New milestone Switch +Go A century of experience · 2018. 11. 1. · longstanding experience in the development of GC systems as well as continuous exchange of ideas with users, Shimadzu

New milestone

The new Nexis GC-2030

Switch + Go

Nexera UC/s – Analysis andevaluation of chiral drugs

A century of experience

Shimadzu celebrates 100 yearsof testing machines

Pharmaceutical

Plastics and Rubber

CONTENT

Clinical

Chemical, Petrochemical,Biofuel and Energy

Harmful substances in textiles 4

Analysis and evaluation of chiraldrugs in biological samples –Nexera UC-MS/MS 8

Simple and fast characterization tool for biosensor development 10

Lighting up – Determination ofquantum efficiency of fluorescencestandards with high quality 12

Food and food packaging: non-intentionally added substances 15

Life can be hard – Contamination ofbread – Particle analysis with FTIRATR single reflection ATR techniqueand EDX for inorganic analysis 18

APPLICATION

One-stop solutions for clinical market applications – Shimadzu has acquired a specialist for high-quality analytical isotope labeledstandards 7

Feed to Food – Strategies for measurement of pesticides in water and foods 20

Brave helper in allergen management – Cleaning validationin specialty foods production 22

Current state of process analysistechnology – 2nd TOC ProcessAnalysis day in Duisburg 24

A century of experience – 100thanniversary of testing machines 26

Wine tasting and wine testing –from sniffing and tasting to measuring 27

New – Materials Testing ApplicationHandbook – 100th anniversary ofmaterials testing equipment 28

LATEST NEWS

MARKETS

The Next Industry Standard –Nexis GC-2030 2

How spectrometers take care of human health 14

PRODUCTS

SHIMADZU NEWS 2/2017

Food, Beverages, Agriculture

Environment

Automotive

The Next IndustryStandard Nexis GC-2030 – a new milestone in precision and sensitivity

I t has been 60 years sinceShimadzu initiated the devel-opment of GC systems with

the introduction of its GC-1A.Since then, numerous innova-tions have made gas chromatog-raphy one of the most importantanalytical techniques. Based onlongstanding experience in thedevelopment of GC systems aswell as continuous exchange of ideas with users, Shimadzu has set a new milestone in preci-sion and sensitivity with its Nexis GC-2030.

demand to learn more about thecomplex composition of prod-ucts as well as their emissioninto the environment and theirresulting interactions.

To meet these demands, an anal -ysis system must fulfil manyrequirements:

• chromatographic solutions forseparation of complex samples• high detection sensitivity forthe smallest traces of toxicimpurities

The Nexis GC-2030 stands for‘Next Industry Standard.’ Mee -ting this demand requires a clearview of present day GC require-ments. Gas chromatography isused in all areas of re search anddevelopment that ultimatelyenrich but also influence ourdaily life through foods, con-sumer goods, electron ics, healthand environment. Grow ing con-cern over the extent to whichtechnological advances negative-ly influence people and the envi-ronment has led to an increased

Figure 1: Milestones in GC development –

Nexis GC-2030

PRODUCTS

3SHIMADZU NEWS 2/2017

• highest precision for reliableresults• simple instrument control andoperation, also for novice GCusers• high reliability supported byautomated diagnostic routines• safety when handling hydrogencarrier gas.

Simple operation via self-explanatory user interface

The new color touchscreen dis-play offers fast and intuitive ac -cess to the Nexis GC-2030. Self-explanatory icons guide usersthrough the entire system opera-tion (figure 2). The same icons arefound in the new LabSolutionssoft ware, which offers optionalconventional or graphically con-trolled operation of the GC.Network-operated control alsooffers all advantages of the WorldWide Web. Direct access viaiPhone or iPad to the status of anongoing analysis as well aslaunching of new measurementsequences is possible at any timeand any place.

Maintenance without tools

Regular maintenance tasks on theNexis GC-2030 such as septum,liner and column replacement nolonger require any tools and canbe performed quickly and effi-ciently (figure 3a). The new ClickTek connector technology also

enables simple column installationby hand without the use of anytools. An optional oven lamp isavailable for illumination andoffers an excellent view inside theoven (figure 3b).

Flexible choice of carrier gascontrol with highest precision

Intelligent and ultrafast carrier gascontrol (AFC – Advanced FlowControl) enables superior repro-ducibility of the Nexis GC-2030through high-precision gas flowrate control. Carrier gas flow canbe adjusted and controlled basedon constant linear velocity, volumeflow or pressure.

Highest sensitivity worldwide

Redesigned detectors such as theflame ionization detector (FID),electron capture de tector (ECD)and flame photometer (FPD) openup new dimensions in trace analy-sis. The unique barrier ionizationdischarge (BID) detector allowssimultaneous detection of inor-ganic and organic components inthe low-ppb range.

Efficiency for fast results andhigh sample throughput

Special techniques such as back-flushing, detector splitting andheartcutting are supported by theGC system and can be integratedinto the chromatographic configu-ration as simple modules. In thisway, the graphical support allowseasy access to advanced chromato-graphic techniques.

Fast GC is the technique of choicefor obtaining fast results or highsample throughput. High-resolu-tion capillary columns allow sepa-ration of complex samples in theshortest amount of time by usinghigh carrier gas flow velocities andhigh heating rates of the GC oven.

This, however, requires an ultra-fast responding detector system inorder to capture sharp signals atfull height and symmetry. Eachdetector of the Nexis GC-2030system can be adjusted optimally

to any chromatographic techniqueby setting the data acquisition rate(up to 500 Hz) and the filter timeconstant (2 - 2,000 ms).

Safety with hydrogen as carrier gas

For most GC applications andparticularly for fast ones, hydro-gen is the ideal carrier gas. How -ever, its use presents risks in theevent of leakages.

The Nexis GC-2030’s automaticgas leak check assures adequatesafety.

A residual risk remains at highergas flows in low pressure ranges.Here, the optional hydrogen sen-sor offers additional safety. TheGC-2030 oven is monitored con-tinuously and the GC system isshut down before an explosivehydrogen concentration can form.

Summary

Over the past decades, gas chro-matography has developed into acomprehensive technique. Withthe Nexis GC-2030, Shimadzumakes this available to every user.Simple self-explanatory operationfacilitates access for ‘GC novices’intending to use GC analyticaltechnologies in their fields ofexpertise. Sensitive detectors alsomeasure in the ultra-trace rangewith highest precision. Sophisti -cated diagnostic techniques pro-vide information on the system’soperational capability at any time.Daily tasks are carried out withjust a few steps. Safety for eco-nomical operation using hydrogenas carrier gas is assured via op -tion al sensors. As the new stan-dard, the Nexis GC-2030 com-bines high performance with sim-ple operating conditions and reli-able long-term operation.

Figure 2: Color display with the

Nexis GC-2030 touchscreen

Figure 3a and 3b: Maintenance without tools: liner replacement of the split injector (SPL) or column installation in the GC oven.

The optional oven light facilitates these steps by providing good illumination of the column connections.

Further information

on this article:

• www.shimadzu.eu/

gc-2030

http://www.shimadzu.eu/nexis-gc-2030

http://www.shimadzu.eu/nexis-gc-2030

APPLICATION

4 SHIMADZU NEWS 2/2017

Harmful substances in textilesDetection of harmful substances in textiles using LC-MS/MS

T here is hardly anything thatwe allow as close to ourbody as our clothing. It pro-

tects us against cold, moisture orthe sun. We choose our clothingcarefully and wear it next to ourskin, and children often like toput pieces of clothing into theirmouths.

Many different chemicals are usedin textiles and clothing produc-tion. These chemicals have specificproperties such as vivid colors,wind and moisture impermeabili-ty, antibacterial effects, crease re -sistance and much more.

Ad ditive chemicals are used asoptical brighteners (optical white)or to reduce creasing and shrink-ing. They are used as stiffeningagents, anti-electrostatics, waterand oil repellents, protectionagainst pests, flame retardants andantimicrobial agents. In total,these preparations include about600 different chemical com-pounds, whereby dyes are not yettaken into account.

Not all of these substances areenvironmentally safe. Some aresuspected of causing allergies.Others do not degrade in the

In the manufacture of clothing,the number of additives and fin-ishing agents is vast. Nearly 6,000preparations are available, e.g. re -finement agents for fibers andyarns such as bleaching or hydro -philic agents for pretreatment, ad -ditives for dyeing and printing, aswell as dispersing agents, protec-tive colloids, dye accelerators, oxi-dation, mordant or resist agents.The list appears to be endless.

children from the effects of toxicchemicals, the so-called ‘silent epi-demic’ [1].

Legal foundations

The European Textile LabellingRegulation merely requires thatinformation is provided on thetextile fibers used. Labelling of theadditives used is, however, not yetrequired. Textiles are consideredto be commodities, and in thiscase the general prohibition ap -plies that the manufacture andtreatment of commodities mustnot be harmful to human health.Compliance with the legal provi-sions is primarily the responsibili-ty of the manufacturer. Monitor -ing of compliance with the legalprovisions is the responsibility of

the federal states. Since there is nomandatory authorization or re -porting requirement for textiles –only information on the fibersused is required – there is no com-prehensive knowledge of potentialrisks [2].

Experts and consumer organiza-tions have long been demanding aprecise declaration of ‘ingredi-ents.’ However, some harmfulsubstances are already prohibitedin clothing, such as carcinogenicazo dyes or certain flame retar-dants. A labelling requirement ap -plies to formaldehyde and a maxi-mum concentration of 5 mg/kgtextile applies to pentachlorophe-nol in accordance with the Ger -man Chemicals Prohibition Ordi -nance. However, these are only

environment and accumulate infatty tissues or are suspected ofbeing carcinogens.

Children are considered to be par-ticularly at risk in their mentaldevelopment. A multitude of com -pounds can potentially harm thebrain, but they are neither regulat-ed nor tested for harmful effectson fetuses and children. Expertsare calling for better protection of

Minutes

(x 100,000)

0.00

0.25

0.50

0.75

1.00

1.25

1.50

1.75

1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0

Figure 1: MRM Chromatogram of a PFC standard with 26 PFCs in a textile matrix (concen -

tration: 1,000 pg/mL, internal standard 13C-PFOA = M-PFOA concentration: 500 pg/mL)

Figure 2: MRM Chromatogram of a methanol extract of the textile GS with internal standard

(M-PFOA, 500 pg/mL). Traces of PFOA could only be detected in this sample.

Minutes Min.

(x 10,000) (x 10,000)

0.00.51.01.52.02.53.0

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.54.04.5

1.0 2.0 3.0 4.0 5.0 5.06.0 7.0 8.0 9.0 10.0


APPLICATION

specific legal provisions. Unlike,for example, cosmetic products,there is no standardized, compre-hensive regulation for textiles orleather.

Over 90 million tons of textilefibers were produced in 2015, alarge proportion of which incountries that do not adequatelyregulate the use of harmful sub-stances. This is a huge problem.The textiles produced are contam-inated with harmful substances,and the release of these harmfulsubstances into the environment is immense. The majority of thechemicals used for finishing aretransferred untreated into rivers,and sooner or later will enter thedrinking water and food chains.

PFCs also get into foods

Just how far-reaching the input ofchemicals into the environmentcan be, is shown by example ofthe so-called PFCs (perfluorinatedand polyfluorinated chemicals).This substance group includesmore than 800 compounds whichdo not occur naturally. Due totheir outstanding water, oil anddirt repellant properties, PFCs areoften used in the production ofhome textiles such as carpets, cur-tains, table cloths and pillow casesas well as upholstered furniture.PFCs are found in virtually allrainproof, breathable outdoor tex-tiles, they are used as anti-greaseingredient in coated paper such aspaper cups or pizza boxes, or asnon-stick coating in Teflon pans.

The problem is: PFCs are ex -treme ly stable. They do not de -grade in the environment but areonly distributed. In 2015, a Green -peace expedition found PFCs inall snow and water samples col-lected from seemingly untouchedand remote locations around theworld (among others, in China atan altitude of 5,000 meters and ina Chilean national park).

Persistent harmful substances aredistributed via the air and flowingwaters throughout the world andfinally also reach our food. Theyaccumulate in the body, bind toproteins in blood, liver and kid-neys and can be passed on to thefetus or the baby during pregnan-cy.

