Obrč28 Protein Z (43363kDa)

intensity

Mass (mz)

darr

darr

darr

darr

darr

53

b) MSMS spektrum vybraneacuteho piacuteku naacuteležiacuteciacuteho peptidu s mz 2457 Da vznikleacuteho enzymatickyacutem štěpeniacutem (chymotrypsinem) proteinu Z (obrč29)

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 MSMS Precursor 242819 Spec 1 MC[BP = 17218 3523]

1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29

Protein Z patřiacute do skupiny proteinů ktereacute pozitivně ovlivňujiacute pěnivost piva Jednaacute se o ječnyacute albuminovyacute protein vykazujiacuteciacute hydrofobniacute charakter kteryacute usnadňuje přechod proteinu do pivniacute pěny Protein Z představuje 10 ndash 25 nedialyzovatelnyacutech proteinů piva a je přibližně z jedneacute třetiny glykosylovaacuten [71] Uacutečast proteinu Z na stabilitě pivniacute pěny s největšiacute pravděpodobnostiacute zaacutevisiacute na charakteru sladu

Mass (mz)

intensity

54

d) Identifikace proteinu Z pomociacute databaacutezoveacuteho vyhledaacutevaacuteniacute v programu Mascot za použitiacute databaacuteze NCBInr

Obrč30

55

533 MS spektra dalšiacutech identifikovanyacutech proteinů ječmene ziacuteskanyacutech z teacutehož gelu

Identifikace proteinů (viz obrč25) byla opět provedena s použitiacutem databaacutezoveacuteho

vyhledaacutevaacuteniacute v programu Mascot

a) α - amylasa subtilisin inhibitor (22378 kDa)

800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358

Obrč31 b) peroxidasa (32151 kDa)

800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56

c) β ndash amylasa (59912 kDa)

800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33

Peroxidasa představuje enzym třiacutedy oxidoreduktaacutez ktereacute pomociacute peroxidu vodiacuteku oxidujiacute různeacute substraacutety Obvykle nevykazujiacute vysokou specifitu pro oxidovanyacute substraacutet

Amylasy jsou nejdůležitějšiacutemi enzymy ve sladu (viz kapitola 312) α a β ndash amylasy se vyacuteznamně podiacuteliacute na zcukřeniacute čili uacuteplneacutem rozštěpeniacute makromolekul škrobu za vzniku různyacutech nižšiacutech cukrů a dextrinů

Mass (mz)

intensity

57

534 Změny proteinu Z v průběhu celeacuteho procesu sladovaacuteniacute (MS a MSMS spektra)

Na naacutesledujiacuteciacutech obraacutezciacutech jsou vyacuteřezy z MS spektr proteinu Z v po sobě jdouciacutech dnech

sladovaacuteniacute ndash od zrna až po hotovyacute slad (viz obrč25) a k nim jsou přiřazena MSMS spektra peptidů 2457 Da a 2619 Da

1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity

4700 R ef lector Spec 1 MC [ B P = 1410 8 12761]

2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 MSMS Precursor 242819 Spec 1 MC[B P = 17218 3523]

1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34

2) odběr z prvniacuteho dne sladovaacuteniacute

2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Reflector Spec 1 MC [BP = 14109 17497]

2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 MSMS Precursor 245743 Spec 1 MC [B P = 14106 2701]

1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58

3) odběr z druheacuteho dne sladovaacuteniacute

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 MSMS Precursor 261945 Spec 1 MC[ BP = 868 1149]

2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36

4) odběr ze třetiacuteho dne sladovaacuteniacute

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 MSMS Precursor 245739 Spec 1 MC [BP = 24599 9722]

2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59

5) odběr ze čtvrteacuteho dne sladovaacuteniacute

2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Ref lector Spec 1 MC[BP = 14108 29977]

2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 MSMS Precursor 245735 Spec 1 MC [BP = 24598 23 688]

2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38

6) odběr z paacuteteacuteho dne sladovaacuteniacute

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 MSMS Precursor 245737 Spe c 1 MC[BP = 14107 19 038]

1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 MSMS Prec urs or 2619 45 Spec 1 MC [B P = 250 02 349 5]

