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Loyola University ChicagoLoyola eCommons
Dissertations Theses and Dissertations
1959
Investigation of Quantitative PolarographicMeasurements of Oxygen Tension in TissueAlbert Frederick KelsoLoyola University Chicago
This Dissertation is brought to you for free and open access by the Theses and Dissertations at Loyola eCommons. It has been accepted for inclusion inDissertations by an authorized administrator of Loyola eCommons. For more information, please contact [email protected].
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.Copyright © 1959 Albert Frederick Kelso
Recommended CitationKelso, Albert Frederick, "Investigation of Quantitative Polarographic Measurements of Oxygen Tension in Tissue" (1959).Dissertations. Paper 571.http://ecommons.luc.edu/luc_diss/571
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.,. AlM .. t ....... ,., l.el_
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UNIVERSITY
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bel' 19, 1917, ... tile fh.t of _Yea cld.lu •• 1. tile fai11 of
Jfewtoa T. aDd DOl'oth, lAidolfKel... lie 1'80el .. 4 Illa .l .... t .... '
.et_atloa i. ,.t Va,.. .. t Ml •• 1' &ad. .... 11 .... p'ad. Mhool., &ad
paduat .. Il'a SOdIl .148 Ill", SC .... 1 i. 1936.
18 tbe fall .f 1931 1M •• tue. Cleo .. ,. .111i_ Coli ....
.... l .. l1ala'la 1afeaa1 ..... tl .. 1. tile fleleta of "<la aad
... .,.10&1 lId1lCatl... I. 1940 be wu ....... et tile »dward D •
• t.loa •• SCIlOl ..... lllp la Realth aa4 ... ,aleal Uacatl-. TU
acJaolu.lllp ,...riel .... 0,,..,,,,,,,, '0.. aallll., la ""aloloal.
cal r ...... ch. ..1£ ...... 1 til ... .-tllu H. .t.i ••••• , ... o..,l."e. a attldy .f the .ff .. ta .1 cold 1l1p .. t ........... l-_t_ per
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alcal ...... tl.. ia 1946 •
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la. aad Ill. _i •• ioDill, .. Li •• tea .. t, J.,. t V.S.N.lt. Be w ..
.. acti ... uty la tile pacific Ar .............. tl0 •• offic ... ; of
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biolo., &ad 1. play.iolo.,. as a ......... of tbe ,acul ti.. of Gt .... ,.
Willi ... COli... Uld Chic ... COlI ... of Osteopathy. He alao
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ia tbe 8pela, .f 19$9.
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affol'c1iDI hi. tit. oppert""'t, to at.ely f ... &d._ced clepee.
ae ia mo.t patefal to Dr. Clue •• N. hi.a aac1 Dr. Walte. C.
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worki., witb tbea. .. .ell a. the ... istance tb.y haft reDcl.re"
ia tbe ..... iou. pb.... ., hi. traiala.. !he iDdirect .. ai.tuce
of bi. f.ily, DI". Sb_ C. All ••• aacs Mr •• lett, Witte aDd
••• joyce elley's ackaewl .... ca f."M i_&auabl. coatr!_-..
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Iv
TADLB OP CONTENTS
Chapter I. G.BNEIlAL INTROmr.:TlON, REVIllW 0' nIB BACKGROUND
AND IDENTIfICATION O' paOJIJi.M
A.
B.
c.
D.
The place of ,i.sue lOa in the relationabip of oXJlea to yital procea .. a • • • • .. '" • • • .. .. Dillus oa 1. QUaDtitatlYe baa'a for diffusi08 • • • • .. .. .. 2. Plck t • aDd "'optacoeflle!fult. .. .. .. ....... 3. permeabilit, coastants • .. .. .. .. • .. .. • .. .. .. 4. Appllcatioaa of D.~, .. 4 k •• • .. .. .. .. .. .. Po1arosrapblc pcincip1ea .. • .. .. .. .. .. .. .. • • .. .. 1. R.elation to Ollie .. types of electrOly.i.
ad e.lecta-oaaal,ais ..... .. .. .. .. .. .. .. • • • Polari.atioa .. • • .. .. .. .. .. .. .. .. .. .. .. .. .. • QualltatiYe Ud quaatitatiye &D&1,S'8 .. • .. • Co.,., ... t. of pelaropaptaic CHrent •• .. • .. IllecUod.e ..... ti.... .. .. .. .. .. .. .. .. . .. .. . Metbod. of a ........ l., i<S a. Absolute _tiled ....... .. .. .. .. .. .. .. .. • b. c..,~.ti .... 'bod .. .. • .. • .. .. .. .. .. .. ..
T. fOlar!lraJbie las tr ument at ion a. cueulta....... .. • • .. .. .. .. .. .. .. .. .. .. b. alectrodes........... .. .. .. .. .. • .. .. • .. c. Modified circuit. ..... • .. .. .. .. .. .. .. • d. Modified electrOdes ...... .. .. .. • • .. .. •
Re'f'lew of polaropaphic literatwe 1. History 01 oxy,.. aDal1sis .. • .. .. .. • .. • .. • 2. Trace ... &1,ais • • .. .., .. • • .. • • •• • .' • " 3. 1l •• pia'atioll ael photoapt"'! ........... .. 4. ADal,ala ia blOOd .... ".. • .. • • • .. • • • S. Aaalyais in tis ... s • .. • • • .. .. .. • • • • ..
11. BXPJIlIMBNTAL PROCIOOllBS
A. 8.
c.
Polarop:apbic lutru.atatioll ..... .. .. • .. .. • .. Diffuion .............. " ......... " .. . 1. Me .. ure ... t of ellff .. ioD coellicleat • • • • .. 2. Discu •• loa of diffusion coeflleieat ..". • Ti •• ue .... ur ••• nt. 1. Hypothe.is.."......"....""..."..." .. 2. A"d.lable •• tbod. for teatiac hypothesis .... 3. Baais for Calculatiag tlaaue ox, •• teasloa • 4. Methods filM" for tiu.e _aaure .. ata " • " • •
.y.
1
3 4 1 , 8
9 9
10 13 16
22 24
25 28 30 3S
39 41 42 48 52
59 64 68 12
15 11 78 81
III. RESULTS! CALCULATIONS AND ANALYSIS O' MBASt.l'UDmNTS Of TI SSUl! oXYGBN 'l'.BNSIOH
II. c. D.
Method of evaluatiD, data for CoapUi60D of p02 obtaiDed by two dlffereat _tilOd. .. • • • • .. • • Metllod for calculation of pO. • • • • • .. • .. • • S_UI of ti... ox}'... t ••• lon .. .. • • • .. .. • .. Anal, •• of polu:ograpbic _ aaure .. at. • • • • .. •
IV. DISCUSSION OP TISSUI OXTGlUf TBNSION MlASt.l\lJ.tiNTS
A. The approxiaatioB obtained throuJb the •• e of Ketr" equatlo. ... • • .. .. .. • • .. • ••• • • •
B. SlsRificlDce of the a::tiC.lto. of Xet,'. equation to polu:opap c __ reaeat .. • • • .. ..
c. .. e1labiU., of ..... euate .. .. • • • • .. • • • .. D. ,....., of e¥per1meatal fi.dla,. • • • .. .. . • • •
V. CONCLUSION
8lBLIO<ItAPHY
89 90 91 92
110
113 116 120
122
124
LIST OP TAJLIS
Tabl.
I. DB'l'ERMINANTS OP TlSSUI AVAILABILITY OP OXl'GJlN • . ... . Pa,.
2
II. ESTIMATION 0' leo PROM DIPPUSlON TIMI ....... • • • 70
Ill. CALCtJLATIONS OP DlPPUSION COBP'ICIBNT O' OXYGBN IN SALIN.! • • • • • • .. .. .. • .. • .. .. .. • .. .. • • •• 1a
IV. VALUl!S POR 'l'RJ! DIPPUSIOH COIPPICIBNT 0' OXYGBN IN WATIR • • • .. • .. .. • • • • • • ... • • .. • • • ... 13
V. TI SSW OXYGEN TBNSl ON IN ltABBIT MUSCL..e .......... 95
VI. COMPARt.," 0' leI,AND pOa POR. RABBIT MUSCLE ••••• 100
VII. 1'JSstm OXYGBN TIN.ION IN R~IT JAIt ............. lOa ' . . '
VIII. COMPARJ~N 0' id AND ,02 POI IWUIIT BAIt • • • • • • ... 101
IX. ANAL Til S O' .BLECTltODB MlASlIlBM.iNTS ... . .. . • • . .. 109
p
LIST 0' PlGUIlBS
pipJ:e
1. POLAROcaAMS 0' OXYGIN AT DIPPEIl.B.NT (x)NCBN'l'IATIONS .. . 2. THE POLAl\OCllAPllIC C-Y CUlVS ....... .. .. .. .. . . • • 3. .. .. . . .. .. .. .
SIMPLB DIAat..AM 0' POLAROGIlAJ.1H1C CIRCUIT .. • .. .. . .. . .. .. .. .. .. .. . .. .. .. . ..
6. DIRIVATIVE POLNtCXItAPHY .... .. .. .. . . .. • • • • • /I
Pap
10
23
2a
as
26
31
7.. PRINClPUS OPSQUAIlB WAY.! POLAIlOGllAPllJ .. .. .. • • •• 32
8. TA&.IS 01' MODIPlliD SOLID AMALUI. BLBCTRODES ........
10.
11.
12.
13.
14.
IS.
. .. DIPPUBION IN A 8YSTBM CON'lAINlNG A MBMaAtUl .....
APPLICATION 011 SOLID '&IiO'faODBS '10 THB STUDY 0, CHANGBS INDU.:JD IN llBSPlRATION ....... .. .. .. • .. .. .. POLAlOC8AfH ...... .. . .. . ...... .. .. .. .. .. .. .. .. .. ANALYSIS BLECTRODES .. .. . . .. .. ........ .. .. .. .. .. .. ,. DIPPUSIOH CHAMBBR. .. .. .. .. . . .. . . .. .. .. .. .. .. . .. . DIMBNSIONS POI. CALCULATION O,"DIPPUSION IN TISSUE .. ..
COMPAJlISON 01' ld AND ,02 POll MJIBIT MUSCLB . .. .....