Sensitive detection of PFCs in textiles using LCMS

Because of these alarming proper-ties, consumer protection and en -vironmental organizations arefocusing on PFCs. The most pro -minent representatives are the per-fluorooctane sulfonic acid (PFOS)and the perfluorooctanoic acid(PFOA), both of which are likelyto be carcinogenic.

While the use of PFOS and itscomponents is already prohibitedas a substance in goods in the EUREACH Chemicals Regulation,PFOA is still on the list of candi-dates. Unintentional trace impuri-ties are exempted and for textilesthere is a threshold of 1 µg/m2 ofthe coated material. A fast andsensitive method for testing textilesamples is presented below.

Figure 1 shows the MRM chro-matogram of a PFC standard in a textile matrix with 26 compo-nents, including 13C-PFOA (= M-PFOA) as internal standard. Themeasurements were performedusing the LCMS-8050 triple quad -

Table 1: Limits of detection and quantification for 26 PFCs, including PFOS, PFOA, as well as reproducibility data (n = 6) for the concentration

levels 50 pg/mL and 1,000 pg/mL

rupole mass spectrometer coupledto a Nexera UHPLC. Separationof the compounds was achieved inless than twelve minutes with anexcellent limit of detection forPFOS (LOD = 5.3 pg/mL) andPFOA (LOD = 5.8 pg/mL).Further important statistic fea-tures of the method are listed intable 1. (Detailed information onthe methods can be downloadedvia the QR code at the end of thearticle.)

Analysis of real samples

PFC concentrations in five differ-ent articles of clothing (designatedas BS, GS, YS, WS and BR) fromlocal stores were determined usingthe method described. As shownin figure 2, traces of PFOA (650 pg/g = 0.65 ppb) were onlyfound in sample GS. None of the26 PFC compounds could bedetected in the other four samples.

Dyes

Dyes not only represent thelargest group, they are also themost important group of sub-

stances used in textile manufactur-ing with regard to health risks.For the assessment of healthaspects, a classification based onthe dyeing process is helpful.Water soluble direct dyes areincorporated in the hollow spaces(cavities) of the fibers. Water solu-ble reactive dyes, in contrast, arefirmly bound to the fiber bymeans of a covalent bond. Thesedyes are usually harmless becausethey are poorly absorbed by theskin.

Disperse dyes are lipophilic sub-stances that are directly dissolvedin the chemical fiber. For technicalreasons, only small molecules withthe addition of organic solvents(dye accelerators, carriers) areconsidered suitable as dispersedyes. In case of incorrect treat-ment like over-dyeing or incom-plete removal of the carrier, healthrisks due to these lipophilic sub-stances, some of which are readilyabsorbed via the skin, cannot beruled out. �

1234567

891011121314151617181920212223242526IS

No.

PFBAPFPAPFBSPFHxAHPFHpAPFHpA1H,1H,2H,2H-PFOSPFOAPFHxSFOEAPFNAPFHpSPF-3,7-DMOAPFDAPFOSH4PFUnAPFUdAPFDoAPFDSPFTrDAPFTeDAFOSAPFDHxAN-MeFOSAPFODAN-EtFOSAM-PFOA

1.3793.0634.2114.2454.3535.0045.373

5.5945.7335.8966.1126.2856.3086.5926.7936.5417.0457.4847.7217.9288.3728.4209.1989.9559.980

10.3695.556

Name RT (min)

50~5,00050~5,00020~5,00050~5,00020~5,00050~5,00050~5,000

20~5,00050~5,000

1,000~5,00050~5,00050~5,00050~5,00050~5,00020~5,000500~5,00020~5,00050~5,00050~5,00050~5,00050~5,00050~5,00020~5,000200~5,00020~5,000100~5,000

500

Range(pg/mL)

1.0000.99930.99910.99950.99930.99920.9992

0.99960.99920.99880.99960.99910.99960.99980.99900.99730.99950.99900.99910.99910.99910.99950.99850.99860.99820.9984

15.611.56.116.62.813.814.9

5.811.8246.313.38.715.39.65.382.35.311.515.315.213.411.61.460.85.032.1

R2 LOD (pg/mL)

47.334.818.550.28.541.845.3

17.735.6746.340.226.346.529.016.1249.316.035.046.246.040.635.04.1

184.115.197.2

3.56.93.55.74.59.19.3

5.910.7N.A.6.39.614.47.812.8N.A.11.110.48.013.57.614.014.7N.A.11.0N.A.

Not available

LOQ (pg/mL)

RSD (%), n = 6(50 pg/mL) (1,000 pg/mL)

3.71.03.43.62.23.95.7

1.32.112.66.73.54.17.26.112.44.83.43.68.38.88.22.56.21.76.0

Carcinogenic amines in azo dyes

By far, the most important groupof dyes are the azo dyes, about500 of them are synthesized onthe basis of carcinogenic amines.Even today, 150 of these com-pounds are still commerciallyavailable. Following absorptionby the body, azo dyes can becleaved enzymatically (for in -

REACH regulation, no textilestreated with such dyes may bebrought on the market if limit val-ues of 30 mg/kg are exceeded.

In the literature, a total of 49 dyesare described as contact allergensin the context of textile-relatedcontact allergies. About two-thirds of them are disperse dyes.In addition, the BfR (Federal In -stitute for Risk Assessment, Ger -

stance by intestinal bacteria, cer-tain liver enzymes or skin bacte-ria) back into the original poten-tially carcinogenic amines.

The German industry already hasgiven up the use of azo dyes for along time which can cleave intocarcinogenic aromatic amines.Imported textiles, however, maywell contain such harmful dyeseven though, as stated in the

APPLICATION


many) specified eight disperse dyesseveral years ago, that should nolonger be used in clothing textiles.

Screening and quantitativedetermination of 47 azo dyes

The following application exampledescribes a method for quick andreliable detection of 47 dyes fromdifferent dye groups, 23 azo dyes,21 disperse dyes and three triph-enylmethane. These also includeseven of the eight disperse dyesspecified by the BfR (disperseblue 35, 106 and 124, disperse yel-low 3, disperse orange 3 and 37/76; disperse red 1).

Separation of the 47 componentsis achieved within twelve minutes,as shown by the MRM chromato -gram in figure 3. To develop theanalytical method, a dye-free tex-tile sample was spiked with thecorresponding standard dyes andthe instrument parameters wereoptimized. The measurementswere performed using Shimadzu’sLCMS-8040 triple quadrupolemass spectrometer coupled toShimadzu’s Nexera UHPLC. (Adetailed description of the methodcan be downloaded via the QRcode at the end of the article).

Table 2 lists the limits of detectionand limits of determination as wellas calibration linearity data andreproducibility measurements.

Analysis of random samplesfrom clothing stores

A total of three light-colored arti-cles of clothing, designated as 0B,0G and 0Y, were purchased inlocal textile stores and analyzedusing the method presented. Oneof the 47 dyes could be identifiedunequivocally in one of the threerandom samples: disperse red,which is regarded as a contactallergen belonging to the eightBfR banned dyes (figure 4).

Conclusion

Fast and sensitive detection meth-ods for harmful chemicals in tex-tiles are necessary to enable ran-dom monitoring of, for instance,clothing. The introduction andmonitoring of limit values notonly serves to protect persons

50 ppb

Table 2: Limits of detection and quantification for 47 dyes, as well as reproducibility data (n = 6) for the concentration levels 10 ppb

and 50 ppb

1234567891011121314151617181920212223242526272829303132333435363738394041424344454647

No.

2,4-toluenediamineBenzidine4,4’-oxydianiline4,4’-diaminodiphenylmethaneo-anisidineo-toluidinep-cresidine2,4’-diaminoanisole2,4’-xylidine3,3’-dimethoxybenzidine4-chloroanilineo-tolidine4,4’-methylene-bis(2-methylaniline)2,6-xylidine2,4,5-trimethylaniline2-naphthylamine4,4’-thiodianiline4-chloro-o-toluidineBasic Red 94-aminobiphenylBasic Violet 145-nitro-o-toluidineDisperse Blue 7Disperse Yellow 9Disperse Blue 3Disperse Red 11Disperse Blue 102Disperse Red 174-aminoazobenzene3,3’-dichlorobenzidine4,4’-methylene-bis-2-chloroanilineDisperse Blue 106Disperse Orange 3Basic Violet 3Disperse Yellow 3Disperse Orange 11Disperse Brown 1Disperse Red 1Disperse Blue 35Disperse Yellow 1Disperse Yellow 49 (Leather)Disperse Blue 124Disperse Blue 26Disperse Orange 37Disperse Yellow 23Disperse Orange 1Disperse Orange 149

1.1481.5061.2681.5831.7332.0493.9753.9753.9334.5144.0944.3954.4844.9915.0015.4725.4576.2826.1396.6126.4796.8887.4597.8057.9217.9368.2948.4898.8158.7898.9788.9389.1289.29.2039.2739.2899.5719.8758.43810.09910.16310.77910.86411.04911.19512.069

1 - 1,0001 - 1,0000.5 - 200

0.5 - 1,0000.2 - 2000.2 - 2000.1 - 2001 - 2001 - 200

0.2 - 2001 - 200

0.5 - 2000.5 - 2001 - 200

0.5 - 2000.2 - 200

0.5 - 1,0000.2 - 5000.05 - 1000.5 - 1,0000.05 - 1005 - 1,0001 - 200

5 - 1,0000.5 - 2000.5 - 1002 - 200

0.5 - 2002 - 2002 - 2002 - 200

0.1 - 2000.1 - 2000.05 - 2000.5 - 2000.5 - 2001 - 200

0.2 - 1005 - 1,0002 - 200

0.5 - 2001 - 5005 - 2005 - 200

0.2 - 2000.2 - 2001 - 200

0.99920.99600.99440.99920.99930.99980.99920.99950.99940.99510.99970.99920.99930.99950.99950.99940.99930.99970.99310.99960.99770.99940.99910.99970.99940.99670.99910.99920.99950.99940.99930.99940.99940.99900.99910.99920.99930.99420.99800.99600.99960.99810.99860.99700.99690.99790.9975

0.680.520.260.350.150.160.060.580.730.200.780.280.210.850.290.100.190.150.020.450.021.980.715.000.280.200.970.501.231.472.000.040.080.020.210.341.000.201.982.000.260.784.094.050.170.120.73

0.220.170.090.120.050.050.020.190.240.060.260.100.070.280.100.030.060.050.010.150.010.650.241.500.090.060.320.150.410.480.600.010.030.010.070.110.330.070.650.630.090.261.351.340.060.030.24

1.924.894.471.284.902.673.086.721.652.013.753.642.623.213.223.542.911.991.221.812.7810.863.1015.165.143.0416.326.1610.4311.044.7710.345.814.423.9711.596.243.1610.5410.433.634.707.084.315.192.574.80

3.733.884.122.312.811.581.362.612.331.121.931.832.773.231.301.722.002.081.932.451.428.181.334.161.912.327.613.783.956.064.593.562.012.452.744.698.335.545.998.292.404.7412.853.351.751.237.42

Compound RT (min)

Range (ppb) Linearity LOQ LOD

RSD (%), n = 610 ppb 50 ppb


LATEST NEWS

wearing the piece of clothing butalso to support sustained protec-tion of the environment. Evenwhen some manufacturers arecommitted to stop the use ofknown harmful substances incloth ing, contaminated clothes arestill found in random samples col-lected by environmental and con-sumer organizations [3]. UsingLC-MS/MS, even small traces ofharmful substances can be deter-mined accurately and quantita -tively.

Literature[1] Focus online 06.03.2014: In Billigländern

produziert – Krebs- und Allergiegefahr:

So giftig ist unsere Kleidung.