2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61

Vyacutesledky poukazujiacute na možneacute posttranslačniacute modifikace ječnyacutech proteinů (ktereacute jsou patrneacute už od druheacuteho dne) po dobu sladovaacuteniacute Ječneacute proteiny jsou v průběhu sladovaacuteniacute štiacutepaacuteny zvyšuje se obsah cukrů ktereacute jsou uvolňovaacuteny ze škrobu a ty spolu se zvyacutešenou teplotou způsobujiacute glykace proteinů Vznikleacute glykovaneacute proteiny byacutevajiacute většinou stabilnějšiacute prochaacuteziacute celyacutem procesem vařeniacute piva a objevujiacute se i v pivu kde ovlivňujiacute jeho vlastnosti jak pozitivně tak negativně (pěna zaacutekal) 54 Měřeniacute intaktniacutech proteinů ječmene

Pomociacute MALDI TOF-MS v lineaacuterniacutem modu byly měřeny a sledovaacuteny změny (posttranslačniacute modifikace) v průběhu sladovaacuteniacute u vyacuteznamneacuteho proteinu ječmene LTP 1

LTP 1 (lipid transfer protein) ndash polypeptid- intracelulaacuterniacute přenašeč lipidů Jednaacute se o albuminovyacute protein kteryacute je během procesu sladovaacuteniacute přeměňovaacuten v minimaacutelniacutem rozsahu Jeho stabilitě pravděpodobně pomaacutehajiacute čtyři disulfidickeacute můstky ktereacute spojujiacute čtyři α-helikaacutelniacute čaacutesti tohoto proteinu [71] LTP 1 patřiacute mezi proteiny pozitivně ovlivňujiacuteciacute pivniacute pěnu protože podporuje stabilitu pěny

Na naacutesledujiacuteciacutech obraacutezciacutech jsou zachyceny změny intaktniacutech proteinů u zrna (obrč42) odběru ze třetiacuteho dne sladovaacuteniacute (obrč43) a u hotoveacuteho sladu (obrč44)

a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62

b) Odběr ze třetiacuteho dne sladovaacuteniacute

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63

d) Pozorovaacuteniacute změn intaktniacutech proteinů v průběhu procesu sladovaacuteniacute ndash spektra LTP 1 seřazena vedle sebe

Na obraacutezku č45 jsou zobrazeny vyacuteřezy hmotnostniacutech spekter intaktniacutech proteinů

extrahovanyacutech ze vzorků ječmene reprezentujiacuteciacutech celyacute proces sladovaacuteniacute od zrna až po hotovyacute slad

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047

zrno 1den 2den 3den 4den 5den sladzelenyacute

slad

Obraacutezek č45 ukazuje změny LTP 1 v průběhu sladovaacuteniacute V průběhu sladovaacuteniacute je tento

protein modifikovaacuten hexosami vaacutezanyacutemi na lyziacuteny Od druheacuteho dne sladovaacuteniacute lze pozorovat postupně rostouciacute piacuteky ktereacute odpoviacutedajiacute aduktům 162 Da

Obrč45

64

6 ZAacuteVĚR

Prvniacute čaacutest teacuteto praacutece byla věnovaacutena literaacuterniacutemu přehledu V teoretickeacute čaacutesti jsou shrnuty zaacutekladniacute informace o pivu o procesu jeho vyacuteroby a o zaacutekladniacutech surovinaacutech pro jeho vyacuterobu s důrazem na sladovnickyacute ječmen Daacutele informace o proteinech nachaacutezejiacuteciacutech se ve sladovnickeacutem ječmeni o vybranyacutech metodaacutech separace proteinů a o hmotnostniacute spektrometrii MALDI-TOFTOF MS

V experimentaacutelniacute čaacutesti praacutece byla provedena analyacuteza proteinů obilek ječmene a jejich identifikace pomociacute MALDI-TOFTOF MS

Tato praacutece byla zaměřena na změny proteinoveacuteho profilu ječmene během procesu sladovaacuteniacute protože se ukaacutezalo že ječmen podleacutehaacute během sladovaacuteniacute vyacuteznamnyacutem změnaacutem ktereacute buď pozitivně nebo negativně ovlivňujiacute kvalitu sladu a naacutesledně i piva Pozornost byla zaměřena na vodorozpustneacute proteiny Změny byly sledovaacuteny pomociacute jednorozměrneacute geloveacute elektroforeacutezy (1D GE) dvojrozměrneacute geloveacute elektroforeacutezy (2D GE) a MALDI-TOFTOF hmotnostniacute spektrometrie (MS)