35
38
45
61
64
66
78
101
16. .. .. .. .. 108
.ylil-
CBAP'tJilt I
GmfBRAL INTRODUCTION, REVIBW OP nm BACKGItOUND AND
IDBNftPICATlON 0' TIUl PROlLBM
This di.sertation pr •• eata tbe i.lors.tion avallable OB
polarographic: _aau_nts of oxYlen cODcentl'atioD !! .!!!! IIIld !!. vitro with a cOIDpuiaoa of the relative accuracy io the two ays.
te.s. The factors _ieb alSbt HCO •• t for acceded accuracy ill -viYO ue discua.ed. ua e.&perillental .resulta are pre.ented as aD
ori,ioal critical teat of quantitatiYe polu-oaraplllc: .uare.nt
01 OX" •• tensioa ia ti..... Tile 'Dtredactio. indicat.s the 1:01.
of aeasure •• at of tis... oxy,.n te •• ioD ia the mo«e se.eral prob
lema 01 relatift, oxyaen a.pply to .ttal proc •••• s. It also iD
cludee a di8c ••• loa of dilf.sioa and the principles of polaro
Il'apb, pertiaeat to the 'a.esti,atio.. .bleh are repor ted and to
tbe material presented ia the text. "
Polaro,raphic .... ur ... nt of the" ti.s"e ox,,.o tensioa repre.
s.ats a receat approach to l'I',. •• ti,ation of tbe relationallip of
oa,,.a to vital 'roc...... Siace the in .. sti,&tloas1 of Boyle,
Nayow. Pri •• tl.y aIld L ... iaier l.dieated tbe fund __ tal role of
OX1,ca in blolos:l.cal 'r.c...... biochemi.ts, biophysicists and
blolo,lst. kave elucidated .'" .. ,eeta of this relatieDshlp.
Tbe preseat atatus of knowled,. in this field i. illutrated la
Table I. UDder vaacular iafl •• ace. are listed the eater. at re
apiratory, traaaport, aad deliyel'Y faetora related to oxy,eD
TABLE I
DBrERMINANTS o. n SSUB AVAILABILITY O' OXYGEN
Compart .. at. IDYOlved aad Proce .... Occuri_, Within Compartaents
VASCULAR
VASCULAR INPLUEJCES
EXT.BltNAL RESPIRATION OxYJeD .ataratioD (VeatilatioD aad Diffualn, capacity)
TJtANSPORT Ox,,.. c&j'aci. ty (1.36 .l/p Bb) (Diaaolved 02)
P.mtfIUSION 0' TI SSUE (.I/IlI/ua)
DBLIVER.Y AT TI SSUE HbOa • HHb + 02 (pH and T effect.) (DifEusi8, capacit,
and rate at tissuea)
INTERSTITIAL
INTERSTITIAL PACTOllS b
DIPPUSIOH • 90-991"
OOIIVECTION (balk transport) Traaacapillary + 0.3 ,. 120 formation - 0.3 ~.peratur. .. 1 fu'l •• tioD .. 1 Acti.. 0
S'fOItAGB
UTILIZATION
CBLLDLa (Mitroc:bondria)
CBLLULAR INPLUENCBS
utILIZATION
(QOa : ai/me/min)
Por exaaple: (Subatrate a DPN(R)
PAD(R) • (Cytocllrome .yatea) a 02)
STORAGi
(... Myoglobin)
..,
3
supply of the tissues. With the exception of the diffuslDg ca
pacity aDd rate between capillaries and tiaaue these lactors bave
been known for several decades. 1,2.3 As indicated in the last
column of the table many of the cellular influences bave been in
tensiYely investl,ated. Respiratory quotients for tlasue. and
cells and respiratory rates for small or,aaisms have been estab
lisbed by various !! Yit~o tecbnlquea. 4 •5 The specific chemical
reactions for tbe metabolism of oxygen are known In general' and
in SOme respects in great detal1. 6 It bas been po.sible to relate
the chemical reactions by which oxidative processes are coupled
with the formation of high enerIY bonds to ~tions of tbe mito
chondria within the cel1.1 In the center column of the table are
the interstitial factors related to oxysen supply to the cells.
August KrOBb8 established diffusion as the fundUlental process
occurring at this level. Other factors and factor. whicb modify "
" diffusion have not been establisbed. A quaatitativ8 method for
meuuzellent ofaxysen cODCeatratiOlls a.t the cellular level would
aid in the development of iDfortaation OD o.xypn tl'anapol't, circu
lation and metaboli •• at the cellular level in vivo. --DIPJlUIION
Aft uDderataDdlo, of diffualon i. baaie to tbe develop.eat of
a quantitative meaauellent of oxy,.. in tia..... Xn the f011owia,
diacuasiona of the diffasloD cur •• t of polaro,raphlc _uure.nt,
the calcalatioa of eatlates of ti .... -1Ie& ten.loa and of
tran.fer of oxygen from the blood to the ti •• ue involve v~io.a
characteristica of diffualon. Tbe fo11owln, para,rapba present
the iaforaatlon e •• ential to the discu •• ioas, iaatrumentation and
metboda entailed in the investigations reported.
Studies of dillasioD ,aiDed quaatltative aianificance throuab
pick'a 1 ... , in which be introduced a diff •• ioR coefflcieat which
aak •• it possible to calculate either the quantit, of material
,u:oa.1I1, a aUl'face in a diffwsiol1 • .,.tem or the quantity of
•• terial acc..ulatio, withia a portioR of a diff •• loD a.,stea.
,lek t • firat law de.cr1~. the flux of a.terial in a aystem wbere
area. cODceatratioD .rad!~ut and tiae bave beeD defiaed,
Q=-DA1sdt d ..
Q - Q1Iantit, dC/dx - gradient
D - Diffusion coefficient dt - iaterval
A - Area
(1)
Pick' •• econd law deacribea the rate of accuaulation of •
aubatance within a given volume of a difluaion a.,atea wbea the
eODceDiration sradient aero •• that v01ue ad tbe time haft been
defined. (2)
dC:15!Cs dt dll
The diffa.loD coeffici •• i, U, haa the .am. uait aad diaeo--.101l&1 va1u •• ia both cuea. Ira tile first la. D haa been defilled -for a ualtol'. coaceatl'atloa padieDt aad ia ideDtifled .. tbe ! .
c1iffel'eatial,g. In the "coad law ! baa been obtained 4.,.i8, a
s
variable concentration ,radient and is identified .a tbe lntearal
D or n. The aasnitude of R aad i ia identical only for con4itioaa -in which D is linear witb respect to concentration ,radienta of
dilferent magnitudes.
Piek'a diffusion coefficients are widely as.d and it is fre
quently aece.sary to ehan,e the unit. of area. le8lth, CODeentea
tion, quantity or time for a particular uae. Accurate cbaD,es
CaD be .ade only Oil the basi. of adequ.te kaowledp of botb the
unita and dimenalooal values involved. Ie the CIS. _,stea tb.
naaerical value of D determined from Bquation 1 ia
D:~X~ x t: :'z x (Q';/~mi))( dt (3)
Upon caacellation tbi. reduces to ca2aec·l or a dimeDaioea! value
of L2t-l •.
Calculation of .&8 diffusion in liquid and biological .yate ••
ia complicated by lack of informatioaoD ODe or ~.of tbe vari
able. required to eatablio a diffuai'on coefficie.t for tbea.
ayat.as. AuSUat Kro8b9 iDtroduced the method of expressin, tbe
,a. concentration in tel'ma of the pre •• ue of tbe , .. in the ....
phase, ~, qUaDtitati .. treat.eat of diffu.1oa 18 aolatioD 01'
biolo,ieal .yst ••• became poaaible. la tbi • .etbod the uait aDd
dlHulonal ftlue fOl' a dllfaaloll coefficient .. appear to be
f •
*Cent!'ete ... era. second system of units.
**lbere ia no adYantage in usln, the tera con.taDt, .ince tbe coDfuaioD between Pick's D aQG Krogh'. D aria •• fro. applicability and dimenaions, rather tban from &6aolute Yalue.
r __ ------------------. 6
nearly identical to thoae used by Pick. KroSh'. unita were
milliliter of ,aa for qdantit" atmospherea of ,aa te0810D for
concentration gradient, _lerona for diffualon distance. and min
utea for diffuaion tise. The unit value of Kro,b' a dlffuaioa
coefflet.at as determined by £quation 1 ia
_ ml I ,,,0-4 em I DK - cmt x at mol )( sO ,Ie
Alpha,. the absorption coeffiCient, CaD be used to convert tenalo
into concentration if tbe 801ubillt, of tbe Cas ia kaowu.** Mul
tiplying the tension of the previoua unit analysis by alpba re
aults in
(S)
This reduces to tbe di .. _.to.s of L2t-l , lIfbich ia the su. aa the
dimensiona of Pick'. D. It is evideat that if both D'. are ex
pre.sed in tbe ega 8ystem, they differ only by the .ffect of "
solubility of the gaa or in the us •• f ten.iOD for calculatloD of
tbe concentration gradi •• t. i.e.
m at
*Defined aa tbe quutity of ,as in solution at equilibria with the partial presaure of a ,as phase at a given temperature. a1/, •• /ml solution.
**The major advantage of ~rolbts diffusion coefficient ia tbat aolubl1ity. wbich in many cases could not be determined, .e.d Dot be kaown.
p
7
some biological .yat ••• aay hay. aD unkaown difl •• ioa dis
tance and/or ditl.aioD .. ea, ia addition to .. uDknown concentra
tion cradieat. Several metbods allow quantitative treatmeat of
the diffuaioa ,ncea. ia spite of the IUlJlber of unknown vuiabl.s.
The aetboda are aimilar in that in each case all of the unknowns
are incorporated into a proportionality constant of diffusion.
Diffusing capacity of the lung.IO ia an example of a case I.
which uea. and di.tance are •• known, add pe ..... sbilit, const .. t.11
are examples of cas •• in wbich distaac. ia •• known. Generally
eoncentration cradient aDd area ate kBowa. bUt the deriy.tioD of
the coutaat atlould ... ex.iRed carefully to determine which un
kDowaa are included with the diffuaion eoeflleieat.
There is considerable v&riatio. in the freedom whieh can be
taken with coaatant. aDd GO.ffiel.ata durlDS their applicatioD.