[2] BfR, Bundesinstitut für Risikobewertung;

Einführung in die Problematik der Beklei -

dungstextilien. Aktualisierte Stellungnah -

me Nr. 041/2012 vom 06. Juli 2012

[3] Greenpeace Homepage, Report: Chemie in

unberührter Natur und Gefährliche Chemi -

kalien in Outdoor Ausrüstung (Greenpeace

Produkttest 2016)

[4] UBA, Umweltbundesamt, Homepage

Themenseite PFC

European Headquarters in Duisburg, Germany

Minutes

(x 100,000)

0.01.02.03.04.05.06.07.08.0

1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 9.0 10.0 11.0 12.0

Figure 3: Chromatogram of MRM-traces of 47 dyes of different dye groups in one run

(concentration 20 ng/mL (20 ppb) each; injection volume: 5 µL)

Minutes

(x 1,000)

0.01.02.03.04.05.06.07.08.0

2.5 5.0 7.5 10.0

Figure 4: MRM Chromatogram of the real

sample 0G with detection of the disperse

dye ‘Disperse Red 1’

Further information

on this article:

• Poster: SAP-ADSC

ASMS 2016 poster

LCMS-8050 for 26PFC

in textile

• Poster: SAP-ADSC – ASMS 2016 poster

LCMS-8040 for 47 dyes in textiles

One-stop solutions for clinicalmarket applicationsShimadzu has acquired AlsaChim, a specialist for high-quality analytical isotope labeled standards

S himadzu has joined forceswith the France-basedAlsaChim company special-

izing in stable isotope-labelledcompounds, metabolites and phar-maceutical related substances.AlsaChim is now by 100 % part ofthe Shimadzu family. The brandname will be kept for the future

complemented by the subtitle “a Shimadzu Group Company.”Through this acquisition, Shimadzuwill further develop and extend itsactivities in the clinical marketwhich is one of the focus areas forthe European Innovation Center(EUIC). AlsaChim technologycomplements Shimadzu’s product

and solution portfolio in the clini-cal market. Now, Shimadzu is ableto enter the market with completesolutions consisting of hardware

and software as well as applicationkits. Clients now benefit fromone-stop solutions of complexanalytical systems.

APPLICATION

APPLICATION


Nexera UC-MS/MS for highly sensitive SFC analysis

Analysis and evaluation of chiral drugs in biologicalsamples

P rojects in drug discoveryand safety are constantlyaiming at the development

of novel and safer drugs, thera-peutics and diagnostics. DuringAPI (active pharmaceutical ingre-dient) development, drug stereo -isomerism is recognized as anissue having clinical and regulato-ry implications.

Enantiomers have essentially iden-tical physical and chemical prop-erties, while potentially showinglarge differences in toxicity.

Stereoisomeric composition of adrug with a chiral center shouldtherefore be well documented. To evaluate the pharmacokineticsof a single enantiomer or any mix-ture of enantiomers, manufactur-ers must develop quantitativeassays for individual enantiomers

early in the drug developmentstage.

One of the challenges of chiralseparations is the fact that enan-tiomers exhibit the same physicaland chemical properties, so theycan only be separated in a chiralenvironment.

For chromatographic separationschiral stationary phases are avail-able that enable the separation ofstereoisomers. In order to deter-mine the stationary phase thatoffers optimum selectivity for thechiral separation problem at hand,screening runs are usually per-formed, testing several chiralcolumns at a time. Normal phaseHPLC is often the method ofchoice, however, SFC has fre-quently been proven to be thesuperior method for the separa-tion of chiral compounds.

This article presents an example ofthe selectivity and sensitivity ofdrug level monitoring in a biologi-cal sample and the evaluationresults of the analytical method asan application to pharmacokinet-ics research of chiral drugs usingSFC-MS/MS, after having selectedan appropriate column.

Analysis of omeprazole in a plasma sample

The applicability of human plasmamatrix to SFC was evaluated usingthe enantiomeric drug omepra-zole, a well-known proton pumpinhibitor, as an example. Figure 1shows the chemical structure of

Figure 1: Structure and chiral center of omeprazole

Figure 2: Nexera UC SFC-MS system

(x 1,000)

0

0.5

1.0

1.5

2.0

2.5

1.00 2.0 3.0 4.0 5.0

1:346.10 > 198.10 (+)

(A)

(B)

(x 10,000)

0

0.5

1.0

1.5

2.0

2.5

3.0

1.00 2.0 3.0 4.0 5.0

1:346.10 > 198.10 (+)

(A)

(B)

Minutes

Minutes

Figure 3: Chromatogram of a plasma sample spiked with a) 2 µg/L and b) 20 µg/L

omeprazole, analysed with the Nexera UC LC/SFC switching system

Mobile phase

Flow rate

Column temperature

Injection volume

BPR pressure

BPR temperature

Detector

Make-up solvent

Make-up flow rate

Column

CO2 / Methanol (80 : 20 v/v)

3 mL/min

40 °C

3 µL

10 MPa

50 °C

LCMS-8050 (ESI, MRM mode)

Methanol

0.1 mL/min

CHIRALPAK®, IC-3 (100 × 3.0 mm, 3 µm)


APPLICATION

omeprazole. For the preparationof a blood plasma sample, 20 µl ofplasma were mixed with 250 µl ofacetonitrile for protein precipita-tion. The supernatant was mixedwith acetonitrile and alkalizedwith 28 % aqueous ammonia.Three µl of the resulting samplesolution were injected for analy-sis. The analytical conditions forSFC-MS/MS analysis are dis-played in table 1.

Daicel’s CHIRALPAK® IC-3analytical column was identifiedas the most appropriate separationcolumn in an initial column screen -ing. Detection was performedusing the Shimadzu LCMS-8050triple quadrupole mass spectrome-ter.

A calibration curve was createdusing human plasma samples con-taining 1, 2, 10, 20 and 100 µg/Lof omeprazole standard. Figure 3shows the MRM chromatogramsof the samples spiked with 2 µg/Land 20 µg/L respectively, and thetwo omeprazole enantiomers (A)and (B) are well separated. Lin -earity of the calibration was R2 >0.99 for both isomers.

The method produced good re -peatability tested by five injec-tions of the 2 µg/L sample withRSD values of peak area of 4.4 %for both omeprazole (A) and (B).Recovery was determined from a10 µg/L sample compared to analy -sis of stock solution. Recoveryrates of 101.1 % and 100.5 %respectively were obtained.

Analysis of rabeprazole in a plasma sample

Rabeprazole, known as a gastricacid secretion inhibitor, has a sim-ilar chemical structure to omepra-zole, suggesting the possibility ofsuccessful chiral separation undersimilar analytical conditions.Rabeprazole was therefore ana-lyzed in a plasma sample using thesame analytical conditions asdescribed for omeprazole in theprevious section. The chemicalstructure of rabeprazole is shownin figure 4, where the structuralsimilarity to omeprazole can easi-ly be recognized.

Figure 5: Structure and chiral center of

rabeprazole

Table 1: Analytical conditions

Table 2: Results for linearity, repeatability and recovery rate for the determination of

omeprazole and rabeprazole in plasma samples

(x 1,000)

0

1.0

2.0

3.0

4.0

5.0

0

1.0

2.0

3.0

4.0

5.0

1.00 2.0 3.0 4.0 5.0

(x 10,000)

1.00 2.0 3.0 4.0 5.0 MinutesMinutes

1:359.90 > 150.10 (+)

(A)(B)

1:359.90 > 150.10 (+)

(A)

(B)

Figure 4: Chromatogram of a plasma sample spiked with a) 3 µg/L and b) 30 µg/L rabeprazole, analysed with the Nexera UC LC/SFC switching system

A calibration curve was generatedusing human plasma samples con-taining 0.3, 1, 3, 10 and 30 µg/L ofrabeprazole standard. Figure 5shows MRM chromato grams ofthe samples spiked with 3 µg/Land 30 µg/L respectively. The tworabeprazole enantiomers (A) and(B) are well separated. Linearityof the calibration was R2 > 0.99for both isomers.

The method achieved good re -peatability tested by five injec-tions of the 10 µg/L sample withRSD values of peak area of 1.8 %and 2.4 for rabeprazole (A) and(B) respectively. Recovery wascompared to analysis of stocksolution as 102.5 % and 100.1 %.Table 2 summarizes the linearity,peak area repeatability and recov-

ery rate for each compound.These results verify the applicabil-ity of this method to the practicalanalysis of plasma samples.

Omeprazole (A)

Omeprazole (B)

Rabeprazole (A)

Rabeprazole (B)

0.99996 (1)

0.99998 (1)

0.99996 (2)

0.99999 (2)

4.4 (3)

4.4 (3)

1.8 (4)

2.4 (4)

101.1

100.5

102.5

100.1

Linearity (R2) % Recovery (4)

Repeatability (% RSD peak area)

APPLICATION


Simple and fast characterizationtool for biosensor development

event occurs, which in the case ofa DNA biosensor means the cap-ture of the target DNA. There -fore, it becomes inevitable to havethe right probe that binds specifi-cally to the target. A controlledfabrication strategy is to monitoreach step of the development ofthe biosensor. Hence, it is impor-tant to know if the probe which inthis case is a single-stranded DNAand the target sequence are bind-ing. Shimadzu UV-1800 withThermal Melt Analysis Systemfinds its place in such a develop-ment phase of the biosensor (fig-ure 1).

Nucleic acids, as single or doublestrands of DNA and RNA, absorbultraviolet (UV) light due to theheterocyclic rings of the nucleo -tides. The absorption propertiesare normally used for quantifi -cation, since all absorb around 260 nm.

Furthermore, the absorption canbe used to detect sample hybridi -za tion in solution as a function oftemperature, for example. InssDNA, the bases stack on top ofeach other, but this conformationis maximized in dsDNA. Thehybridization stabilizes the helicalstructure of DNA and also RNA.

UV-spectroscopy with Thermal Melt Analysis System – microvolumes in spectral focus

O ne of the key aspects in the development of DNA-based biosensors relies on

understanding the molecular bind-ing event to be detected. DNA is apolymer comprised of nucleotidesforming four bases (adenine, thy -mine, guanine and cytosine).These nucleotides are bonded to -gether via phosphodiester bonds.In a double helix, two strands ofDNA are held together via hydro-gen bonding between adenine andthymine with two bonds and gua-nine and cytosine with triplebonds respectively. The study ofDNA is central to the understand-ing of many biological processesthat have been identified in manyhuman diseases.

At the University of Bath, theBiosensor Research Laboratoryhas been pioneering the develop-ment of DNA-based electrochem-ical sensors for the detection of awide range of biomarkers rangingfrom DNA, microRNAs, proteinsand cells. The addition of the UV-Vis spectrophotometer togetherwith Thermal Melt Analysis Sys -tem has been a very useful tool tounderstand the biosensor develop-ment approach.

Biosensor, signal and target DNA

A biosensor is an analytical devicethat can be used for detection ofan analyte (pollutant, viral DNA,antigen or protein) by combining a biological component (enzyme,DNA, antibody or aptamer re -spectively) that acts like a probeand binds to the target. Such abinding event gives rise to a signalwhich is detected by a transducer(mass sensitive, optical or electro-chemical) [1].

In order to get a signal, it is im -portant that the specific molecular

Figure 1: Shimadzu UV-1800 with Thermal Melt Analysis System (TMSPC-8) consisting of eight

series micro multi-cell cuvette

Figure 2: Schematic of a DNA biosensor. Left shows the co-immobilization of thiolated DNA

probe with a spacer molecule (6-mercapto-1-hexanol). Right represents post capture of target

DNA resulting in the formation of a duplex strand.

Such a feature has been proven tobe useful in understanding of thebase-pairing of double-strandedDNA [2]. Such a property wasused to understand the meltingcurve of the sequences of DNA-based biosensor development.Figure 2 demonstrates a schematicof a gold electrode surface afterthe modification with thiolatedsingle-stranded DNA probe with a spacer molecule (6-mercapto-1-hexanol) to control the probe sur-face density.

Methods

For characterization of the natureof hybridization of the probeDNA and target DNA, an 8-seriesmicro cell was used with siliconeplugs, which fits inside theTMSPC-8 temperature controlledaccessory. A Shimadzu UV-1800spectrophotometer equipped withthe TMSPC-8 was applied for thethermal melt experiment.

1 µM concentration of probeDNA (5’- SH(CH6) -TTTT-TATTGTGACAGACCATTGC-TACA-3’), target DNA (5’-TGTAGCAATGGTCTGTCA-CAAT-3’) and the mixture ofprobe and target DNA was pre-pared separately in 10 mM phos-


APPLICATION

to changing of the color to deepred.