Jako vzorek byly použity pomleteacute obilky ječmene (zrno zelenyacute slad hotovyacute slad a vzorky odebraneacute v průběhu sladovaacuteniacute ndash prvniacute až paacutetyacute den) u kteryacutech byly sledovaacuteny změny proteinoveacuteho profilu mezi jednotlivyacutemi faacutezemi sladovaacuteniacute Pro uacutečel teacuteto praacutece byla zvolena odrůda Jersey

V prvniacute faacutezi byla provedena optimalizace daacutevkovaacuteniacute vzorku na gel pro jednorozměrnou gelovou elektroforeacutezu optimalizace vizualizace a optimalizace stupně zesiacuteťovaacuteniacute gelu Pro naacuteš uacutečel se jevil optimaacutelniacute 15 TRIS-HCl gel na ktereacutem jsme mohli pozorovat proteiny v rozmeziacute molekulovyacutech hmotnostiacute 6 - 200 kDa Vzorek byl na gel nadaacutevkovaacuten v množstviacute 10 microl a gel byl obarven barvivem Commassie Brilliant Blue G-250 Tento gel byl daacutele posloužit k identifikaci proteinů pomociacute metody peptidoveacuteho mapovaacuteniacute (peptide mass fingerptinting) a MSMS

Na 15 TRIS-HCl gelu byly pozorovaacuteny změny v elektroforetickyacutech profilech mezi zrnem vzorky odebranyacutemi během procesu sladovaacuteniacute (1až 5 dnem) a jeho zelenyacutem sladem a hotovyacutem sladem Proteiny ktereacute se nejvyacuterazněji po dobu sladovaacuteniacute měnily byly z gelu vyřiacuteznuty a poteacute naštiacutepaacuteny enzymem (chymotrypsinem) a identifikovaacuteny pomociacute MALDI-TOFTOF MS Jako matrice byla použita kyselina α-kyano-4-hydroxyskořicovaacute V raacutemci teacuteto praacutece byl metodou peptidoveacuteho mapovaacuteniacute a MSMS identifikovaacuten protein kteryacute pozitivně ovlivňuje stabilitu pivniacute pěny - protein Z ale i dalšiacute proteiny ktereacute byly ve vzorciacutech pozorovaacuteny (α-amylasa inhibitor β-amylasa peroxidasa)

V dalšiacute faacutezi byla provedena dvojrozměrnaacute gelovaacute elektroforeacuteza kde byly rovněž pozorovaacuteny rozdiacutely mezi jednotlivyacutemi vzorky Pro ilustraci změn byly vybraacuteny gel zrna gel vzorku odběru ze třetiacuteho dne sladovaacuteniacute a gel hotoveacuteho sladu Na gelech jednotlivyacutech vzorků se ukazovala řada proteinovyacutech spotů v rozmeziacute molekulovyacutech hmotnostiacute 10 ndash 70 kDa

Daacutele byly pomociacute MALDI-TOF MS analyzovaacuteny intaktniacute proteiny extrahovaneacute ze všech vzorků (od zrna až po hotovyacute slad) Extrakty byly podrobeny odsolovaacuteniacute pomociacute techniky bdquonanosep centrifugal deviceldquo Hmotnostniacute spektra byla ziacuteskaacutena v pozitivniacutem iontoveacutem moacutedu při lineaacuterniacutem uspořaacutedaacuteniacute a jako matrice byla použita kyselina sinapovaacute Ze spekter bylo patrneacute že během procesu sladovaacuteniacute dochaacuteziacute ke změně proteinoveacuteho profilu ječneacute proteiny jsou modifikovaacuteny dochaacuteziacute ke glykaciacutem proteinů Vznikleacute glykovaneacute proteiny jsou potom stabilnějšiacute prochaacutezejiacute celyacutem procesem vařeniacute piva a naacutesledně ovlivňujiacute kvalitu piva ndash jeho technologickeacute a senzorickeacute vlastnosti Pozornost byla zaměřena na protein LTP 1 (lipid