The dilE.sloa coefficieat re,~e&ent. propertiea of bOth tbe aat •••
ial wbicb ia dil, •• '., .. d tbe .,atell·' la wldeb tile diff .. l_ oceura. Coaaeqaentl, tbe application of dlll •• !oa coefficie.t. ia
l.i.Dlited to the ..,.tea f. _leI! th • ., Ita .. been deflDed.. III tbe
e ... of Pick t • 0 and to .... exieat .f lZo,b'a Dr, •• ef.l app11-
catioD ia ext .. ded to otbe~ a,ateaa bee .... facto~. bawe bee. de
ieraiDed vbi.b allow correctio.. '0 be aade. Usef.1 correctio ••
baft He. eatabllshed for temper atce, viacoal t,.. and COllceat ....
tloa differeac... It is alao po.aible to prediet dilla.lvii, for
a give a _,atem 08 tbe baa'. of tbe ia¥erae ... tio of tbe molecular
.el,bta if the diffUSion coefficie.t of ODe of the .olecules I.
jP
b
8
known. Tbeoretically, proportio.ality coast .. t. of dlff •• ioD are
1i.ited to a .ystem identical with the oae for ~ch tbey are de
teraiaed. Aa the, contain .. taow. 9ariabl.8, tbe exteaaloa .f
their application to otbet .y.te .. lapli •• that tbe vakDowa factor
are ideatlcal ia tbe two .,at ••••
POLARO<ltAPJtlC PIlINCIPUS
The priaci,le •• f poluopapb., od the f,,_ .... tal. of polar-
opapld.c i .. tr .... tatl.. ... , ...... t.4 ia thia Hctioa. A cU."_. alol1 of 81ect .. od ...... tio.a, aoctificatiou .f el •• trOd •• &Il<l eir-
cai t. applicable to tbe •• al. ,cobl.. la laclUded. A review of
the Ii teratun and a _"y of polU'opapM,c ........... t. of
oxy ... coDCeat .. atlo. ccaplet •• tae •• Ctl08.
POluo .... .,b' 'a &J1 last ..... _t.l .. tJaod of 81ectrochellical
_at.,.l. of the qaalitatl .. aad ._:,titati •• aature of _batucea
in _18tloa.12 It differs fr08 othel','fo ... _ of electrolyal •• sed
.alyti •• ll.,. I •• Gada.tOllet .. !C aDd ele.trepllo .. etic 88alysla the
so ...... of charJed puticl.a ia tile electI'01.,.18 cell ia &Da-
1,..4.13 'GteatIOMetr! ... 4 electrolytic .. tbOda are .ia11ar to
polu-oS!'apby la tbat tile., depead upea oxidatf.oa-l'eductioD reac
tio... III til. pote.tioM'Ulc _thod til. _&1,.. 1. b_d 011 the
pote.tial8 .atablisbed by the .. eacti.a, wldle i8 til. elect.-ol,ais
lIeth04 the aa&1,.l. 'S ., ... d OD tile , ... ti t, or .. t • .-ial 1' ..... 4
f.-OlD the eo1utioa by tile "eaction. 1ft botb .f th ••• _ttloda tbe
bod, of the solutioa of the electrol,sis cell 18 &8al,.e4. 12 In
I
,.. 9
poluopap • .,. boib tbe ~e .. iloll _d quanti iy of .. b.imce are la
volYe<1. and ibe body of the 801uiloD .... ala ..... ntl&111 ullchanpd
by tbe reaction, wIllch 1. limited io the •• &11 .. e.ion ill the
lamediate vlciDity of the electrode. 12
Poluopapblc aD&1,si. la aee .. ,lls"ed by eaiabll.biDI COD
e.auatloa polubatioD of tbe aD.l,sia eleotrod., nlatill, the
poteatlal at Which CODceatr.tioD poluizatioa is establlshed to
the Datue of the r.act_" aad the cuueat flow followin, COD
ceatratloD polarl&atl08 to aoaceairatioa.
Darla, electrol,sis, conditio.s ma, c .... ODe of the '1'0-
c ..... to bee ... 810wer tbaa otbel' '1'0 •••••• I'elated io the elec-
tl'Ode l'.acti .... IS _hea this rate limitiD, p~oc ••• occurs, th.
electrOde will .. swae AD iIIpre ... d YOlt ... i .. tead of the volta,e
charactel'l.tic of tbe .1.Gt~olJ.l. reaction, aad it is said to be
,olaiaed. ne cu ••• of poluis ...... oD iDelude .1OWDe.s of the
formatioa .. d/~ dl.c~,. of 10", the .lowae •• of tr..afer of
re&Gtut 01' PI'MUCts ia the I'esioa of tu .l.ctl'ode, aDd tbe.low
ae •• of reactloD rat •• d~l., electrolysia. 1S
III pelaropapby coad! tio •• "'41 contzol1ed to cu .. c ......... ..
tion polarization (.lown ••• of i .... t .... of .... aciant to the elec
irode) by .&kiaS tile &Dal.,.l. electrode ft ... ,. a.all and te.pi., It
eitber motiOD1 ••• Or ill con.taDt .otio. relative to tbe ac1a
tl08. 14 At tbe .... tt.. the ot ...... 1 ...... 04e 1. tept at co.atant
poteatial, tbe .. e'ereace potelltial of tbe cell, by pre •• atioa of
polarlaatioD at this electrode.
p 10
In practic •• a relativel, poaitiYe potential ia .atablished
at the anal' ... electrode and tbe current flow i~ the electrode
circuit is recorded as the analysis electrode ia made pro,re ...
1 .. 11 acre ae,ative with r •• pect to tbe reference electrOde. 14 A
plot of the current observed relative to the .. 81yeis electrode
poteatlal i. obtained, .. illu.trated in Pip ... e 1. 18 the fiaure
35.0
I! '00" - 30.0 "i 0
• 2 5.0 a.. 2 cI 20.0 -
eo .. .... z 15.0 l&J a: a: ::> I 0.0 u IIOOM AI"
5.0 , 0 'It
T ANIC Nt
OL-__ ~~ ______ ~------~--------~~~ -0.7 -0.8 -0.9- - -1.0 1 o - 0 I -0.2 -0.3 -0.4 -0.5
I I
VOL T S V S. S. C. E.
PIGURB 1.
POLAROGllAMS 01' OXYGBN AT DIPPBJl1!NT CONCEHTRATIONS
tile portioa of tbe c-v curve to tbe left of the ae,ati Y8 t.o-ten t
I
I
f 11
volt 1. ideatieal for all c-. cur ... of differeDt cODoeatratio ...
The el.ctrode i. pol~i •• d aiaee DO oxld.tioD-reduetloa reactio ••
occU" at tbe applied 'f"Olt.,... Bltweea a ae,ativ. two-teatha .. d
four-teaths applied YOlt..-, tbe ox,,.. ia aolati08 ia reduced at
tb. electrode, aRd the electrod. i4 depol&r'iaed by tbe pr ......
of tbe .. eactioa. * Tbe laiti&! ri •• Oil tile c-v curve representa
the "ecOIIpOaltioa pot •• tial. TIle peak lalleetioD .:epr.Mllta the
liaitatioD of cur:reDt flow due to the lillitation of the teactio.
rat. b, .low rate of troar .... of reactant fro. tbe body of the
solutioD to the electrode .arface. Tbe YOlta,e corra.poDdiDg to
the POiDt balfwa, betw.e. the lid ti a1 ria. &ad tbe terminal in
fleetloD i. the h.lf ...... poteDtial of tb. polarographic reaction.
Tid. ia chuacte ... iatic of tbe .. eactia ad ia lDelepeadeDt of oeD
ceatl'atloa. It ia clepea4eDt .pe. tile ell_act.rut!c. of tile
ayat •• aiaee tile Daftre of tile a.,"rUD, electl'ol,te .olutioa,
the _terial of the electrode aad til.· eoadi tiOR of the electl'Oft
a.l'face will modif, the balf.waye ,ot .. tlal. 14 qaaatitative
po1uop'aphic ualyua la based upGI1 tile fact tIlat f. a Ii"e.
a,ate. tbe half ..... poteattal Cbuacterlaea lbe oxidation.
reemetioD I'eactioa.
Ie 'ipl'e 1 tbe ... loa. ourfta ve labeled wi til the vol_
per ce.t of the ,aa llixwre which wu used to equilibrate the
*Xt ia 01l1, put!al depolarizatioD, aiDCe there eould be 110 chaafe in pot.atial with iacre ... d c~reDt flow tor coap1.t. depolu: aation.
jP
12
801utloa. Bach eu .... epr.a.ata tbe total cu .. eat flow, "r. for
tbe putieul .... cODeentratlOll. Qautl tl ... pol .... oll"aphlc aDat,.sis
d.pendS UPOD ..... 1 da, the total poluopaplslc ....... Ilt. iT t iato
cOIIPoa.ats and tb. lel.DtllieatloD of ODe of tiles. co.,.. •• ,. witll
th. diffaaloa.llalted coaceatratioa polarl •• tloa of tbe aual,sis
eleet...... Tbe eompo.eats wbleh .... e ... epr.seuted In ,l,ure 1 CaD
al80 be .... r.aeat.d ayaabolJ.c&11y aa ID4f.cated ia Iquatioll ,.
iT = ir+ id ±pm+ia+ik] ! (7)
iT - total current
11' - reaidual curreRt
Id - 41fluai08 ... rent
i. - mi,ratioa current
ia - adaorption curreat
it - kinetic curreat
The valuea wIllob. appeu i. tile .aoke'. dO Dot appear ill Pipce 1.
n. " .. aclteted .&1 ... "., ...... , fact.e whicll andel' cal'tain coildi
tio •• modify the polaJ'oa.raphic curl".,at. "
The ...... d.&1 cureat i. 4eflu. b tile curent whicb would
be pr .... , ia ... ableD.e .f tbe oxidatl ...... ductloD .. eaction re-
lated '0 tbe ... 1,.'8.12-14 This ..... e.t v.rlea ia differeat
a,a' ••• aad ... le. witb ti.e fol1owl., tke applicatioll or cbanse
ift pote.tial dIt~bJ ualyaia. It i8 tbuefo .. e .. ce ..... ' to de
teral.. tbe .... ldual ."'1'.8t of aacla a,stea &lid to make allowallce.
to .. tbe variatioD with time.