Thereafter, the synthesized AuNPwas modified by polyethyleneglycol (PEG). 1 mL of AuNP wasincubated with 100 uL of 0.5 mM(O-(3-Carboxypropyl)-O’-[2-(3mercaptopropionylamino) ethyl]-polyethylene glycol (PEG) fromSigma-Aldrich [MW 3000 Da]) for16 hours at 4 °C with constantstirring. The modification ofAuNP before and after the modi-fication was monitored usingShimadzu’s UV-1800, looking atthe absorbance spectrum in thewavelength from 400 to 800 nm.

Results and Discussion

Figure 4 shows the real-time melt-ing curve obtained for the DNAprobe and target DNA from 19 °Cto 90 °C and back. Two DNAstrands hybridize at room temper-ature and with the increase oftemperature, the conformationchanges until 60 °C. Up to this

reducing necessary resources andcost.

Instrumentation

Shimadzu UV-1800 includingcomputer control softwareUVProbe, high precision cellholder: The TM analysis system (TMSPC-8) is an accessory forUV-Vis, comprising of a thermo-electrically temperature controlledmicro multi-cell holder employinga Peltier element and PC softwarespecially designed for the purpose.

Figure 3: Synthesis of gold nanoparticles

(AuNP) followed by modification with poly-

ethylene glycol (PEG)

0.210

0.220

0.230

0.240

0.250

0.260

0.270

20.0 30.0 40.0 50.0 60.0 70.0 80.0 90.0

Abs.

Temperature in °C

Figure 4: Melting curve of DNA probe and target DNA in 10 mM PBS buffer, pH 7.4

Figure 5: Absorbance curve of gold nan -

oparticle (AuNP) before and after modifi -

cation with polyethylene glycol (PEG)

value, the double helix denatures,increasing the absorbance. Whenthe temperature decreases, the sin-gle strands hybridize again andthe absorbance reduces. The tem-perature at which half of DNAstrands are in the single-stranded(ssDNA) state is termed as melt-ing temperature (Tm). From thesoftware analysis, the Tm in 10mM PBS (pH 7.4) was found to be66 °C.

The analysis confirmed that thetwo strands obtained hybridizedwell due to their complementaryproperties. Different nucleotidesequences lead to different denat-uration temperatures which can beanalysed with UV light. Similarly,hybridization events can be con-firmed using this technique.

Figure 5 shows the absorbancecurve obtained for the synthesisedAuNP. The graph shows charac-teristic peaks of 20 nm AuNPs at520 nm. After the modification ofAuNP with PEG molecules, thecharacteristic peak shows a 3 nmdisplacement which is well inaccordance with the reported lit-erature [3].

Conclusion

In conclusion, the impetus of thisapplication news was to demon-strate how the Shimadzu UV-1800together with Thermal Melt Anal -ysis (TMSPC-8) system can beutilized as a characterizing tool to develop a biosensor. The UV-1800, when incorporated withthe TMSPC-8 System is capableof monitoring the changes inabsorbance of the dsDNA meltingas a function of temperature anddetermining Tm values for thedsDNA in the buffer applied. Itwas also used to understand thenature of hybridization and toconfirm the use of right sequencesfor the development of DNA-based sensor. UV-1800 was alsoapplied to monitor the changes inabsorbance on the modification ofgold nanoparticles with PEG.With the 8-channel micro cell,multiple curves can be capturedsimultaneously for the analysis ofrepetitive sample. Another advan-tage of using such a cell is theneed of low sample volume (100 µL) enabling analysis of sam-ples in a most efficient way by

Abs. (a.u.)

� (nm)

0.0

0.2

0.4

0.6

0.8

1.0

400 500 600 700 800

3 nm shift

–– AuNPs–– AuNPs + PEG

AuthorsPawan Jolly 1, Marina Batistuti 2, Pavel

Zhurauski 1, Pedro Estrela 1

1University of Bath, UK; 2 University of

São Paulo, Brazil

Literature[1] Bhalla N, Jolly P, Formisano N & Estrela P.

“Introduction to Biosensors”, Essays in

Biochemistry. Volume 60 (1), p. 1. 2016

[2] Boyer, Rodney F. “Modern Experimental

Biochemistry.” Second Edition. The

Benjamin/Cummings Publishing Company.

1993

[3] Gao, Jie, et al. “Colloidal stability of gold

nanoparticles modified with thiol com-

pounds: bioconjugation and application in

cancer cell imaging.” Langmuir. Volume 28

(9), p. 4464. 2012

phate buffer saline (PBS, pH 7.4).The first cell was just used tomeasure the blank with PBSbuffer, followed by the use of twocells each for probe DNA, targetDNA and mixture (probe + tar-get) respectively. The programwas set with a complete cyclewhere the temperature was raisedfrom 19 °C to 90 °C and back.Using a ramp rate of 0.5 °C/min,the absorbance was recorded at awavelength of 260 nm.

Figure 3 shows the schematic ofgold nanoparticle (AuNP) beforeand after modification with poly-ethylene glycol (PEG). Synthesisof 17 nm AuNP was adopted bythe protocol reported by Gao, Jie,et al [3]. Briefly, 5 mL HAuCl4gold (III) chloride hydrate (0.2 %w/w) from Sigma-Aldrich wasadded to 90 mL water and washeated to 80 °C with constantstirring. Later, 5 ml of trisodiumcitrate dihydrate solution (1%w/w) from Sigma-Aldrich inwater was added quickly leading

APPLICATION


Lighting up Determination of quantum efficiency of fluorescencestandards with high quality

O ne of our first experienceswith materials lighting upat night is definitely the

glowing dial of a wristwatch. Awide variety of materials exhibitthis property in which energysupplied from an external sourceleads to the emission of light (flu-orescence or phosphorescence).This physical principle of fluores-cence is used in the developmentand quality control of lightingsystems, displays or monitors, as well as in biotechnology andmedicine. For these applications,modern instrumental analysismakes use of reliable and precisemeasurement technologies tocharacterize this fluorescence.

In fluorescence spectroscopy,there is an interdependencebetween instrument technologyand the wavelength of the light.Modern instruments can performcomputational spectra correctionin order to reliably compare dif-ferent instruments. Such a correc-tion takes into account the charac-

teristics of the emission optics aswell as the excitation optics. Thespectra corrected for these charac-teristics are subsequently used todetermine the so-called quantum

yield of the fluorescing materials(fluorophores). The quantumyield describes the probability ofhow effectively a material canconvert the irradiated energy bythe excitation light into the emit-ted fluorescent light.

Quantum yield: relative andabsolute method

In the past, this determination wascarried out using a relative meth -od, in which a known referencestandard was measured and its flu-orescence intensity was then com-pared to that of a sample. Forunknown compounds, however,these quantum yields must bedetermined in a different way viaan absolute method – known as‘quantum efficiency’ (QE).

In this case, the quantum yield iscalculated directly from a spec-trum measured from the fluores-cent substance using an integrat-ing sphere, which effectively col-lects all fluorescence quanta andpasses these to the emission detec-tor. Figure 1 illustrates the approachand mathematics for the determi-nation of the quantum efficiency.

Figure 1: Determination of the absolute quantum yield

SHIMADZU NEWS 2/2017 13

L

are prepared are important. Sol -vent concentration, pH value inaqueous solution and also temper-ature must be taken into account.Different solvents used to preparethe fluorescent standards lead todifferent quantum yields.

In addition to these aspects, theIUPAC report also discusses themeasurement technology applied.Two variations are the rule:• relative measurement of quan-tum yield using a cuvette in a

standard sample holder of a flu-orescence spectrometer. • absolute measurement of quan-tum efficiency using an integrat-ing sphere.

The present application not onlydemonstrates the absolute deter-mination of the quantum efficien-cy of three fluorophores usingShimadzu’s RF-6000 with integrat -ing sphere (figure 2) but also thegood correlation of these resultswith corresponding literature val-ues. In this application, the quan-tum efficiencies of fluorescein,quinine sulfate and tryptophanwere determined as examples.

Carrying out the measurement

Prior to measurement, the appro-priate concentration of the solu-tion is adjusted and the dilutedfluorophore is transferred to acuvette with four polished sides.This cuvette is placed into theintegrating sphere of the RF-6000and measurement is subsequentlystarted.

The measurement parametersselected in the LabSolutions RFsoftware are listed in table 1. Ameasurement was carried out withan average speed of 600 nm/min.The instrument was set to a slitwidth of 5 nm for excitation aswell as for emission. Detector sen-sitivity was set to low.

The ‘Quantum Efficiency’ func-tion of the LabSolutions RF soft-ware was used to evaluate quantumefficiency. The areas used for sig-nal integration are listed in table 1.As an example, the quantum effi-ciency measurement of quininesulfate is shown in figure 3. Table2 compares the measured absolutequantum efficiency with the dataobtained from the literature.

All three QE values of the fluo-rescence standard examined lieexactly within the range of thedescribed target value.

Conclusion

The RF-6000 is excellently suitedto quantum yield measurements.With its high signal-to-noise ratioin combination with its high speedand quantum yield/efficiency soft-ware, absolute quantum yields canbe determined quickly and accu-rately.

Literature

[1] Standards for photoluminescence quantum

yield measurements in solution (IUPAC

Technical Report)*

Albert M. Brouwer; Universiteit van

Amsterdam, P.O. Box 94157, 1090 GD

Amsterdam, The Netherlands, Pure Appl.

Chem., Vol. 83, No. 12, pp. 2213-2228,

2011; doi: 10.1351/PAC-REP-10-09-31; ©2011 IUPAC, Publication date (Web):

31 August 2011

[2] Reevaluation of absolute luminescence

quantum yields of standard solutions

using a spectrometer with an integrating

sphere and a back-thinned CCD detector

Kengo Suzuki, Atsushi Kobayashi, Shigeo

Kaneko, Kazuyuki Takehira, Toshitada

Yo-shihara, Hitoshi Ishida, Yoshimi Shiina,

Shigero Oishic and Seiji Tobita; Phys.

Chem. Chem. Phys., 2009, 11, 9850-9860;

DOI: 10.1039/b912178a

In 2011, IUPAC published a tech-nical report [1] in which the quan-tum yields of various fluorescencestandards were compared basedon the existing literature. As asmall set of well-established stan-dards, quinine sulfate, fluoresceinand rhodamine 6G are listed. Inaddition, the review also includes9,10-diphenyl anthracene, beta-carboline, (norharmane), harma-line, rhodamine 101 and cresylviolet as well as 28 other frequent-ly used substances [1].

Fluorescence standards dependent on solution conditions

When considering the selection ofthe standard, the conditions of thesolution in which the standards

APPLICATION

Quinine sulfate

Fluoresceine

Tryptophan

5 x 10-5 M

10-7 M

10-5 M

0.05 M H2SO40.1 M NaOH

Destilled water

350 nm

488 nm

270 nm

330 - 365 nm

470 - 500 nm

250 - 290 nm

370 - 635 nm

500 - 700 nm

300 - 480 nm

Concentration Solvent Excitation atExcitation Emission

Area range evaluation

Table 1: Conditions for evaluation of the measured spectra for quantum efficiency determination

Figure 2: Integrating sphere for the RF-6000; left: integrating sphere; middle: cuvette

in the opened sphere; right: graphical representation of the light path of excitation and

emission within the sphere.

Table 2: Comparison of absolute quantum yields (QE, quantum efficiency) for fluorescein,

quinine sulfate and tryptophan measured with an integrating sphere (data obtained from the

literature) with quantum yields obtained using the RF-6000 with integrating sphere

IUPAC [1]

Reevaluation [2]

LabSolutions RF

Source of the QE values

0.91 ± 0.05

0.88 ± 0.03

0.9045

0.60 ± 0.02

0.60 ± 0.02

0.5898

0.15 ± 0.01

0.15 ± 0.01

0.152

QE value QE value QE valueFluoresceine Quinine sulfate Tryptophan

Intensity

Excitation wavelength [nm]

0.00

20,000.00

40,000.00

60,000.00

82,908.87

300.0 400.0 500.0 600.0 650.0

Figure 3: Determination of the quantum efficiency of a quinine solution at a concentration

of 5 x 10 -5 M; QE was calculated as 0.5898

External view of integrating

sphere attachment

Inside of integrating sphere

attachment

To emis-sion mono-chromator

Excitation light

Cell

CellTo emissionmonochromator

Excitation light

Behavior of light

in integrating sphere

attachment

PRODUCTS


I n the last issue of ShimadzuNews, the first part of thisEDX article series focused on

safety application in aircraft, auto-mobiles and railway locomotives,in particular the analysis of engineoils containing fine wear productsas well as particles of metals andalloys in the form of shavings.Testing laboratories of manymajor airlines use X-ray fluores-cence spectrometers for fast analy-sis of small metal shavings. Thissecond part of the EDX articleseries covers food packagings.

packaging. For a typical measure-ment, the analyst breaks the sam-ple and dissolves part of it withinclusion in the appropriate re -agent. Such sample preparation istime-consuming and dissolution is usually associated with the useof hydrofluoric acid, requiringspecial utensils and a separatework area.