65

transfer protein 1) kteryacute se pozitivně podiacuteliacute na tvorbě pivniacute pěny Na MS spektrech LTP 1 byly pozorovaacuteny změny už od druheacuteho dne sladovaacuteniacute

Sladovnickyacute ječmen je specifickou plodinou na kterou jsou kladeny mimořaacutedně vysokeacute požadavky ze strany sladovnickeacuteho a pivovarskeacuteho průmyslu proto by se v budoucnosti dalo na praacuteci navaacutezat pozorovaacuteniacutem změn u jinyacutech odrůd sladovnickeacuteho ječmene srovnaacutevaacuteniacutem odrůd mezi sebou detailnějšiacutem studiem změn mezi jednotlivyacutemi faacutezemi sladovaacuteniacute nebo třeba identifikaciacute většiacuteho množstviacute proteinů Proteomickaacute analyacuteza ječmene maacute vyacuteznam pro šlechtěniacute novyacutech kvalitnějšiacutech odrůd sladovnickeacuteho ječmene

66

7 LITERATURA [1] Vodraacutežka Z Biochemie Praha ACADEMIA 1999 [2] httpvydavatelstvivschtczknihyuid_es-002heslabilkovinyhtml [3] Chmeliacutek J Řehulka P Mayrhofer C Allmaier G Proteomika ndash novyacute a rychlyacute naacutestroj pro identifikaci odrůd ječmene Kvasnyacute průmysl (2001) [4] Kellner V Pivo vitaminy a dalšiacute důležiteacute laacutetky pro vyacuteživu a zdraviacute člověka Pivovarskaacute uacutestav Praha [5] Čepička J Karabiacuten M Polyfenoloveacute laacutetky piva ndash přirozeneacute antioxidanty ChemListy

(2002) [6] De Keukelaire D Pharm Pharmacol Lett7 83 (1997) [7] Zoufalyacute T Brynych P Českeacute pivo Každodenniacute fenomeacuten ChemListy (2006) [8] Domeacuteny Z Šmogrovičovaacute D Šturdiacutek E Kontinuaacutelne fermentačneacute precesy při vyacuterobe

piva pomocou imobilizovanyacutech kvasinek Chem listy 92 (1998) [9] Čepička J Fermentačniacute technologie kapitola Sladařstviacute [10] Prokeš J Pivovarskaacute škola Vyacutezkumnyacute uacutestav pivovarskyacute a sladařskyacute

httpwwwpivovarskaskolacz [11] httpwwwvschtczkchkestazenisylabysladarstvipdf [12] Čepička J Fermentačniacute technologie kapitola Sladařstviacute [13] Ťopka P Čejka P Kvasnyacute průmysl 333 (1987) [14] Evans D E Sheehan MC Steward D CJ J Inst Brew 105 171 (1999) [15] Gray P Stone I Am Soc Brew Chem Proc 1956 83 [16] Kosař K Prochaacutezka S a kolektiv autorů Technologie vyacuteroby sladu a piva 2000 [17] Hardwick W A Handbook of Brewing Marcel Decker New York 1995 [18] Čepička J Fermentačniacute technologie kapitola Pivovarstviacute [19] Naučnyacute slovniacutek zemědělskyacute diacutel 2 e-j Praha 1968 [20] Zimolka J a kolektiv Ječmen ndash formy a užitkoveacute směry v Českeacute Republice 2006 [21] Rostlinnaacute vyacuteroba 3 Rybaacuteček a kolektiv 1965 [22] Sborniacutek referaacutetů ze seminaacuteře Genofond zeměděl skyacutech plodin a jeho využitiacute pro rozšiacuteřeniacute agrobiodiversity 2002 Českaacute akademie zemědělskyacutech věd [23] Současneacute trendy a perspektivy šlechtěniacute sladovnickeacuteho ječmene Uacuteroda 22007 [24] Basařovaacute G Hlavaacuteček I Českeacute pivo 1999 [25] Českaacute staacutetniacute norma 46 1100-5 Ječmen sladovnickyacute 2006 [26] Metodika pěstovaacuteniacute jarniacutech obilovin Zemědělskyacute vyacutezkumnyacute ustav Kroměřiacutež 2001 [27] Časopis pro rostlinnou produkci Uacuteroda 22007 Ing Kateřina Vaculovaacute Agrotest fyto

sro Dana Gabrouskaacute Vyacutezkumnyacute uacutestav potravinaacuteřskyacute Praha [28] httpwwwscienceworldczswnsfperlicky3A7D387BBAD74618C12572AD006D