The I'eaiclual hr .... eat 'acltad.. the eNl'&'eat Ilowdue to tbe
re.ia.aaee of tile 801 ....... t electI"01,..'8 of .. bat_e.. ..educed at
.ore poal tl Ye applied pot •• tia18 tbaa tbe poteati a1 uaed during
13
aDalysi., aDd a char,iA, current, lc.12-14 The total current will
be a fuactioD of the electrode earface area aad will reaain COD
staat provided electrolysis curreat r.mat .. conatant. The charl
in, current or c •• denMr curre.t, u it i. called in polaro,raph"
is a coaatut for a ,i .. R electrode surface uea ard external
realataae. of t_e circuit.*
Oft the baal. of thl. aaal"ai. It ia •• ideat that tbe resid.
ual curent, i r • CaD be tl.'eated as a conat .... ullder the follo.in,
cont.U. tiGHt
1. It i. deter.ined lor the eal.,ala .,at.e in .ae.
2. MOdification of ir by extraaeoua oxidation-reductioD react!o .. is ruled out.
3. The r.aidual Clu!'rellt ia dete,..l1ined after tbe cowl.IIMr curent influence la ••••• tlally Bero.
Slac. the residual ourreDt atabilla.s wi thiD a few tn tlls of a
a.eoad for moat circults,1C. 16. 17 :,tbls COIllpODent 01 the total
polu:osraphlc current 1 ..... il' r.801"d.
The dillua10D current, 1<1. i. defined as that portion of
the tetal current _oclated with electrolyata of tbe subatance
traaa'erred to the electrode by diffusion. 12- 14 The gradient lor
diffusion is •• tablished by depletioD of the concentration of
substance at the electrode aurface by electrolysis. 'If DO otller
*A Bel.mbolu-Gouy-Stera layer (iOD Cloud) ia formed at the electrOde surface. of electrolysi. cells vben a potential exiat. oa th. electrode. Thia layer acta like a co&4e •• er aDd the char,ing tl.e 1. a fUllotio& of the external circa! t reaiatance. Tbe capacity ia a fUDctlon of the .ffective surfact uea of the electrode aDd a value of 20 u'/ca2 haa been cited.'
14
forees are responsible for tbe tranaport of a.batance to tbe elec
trode, the polarosrapblc current beeo.a diffusion li.iled and
will YU, witb the concentration of tbe a.batanee in the body of
tbe solution. Whe. tbe diffusion component ia resolved aDd tbe
otber factora diacu.sed above ..... c08trolled, the diffuaion car
rent yari •• directly witb coocentl'atioa. Tbe ooly requirement to
be couidered wi tta reapect to tile diff.a.loa cUI'rent ia that It
.uat be • difl.sio. 1ialtatloD of the electrolysis proces ••
A naaber of otber forces supplement the polaroarapbic cur
reDt. Mecbanical, ll'avitatloDal. ther •• l &ad electrol,tic forc.s
tend to .edtfy the diff •• loD of .. balance to tbe electrode.
TraDe'er of .abetaa... to tbe .1ectrode frequently occur as the
r.sult of coa .. ctioa associated with .. chaD!c.1 .o ... eat of tbe
electrode 01' aolatloD, ... reault of tberaal currents ... ociated
with electrol, •• s or temperature d.f~ereace. io the .,.te., aDd 'j
.. a re •• lt of co.ftctiYe ChUreata .... eiat.4 witb di'ffereat
deDalti •• of tile aolutioD daria, el.ctr01.,.I.. Thes. factors u •
• ulable. Uale •• they ue cODtrol1ed, they deer ... e the accuracy
of polarop'apla.lc .. alysi.. Durla, electrolysis the ch .. ,e 00 the
electrOde .. ppl .... t. the dilf •• io. proce.. aft4 deer..... tbe
traesport 01 ioa1zed su'atances to tbe electrode. This effect,
known .. tbe alaratlo. carr •• t, i r ,I3-14 1 •• uppr .... d for iODIZed
aubetuce. by additioa of .. iruU.ffereat electrolyte.
Tbe effecta of alaratioD carrent aDd coaftct!o. require tbe
fol1owia, additional coaaideratioD. during &Daly.lal
r...------------------. 1. Minimization of density 1Allaeacea by poa1tioB1a,
tbe electrode.
2. Vaia, water bata. aDd pr.ct •• t.aperature control for the electrolysis cell.
3. Batabliebmeat of a constant diffuioa aradieat.
4. MiDi.iaiDI the laflueDce of tbe coadena ... carreat.
15
sey •• al physlco-db.miCa! 'roc ....... p'l .... t Ibe pol~o-
graphic curr.nt. SUb.tance. adaorbed on lb. electrode iacrea ••
tb. polU'opaphlc curreat WheD they ue dischuted dar ID' elee
trol,.1 •• 12- 14 TIle dischar,e aa, be relatiftl,. "a,id foe surface
adaorptl ••• Hi aa, be quite protracted ill 80M CU.s. In this
discus.ioa the cb.al.al infl.eaces ue incorporated iato a aia,le
constaat, the kia.tic CUI'HIlt. i1&:, of the equatioa. Daria, elec
trol,sis sou p&'oducta aa, daco.,." .,o_t ... o •• ly e.aia, ia-
cr .... 4 availability of r.actant and .ollaao.meat of tbe polato
J1'aphic cUl'eat.12 Bahaace_at al.o Gecus i. tho .... eactl0.8 III
which so .. put of tbe reacti •• wllica.,'l'eri.o.ly baa beell .. ate
lil11tla, i. IIOdlfled by tbe electro1,als. l'.s.ltia, i8 iacreased
ayailabilit, of reactaat. 12 OCCasioDally .lectl'olytic products
01' tbe poducta .. leb they lora with _batallcee 111 solutioa will
act a. eatal,a" for the r •• ctl08 being analyaed.14 The pbyslco
ch •• lcal iafluences dlscu •• ed above require the followl., COD
aideratloDa to be a.de in order to luue ..-alld poluo,...pbie
.&.aue_ata,
1. Recopi tiOD and allOWMC. for the preNnc:e of ad.orpti.D ph ...... o ••
I:
I
:1
rr----------. 16
2.
3.
Analyais of ~eactloDa aa.oclated with electrolyais for tbeir influence.
Aware.... of poaaib1e triaetic effect. ill tbe I'eactioas, and aDalyal. of data for this Influellce.
po1arograph1c analyais of oxypn t.aloa In ao1utioa or ill
tissaea depea4a upoa the .1ect~01,tic reductioD of oxy,en in the
presence of wate~. Por neutral or alk&1ille aolatlolls tbe reactloll
1413
-L 0 + 2 H2 C + 2 (} - - Ho 0 . -t 2 0 .. 2 2 . • (8)
(9)
Hi of re&4ti08 III EquatieD 8 ia about -O.OS ver._ S.C.B. * fo~ a
drop,!D, mercu., electrode, 4.e.,.* aad i. &ppl'ox,iaatel., oae-tenth
volt _re ne,atift f. bl'ipt platin_ electrode. in phyaiolo,ic
aaliae aolutioD. The It of the reaction 1ft Equation 9 for the
abOve a,ate.. ia -0.9 'fOlta .Ul4 ia riot. Au,1y ..... lyed lid. til
either the .ercury O~ platl ... electrode. bee .... of the extr ...
b"eadth of the waft.
quaDtltatl ..... _ ••• t of oxYlea t.aloD by the ,01 ... 0-
Il'apbtc 41"0108 cul're.' 4.pe84a IlpoD tile lIWIbu of e1ect~o •• io
YOlft. iD the .. eductioD of oXl,e.. If the first reactioll a10De is
.S.C.E. will be uaed to indicate the Saturated ~CL calomel reference electrode iR the s:ewnder of tbe text •
•• In the remainder of tbe dissertation tbis eleetrfHle will be designated d.e. ,
I'
~ -~----------------------------------------------------------~
11
involved It ia a two electron traDafer. while if both reactions
are involved, it la a four electroa tranafer. Tbere 1. atl11 80m.
questioD cOllc.l'ainl the reduction.l3.l8-20 Dayies aad kink21
calculated t4e electrons involYed in oxygen reduction and found
evidence for a two electroD transfer with ODe electrode and a
four electron transfer with aoother. The two electroD tranafer
would be expected at the .ore poaitiy. applied potentials except
for tbe fact tbat the breadth of the hydroteD peroxide wave places
tbe deCOIIpOsitioD potential at applied potential. uaed for oSYI.a
analysi.. If it ia asaumed tbat the electrode sarlace determinea
either the decoapoai tion poteDtial of hydro,e. pe.roxide or the
tranafer of tbe second pair of electrons the current-concentratioD
relationablp for • liven electr04e Is conatant.
It is pnerally accepted that below pH 9.0 the oxypn-water
hydro,eD peroxide couple described by Equations S ad 9 i. Dot
reftrsible. 18,19,22-24 COllsequently,:. -Paraday's laW5 predict oa1y
the equ.IYa1ent cureDt for tlte reduction of oxypnl the actua.1
current will be los.-hat more. Tbe following equatloD 1. the re
sult of iacorporatioll of the diffusion limitation of po1aroaraphlc
currents into a mathe •• tical expres.ion for predicted current
flowl
(10)
i4 - diffusioD current
n - PUaday/mole
P - Puaday
t - ti ••
18
c: - moles
A - electrode area
D • diff.slon coeffieieat
Krogh's coelficieat CaD be ... d if a aad C are converted to tbe
.,stem baaed 08 te •• i08, boweyer, it is easier io use Pick's D,
as solubility musi be knOWD in either case.
The a.JDitude 0' tbe curre.t froa the reactions 1. EquatieD.
8 aDd 9 Is independent of pH effects, but depeDda upon tempera
ture, vi.coaity, aD4 coapositioa of the supportiD, electrolyte,
aince all of tbese affect the diffusion coefficieni, and in some
case a , the electrolysis ~action. A temperature correction of
1.3 - 1.6S for each 4eP' •• ceatip-ade iemperature chanp at phya
iolo,ic iemperatues isreeo_eaded by LiD,ue. 13 In studies of
the iall.ence 0' "iscoai ty, i4 haa bf!~fl show. to vary inversely
with the square root of the viscosity of the supportin, solu
tion. 20,25,26 The concentratioD iDllueace on oxyseD haa DOt been
studied, howenr, otbel' oall io.. i8 &olution bave bee. studied
wi th no l'eco_endatioD .ade I~ cOI'reetioll of id Oil tbe baai. of
cODeeairatioa of ",portill, solutioa.