All this can be avoided by usingthe energy dispersive X-ray fluorescence spectrometer EDX-7000P or EDX-8000P forthe analysis. These are used fornon-destructive elemental analysisof solid, powder and liquid sam-ples while offering excellent main-tenance performance. Both instru-ments are BfS (safety standards ofthe German Federal Institute forRadiation Safety) type approvalcertified. The sample observationcamera and automatic collimatorswitching system allow localanalysis of different parts of thesample.

Standardless quantitative fastanalysis of single extraneousinclusion in glass jar

In an experiment, single extrane-ous inclusion on the outer surfaceof glass jars was measured usingEDX-8000P. A glass jar was

Contaminants in packagingmaterials

Human health can be influencedby the choice of food – in a posi-tive as well as in a negative way.Despite their apparent quality andhealthiness, food products stillhave to be analyzed for the pres-ence of harmful and dangerouscontaminants. Such impurities orresidues can get into the productsfrom packaging materials – forexample, bags cans, jars, and bot-tles. Although glass packagingmaterials seem to be the safest,they can also contain contami-nants that have entered the glassduring the manufacturing processfrom furnace, piping or meltingforms.

These inclusions can be hazardousto human health. In addition, theycan influence the durability of the

How spectrometers take care of human healthInclusions in packagings analyzed with EDX-7000P/8000P

placed in the sample compartmentso that the inclusion was in thecenter of the area to be analyzed.A collimator of 1 mm diameterwas selected for the analysis (fig-ure 1). A routine measurementprocedure of un known sample byFundamental Parameters (FP)method was used, as included inthe standard spectrometer soft-ware. The spectrum of the sampleis shown in figure 2.

Conclusion

The results of the sample analysedare listed in table 1. They showthat the analysis of extraneousinclusions by EDX-7000P/8000Pcan be done successfully withouttheir separation from glass, whicheliminates time-consuming samplepreparation and use of chemicalreagents.

Moreover, it greatly reduces timeof analysis. Data acquired helps todetermine the possible source ofcontamination (cast iron meltingform, furnace lining or other).

Figure 1: Photo of the inclusion at

PC screen

Table 1: Results of analysed example

[cps/uA] Na-U

5.00 10.00 15.00 20.000.000

0.005

0.010

0.015

0.020

Figure 2: EDX spectrum of inclusion on outer surface of glass jar. Total analysis time

including placement of the sample in the cell was approximately three minutes.

Concentrationwt %

68.624

12.534

8.048

3.570

2.987

2.766

0.999

0.129

0.123

0.089

0.067

0.064

SiO2Al2O3SO3CaO

K2O

Fe2O3TiO2ZrO2SrO

MnO

CuO

Rb2O

Element

APPLICATION


Food and food packaging: non-intentionally added sub-stances FTIR, EDX and LC-GC-online Technique for colored, non-transparent plastic packaging

F ood can be contaminated notonly through direct contactwith its packaging, but also

along the entire production andcommercial chain. So far, legisla-tion on the migration of substanc -es from plastic packaging intofood in particular has covered thestarting materials used in the plas-tics manufacturing. Regardinghealth risks related to the transferof substances, the so-called NIAS(non-intentionally added sub-stances) should be focused onmore strongly; these substancesare added non-intentionally dur-ing manufacturing of plastic pack-aging materials. These can includeimpurities originating from rawmaterials, reaction and degrada-tion products in manufacturing aswell as impurities arising fromtransport and production [1].

Contamination during the production process

Possible sources for impurities canbe found in the production pro -cess, including preservation and

bottling, in which metals can mi -grate into food and beverages. Of -ten, raw as well as processed prod -ucts are in prolonged contact withmaterials such as stainless steel,cop per, glass and other equip ment.

Impurities in the packagingmaterial

Another source is the packagingmaterial. It is in close contact withthe food – during the entire trans-port, storage in retail as well as inthe consumer’s home.

Currently, there is much discus-sion on food packaging with satu-rated and aromatic mineral oilhydrocarbons (MOSH – mineraloil saturated hydrocarbons andMOAH – mineral oil aromatichydrocarbons).

The MOSH fraction consists oflinear and branched alkanes aswell as alkyl substituted cycloal -kanes, whereas the MOAH frac-tion consists of alkylated polyaro-matic hydrocarbons with up tofour aromatic rings. The mainfocus is on the aromatic fraction,which is suspected of being poten-tially carcinogenic and mutagenic[2]. The proportion of the aromat-ic fraction is approximately 15 -30 % of the total mineral oil frac-tion.

In addition to the mineral oil frac-tions MOSH and MOAH, the so-called POSH (polyolefinic oligo -meric saturated hydrocarbons) arefocused, i.e. oligomers that canmigrate from plastic packaging(PE, PP).

Scope of the new plastics regulation

In principle, the CommissionRegulation (EU) No. 10/2011 onplastic materials and articles thatcome into contact with food stip-ulates that impurities as well �

Plastics in food packaging

Occurence

0

2

4

6

8

10

12

14

16

18

PP PE

14 14

7

1

16

PET PA PS

Figure 1: Occurrence of various plastics among the 43 samples investigated

APPLICATION


and inner surfaces of the plasticpackagings could be identified.The absorption was measuredusing Shimadzu’s IR Tracer-100equipped with a diamond ATR. In this method, the IR beam pene-trates the sample to a depth ofabout 2 µm, whereby the intensity

as reaction and degradation prod-ucts must be assessed by the man-ufacturer in accordance with in -ter nationally accepted scientificprinciples of risk management.

As of 1 January 2016, the amend-ed test conditions described there-in fully apply. In addition to over-all migration (migration limit: 60 mg/kg food), there are manyother specific migration limit val-ues that must be adhered to.

‘Multi-material multi-layer mate-rials’ are also included inthe new plastics regulationas well as materials or ob -jects consisting of two ormore layers of differentmaterials of which at leastone consists of plastic [3].As reported in theShimadzu News 1/2017, a total of 32 transparent,colorless and 50 imprintedfood packagings were in -vestigated. This articlenow focuses on colorednon-transparent plasticpackagings. The plasticswere analyzed using FTIRand EDX spectroscopy aswell as an LC-GC onlinesystem. Spectroscopicmethods offer the advan-tage of non-destructivesample analysis with littletime expenditure. Thechro matographic method can befully automated.

Within the scope of the investiga-tion, 43 samples of different originwere analyzed. This included, forinstance, caps or closures of bot-tles and cups, vegetable nets and

trays. First, the samples wereidentified using FTIR spectrosco -py and later analyzed for non-intentionally added substancessuch as heavy metals and mineraloil hydrocarbons.

Identification of plastics usingFTIR spectroscopy

All samples were analyzed in thefirst step using FTIR spectrosco -py. This enabled a non-destructiveinvestigation of surfaces in whichthe main components of the outer

The microplastics particles foundconsisted mainly of PE, PP and PS [5].

Quantitative analysis of heavy metals using EDX spectroscopy

EDX spectroscopy allows theidentification of fillers and ROHSelements that may be present inthe sample. Most samples containlarger amounts of the elements sil-icon, calcium, titanium and alumi -num. This confirms the presenceof fillers such as kaolinite (SiO2/Al2O3) and calcite (CaCO3). Thetitanium is present as titaniumoxide and serves not only as awhite pigment but also as a stabi-lizer. By absorbing UV irradia-tion, it protects against photo-oxi-dation of the polymer [6].

PET is produced using an antimo-ny trioxide catalyst [7]. For thisreason, PET-containing sampleswere screened for antimony resi -dues using a fast screening methodspecifically optimized for ROHSelements as well as for antimonyand chloride. Table 2 shows theanalysis results for sample 08. It contains 311.3 ppm antimony.In other PET-containing samples,antimony concentrations of sever-al hundred ppm were found.

Sample 08 from table 1 still con-tains antimony residues. Many ofthe food packagings investigatedconsist of an outer PET layer andan inner PE layer. In this way, thefood contained is not in directcontact with the antimony-con-taminated PET layer. The exam-ples show that a combination of

of the reflected light is attenuatedwith respect to the irradiatedlight.

In a second complementary step,an EDX analysis was executedusing Shimadzu’s EDX-8000P.Upon suitable excitation, each ele-ment emits characteristic X-raysallowing the element’s identifica-tion. Using energy dispersive X-ray fluorescence analysis, elementsfrom carbon up to uranium caneasily be detected in the lowerppm range.

To analyze the samples, atotal of 48 IR and 40 EDXspectra were recorded.Table 1 lists samples thatinclude packagings consist-ing of one or two mainpolymers. Most of themwere black or white, whileotherswere blue, yellow,orange, green or gold.

The most frequently usedpolymers are polypropy-lene (PP), polyethylene(PE) and polysty rene (PS).Due to its high heat resist-ance, PP is used for pack-aging hot-filled or steril-ized products while PS isused mainly in disposableitems due to its high sus-ceptibility to breakage [4].

Figure 1 shows polymer composi-tion of the investigated sampleswhich shows great similarity tothe composition of maritime mi -croplastics occurring on a remoteMaldive island, as investigated in a study by the Germany-basedBayreuth University.

Figure 3: EDX spectrum of sample 08 (excitation energy 50 kV). The sample contains large

amounts of calcium, titanium and iron.Figure 2: PET tray for tomatoes; Sample 08

[keV]

0.0

2.0

4.0

6.0

10.00 20.00 30.00

[cps/uA]

Figure 4: MOSH/MOAH application system


APPLICATION

Figure 5: Chromatogram of the MOSH and MOAH fractions

FTIR and EDX spectroscopyoffers access to a multitude ofinformation on plastics and theircontents. Both methods are com-pletely non-destructive andrequire minimum time.

MOSH/MOAH analysis usingonline LC-GC technology

Undesirable mineral oil residuesoften enter products through theuse of mineral oil based printinginks which can be found in foodand food packagings. This effecthas been increasingly detected notonly in recycled materials but alsoin packagings consisting of freshraw materials. In many products,the concentration of saturated(MOSH) and aromatic (MOAH)hydrocarbons is particularly high.

For this analysis, a MOSH/MOAH application system is nowavailable based on LC-GC cou-pling and was developed in accor-dance with EN 16995:2016 ‘Deter -mination of mineral oil saturatedhydrocarbons (MOSH) and min-eral oil aromatic hydrocarbons(MOAH) with online HPLC-GC-FID analysis; German and Englishversion FprEN 16995:2016.’

The system consists of the follow-ing components:• Shimadzu’s LC-20ADXRpumpunit with UV detector anddegasser• Shimadzu’s GC-2010 Plus withtwo FIDs

• CTC autosampler • Semrau CHRONECT® LC-GC(figure 4).

The method offers high samplethroughput, reproducible resultsand good sensitivity. The systemenables LC-GC measurementswith a reproducibility comparablewith conventional split/splitlessinjection in a gas chromatograph.Direct coupling reduces the riskof contamination in manual meth-ods. The advantage of the systemis the configuration of two FIDdetectors, allowing parallel deter-mination of MOSH and MOAHwithin a single run and leading tocharacteristic chromatograms asshown in figure 5.

Conclusion

The Commission Regulation (EU)No. 10/2011 on plastic materialsand articles that come into contactwith food (also referred to as‘Plastics Implementation Mea -sure’, PIM) has been in force since4 February 2011. As of 1 January2016, the test conditions pre-scribed therein apply fully. Inaddition to overall migration(migration limit 60 mg/kg food),many other specific migrationlimits must be adhered to.