4334OpenDocumentampcast=1 [29] Kirkman M A Sherry PR Milčin BJ The effect of nitrogen nutrition on the lysine

kontent and protein composition of barely seeds Journal of the Science of Food and Agriculture 33 115 (1982)

[30] Shewry P R Barely Genetics Biochemistry Molecular Biology and Biotechnology 1992

[31] Chmeliacutek J Protetickyacute průvodce Chem listy 99 883 (2005) [32] Sommer L a kolektiv Zaacuteklady analytickeacute chemie 2 Vutium 2000

67

[33] Lilley KS Razzaq A Dupree P Two-dimensional gel electrophoresis recent advances in sample preparation detection and quantitation Current Opinion in Chemical Biology 2002

[34] Vysokouacutečinneacute analytickeacute separace biolog aktivniacutech laacutetek Hudeček J Separace proteinů a proteomika Praha 2006

[35] Firemniacute materiaacutel firmy BIO-RAD ReadyStrip IPG Strip Instuction Manual [36] Shimura K Recent advances in capilary isoelectric focusing 1997-2001

Electrophoresis 2002 [37] Winkler Ch Denker K Wortelkamp S Sickmann A Silver- and Coomassie-staining

protocols Detectionlimits and compatibility with ESI MS Electrophoresis 28 2095ndash2099 (2007)

[38] Shevchenko A Wilm M Vorm O Mann M Mass Spectrometric Sequencing of Proteins from Silver-Stained Polyacrylamide Gels Anal Chem 68 (1996)

[39] Yan X J Wait R Berkelman T Harry RA Westbrook JA Wheeler CH Dunn HJ A modified silver staining protocol for visualization of proteins compatible with matrix-assisted laser desorptionionization and electrospray ionization-mass spectrometry Electrophoresis 21 (17)3666 (2000)

[40] Zdraacutehal Z Plocek J Konečnyacute P Chmeliacutek J Characterization of Biomacromolecules Using Mass Spektrometry Chem Listy 9 811 (1997)

[41] Nguyer D N Becker G W Riggin K M Chromatogr A 705 21 (1995) [42] Carr S A Hemling M E Bean M F Roberts G D Anal Chem 63 2802 (1991) [43] Harvey D J Mass Spectrometry Rev 18 349 (1999) [44] Feng R Konishi Y Anal Chem 64 2090 (1992) [45] Hillenkamp F Karas M Beavis RC Chait BTMatrix-Assisted Laser

DesorptionIonization Mass Spectrometry of Biopolymers Anal Chem 63 1193 (1991)

[46] Cotter R J Time-of-Flight Mass Spectrometry for the Structural analysis of Biological Molecules Anal Chem 64 1992

[47] Kadlčiacutek V Kodiacuteček M Hassman M Využitiacute hmotnostniacutespektrometrie na principu MALDI-TOF pro studium prostoroveacute struktury proteinů ChemListy 96 618 (2002)

[48] Lamer S Jungblut PR Matrix-Assisted laser desorption-ionization mass spectrometry peptide mass fingerprinting for proteome analysis identification efficiency after on-blot or in-gel digestion with and without desalting procedures J Chromatogr B 752 311 (2001)

[49] Franzen J Frey R Holle A Krauter K O Recent progress in matrix-assisted laser desorption ionization postsource decay Int J off Mass Spectrom Ion Processes 206 275 (2001)

[50] Fitzerald M C Parr G R Smith L M Basic Matrices for the Matrix-Asisted Laser DesorptionIonization Mass Spectrometry of Proteins and Oligonucliotides Analytical Chemistry 65 3204 (1993)

[51] Mazanec K Chmeliacutek J Využitiacute MALDI-TOF MB při analyacuteze směsiacute polymerů ChemListy 99 175 (2005)

[52] Makešovaacute DStudium glykokonjugaacutetů v potravinaacutech hmotnostniacute spektrometiiacute MALDI MS Diplomovaacute praacutece Brno Fakulta chemickaacute 2007

68

[53] Harvey D JMatrix-assisted laser desorptionionization mass spectrometry of carbohydrates and glycoconjugates International Journal of Mass Spectrometry 226 1 (2003)