Several of the pbysico-cbemical lactoes me.tloned earlier are
kDown to l.fl.ace poluopapbic _oul'e.at of ox, .. a coacentra
tioa. Adsorpt!oa i.flueaces the cUl'l'ent ia two way.. laactive
substances adsorbed or precipitated on the electrode decre ... the
. I
I
I
~----------------19
.ffectiv. aurface ar.a,27,28 causiag decr ... ed .. asitivlty of tbe
electrode. Thia 18 a variable lallueace, at praseat it appears
best to diecouat re •• lta obtained .ttb .uch electrodes. Becau.e
oxygen ia atroa,l, adaorbed 08 platiDu. electrodea, it coatribet ••
conaiderably to tbe initial polarosrapbic curreat. While studyia,
tbis pbeDO .... for allo&'t period. of adeorptioD, 01.08 foulld tIlat
tbe lIajot' 41.Kllar If! is wi thin the peC'iod of the coRde.eer cuC'reat.
16 Altbeulh quaDtitativ •• tudi •• of 10.,.1' period. of abaor,tioa
have beea mad., It i. poaaible tbat part .f the laflueace of de.
' .... la' electl:04 ••• ul of tile .,illl effect la the r.aoYal. of ad
sorbed oxy,... S.Yel'al ... 41.a !! ... ft..t_I' .... ha •• iadieated tIlat
quaatltatl ....... _.at of oxv,ea 1. modified by tbe ........ ce of
hydrogen peroxide.29,13 Ja ti ... e. thl. cas probably be di ••
couated. The bJd!'o,e. peroxide formed duela, tbe eleetrolysia of
OXYP. apoat ... ou.l, decollpos.a, aD~ l. tla •• tbe proceaa i.
catalyaed by peroxid ..... &D4 "tal .. e. 30 It Ie doubtful "tIlat de.
coapoaltioa i8 tl ..... io pc.se.c. of cataly.ts !acre •••• tbe oxy-
sea a •• ilability to tbe electrOde, .iace tbe oxy,e. 1. probably
iacorperated iato lIetabolic reactiona.Tbia _aId cau.e .. reac
t! .. errol' i. tbe .etaboll_, lNt would DOt lacreaae tbe ,oluo
.. apblc curreat.
MetabOlla CaD alao be aod.ifled b, otber aspecta of the ox,.
lea aaal,sl. reactioas. The electl'ol,.!. pcodDce. bydMoxyl loa
wblch by .ht'tla, tbe pH could markedly alter tbe atate of the
ti •• ue. Thi. occura becauee tbe eaayme .,at ••• are ••• aitive to
20
pH aDd &180 becaua. _tala and alkaline euths tend to be precip.
itated at high pH' ••
DUed upoa the pzececJi.n, cU .• euaioll potaroll'apbic .u.ementa of oxysea iD tl •• ue are subject to the additional COD.i4e~
atioa. liated below.
1. There i. eo_ uaaeJ.'taint, aa to tbe umber of electrona involved ill the ... duCtiOD of ox,,.a.
3. Temperat ... and Yiacoait, variatio •• will modify 14.
Jilect~ode .... it Iv! ty .... be JIOaitol:e4 to det •• t decr .... d effective surface area.
4. AllowlIDce ... t be IUd. for pa adsorbed dariD, l ... tive ,.l:l04e.
J. 11M ll1fl ••• ce of hyc.ko". ,..oxide UJH1t1l the i4 obaerved for ox,,.. a., be different in ii ..... tbaa i. aolatiea.
6. The pH ell... u4 the Pl'odtaotl08 of bydJ:op. peroxide 4111'18, polarop'apl\ic &IIalyaia uy cause I:eacii.. ~rol'. la tbe tl ......
"
MlTJI)OS O' OBrAlNlNG pOaPROM POLAItoawcs
COKeatratioa 01' ten.on of ."en ia .olutioa is obtaiDed
fro. •• polu-ographic CU~l'ent ill one of ..... at wa,a. 12- 14 A
4iac •• aioD of tbe baaic aethoda involved in obtaiaiD, reliable
COn.ceRt .... t!O ... &1Y8i8 froa pol .... oll'.pld.c ...... meat. baa been
_iitell b1 Joha Ce._ Tayloe of the Natioaal Bureau of Staad-
... da. 31 Thia 4i .... loa llaa beeD aHd .. tbe .... atlal baaie of
the fol1a.ia, pce ... tatl... '1...... a cepreaeai. typical C-y
cur... for oxr... at v ... l... coDcentrationa aDd ill •• trate. the
~----------------~ 31
'Variod •• tboda.
In the abaolute method, a c-y cur .... fo~ each of .eftral
known and unkDowa coDcentrationa ia recorded. Tbe plate .. s before
deeompo.ltioD OCCal'S aDd tbe plat ... after conceatratloD polacla ..
tiOD has been e.tablished are extrapolated and iaterpreted i.
term. of tbe curre.t ordinate of the ,.laro~am. Tbe CODeeDtratlo
can be obtained from a Sl'apb of id ,lotted as a fuctioD of co.
ceatra,lOll. It i. alao pouible to .s. a constul (JttIid/C) if it
is establisbed that there 'e & li.ear rel&tl0 •• hlp bet.... Id .. d
concentration. Ia this .. tbod It ia ~es .. ed that oa1, diffualoD
ia cODtrlbutia, to tbe curreat uaed for eooc.aeratioD aDaly.i ••
The ,reciaioa of the .. tbOd is exeelle.t aiaee Ca) the eoadltio88
haye been coatrol1ed. (1» the a,&t •• 18 tile ... for trDOWll aDd
ultD01fll ... lyal. t aa4 Ce) the c-y e.WI ia a.dla.,1. for de'ectioD
of extrauao •• 1afl ... c... T.,lOC31 ~t.t •• that aD .~ror of lea • . .
thaD 1." Gec ... s 'a tlledet_ •• atio.".' eoace.ratioa .e. tltue
ia <a> Ie •• tb .. n error 1. tbe callbc'atloa, (a.) 1 ••• til .. 1t.
cbaap I.n electrode s.face &l'ea, (c) 1 ••• t.lta 0.5OC. chaa,. 11l
temperatu~e, 01'(4) le •• tbaA 0.'" a1 ter.tioD in parametera of tile
electl'l.cal circuit.
There ... tltte. ...,....tl Ye _thode baaed Oil tile ,l'illci,l.
tbat carrent at a constat YOl tap. rather titan CUl'reat OYer aD
applied YOlta, •• ,an UDder certal. COnditio •• , can be uaed to ob
tai. cODeeatratioD aa&1y."s. Tile _tIl04 ia illustrated. ia ,lpre
a, b, the line dl'aa per,eadiculu to the abCla •• at flve.t_tba
32
AMP JI, I 0-7 30 ___ , _____ __ ~(...::4)~_r-__ --
25 .
20
(!) 15 " ,',
r .,1
I \
I' 10
5
O~~~~~~~~~;=~~~ o -0.1 -0.2 -0,3 -0.4 -o.e -0.6 -0.7 -0.8 -0.9 -1.0 V LTS i'
PIGfMB a. 1H1 JIOL.Ai'tOOUfH1C a-v CURVB
'Ol&1'OBl''''8 of oxyp. "8.etloft at solid e1ectl'Odea. The dotted 11a •• ludicate *be method of extrapolation. Tbe Yel'tical line at .0.5$ YOlta ladieatea the YOltale unci fOl' coapuatlft _&1,.sis aad tbe urowa ladle .. te id for the co.paratiy. &Aa1ys1.. Bqaill~ated oxyae •• (1~ 11:1 residual CUl'l'cuat, 0 _ H, ox,.,.. teuion. (2 13;')" as o.xrp. te.aloa (al YOl~ oxy •• a) (3 380 _ H, uy,.. teaalOn(SO yolS oxy,eD) " (4) 760 _ H, ox,,.. t_._(l00 volS oxyaea)
35
It 30
~25
• 20 ~15 cliO
5
O'0~~~~2~0~o--~aO~O~~4~o~o~~e~O~~~~1~o~o--:h800P02 PICIIU 3.
ClIUUINT-COJfC.BN"lRATlON ULATlONSRX'
(1) Plot of 14 &,.l.st coDceDtl'atioe (fro. PilUre 2.) (2) Plot of ir acal •• t GODceatl'at!o.. Note that the
two plota differ by tbe alope of the residual current.
I
23
of a volt. At this 901tag., sligbt variations in either the ref
erenee potential or in applied potential will Dot appreciably
al tel' tbe current flow. Under tbeae coDdI tiona polaro ..... Mc
current for known concentratiolls can be •• ed to eatablbb the
basia for unknown concentration aDalysis. Tke slmpleat procedure
1. to plot iT a. a funetion of several concentrations in the
rang. of tbe expected unknown concentration and to analyze the
results by graphic interpolation. This method i. presented as
curve (2) in 'lpre 3. It is &180 posdble to add a known a.
mount of SUbstance to tbe • .,at •• aDd to .M tbe conatant Obtained,
Ie' • iT/Cadded, for evalutin. the currents obtained in analyala
of unknown coacentrationa. Correction for residual current aDd
non-linearity of tbe current-concentration relationship is'n.c
••• ar,. f~ satisfactory precision. Tbe third method depends OR
the fact that i4 of two substances 1. ~etermiDed by their electro-"
chemical equivalence and diffusion rate. Consequeatly it has
beea pOSSible to add a tnowa mount of SOlIe subs t&nce to the .,s-'--
tem. 1idlich ia reduced (or oxidized in anodic analysis) at tbe
applied poteatial. A factor k" similar to kt is obtained. k" is
converted to k' on the basis of the difference between electrical
equ!valeace and diffusion of the two substances • • , 0'
" k' = k"'X !L x -n" 0"
In the pilot ioa addition .. thod Just described above. the same
Uai tatloas are implicit as in the USe of let. In the three
I'
24
comparative methods the precisioD could be aa accurate .. in the
abSolute method provided the conditions of calibration and .. aly
sis are similar. It is advisable to obtaJ.n oeca.ioDal Cay curve.
witb comparative methods to insure a,ai.st i •• trumen"s1 &ad elec
troehemica1 artifact ••
Additional cO.BideratioDs ate raised wileD concentration is
obtaiaed through tbe use of poluogranuu
1. PrecisloD of &Daly.is i. reflected from the preci_ioD of ca11bratloll, _uuellent 01 iT or 14, aceur &0, .1 latl" .... tatl_. aad i. control of parameters of the aaalysis .,8t •••
a. COllp&rative Mth048 .e likely to be le •• pa'eel.. tIl_ "'solute _thoda ale.. add! tloUl cOIlude .... tlo. 1_ Ii yea to the aature of the Cay cvve aIl4 to tbe ,lateau repon at tbe applied YOltap.
l. AttentioD ahould be 'i ..... to linearity betweeD CUl"eat aad coace.tratl .. .