Shimadzu offers a comprehensivesolution for the analysis of non-

Table 2: Results of EDX screenings for ROHS elements, antimony and chlorine of sample 08.

ND = not detected; 3-sigma = triple standard deviation.

Table 1: Organic and inorganic main components of ten selected food packagings

01

02

03

04

05

06

07

08

09

10

No.

Fast Food

Cutlery

Soft Drink

Coffee

Milk

Sweets

Potatoes

Tomatoes

Pears

Sausages

Sample

white

gold

silver

black

blue

black

yellow

black

black

black

Color

PS

PE

PET

PS

PE

Polymer(inner surface)

PS

PP

PE

PET

PE

PP

PE

PET

PS

PA (Nylon6)

Polymer (outer surface)

06 PS

––

––

––

––

01 PET

––

01 PET

06 PS

––

Recycling symbol

Si, Al

Si, Al, Cu, Ti, Zn, K

Ca, Ti, Al, Si, K

Al, Si, S, Fe, K, Ba

Ti, Al, Si, S, Ca

Ca

Ti, Al, Ca, Si

Ca, Ti, Si

Si, Al, Ca, S, Ti

Si

Additive

Cd

Pb

Cr

Hg

Br

Cl

Sb

Element

ND

ND

3.9 ppm

ND

ND

53.5 ppm

311.3 ppm

Result

[18.0]

[10.8]

[4.4]

[2.0]

[2.1]

[59.5]

[62.0]

3-sigma [ppm]

0

250,000

500,000

750,000

1,000,000

7.5 20.0 22.5 min.17.515.012.510.05.02.50

0

250,000

500,000

750,000

1,000,000

7.5 20.0 22.5 min.17.515.012.510.05.02.50

uV

intentionally added substances(NIAS) including heavy metals(antimony trioxide) and mineraloil fractions (MOSH/MOAH).

Literature[1] www.vzhh.de/ernaehrung/368577/

lebensmittelverpackungen-unbeabsichtigt-

eingebrachte-stoffe.aspx

[2] R. Lorenzini, M. Biedermann, K. Grob,

D. Garbini, M. Barbanera, I. Braschi, Food

Addit Contam 30 (2013) 760-770

[3] VERORDNUNG (EU) Nr. 10/2011 DER KOM-

MISSION vom 14. Januar 2011 über Mate -

rialien und Gegenstände aus Kunst stoff,

die dazu bestimmt sind, mit Lebensmitteln

in Berührung zu kommen

[4] Dr. H. Saechtling: Kunststoff Taschenbuch.

31. Ausgabe, Carl Hanser Verlag, München

2013

[5] Hannes K. Imhof et al., Spatial and tempo-

ral variation of macro-, meso- and micro -

plastic abundance on a remote coral

island of the Maldives, Indian Ocean,

Marine Pollution Bulletin (2017)

[6] Dr. H. Zweifel: Plastics Additives Hand -

book. 6. Ausgabe, Carl Hanser Verlag,

München 2009

[7] DLG: www.dlg.org/verpackungsmaterial_

pet.html

Further information

on this article:

• Flyer MOSH/MOAH

Analyzer

http://www.dlg.org/verpackungsmaterial_ pet.html

http://www.dlg.org/verpackungsmaterial_ pet.html

http://www.vzhh.de/ernaehrung/368577/lebensmittelverpackungen-unbeabsichtigt-eingebrachte-stoffe.aspx



W ith a bright smile, theauthor has boughthealthy full grain bread,

persuaded to do something goodfor the body. Being hungry, he atethe bread straight away and may -

Let’s name this particle (figure 1).It has a size of 3 to 5 mm and ashape like a rice corn. Under thestereo microscope, it showed anorange color. Under normal daylight, it appeared white. Having

thing okay. But a hard particleneeds investigating ...

Once isolated, the particle lookedlike a rice corn which had dried tobe hard as a stone.

be a little too fast. Suddenly –a cracking sound, he has bitten on something incredibly hard. Ohno, the teeth! Is one broken?Repair is so expensive, it would bea disaster. Quickly checked, every -

Life can be hardContamination of bread - Particle analysis with FTIR-ATR single reflectionATR technique and EDX for inorganic analysis

APPLICATION


Abs.

cm-1

-0.0050

-0.0025

0.0000

0.0025

0.0050

0.0075

0.0100

0.0125

0.0150

0.0175

0.0200

4,500 4,000 3,500 3,000 2,500 2,000 1,500 1,000750 500

––– Contaminant in bread 1

Contaminant very hard in bread, Quest

Figure 2: Low-pressure: infrared spectrum from the surface of the particle, which is

dominated by cellulose

Figure 1: The contaminant in wholegrain bread has a size of 2 x 5 mm; this photo was

obtained with a camera integrated in a stereo microscope


APPLICATION

being hurt by this particle, it wasof interest to analyze the realchemical consistence.

Particle measurement techniques

For a rough screening and identi-fication of materials, two non-destructive analysis techniques areavailable: EDX and FTIR. Non-destructive means that the samplekeeps its form and that a chemicaltreatment is not needed.

EDX (energy dispersive X-Rayfluorescence spectrometer) makesthe analysis easy due to the “placeand measure” nature of its appli-cation. Infrared spectroscopy(FTIR Fourier Transform InfraredSpectroscopy) is a measurementtechnique which helps to identifyvery hard material using the singlereflection ATR (Attenuated TotalReflection) tool without any otherphysical treatment. This is possi-ble because the ATR unit SpecacQuestTM has an integrated dia-mond window which makes itpossible to analyze very hard solidmaterials at their surfaces (pene-tration depth of the infrared beamof ~2 µm). The hardness of thediamond will help to do so.

Sample preparation and spectral analysis using FTIR

No special sample pretreatmentwas used. The particle had a rice-corn shape of 3 by 5 mm. It wassimply placed on the diamond

Abs.

cm-1

0.00

0.01

0.02

0.03

0.04

0.05

0.06

0.07

0.08

4,500 4,000 3,500 3,000 2,500 2,000 1,500 1,000750 500



Figure 4: high-pressure: reconfirming the result from 2, a collection of fine crystals from

the particle separated powder

window of approx. 2 mm size andthe anvil (accessory) was screweddown to press the corn onto thediamond window, breaking it intosmaller, still very hard pieces. Atthis moment, it became obviousthat an inorganic material waspresent. FTIR measurement wasdone in a range of 400 to 4,600 cm-1.Infrared reflection spectra are pre-sented in figures 2 to 4.

Anvil pressure was set in threelevels. With low pressure a cellu-lose spectrum was measured at theeffective top surface. This is ac -ceptable because the environmentof this specific corn was previous-ly bread. Traces of cellulose re -mained on the surface of the parti-cle.

Figures 3 and 4 show measure-ments of fragments from the cornand of the crystal dust. The dustappeared after the particle crackedand was collected from the meas-urement crystal. Both measure-ments with different pressure –middle (fragment) and high pres-sure (particle dust) resulted in aninorganic spectrum. The spectrumis most probably the SiO2 spec-trum. A library search resulted ina good match dedicated to diato -maceous earth.

Sample preparation and X-rayanalysis with EDX-8000

In addition to FTIR-ATR meas-urements, EDX technique wasapplied to obtain elemental com-

position of the sample in the ppmto percentage level without theneed of sample preparation. Stan -dards for calibrating the instru-ment are useful to increase accura-cy of measurement results, but arenot mandatory. The analysis isnon-destructive, making the tech-nique very useful for screening ofunknown samples.

For this measurement, the particlewas transferred to a special plasticcup. With a collimator (standardaccessory) the measurement rangewas narrowed down to 1 mm.Using a camera (standard accesso-ry), the exact position of the sam-ple within the sample chamberwas confirmed. �

0.6

0.4

0.2

0.0

4,000 3,800 3,600 3,400 3,200 3,000 2,800 2,600 2,400 2,200 2,000 1,800 1,600 1,400 1,200 1,000 800 600cm-1

1.00

0.75

0.50

0.00

0.25

4,000 3,800 3,600 3,400 3,200 3,000 2,800 2,600 2,400 2,200 2,000 1,800 1,600 1,400 1,200 1.000 800 600

cm-1Duatomaceous Earth, Granular /SiO2 DuraSamplIR

Abs.

Abs.



––– D SiO2 1

Figure 5: Library search and hit with diatomaceous earth, granular / SiO2 DuraSampllR

Abs.

cm-1

0.000

0.005

0.010

0.015

0.020

0.025

0.030

0.035

4,500 4,000 3,500 3,000 2,500 2,000 1,500 1,000750 500



Figure 3: Middle- pressure: infrared spectrum from a collection of particles from the corn,

mostly identified as SiO2

APPLICATION

LATEST NEWS


Feed to FoodStrategies for measurement of pesticides in water and foods

W ater is not only the basisfor all life, but it is alsoan important resource

for many types of productionprocesses, particularly in agricul-ture. Fresh water sources especial-ly in surface and ground water areof vital importance to humans, butthey occur in low amounts only.Water plays a role in various ap -plications, for instance as a food-stuff in the form of drinking wateror as a basis for food and beveragepreparation. Water is also impor-tant in the production or supplyof raw materials in industry andagriculture [1, 2].

This is why water plays a centralrole in the food chain and whycontaminations have a direct effecton human health. Plant protectionproducts (PPPs) are used exten-

sively and comprehensively inagriculture to increase production.As a result, PPP residues can per-sist on foods and feedstuff orleach into surface and groundwater via various routes. In addi-tion, PPPs are prone to conver-sion and degradation pro cesseswhich may lead to the formationof many different meta bolites overtime. In monitoring programsthese metabolites, with their oftenwidely varying physicochemicalproperties, must also be taken intoaccount, resulting in highly com-plex requirements on analysisstrategies.

Method: Analysis of plant protection products

In food analysis, the QuEChERS(quick, easy, cheap, rugged and

[cps/uA] AI-U

[keV]

0.20

0.00

0.40

0.60

10.0 20.0

Figure 6: EDX spectrum of the particle

For the measurement, a simplestandardless FP (fundamentalparameter) method was used.Calibration of the instrument wasnot needed. No filters (standardaccessory) or vacuum condition(optional accessory for measure-ment of light elements) were used.

Results

It was clear that the main com-pound of the sample was SiO2, i.e.

a stone. Other elements presentsuch as Fe are typical elementsoccurring in stones and minerals.

Instrumentation

• IRTracer-100 with LabSolutionsIR software• Single reflection unit – SpecacQuestTM

• Shimadzu Libraries• EDX-8000• EDXIR Contaminant Library

Conclusion

Infrared spectroscopy can identifythe major components in naturaland complex sample material. Thedomain of infrared is the identifi-cation of substances from organicor inorganic characteristics. Asinorganics have more unspecific

signal groups, EDX is a goodcomplementary analysis for theelemental distribution in a smallparticle such as the corn in thiscase.

Table 1: EDX results using Fundamental Parameter (FP method)

SiO2K2OFe2O3CaOSO3CuO

Analyte

91.8516.6010.9490.3710.2130.016

Result[%]

[1.891][0.246][0.016][0.018][0.029][0.002]

[3-sigma]

Quan-FPQuan-FPQuan-FPQuan-FPQuan-FPQuan-FP

Proc.-Calc.

SiKaK KaFeKaCaKaS KaCuKa

Line

3.01730.74653.78340.33550.15330.3445

Int.(cps/uA)

IMPRINT

Shimadzu NEWS, Customer Magazine of Shimadzu Europa GmbH, Duisburg

PublisherShimadzu Europa GmbHAlbert-Hahn-Str. 6 -10 · D-47269 DuisburgPhone: +49 - 203 - 76 87-0 Fax: +49 - 203 - 76 66 [email protected]

Editorial TeamUta SteegerPhone: +49 (0)203 76 87-410 Ralf Weber, Maximilian Schulze

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Circulation German: 5,850 · English: 3,595

CopyrightShimadzu Europa GmbH, Duisburg, Germany – July 2017.

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Apple Inc. All rights reserved. Apple, the Apple logo, Mac OS and iOS are trademarks of Apple Inc.