[54] Wang J Sporns P Low N H Analysis of Food Oligosaccharides Using MALDI-MS Quantification of Fructooligosaccharides Journal of Agricultural and Food Chemistry 47 1549 (1999)

[55] Bahr U Karas M Hillenkamp F Analysis of Biopolymers by Matrix-assisted Laser DesorptionIonization (MALDI) Mass Spectrometry Fresenius Journal of Analytical Chemistry 348 1994

[56] Cohen S L Chait B T Influence of Martix Solutions Conditions on the MALDI-MS Analysis of Peptides and Proteins Ana Chem 68 1996

[57] Karas M Hillenkamp F Laser desorption ionization of proteins with molecular masses exceeding 10 000 Daltons Analytical Chemistry 60 2299 (1988)

[58] Vorm O Roepstorff P Mann M Improved Resolution and Very Hight Sensitivity in MALDI TOF of Matrix Surfaces Made by Fast Evaporation Analytical Chemistry 66 3281 (1994)

[59] Shevchenko A Wilm M Worm O Mann M Mass spectrometric sequencing of proteins from silver stained polyacrylamide gels Analytical Chemistry 68 850 (1996)

[60] Fernandez-Patron CCalero M Collazo P R Garcia J RMadrazo J Musacchio A Soriano F Estrada R Frank R Castellanos-Serra L R Mendez E Protein reverse staining Hight efficiency microanalysis of unmodified proteins detected on electrophoresis gels Analytical Biochemistry 224 203 (1995)

[61] Bouchal P Kučera I Dvourozměrnaacute elektroforeacuteza v proteomice Principy a aplikace Chem Listy 97 29 (2003)

[62] Collinsovaacute M Jiraacuteček J Současnyacute vyacutevoj v proteomice Chem Listy 98 1112 (2004) [63] OrsquoFarrel PH J Biol Chem 250 4007 (1975) [64] Klose J Humangenetik 26 231 (1975) [65] The Nobel Prize in Chemistry 2002 ndash Advance Information

wwwnobelsechemistrylaureates2002chemadv02pdf [66] The Nobel Prize in Chemistry 2002 ndash Advance Information

wwwnobelsechemistrylaureates2002 [67] httpbiomikrovschtczmaldimancztheorypmfphp [68] Spina E Cozzolino R Ryan E Garozzo D Sequencing of oligosaccharides by

collision-induced dissociation matrix-assisted laser desorptionionization mass spektrometry Journal of Mass Spektrometry 35 1042 (2000)

[69] Osborne T B The Vegetable proteins London Longmans Green and Co 1924 [70] Štuliacutek K a kolektiv Analytickeacute separačniacute metody 2005 [71] Čiacutežkovaacute H Dostaacutelek P Fiala J Kolouchovaacute I Vyacuteznam biacutelkovin z hlediska pěnivosti

a stability pěny piva Chem Listy 100 478 (2006)

69

8 SEZNAM SYMBOL Ů Seznam analytickyacutech metod a přiacutestrojů DD Dried-droplet (technika nanaacutešeniacute vzorku v MALDI-TOF spektrometrii) TL Thin-layer (technika nanaacutešeniacute vzorku v MALDI-TOF spektrometrii) TOF Analyzaacutetor doby letu (time of flight) MALDI Laserovaacute desorpce a ionizace za uacutečasti matrice (Matrix-assisted laser

desorption and ionization) MS Hmotnostniacute spektrometrie 1D GE Jednorozměrnaacute gelovaacute elektroforeacuteza 2D GE Dvojrozměrnaacute gelovaacute elektroforeacuteza SDS-PAGE Elektroforeacuteza v polyakrylamidoveacutem gelu s dodecylsulfaacutetem sodnyacutem IEF Isoelektrickaacute fokusace