The .. e ue a lI""r of d.cu!ta Ie u. ia polal'op'apllic 0-
atnllents. The tlUeeeu.ti.1 COIIpOaeataare ". .... t •• "beat.
ica11y tD Pi ... e 4. In ttll. flpre, B ".,..Mllta 'ttle 80urce of
applied e ... '. t P .. epr ........ polarep'apbic eell coasiatla, of
&D &D&lysis elect!'ode and a .... eleJ:eace electrode. aad 0 I'eps'e.eata
tbe meana of .u"rin, the current flow In the polaro,raphlc eell.
II
II I
I
VOLTAGE
8
SOURCE
- POLAROGRAPHIC , CELL
uALVANONETER ~ --- •
PIGlllll 4.
SCHBMATIC DIACIlAM 0' 1'D POLAI\OCItAIHJC CIRCUIT
25
I. .isure S 1. & dla..... .f ODe of the .!apl •• t cueui ta. B
io this Ii,. •• coaaiat. of a batter" YOltace d1y14.~ aDd yolt
.etel't &lid po .. lbl., a poluit, ... _.aiD, awltcll. 'lublipt
batt .... i •• ue _equate souc •• of •• a.'. f ad a •• itable .. adi.
pote.tloHt ... will "1''' U & YOlta,. dlYid.... The YOltatel'
aboald 1M a ... a.. tuM t". of hllh taped.... to ... " •• t 10&CU.D,
the circali &lid 81'I'OM." ladleailea,.f tile Y01ta,.. * The •• I'.e.t ..
l'e.01'481' It. .u be • dl ... ct ,.eadi.,. o,tlcal .. .800l'dI8, .... tel'.
SI... the aax'", "'1' •• ' Ie •• 1, a few micro .... re. tbe meter
abou14 b ... a .... ltlvii' of at leut a teatb of a microvolt pe ..
acale cSi.i.i.. aad ahou14 be readable to oae huDdl'edtb of a .iel'o
upere to ' ...... e Ill<ha_ preci8101l.
n J
*A pcecialon pot.ati ... t." may be subetit.ted for the .olt&,e 41.14er &ad voltmeter if circuit •• aietaaeea are calculated aDd tbe applied vo1tace ia calculated.
26
B p
'IGUU 5.
SlMPU POLAIl()(llAJHlC ClltCUlT
The ref.eDce electrOde _sed ill tile polarographic cell "8t
(~fti.b a reliable refer.ace pot.atlal aad ha.. .. minimum teDd.n.y
toward polaris .. tioa. 1ft biolopeal IDftstip'llo •• the calo_l or
ailver-silftl" chl.soide refer.ace electrode la co80aly uae4. The
chloride coateat of the media or tl ...... akes thea. electrodes
ideal .. leoti ... as tMre ia _fflol~at _loa to pre .. Dt coueD-"
iratloD pola.laati.a aDd to .. pte... lbe -'1J:&tiOD curreat. Tbe
charaeteriatiea of t~a. electrOdes are.
(12)
.0.2415 t 0.00016 (1:.as) YOlt
1(13)
-0.2224 volt
Interface effecta betweeD the media of tbe refereDce electrode and
the an&ly.i8 portion of tbe cell, .. well a. Y&I'1&tl08 in pH and
7
aT
teDlperatve, introduce etl'&, poteatiale which affect tile celereH.
potential. POI' cODce.tr&tioa aDal,si. aa4 bl010,leal applieatioD,
.. refereace potential .. curate to two eisallie .. t fl,.re. 1. ade
quatea it ia )eobable tbat tbe 1.ll •• ace. a.alioGed do .ot alter
tbe firat two .1,.ille .. t lilUrea cited. The refereRce electrode
.surface area aIlOuld be tea t.laes tbe ana .f tbe .. al,als .1ec
trodeS to pre.ea. cea .. ntrattOli polubatt ... 14
".......... klel ..... twated wi tb potaui_ .ltloride oS' .01ute
of tile aa&1y.ls .el1 U'e ueel to CODect tbe two portio .. of the
pol ..... 8I'aphic cell Ud preYeat cOIl, .. iaatioa of tbe refereace
half .ell. Tlds laor ..... til. iatuaal realstance of th. circuit
b, a ... ral thou .... obaa and introduc •• the pos.ibilit, of addi
tloaal iate.face poteatlal. aDd ~iff •• loD ,oteotlal&.12 o.e la
.. atl,ator repected that a .&aute fl.. of .01utioD out of the
refHe_electrode tuoap the wid,. pr ...... ted the padu&1 de. "
eliDe in tel wJaiclt i •• _U ... ob ...... Ci _d J!'e •• 1ted ,. lllcl'eued
.tabillt, of tbe ........ at •• .12 Witb tbe exceplioa of tile laat
factol". the i.11ue80e of the '-14,. Oft tbe I'efuence poteatial ia
81Waated if the .aIU.-.ti __ d ...... e •• t. ue c..,leted e.
alail_ ay.t.... ta otber c .... it _at be eyala.ted.
ADal,aie electrod.s ., be either the dI'Oppill, .ercury or
aolid •• tal elec:t .. OCS ••• IJ AltbouJll platia_ I. tile _tal uMd
.at fl"equeat1,. p14 elect .. O<I •• laaYe .... at_ uHfulne •• III 80_
app1icatioa •• at Other .. tala, ill.1ud!., t •••• tOD t .11.er, .taln-
1 ••• atee1 &ad .. tiaoa,. have beea tried but the 1' ••• 1t8 bave BOt
28
beeR prodsia,_ The electrode .ia. yuie. from seYeral square
.illl .. teI'8 surface area to the tea .. icl'a-dlametel', ,old-plated
electrodes ueed by NiHall,.33 CODceatl'atiOll polarisation is ea
tabliabed -I'e rapidly aDd appeua to be IIOl'e stable with the
smaller surface areaa.
The reaction. whicb occur at the analysis electrode are iD
fluenced by a atlllb .... of factol'a. Pol' the reactioD to be aaalyaed,
DO ioterf.l'i8, subatancea •• y cau •• depolarization of tbe elec
t1'04e. Daria, analysia baaed OD reductioa, bydrogea evolutloa
f!!ftatually caaaes depolarization. Theoretically tbis should. OCCUI'
at Yoltagea tbat were more positive thaa thoa. actually observed.
The differeace between the theoretical and observed voltage at
which hydl'ogea ewlves i. called oyervoltage. Acid lIedia, natue
of tbe electrode sufaee. current deaal ty, and duration of elec
trolydelS ue., .. of tile lactora ~whlGh decreue oyerwltap. "
Investi,.torl baTe alao reported influence of previous ua. of the
electrOde &ad filii forMtiOll oa the electrode surface ... factora
wbich aodify tbe electr04e zeactioD. 34•35 The reaction for ox'S ••
reductio. i • .,uticulM1, auaceptlble to .ul&tloo. Davi.s and.
Brink have zeported a two-electro. &Dd a four-electroD tranafer
for the a... t,pe electrode UDder ideatical CODdi tio.s. 21 Thea.
variation. and the fact that to a peat ext.at tile" oripA ia
i.perfectly uaderstood iDdicate that a sreat deal of cautioa
should be attached to DeW interpretationa of v&riatioaa ia r •• ult
29
Solid electrodes have additioaal dlaadvaata,.. which are
not preseat wltb tbe d.e. The abort-tera accurac, with moat elece
trode. haa beea lea. thaa five per eeat. 184'1 10D,-tera accurac, is
betweea fifteea and , ... t, five per ceDt for .elected electrode ••
part of tbe ... labilit, i8 due to modified 41ff .. loo ooadltioDa iD
tbe re,loD of tbe aolid electrode wIlieh ue alai.al iD tbe .. e,1oD
of a d. e. lOti, tber_l and Mehuteal effects hay ... re iafla
eDce OD aolid electrod •• thaa oa the d... In additioa, the .lec
trode ..... fac. i. aot coututl, r ..... d U4 tead. to becoM COD
t.-la&ted e.-. tboG,1l *be reaction take. place in aolution. SOme
of til ... factora will be .phulaed in tile review of the polaro
p.plItc Hasare .. at. of pOa 1. tt .... a aDd blOOd.
la.tr_DtaUoa latrod.C •• tile foUowia, factors f. attea.
tioa durin, quaD'lltatl ... COlle_tratloD _al,slal
1. The electrical coapo.eDt •• botJ14 be selected to o.llow an acew:ac, of at ledt o. S% ill tbe applicatioa or •• &aU ..... of \'01,.,. or current. "
2. Pr •• iaioa abould be made to read the polaroarapbic current to 10-8 aapere.
The r.fe~eAce peteAtla! aad factor. which mo4ify it 1"eqol ... 1I08it.l'la" putle.lul,. wi tb the comparative method of analysis.
The ADaly.ia electrode $~fac. &rea should be ainia1aed if at all po ....... 1e.
5. The aa&lysis electrode a.rfaee sbeald be cleaa at the be,iunina of analy.i. and checked for cont .. -1aatioD 481"18, and/or alte. experi ... , ••
6. Pactor. diaturblD, the 4iflO8l0. abell of tbe anal".ia electrode should be mini.laed, particularly with 801ld ele.trode ••
II
I!
ill
It I
l Id,:, ,I,'.
'" "
rr-----------. 7. Maxia .. accuracy with 101i4 electrode. will de pella
upon frequent calibratioD.