LATEST NEWS


Online-SPE-HPLC-ESI-MS/MS

safe) method as described byAnastassiades et al. has becomeincreasingly popular in recentyears [3]. In this method, the sam-ple is extracted with acetonitrile,the extract is subsequently mixedwith buffer salts and purified bythe addition of suitable additives(PSA, C18, GCB). The extract canthen be measured directly usingGC-MS or LC-MS.

In recent years, much work hasbeen carried out to validate thismethod for many different foodmatrices [4 - 11]. The developmentof multi-analyte methods in thefields of GC-MS and LC-MStechnology has advanced consid-erably whereby it is now possibleto simultaneously determine morethan 100 analytes within a singleanalytical run. If a limit of deter-mination of 10 µg/kg has to beachieved, a system determinationlimit of 10 µg/L is usually suffi-cient for these methods and is, ingeneral, easily achievable usingstate-of-the-art instrumentation.

The situation is different in resi -due analysis of water intended forhuman consumption in accor-dance with EU Council Directive98/83/EC [12], where a single sub-stance must be quantified with alimit of determination of 0.1 µg/L.For pesticides in particular, whichmust be determined using LC-MSin the ESI (electrospray ioniza-tion) negative mode, this limit ofdetermination often cannot beachieved without additional sam-ple preparation, such as an enrich-ment step.

It is also known that the ESI ion-ization source is highly sensitiveto matrix effects [13]. Especially,fluctuating salt concentrations andorganic compounds such as humicsubstances very often lead to ion-ization suppression but also toionization amplification [14]. Thiscan subsequently lead to false-lowor false-high measurement resultsin pesticide analysis. The requiredsample enrichment step and, inparticular, the removal of undesir-able matrix components, oftennecessitates unavoidable samplepreparation methods such assolid-phase extraction (SPE) inwater analysis. Such steps usuallyrequire expenditures in time, per-sonnel and resources.

One possibility to achieve com-plete automation with minimalsample and solvent amounts is theuse of online SPE-LC/MS sys-tems. Water samples of just a fewmilliliters can be simultaneouslyenriched, purified and measuredusing multi-analyte methods.HPLC buffers are simultaneouslyused for sample extraction as wellas for analysis. The total analysistimes for sample preparation andmeasurement can thus be reducedto just a few minutes, as demon-strated by ESW ConsultingWruss, a company in the field ofenvironmental analytics, usingShimadzu’s LCMS-8040 [15].

Conclusion: Automated online analysis of water samples

Particularly in view of the largenumber of PPPs (> 1,000) usedworldwide, analytical systemsmust be very flexible and easilyapplicable with regard to the sub-stances to be analyzed. Samplethroughput and analysis time playan essential role in routine labora-tories. Economical use of materi-als and solvents and the requiredsample amounts are of great im -portance in the context of exten-sive monitoring programs.

By using such automated onlinesystems, errors in the sample pre -paration process can be reduced toa minimum and analysis becomesmore robust with respect to awide range of matrices in the ex -traction of water samples of dif-ferent origins.

AuthorOliver Mann, Wruss Umweltanalytik

Literature[1] R. Mull, H. Nordmeyer, Pflanzen schutz -

mittel Im Grundwasser – Eine Interdis -

ziplinäre Studie, Springer-Verlag, 1994.

[2] A. Reller, L. Marschall, S. Meißner,

C. Schmidt, Ressourcenstrategien, WBG,

2013.

[3] M. Anastassiades, S.J. Lehotay,

D. Stajnbaher, F.J. Schenck, J. AOAC Int.

86 (2003) 412.

[4] S.W. Lee, J.-H. Choi, S.-K. Cho, H.-A. Yu,

a M. Abd El-Aty, J.-H. Shim, J. Chroma togr.

A 1218 (2011) 4366.

[5] D. Tomasini, M.R.F. Sampaio, S.S. Caldas,

J.G. Buffon, F.A. Duarte, E.G. Primel,

Talanta 99 (2012) 380.

[6] R.P. Carneiro, F.A.S. Oliveira,

F.D. Madureira, G. Silva, W.R. De Souza,

R. Pereira, Food Control 33 (2013) 413.

[7] M.R.F. Sampaio, D. Tomasini, L.V. Cardoso,

S.S. Caldas, F.A. Duarte, E.G. Primel, Anal.

Methods (2013) 2028.

[8] U. Koesukwiwat, K. Sanguankaew,

N. Leepipatpiboon, Anal. Chim. Acta 626

(2008) 10.

[9] S.C. Cunha, J.O. Fernandes, a Alves, M.B.P.P.

Oliveira, J. Chromatogr. A 1216 (2009) 119.

[10] S.J. Lehotay, A. de Kok, M. Hiemstra,

P. Van Bodegraven, J. AOAC Int. 88

(2005) 595.

[11] S. Walorczyk, J. Chromatogr. A 1208

(2008) 202.

[12] The Council of the European Union, Off.

J. Eur. Communities (1998) 32.

[13] C.R. Mallet, Z. Lu, J.R. Mazzeo, Rapid

Commun. Mass Spectrom. 18 (2004) 49.

[14] A.C. Hogenboom, M.P. Hofman,

D.A. Jolly, W.M.A. Niessen,

U.A.T. Brinkman, J. Chromatogr. A 885

(2000) 377.

[15] O. Mann, E. Pock, K. Wruss, W. Wruss,

R. Krska, Int. J. Environ. Anal. Chem.

(2016).

http://www.shimadzu.eu

LATEST NEWS

W hile the TOC serves as auniversal parameter forthe analysis of the level

of cleanliness in a food productionplant, the TNb is a more selectiveassessment parameter with regardto contamination by proteins.Together, they serve as a measureof food safety, and thus aid con-sumer protection, which is partic-ularly important to food allergysufferers.

The German Allergy and AsthmaAssociation (DAAB) estimates thenumber of food allergy sufferersneeding treatment to be aroundsix million. While in children andinfants, cow milk, soy, wheat, pea -nuts and hazelnuts are the maintrigger, adolescents and adultsgenerally react more strongly toraw vegetables and fruits, nuts,fish, shellfish and mollusks.Allergic reactions often occur onthe skin and mucous membranes,in the neck and around the nose,in the bronchi or in the gastro-intestinal tract. The most severeand life-threatening allergic reac-tion is the so-called anaphylacticshock, which can lead to circula-tory collapse and even death.

Persons with food allergies aresensitive to certain food ingredi-ents, so-called allergens. To pro-vide information on the consump-

organic carbon). It detects thetotal amount of carbon originatingfrom organic compounds.

TOC as universal parameter

For TOC determination, a miner-al acid is first added to the rinsingwater, converting the carbonatesand hydrogen carbonates in thewater to CO2. A rinsing gas re -moves the carbon dioxide fromthe sample. Subsequently, analiquot of the prepared sample isinjected onto the hot catalystwhere the organic substances areconverted to CO2. A carrier gastransfers the CO2 to an NDIRdetector and the amount of car-bon dioxide is measured.

Modern analyzers such as theShimadzu TOC-L series carry outautomated sample preparation(acidification and degassing). Thesystems use a highly effective plat-inum catalyst and operate at acombustion temperature of 680 °C.A special injection unit enablesautomated dilution of the sampleupon exceeding the calibrationrange of the samples. This is alsothe case for the preparation ofstandards to create calibrationcurves in equidistant concentra-tion steps.

As nearly all foods consist oforganic matter, the TOC is a uni-versal parameter in cleaning vali-dation and is suitable for a varietyof products. In addition, TOCdetermination also detects clean-ing agent residues. An additionaladvantage of TOC determinationis its simplicity and speed. A tripleTOC determination of final rinsewater generally takes less than 15minutes. If a corresponding limit

Cleaning validation

Many foods are produced discon-tinuously in multi-purpose pro-duction plants. Following produc-tion and packaging, foods areready to enter the market and thefood production plant is subse-quently cleaned in order to pro-duce new batches of the food orother foods of different formula-tions. In addition to optical in -spection and microbiologicalswabs and to confirm the effec-tiveness of this cleaning process,the final rinse water is examinedfor possible residues and ingredi-ents. For this purpose, a widely-used parameter is the TOC (total

tion of allergen-containing foodsto those persons affected, the 14most frequent food allergy trig-gers (for example mustard, eggs,celery, peanuts etc.) are subject toappropriate package labelling. If afood contains one of these ingre-dients, this must be clearly indi-cated on the packaging.

During food production or pro-cessing, traces of allergens caninadvertently get into foods viapreliminary or intermediate prod-ucts, without being labeled on thepackaging as an ingredient. Toavoid such cross-contamination,many food manufacturers rely oncleaning validation.


Brave helper in allergenmanagementCleaning validation in specialty foods production

100

Product concentration [ppm]

TOC [ppm = mg/L]

1

2

3

4

5

6

7

8

0 20 30 40 50 60 70 80 500,000


TOC [ppm = mg/L]

10,000

20,000

30,000

40,000

50,000

60,000

70,000

80,000

0 150100 200 250 300 350 400 450

Figure 1a and 1b: TOC results of various allergen-containing foods in different concentrations

LATEST NEWS


value is not exceeded in the finalrinse water, the cleaning of thefood production plant has beenvalidated analytically.

The TNb

In a TOC analysis of the finalrinse water, combustion does notonly produce carbon dioxide fromorganic substances. At tempera-tures above 700 °C, nitrogen-con-taining compounds are convertedto nitrogen monoxide. A carriergas transports the NO producedto a chemiluminescence detectorwhere the measuring gas is broughtinto contact with ozone.

Ozone is a strong oxidizing agent,which oxidizes NO to nitrogendioxide. In this reaction, lightquanta (photons) are emitted(chemiluminescence) and aredetected. The TNb sum parameter(total bound nitrogen) is a meas-ure of the organic and inorganicnitrogen compounds present in asample. TOC and TNb determina-tion can be performed simultane-ously using Shimadzu’s TOC-Lsystems – one injection yields twomeasurement results. The com-bustion temperature is set to 720 °C and both detectors areconnected in series.

Cleaning validation results canalso be used to estimate a possibleallergen contamination. This isperformed via a ‘worst case’ sce-nario that assumes that all organicsubstances are allergens.

As allergens present in foods arealmost exclusively proteins con-taining nitrogen atoms, the TNbparameter provides considerablymore information for the assess-

ment of allergen carryover in aworst-case scenario than the TOCparameter.

Specialty foods manufacturertests TNb parameter for aller-gen carryover determination

Employees of the Germany-basedDeveley Mustard & SpecialtyFoods company have carried outextensive studies with differentallergen-containing foods anddetermined that the TNb sumparameter is suitable for theassessment of allergen carryoverin the production of foods.

For these tests, contaminated rinsewater samples were prepared, i.e.defined concentrations of an aller-gen-containing raw material or aproduct were prepared in tapwater. Subsequently, the sampleswere analyzed for their TOC andTNb content. As expected, it wasconcluded that both the TOC and

100


TNb [ppm = mg/L]

0.5

1

1.5

2

2.5

3

0 20 30 40 50 60 70 80 500,000


2,000

4,000

6,000

8,000

10,000

12,000

14,000

16,000

18,000

20,000

0 150100 200 250 300 350 400 450

TNb [ppm = mg/L]

Figure 2a and 2b: TNb results of various allergen-containing foods in different concentrations.

the TNb concentrations increaselinearly with increasing productconcentration.

While a TOC value from an un -known rinse sample could origi-nate from innumerable com-pounds such as carbohydrates,fats, surfactants etc., the TNb pa -rameter provides significantlymore selective information on thepresence of proteins. This allowsfor a worst-case scenario assess-ment with respect to allergenscontained in the final rinse withrespect to medical referencedosages. This calculation ensuresthat no relevant amounts of aller-gens can be carried over to subse-quent production. The assessmentobtained here is intended to ex -clude allergen carryover within afood production plant and to con-firm cleaning validation. In addi-tion, the method cuts costs be -cause complex allergen testing isavoided.

Figure 3: TOC-L with autosampler for TOC and TNb determination

Conclusion

Cleaning validation is a usefultool to verify the effectiveness of acleaning procedure in food pro-duction. The TOC serves as a uni-versal parameter for the analysisof the level of cleanliness of afood production plant.