ZMĚNY PROTEINOVEacuteHO PROFILU V PRŮBĚHUSLADOVAacuteNIacute JEČMENECHANGES OF PROTEIN PROFILE IN BARLEY DURING MALTING



VEDOUCIacute PRAacuteCE ING JANETTE BOBAacuteĽOVAacute CSCSUPERVISOR

BRNO 2008

3

Abstrakt





Abstract





4


5





podpis diplomanta


6




32 Ječmen 15



33 Proteomika 20












42 Materiaacutel 33




7













8

1 UacuteVOD




9





10








11








darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr





12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



3

Abstrakt





Abstract





4


5





podpis diplomanta


6




32 Ječmen 15



33 Proteomika 20












42 Materiaacutel 33




7













8

1 UacuteVOD




9





10








11








darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr





12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



4


5





podpis diplomanta


6




32 Ječmen 15



33 Proteomika 20












42 Materiaacutel 33




7













8

1 UacuteVOD




9





10








11








darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr





12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



5





podpis diplomanta


6




32 Ječmen 15



33 Proteomika 20












42 Materiaacutel 33




7













8

1 UacuteVOD




9





10








11








darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr





12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



6




32 Ječmen 15



33 Proteomika 20












42 Materiaacutel 33




7













8

1 UacuteVOD




9





10








11








darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr





12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



7













8

1 UacuteVOD




9





10








11








darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr





12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



8

1 UacuteVOD




9





10








11








darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr





12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



9





10








11








darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr





12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



10








11








darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr





12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



11








darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr


darrdarrdarrdarr





12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



12








13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



13










14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



14





15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



15

32 Ječmen















16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



16



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



17








18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



18







Vlaacuteknina

nerozpustnaacute

()

Vlaacuteknina

rozpustnaacute

()

Sacharidy

()

KM 1910 stand 113 79 94 583

KM 2084 stand 119 94 87 55

KM 2283 stand 110 92 97 555


α-amylaza

(DU)

β-amylaza


KM 1910

ječmen 0 283 598 232



Tolar

ječmen 0 622 606 1696



Jersey

ječmen 0 405 596 19



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



19

















20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



20





21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



21













22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



22











23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



23










24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



24



(MALDI-MS)




25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



25





Vez

mLt

2= (2)




26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



26









27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



27



354 Matrice





28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



28






29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



29









30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



30


Typy fragmentace








31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



31


32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



32


33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



33

42 Materiaacutel








34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



34








35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



35









36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



36












37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



37




- Extrakce peptidů





38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



38


39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



39









A A

40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



40




A A

41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



41






42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



42









43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



43




44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



44




45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



45






46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



46






47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



47





a) Zrno (obr č22)

48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



48



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



49






50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



50



hmotnost




inhibitor 22378 kDa





sladovaacuteniacute





800 1120 1440 1760 2080 2400Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1002510

13246421683828

1492730

1371649 1725787

1699830134064097852015237771356636 1731827

Obrč26

intensity

Mass (mz)

51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



51


800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1052569

12297271282572 1625643

1296691

1341515 1602819951486

15238201312692 2428190

25161562232024

Obrč27

intensity

Mass (mz)

52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



52





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410802

959521

1133561 1518807

1213694

1086597 1523832 1859872

2457269


intensity

Mass (mz)

darr

darr

darr

darr

darr

53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



53


600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Obrč29


Mass (mz)

intensity

54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



54


Obrč30

55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



55





800 1200 1600 2000 2400 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


872402

902497

1870977

897447 15238291333640 22751132519358


800 1060 1320 1580 1840 2100Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1523786

1349717980489 1319680

947486 1416769

1857020

Obrč32

Mass (mz)

intensity

Mass (mz)

intensity

56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



56


800 1140 1480 1820 2160 2500Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1166587

1377635

1195570983537 1423735

1420721

1107514

1128641 14256918733851583778

13066801523816

2257083

Obrč33



Mass (mz)

intensity

57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



57




1) Zrno

2400 2480 2560 2640 2720 2800Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

tens

ity


2457269

600 5606 10612 15618 20624 25630Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1720786

1362729

1291121692849

84089 1265504186143

14777011807882

244131

Y

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč34


2400 2460 2520 2580 2640 2700

Mass (m z)

42362

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457433

2579614

2512497

24364072462439

2435417 246339125174662419302 26034542442345

25543792530483 2639552 2683505

0000000000000000000

Mass (mz)

in

tens

ity

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410671824601409

814138

1160538721400449

15247196

2270941

Y

MSMS 2457 Da

Obrč35

58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



58


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457391

25144162412429 2435344

26204622479367 2727560

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

1000 1360 1720 2080 2440 2800

Mass (mz)