30
Several circuit lIIOdificatioD' iacrea.e tbe utility of the
polarop:apb. ODe of the moat useful is tbe automat Ie atep..wiae
increa.e ill applied volta,. over a selected YOlta, •• pea wi til
automatic recording of e-y curye •• l~ Provi.ion CaD alao be made
to paaa a bucking currat througb the recordill' circuit to reduce
the --,aitude of the recocded deflection. l3
Seyeral adYaDta... aad '88re ... 4 type. of analysis are ia.
troduced whea alteraatin, currents are uaed iaatead of direct
current polarization or whe» tbe d.c. ia modulated -ttb an a.c.
potential. This modulatioD allows an almost iallnite number of
type. which CaR be classified &a derivatiye. $QUAe. wave, oaeil
lo,raphic &Ad interrupted pola$0lZaphy. The la,t aodiiicatiOD
will be discussed .10 4etall, as it is uaed in the reaearch. Tile
other .. thode will be presented to ci~If, the differ.llce'· in the ..
methods compared with interrupted polarography aDd to poiDt op
their potentlaluae.
Derivative polarogram. are obtained by record! •• the alter
natl •• curreat flow which results when a sla0801441 modulatioB of
the applied pot •• ttal t. used. While tbe applied potential ia
swept tbrough 4 selected voltage span, a .. all aiausoldal varla
tioD of a few hondredths of a volt at a frequency of sixty cycles
per secoad t. superlapoaed OD the applied potential. 31 Pi,ure 6
pre.enta tbe priDCiples involved.
u o -
6 A.
o ..;0.1 -0.2 -0.3 0,4 -0.5 -0.6 v. A. d. c. current vs.
d.c. voltage
~ z ~
a:: a:: :> u
o -0.1 -02 -0.3 -0.4 -0.5 -0.6 V.
B. a. c. current vs. d.c. voltage
PIGOIli 6.
DERIVATtVB POLAROGRAPHY
Horiac6tal arrow. indicate 0.05 YOlta,e ·variation for a given value of tbe applied .oltage. The wertleal arrows iadicate the carreapoadiD, alternating current.
6 B. A continuous record ot alternating curreat derived dutiD' a sln,le sweep of the applied Y01iage .paa with superilflPO •• ct a. c. modulation. The maximum CQrrent indicate .• the balf ... ave potential and the II&pi tude of the .. c. at .. ., point iadieatea tbe d.e. slope.,
31
The method quickly locates ihe balf.wave poteatial wb.ich ident
ifies tile reactin, subatance. The alo.,. of ttle ... ealdual curreni
I
I I
I
0 ... plateau i. readily obtained f ... om the m&lDiiude of the alier- I
aating cUJ:l'ent. The mei1lo4 would _ita it poaaible to check tbel
realdualeurreat. idelltify the l'eactinS 8.bataaCet Uld to deter-
Idne the extent to which the alope 1. ellaDling in the region of
comparative ana1y.is in a maiter 01 .. few a.eoad. aDd without
aerious lnterraptloa of ... lyeia. Tbe .. thod haa been reviewed
by Lin .... aDd WiI11ama. 38
aue wave
~-----------, 32
0, C. LEVEL: "":::-'-~=~---_d--: I
A. Square wave modulation B. Charging current
C. Diffusion current D. Polarogram
PIGUIJI 1.
PUNCltLBS 0. SQUAD WAVE POLAJtOGRAtHy
f A. Tile peJ'iOClio llOdulatl.. of' tbe applied pot .... ,i&1 by .. ,.rl.,.8.d .. _are .ave potential •
"
., L Tbe I"elatlve ti_ COHM of the Cbarpa, Ctll'l"eat d .... ia. *I...... .... ,., .• ,1&1 ••
? c. fte .elatlft ti .. 00 ...... of aae 6iff.aloe CDr .... t darial squar., •• v. poteatial ••
1 D. OKil1offapllic record of tbe cureat flow ill a ,. .. iOd 0' modulated potential.
a metbod fOI" a.paratill, tbe eberlin, curl" •• t fro. tbe diffusioa
CUl'reat. Beca.ee the tiM. cours •• of the charJiD, aDd diffusion
curl'ent. are differeDt. It is po.alble to obtaill tbe diffualon
curreat after tbe char in curl"e.t baa deca ed. 18 uare wave
~-·--------------------33~ polUO,rapb, til. applied pote.tial may I'euia co •• tut 01' ., be
swept tueup the wi ta,. .p... .. i. 4iff ...... tia1 po 1&1'0,1' apb, ,
.. d it ma, be ... d wit .. eitbel' the d.e. 01' aolld electrod.a. I •
• N witta aitbel' type of .1ecuo4., Ita •• J- advantap is tbat tbe
opel'&tOI' .aa take baediate I'eadla, of tile ,.laro .... apllie CUl'l'e.t
laatead .f waitlll, f •• apletioa of tile .... cur, dl"o, 01' tOI' .t ...
billaetioa of the cure.t with tIM aolld eleetrode. SOlid .lec
trod.. asy take •••• 1'&1 aia.t.. for .tabl1laetio. to be •• tab
li.bed. BY •• thea, tb. e"'l"oee &1'. 11k.ly to be ,..atel' thaa flf-t... p~ c •• t. 01 ... 16 aad ..... t.l'39 bay. abowD that 14 obtaiDed
witb eol1d .1.ct.I'ode. i. ,ropo ... tl ... 1 to ooace.tl'atioo of .. _&Otia,
•• bat .... witbia • fflttl .atu of a .... 1Id. l01lo.in, pol.laatioD
of tb. ..lid elec •• ode. 18 acta... wave poluo .... apb, tile clloice
baa be •• to ulce tile •• &41_, _ria, tbe _.1y pita .. of polu"'",
tloa. Vade .. th... coadl tiou .. I4C~U' of withiD two pel' ••• t
of tbe ,.obabl. val_ i. clalaed f __ .. eat ... atiou of lr.6
IlOlu.40 TIle aetlaod off.ra tbe a4.,.uta,.. of iacl'.".d .... itiv
it, &I'Jd a.cuM., aacI baa DO outat_cU .• , ellewyant.,. ••
oacll1opapble potuopoa,lly .... de .. loped fn. kaowled,. obe
tdIMeS with .. RYe.tl ••• 1 _ .. bods .f pol .......... y fo~ .."lieatlea
to tbe study of .... actiOll kiDeti... TIle .etbo4 utili ..... Ili ....
re.lataD.e cleeaii aDd .... c .... d. pot.a.tal-tl .. cu ...... with aaall
electrod •• wbicla ue polubed by 'be .... , ¥Olta,. t.cbDlque. It
It ... tbe adYaRta,. of be!., I'apld a1l4 off .... i., .. oPpol'tualt, to
alter rate 1Wtia, futol'a iD the reactl08 with vlauallzet.lo. of
34
tbe induced cbaap in 'the oscil1oaraphic po1ar08l:_.12
.Althoap a BUIlber of lnft8ti,.tol'.'8 llact obauvecl that the
801id electrode was more .table after a period of shortiD, to the
ref ••• ace .lec'tl"ode, tbe first tboroup illv •• ti,atioll of tile fac
tora and .. evaluation 01 their lalluenct wa. •• ported fro. tbe
LabOratOl'Y of Pbysical BioIO.." Natioaal Institutes of Health. 16
In tile iaitial repo ... t at .... coad cycle ... u •• d, and the aDal,
Ida electrode in alte .... t. periOds of .. cyole .... c.i .. d 'll'at a
positiye. tbe. a De,.tive applied poteatial relative to i •• re'.",.
ence eleotrode, ad waa _orted to the refereace electrode be
tween applied poteatlals. The current recorded oaly duria, the
t .... aiDal pllue of Deptive applied poteatlal, .u foud to be
proportional. to OXy,.D co_eat ... iloD. 18 a late .. atad".1 tbe cir
cui t va. modified to a-edu.oe tbe cycl. to t.e. ..coll4. aJld to i.elude a .,at •• for rec_diD, tbe I"at~ of cban,e i. ox."en CODcen ..
"
tratioD based apoa two co ••• c.ti ............ t.. TIle _Ibod Ilaa
tb. &eI ...... t .... f laenued .tul1.tt, Ud Mc.a • ., aad probably
teDde to tee, tbe electl"ode aurlace c1e ...
.lui, atta,ta '0 UM 801J.d e1e.troeS •• '01" _ .......... t. 1.
aalmal ti .... 8 and blOOd 1" ... lt.. ia unreliable .atlaate. of ox?
,ea tension. ~.e have be •• attributed to 8Od111catloll of the
electrode aurface aDd !leterop.eGua ... tue103 of ox.,. •• distribu
tioD c •• ..s by tile ".e ..... of re .. bl004 cel1e. As a re •• lt
.amero.. elect.04e aodiflcatloaa bay. be.. latrOduced to eiccu.
.. at the •• difficulti... 1ft th. ori,inal article on tbe •• e of
35
solid eleets-odes for .uuement in ti •• uea21 two uth04a were
proposed for .odifiaatioD of the bare $Olid electrode. Tbe re
cesaed and covered electrOdes which were deacribed are pceaeated
in Pi,are 8 with other .odilicationa introduced b, Drenth .. aDd by
Cluk. TIle fou typea of elect.rode raodilicatiou lIIui the 01508
A. B. c. D. '1<JUU S.
TYPES OJ )I).DlPIBD SlLJD MALYSI I BLIC'I'llOD.BS
A. Itee .... d (lJI'lak) C. MembrUle (Brink, Clark) B. Covered (Dreakllaa' D. I_iated (Clark)
"
circuit repreaent tbe .at effectly. _tbods of coabatia, the
d1fficulti.a eacowatered i. t1a ••• Me! blood raeUUl'eaents.
The recea ... eleotrode atabillaes the dilluaioa coaditioaa.
thua iaor.asia8 reliability &ad reproducibility of oxYJen teasl0.
_aaure .. at.. Iteepill, the ree ... beJ:e ... 11 or fil1il'l' the rece ••
with a ,el reducea tbe r.b.c. lafluen4e. Tbe electrOde will
eYeDt\1al1, be colltaalaate4, but the process Is delayed.
lkeakhaa fou_ that WeD a tbick collOdiOfl atoctlD, OftI' the
8011d elecuod. "as uae4 "itll 01sont • chou! t t the .aeureMnta
"1'e inae.s! t1 v. to 1'., b. c. • a. He 41180 observed lacreued
36
stabilit" re~oducibl1It, and precisioR oYer otber .olld 81ec
tl'odee. 42 Wi ttl the ".co_ellded cue tbe .1 •• t,,04.8 will aot abow
aipe of cont .. l .... tieD for •• "1'8.1 we."_.