In addition, the TNb parameterenables a more selective assess-ment regarding contaminations byproteins. A worst-case scenario isuseful in assessing possible con-tamination by allergens, helpingto ensure food safety and, conse-quently, protection of affectedconsumers. Moreover, this type ofmeasurement provides an objec-tive, analytically valid measure-ment result, which is suitable as abasis for the assessment of con-sumer risks in order to ensure thatthe cleaning process safeguardsconsumer protection. Thus, if aproduct label contains the follow-ing information: ‘May containtraces of allergen XY’, this consti-tutes a quantified assessment ofthe allergy risk and does notmerely provide precautionaryinformation for reasons of liabili-ty.

Read for you in Laborpraxis 12 /16

LATEST NEWS

equipment such as pumps, pipesor valves. This can lead to mobi-lization of different compoundsfrom these components, whichcontaminate the water. The magni-

tude of these contaminations mayonly be in the lower ppm rangebut these contaminations becomeimportant when using ultrapurewater and most certainly in regard

Current state of process analysistechnology2nd TOC Process Analysis day in Duisburg features lectures and discussions between experts

I n mid-March 2017, Shimadzuhosted the TOC ProcessAnalysis day for the second

time. Users, planners and engi-neers from industry and academiamet to discuss the current state ofprocess analysis technology.

TOC (Total Organic Carbon)determination is of great impor-tance in many industries. TheTOC sum parameter specifies thetotal content of organic carbon ina single analytical measurementvalue. TOC determination can becarried out safely and simply inthe laboratory as well as duringonline process analysis. With itsTOC-4200 series, Shimadzu cur-rently offers online TOC systemsfor continuous process analysis.

Two years ago, Shimadzu organ-ized the TOC Process Analysisday for the first time as a platformfor sharing experiences and net-working for users of process anal -ysis technologies. It now takesplace in alternate years with the

TOC User Meeting that hasmeanwhile existed for 15 years.Just like the first TOC ProcessAnalysis day in 2015, speakersfrom industry and academia pre-sented application-related lecturesand shared their experiences withparticipants.

Monitoring purified water

In many applications, the TOCparameter is a measure of undesir-able contaminations. Monitoringof water quality is especially im -portant when using purified water.Dr. Raoul Schröder (BürkertWerke, Germany) showed that theuse of peripheral equipment alsoinfluences water quality.

Bürkert, manufacturer of valvesand valve technologies, investi-gates the influence on water pollu-tion caused by components offluid plants. When flowingthrough a fluid plant water comesinto contact with various types of


Figure 1: Lively discussions and interesting lectures at the second TOC Process Analysis day in Duisburg, Germany

Figure 2: Seminar speakers (from left to right): Dr. Raoul Schröder (Bürkert Werke GmbH & Co.

KG), Bettina Gierszewski (Shimadzu Europa GmbH), Mischa Dings (Shimadzu Deutschland

GmbH), Ralf Kienle (Clariant Plastics & Coatings Deutschland GmbH), Dr. Tomas Sauter

(Covestro Deutschland AG), Andreas Gräfe (Lanxess Deutschland GmbH) and Sascha Hupach

(Shimadzu Deutschland GmbH).

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to quality assurance for manufac-turers of water-bearing compo-nents.

Higher quality requires higher data density

Demands on modern productionprocesses are steadily increasing.Not only is there a growing de -mand to cut costs as well as tosave energy and raw materials, butproduction capacity, efficiencyand flexibility should also be in -creased. And all this with everhigher quality requirements. “In order to meet these demands,it is necessary to generate higherdata density for automated pro -cess control”, says Andreas Gräfeof Lanxess Germany, when dis-cussing the general benefits ofprocess analysis technology (PAT).

The use of laboratory analysis ismore and more taking a backseat.Nowadays, hardly any large-scaleplant is started up without havingto supply corresponding processanalysis data for process control.The demands placed on the sys-tems used are based primarily onachieving the highest possibleavailability as well as on increas-ing data density. In this way, PATis of benefit to plant and processsafety and also to employee andenvironmental protection.

TOC data density allows for more precise process control

Modern online measuring systemstechnology features availability of99 % or higher and thus offers therequired reliability for automatedprocess control. Andreas Gräfeillustrated this with a practicalexample: Prior to the use of pro -cess technology, wastewater moni-toring had been carried out byappropriate laboratory analyses. A TOC analysis took approxi-mately 40 minutes including sam-pling and sample transfer to thelaboratory. By using Online-TOC,analysis results are delivered auto-matically in ten minute intervals.Because of the increased amountof data, the wastewater is moreclosely monitored. The companythus obtains more information onits wastewater and can control itsproduction process more precise-ly. Cleaning costs incurred in the

wastewater treatment plant cannow be more efficiently allocated(in accordance with the ‘polluterpays principle’). Based on thehigher data density, reporting tothe authorities is also more pre-cise.

MCERTS certified performanceand reliability standard

In order to verify the high avail-ability of process analyzers, theEnvironmental Agency of Eng -land and Wales (UK) has devel-oped a certification system formeasurement equipment – theMCERTS accreditation. It pro-vides the framework conditionsand quality objectives for compa-nies to meet in order to complywith authority requirements forenvironmental monitoring.MCERTS is quickly becoming arequired performance and reliabil-ity standard by organizationsaround the world. With accreditedequipment, companies and author -ities ensure that they are doingeverything possible to protecttheir environment.

Requirements for obtainingMCERTS certification includevarious laboratory tests and a 3-month field test under real condi-tions. Measurements are per-formed by an independent testingfacility according to predefinedtesting methods. MCERTS accred-itation has been issued to meas-urement systems in Shimadzu’sTOC-4200 series. TOC productmanager Bettina Gierszewski atShimadzu, in her presentation onMCERTS, discussed the testmeasurements performed as wellas the results of the TOC-4200field test.

TOC content of waste-water a decisive criterion for disposal

Ralf Kienle, Clariant Plastics &Coatings Germany, reported onthe use of laboratory and onlinemeasurements for monitoring ofindustrial wastewater effluents.The pigment production processgenerates wastewater during vari-ous purification stages, which isdischarged at regular intervals intothe wastewater treatment plant ofthe Hoechst Chemical Park,Germany.

TOC content of the wastewater isconsidered as a decisive criterionfor disposal. Until recently TOCanalysis was performed reliablyusing a laboratory system. The useof an online measuring systemnow saves time. In addition, byconnecting the analyzer to theprocess control system, untimelydischarge of wastewater due tohuman error can be prevented.During the analysis, the processcontrol system automaticallylocks all pumps.

Planning stages and project management forprocess analysis

The final speaker on the 2nd TOCProcess Analysis day was Dr.Thomas Sauter of Covestro Ger -many. He presented an interesting

insight into the various planningstages and project management forprocess analysis in investmentprojects. He showed the tried andtested project phases for the con-struction of ‘plug and play’ analy-sis containers. These were de -signed, built and transported totheir places of destination, forexample a chemical factory inChina, by Covestro PAT special-ists. Thomas Sauter pointed outwhat needs to be taken into con-sideration and how many differentcontacts are involved as well asspecialist departments that need tobe included.

Read for you in LABO Mai 2017

Figure 3: Shimadzu TOC-4200 with MCERTS accreditation

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A fter 100 years of experienceand continuous develop-ment in materials testing

technology, Shimadzu’s productrange today includes static anddynamic universal testing ma -chines, hardness testing instru-ments and capillary rheometers aswell as the HPV-X high-speedcamera, the most powerful in itsclass – with 10 million frames persecond. Shimadzu’s testing ma -

testing system as well as a particlesize analyzer.

Testing innovative and increasingly high-performancematerials

Material characteristics vary wide-ly and behave differently depend-ing on ambient conditions andforces exerted. Research and de -velopment departments need

chines are at the cutting edge oftechnology regarding control sys-tems, sensor technology and in -formation processing, to reliablysupport developers and users.

What started in 1917 with fiberand cement testing machines, con-tinued in the following decadeswith hydraulic, dynamic and fullyautomated testing instrumentsincluding an ultrasound fatigue

A century of experienceShimadzu celebrates 100th anniversary of testing machines

100 years testing ma chines

highly precise and reliable data forproduction and quality control ofinnovative and increasingly high-performance materials, for in -stance composite or jointing mate-rials.

Precision universal testingmachine for research and development

Just one representative example of the versatility and performanceof Shimadzu’s materials testingsystems:

Shimadzu’s AG-Xplus series in -cludes especially powerful andversatile electromechanical testingmachines. With their rigid framesand precise load cells, they areoptimally equipped for all require-ments, in particular for researchand development.

For customer-specific require-ments, numerous solutions areavailable, for instance a dual spacetesting system, without the needto switch testing accessories forcompression and tensile testing.

With its user-friendly Trapezium Xsoftware, all settings required canbe applied in the shortest possibletime.

Further information on Shimadzu’s testing

machine expertise is available on page 28.


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W hich substances in redwine promote health andprolong life? Which aro-

mas are responsible for the wine’svarietal character? Which aromaconcentration range can the hu -man nose detect? How can illegalaromatization be identified? Dohistamines play a role in wine in -tolerance?

In wine, there seems to be morethan just one truth. Sometimes,there is also true criminal intent:counterfeit foods, such as labellingfraud, product counterfeiting orfalsified content generate fat prof-its for the fraudsters and causeconsiderable financial harm to theoriginal manufacturers. In addi-tion, there are economic and oftenconsumer health damages.

Professor Erich Leitner of theGraz University of Technology,Austria, Head of the Institute ofAnalytical Chemistry and Food

Chemistry, has been focussing foryears on food quality from a sci-entific perspective. He has ledShimadzu’s exclusive two-day se -minar ‘the Secrets of Wine Analy -sis and Wine Tasting.’

In addition to the determinationof species-relevant flavouring sub-stances via GC analysis, theoreti-cal and practical aspects were dis-cussed on other methods to char-acterize wine and its ingredients(metals, polyphenols, dyes as wellas residues). In a wine tasting tu -torial of the wines analyzed, theparticipants were able to establisha direct link between sensoryquality and analytics.

Furthermore, they compared theperformance of the human nosewith the instrument’s sensitivity.

This seminar series will be contin-ued, again by personal invitation,in order to give all participants

Wine tasting and wine testingSecrets of wine analysis: from sniffing and tasting to measuring

sufficient time to be introducedonsite to the various analyticaltechniques using real samples.

28

Euroanalysis

Stockholm, SwedenAugust 28 - September 1,2017www.euroanalysis2017.se

MSACL

Salzburg, AustriaSeptember 10 - 14,2017www.msacl.org

Weurman Symposium

Graz, AustriaSeptember 18 - 22, 2017www.analytchem.tugraz.at/weurman

ISSS

Wien, AustriaSeptember 19 - 22, 2017www.isss2017.at/symposium/welcome

Composite

Stuttgart, GermanySeptember 19 - 21,2017www.composites-europe.com

Ourcon

Doorn, NetherlandsSeptember 25 - 28, 2017www.ourcon.org/ourconV

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New – Materials TestingApplication Handbook100th anniversary of materialstesting equipment

S himadzu’s Materials TestingApplication Handbook isnow available and can be

downloaded via www.shimadzu.eu. It covers 112 applications ineight industries such as automo-tive, aerospace, biomaterials andmedical, composites, food, metal,railroad, rubber and plasticsindustries to support consumerand environmental protection aswell as product reliability.

The Materials Testing Applica -tion Handbook is hands-on andsolution-oriented. The applicati-ons described cover most-modern technologies, e.g. univer-sal, fatigue and hardness testingand also high-speed video cameraapplication.

Since 1917, Shimadzu has a pro-ven track record in manufactu-ring testing machines designed tomeet the diverse needs of custo-mers worldwide.

To date, the company has soldtens of thousands of testing ma -chines. In addition, Shimadzu hasmarketed thousands of applicati-on-specific systems tailored tothe unique needs of clients, andremains committed to providingcustomers with this same level ofservice in the future.

Please see also other ApplicationHandbooks on• Liquid Chromatography• Food & Beverages• TOC• Clinical• GC/GC-MS.

Further information

on this article:

• Application Handbook

Materials Testing

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New milestone Switch +Go A century of experience · 2018. 11. 1. · longstanding experience in the development of GC systems as well as continuous exchange of ideas with users, Shimadzu