5886

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457142

2617888

2499178

1023384

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč36


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457315

2514342

2619354

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460067

2499142

1572769

1410725

50 560 1070 1580 2090 2600

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2459694

1410686

1160549

21400631524756

110080 2310024127363970074 227108 478225374165

Y

Y + Hex

MSMS 2457 Da

MSMS 2619 Da

Mass (mz)

in

tens

ity

Obrč37

59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



59


2400 2480 2560 2640 2720 28000

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457351

25143752619413

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2460314

1410729

1160592

1524775 2140098110082

2310086127367147824270083 227126 374169 1913978807370

129107

1000 1360 1720 2080 2440 2800

Mass (m z)

35221

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457203

2461113 2621058

1572788

141074016868341322647 1554789

17809261184499

Y

Y + Hex

MSMS 2619 Da

MSMS 2547 Da

in

tens

ity

Mass (mz) Obrč38


2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457374

25143972619431

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410734

2459557

1160595

214006915247711100902310037

1273680227133 47826437419570088

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499160

2457179

1572787

14527581322651 1686784

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz)

Obrč39

60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



60

7) zelenyacute slad

2400 2480 2560 2640 2720 2800

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457393

2515414

2619448

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410750

2459720

1160611

2140100

1524805

2310070

110092 12736861913999

227140 47827137420270087

100 633 1166 1699 2232 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457207

2503181

2621721

1572839

1410794 1554838 2302207110100 1686865478313 1184503 1322655807408374181227133

Y

Y + Hex

MSMS 2619 Da

MSMS 2457 Da

in

tens

ity

Mass (mz) Obrč40

8) slad

2400 2460 2520 2580 2640 2700

Mass (mz)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2457328

2515348

2619377

Y

Y + Hex

59 566 1073 1580 2087 2594

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


1410699

2460223

1160558

2140049

1524743110086

2310052127364422712870084 374174

1000 1353 1706 2059 2412 2765

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity


2499168

1572786

14527571594678

1410713

16867781023444

MSMS 2457 Da

MSMS 2619 Da

in

tens

ity

Mass (mz) Obrč41

61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



61





a) Zrno

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

8800518

13398341

10283566

9885712

1310593415592146

71096989016636

LTP 1

Obrč42

62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



62


3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99815 3876]

9982563

4032718

7108455

8793883

10145480

1309025310186300

13304434 155796479819869

LTP 1

Obrč43

c) Slad

3000 6400 9800 13200 16600 20000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 11513 5451]

4033795

9988826

10150078

4196850

10304555687601213396372

7111046

10467047 13096947 15591535

LTP 1

Obrč44

63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



63




9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 10823 11404]

9998111

10283566

9885712

9 200 956 0 9920 10 280 10640 1 1000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L inea r Spe c 1 MC[B P = 1 0830 363 ]

9979 9316

100 166084

1 005581 05

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99829 1246]

9981937

10018491

10053771

10091306

10138382

9686410 10305047

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC[BP = 99868 1350]

9987345

10150677

10311371

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [BP = 11514 2085]

9990489

10149925

10284403

9200 9560 9920 10280 10640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spec 1 MC [B P = 99771 532]

9980121

10015456

10143074

10180372

10305037

10344299

9200 9 560 9920 10280 1 0640 11000

Mass (m z)

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4700 Linear Spe c 1 MC [BP = 11 527 35 17]

9989083

1 0151122

1030854 1

1028388 6

104 57798

982653 2

9200 9560 9920 10280 10640 11000

Mass (mz )

0

10

20

30

40

50

60

70

80

90

100

In

ten

sity

4 700 L ine ar Spe c 1 MC[ BP = 115 13 5 451 ]

9988 826

10150 078

10304 555

10467047


slad



Obrč45

64

6 ZAacuteVĚR









65



66










67


















68
















69



64

6 ZAacuteVĚR









65



66










67


















68
















69



65



66










67


















68
















69



66










67


















68
















69



67


















68
















69



68
















69



69



Documents

VYSOKÉ UČENÍ TECHNICKÉ V BRNĚ - COREcore.ac.uk/download/pdf/30295600.pdf · vysokÉ uČenÍ technickÉ v brnĚ brno university of technology fakulta chemickÁ Ústav chemie potravin