Clark ha. I'eported electrodes partiall, 1"1&ted42 &ad co ••
pletel, .eparated43 fro. tbe ti.... 01' .. l.tion b~ a meabr ...
permeable to oxysen, but itiper •• able to ,_ water sol.ble sult
.taDc.s and water. The •• electrode. deteraine the ox18en CODCea
tratlon by tbe ..... 1' .... , of ,he ox,,.. whleb diffua.s throu,h
the aembrane. 44 In the par~ial1y Isolated 'electr04e, contamiDa
tia ey.anal1, GeC.8 by diff •• loft of materlals throup the
'bri4,et beneath the aeabraae which preYldes for electrolytic
cOBnecti .. with the refereDce electrode. Ie the completely lao
lated electl'ode .tth .. ta'eraal I'elenac ... electrod., the oDly
suba,lUu: •• which re&eb the ,electrol,s! •• a •• rift, cell are thoae
soluble ia the MIlbrane. "
M, s.,.'el:l ""lell r.8nes oXYp. 'ct_iftl the proc... of me_
are •• at red.c.. tbe _.,,en cODe •• uatioa ill the re pOD of IHaa
VeM.t uti lweI'S tile quaatlty present. Oft1., tbe droppia,
mercur, electrode aad bare .olld electrode with aD aareatricted
dil, •• loa field are depeDdeat apea tbe coacentratloD representa
tive of the bed, of th. $Olutiol1.lf tbe ratio of the 'VOIUlle of
8011ltlon to the .. lume of the diffusion field ia lar ,e and the
duration of ... .are ••• t Dot too protracted. these two electrod.s
validl, meuure tile oxypn tension of the s,.stem. In the us. of
01soa'a eire.it or with covered or isolated electrodes, the
37
aeasuremeDt i. a function of the CODcCtDuatioa in the .ala body of
.olutton, but ia lower tbaa the r.~e.eDtatl .. cone •• tratioa.
nuria, .... u ... at. wltb 01aoa'. circuit. ad.orbe4 oXYJen aDd ox,
,.n withi. tbe diffua10D field 1. COD .... d. Thl. 10 .. will be
partially I'eplaced 4urlD, the ,.riod of alte ... _te pol ..... i •• tioa &ad
tbe two ahOl"tlaa periods, but the t1_ ia ia.fflcieDt for eClui.
libria to be obtain.d betwe.D the dil, ...... ..... n .. d the body
of the aolutioa 10 the abaen.e of eoD .. ctl ... eff.cts. III the CaM
of coy ..... d electrode. 01.' ._bruea. a dill •• 1011 1I'&418.t i ....
tabllalle4 dalla: to 0" of tIl_e 111 ..... at.. la Pisue 9.
It 1. eYleSeat tba. the cOD.eatratiOR or te.sloa la 1... at
the s ... face of .... brue .. electrode cowri., thaa it is ia the
bod, of 801atioa. Ox".. is les. sol.ble i. ""u.. _d COY.". in,_ i. ... .t pr .... t than 1n solatlou, &ad OX",,1l eolubil:i t, iD
the ._._. ,. the lWtia. lactoJ!' ,11l the quutlty of ox,p. "
electrolyaed i ••• t .,.t •••. * UDder" the .. coAdition. l.t ie tile
ox,,.. at the aol.t1oa-_.b~ .. e .taterf ace which i. aeaaued, &ad
1t is ev1d •• t tbe OX" •• at tbe 1aterface is le •• thaa that ill tile
body of the ao11:ltloa. Wi til co.tl .......... eaeat uia, the O~ao.
c1~cuit or the modified electrod.a. equilibrium ia e_tabliShed &ad
a conat_t valae for ox,.en cOllceatratiol'1 is obtai.ed, but it ia
ld J I
*It i. imDo.aible to 4eterat .. the .. lUbillt, for ... 10 •• ••• bl'a.s which bave beell .s .. , but thie atat_at i. supported bI tbe fact that tbere ia decreaaed current obtained with modif ed electrode. in .pite of aborter d111.sl.D. distanc •••
38
O2 ' - I -- .... /1.. . -------- 240 '!
E
o s s
1. OXypa ..... 801able ill 2. ax.,,.a Ie .. solubl. in •• mbraae tllaa ia .latioa. .abrue tbAa in 801utioa.
E
A., SYSTIM ILLtJa1IlADD IN TlDtNS OJ' CONC.BNfRATION
- 156 MM HG PO Z
o ': , .
E.. If. S.
Electrode Me.brane Solution
a. srSTBM IU,USft.ATlm IN TlIUt, OJ' TJlfSXOH
'1 .. 5 9.
DIPPUSIOJf IN A RftIM CONTAlNJJrfG A MlMawm
•• i4.nt fro. tbe above anal,eie that the concentration at the
electrode will be 1 ••• th&fl tbat whiell exi.t. in the body of the
801utioa.
MOdilleatioD of drcult. or electrOde. u.ke. it po.sibl. to
coaslder addltioDal opportunities aad iDtroduc •• f •• alble cbaaps
1 .. polaro ..... pbJ.c _uue ••• t of oxy __ • aCh u.
,
: I
i , I
"
I
1. ConvenieDce featur •• CaD be added to a cireuit.
2. Additional feature. cou14 be added to a circuit to peralt 1I00itol'iD, of a reaction, iI', aDd the plat ..... e,ioa of coaparat!v. aaa1yal ••
3. Modified circuit. aate it po.aible to r.ad i4 or ite equivalent conlinuoualy aD4 directly.
4. Modificationa .... rally deereaae current aeft-ai ti vi t,. but i8 ... ca •• , at le .. ' t iHr.... it.
,. Se.altl.1tl' atabilitI' apeed of r.spo ... , reproduclbil ty, preei. OD, reliability aDd validity ...... lat8l'relat84. .All a ..... .,. to obaa,e ... will .o4if, one 01' .ore of the otbera.
6. BlectrOde .odificationa iatroduce ,robl ••• in relati., tbe .... 111'..... to tile .... alia! pC)2.
JlBVIJW 0' TIll LlURATtaI
39
ADaly.i. for oxypa .as ..,Il, the ..... 1' deftlopmea'a of the
poluop'apbic metbod. Two 14I ... a aft.r be iatroduced tbe techalque
.f poluolI'apbic &aurals ,12 Ju:osl~. Beyroya.,'" I'epor ted the "
I'ed"ctioa of ."e. at a "eNd.D, "I'cuy catbode. These initial
obs.rvatioDs Oil ox"en analyala bave bee. expanded ., He,rova.y46
and Vitet47,48 ia Herrova.,'. laborator, at Charle. Qai.e .. ait,.
toltboff aDd Ilia aaaociat •• 49-'1 at 'OBivers!t, of Minn •• ota ia
this couatry bave verified aDd amplified .aDy of the earlier flnd
iDI. cOIICerain. poluo .... hic ox,,.a anal ,ai.. The polU'op'apbic
literatare 08 ox, .. a iaclad •• .aa, re~ta by aaalytic Cheal.ta
latere.ted i. the kinetic. of OKY... redactioa or th. i.fl •• ace of
cliuolyed _,geD UPOD other types of ual,aia. OAly a few of
thea. uticl •• haft bee. iacludeel i. tbe 4i ... rtati08. Tile
40
remainder of the literature repre.ents .pplicatioD of tbe polaro
srapbic method to problems encountered in other fields. Tbe re
yiew indicates tbe proble.s and tbe coatributions of polaroFapllie
analysis to their 801utioa. Tbe biblio,raphy contains the Ilter
.tare OD tls.ue aDd blood axy,.n teaaioa aeaaured polar:osraphical-
1y throulb Septeaber 19S8, aDd mach of the related literature on
mea.ureaeat of _,,.a In 801utloa.
The first 18"eati08 for: applicatioD to biololical problema
was made by Prat. S2 Howeyer, it was Dot until iadependeat au,
g.atloaa for ita application bad beeD made by BridckaSl and by
Bawaberser aDd •• 11er,12 tbat iav.atisatioD was besun .sln, 02
medUrelleDt In blolo". Brldcka'a work df.acusses tile broad
application iaclUdia. Gr,aAlc analysie with •• ntioD of the analy
sis far diaaolYed oay,ea. In the Staaford uniYersit, Laborator,
BaWlber,er and .uller ,,"e .. ated a rctpc)rt and de_natratloa of "
awllcatin of tile polaropapble .. a •• eme.t of ORYS- 10 a S.8-
peasion of ,..aat cell. .. a mean. of deierainlB, respirator, rate.
Daria, the applicatioa of tbe polarosrapbic metbod to their
proble •• , la ... ti,ators ia vario •• fields of eadeaTOr baye co ..
pared their re.ult. with the staDdard lIletbods used in their field.
The.. compulaoas ladie-ate the pote.tial value of tbe polaro,raph
Ie method, .a well as Ita limitationa, and at tbe same time reyeal
the many cODtributions Wblch bave been developed throu,b its
.pplica tion.
Ia the field of en,ineeria, aiaute trace. ofaxy,en In the i I
41
bOiler feedwate .. of bigh pres.ure steu bOilers and iabiDes lea.cs.
to extensiYe corrosion. The eaaiaeera are latereeted in coatinu
ous _ai ioria, and a.chaieal coairol of the quallti t, of coat_i
catia, OXJ,ea. In a rece.t s,aposium" the ayallable meiboda .er.
presenied and .. attempt .a. made to set up a ataadard .. the
basi. for cOilparatlft stueli.a. If the reault ..... e indieaiiye.
the 014 _thoda baYe v.ry 11 ttl. preciaioD ia tbe ruse of tea to
ttIO b1llldl"ed pal'te per billion. III coatraet. inatruM.tal .etllo4e
wbicb •• re repeeted indicated that the po1arosrapbic •• thod waa
coaai4erabl, better tbaa o14er metbods and aa SOOd a. ...... aetb
ode. The newer metbod. were coatinuous or ••• 1-contiauou. with a
precia.o. of wiihi. two to t.. per c... iD the r&8.e of three to
one bua4red parta per billioD.
BlOIo,'st. and .aaiaeera bave compared tltratloD aetbOda with
poluo,..aphic aethocla 1ft the i.v •• t~.atlo. of the d!aaol"" .,_
pr ... at ia natural watera and iaduatzial .lf1ue.ta aDd w.at.a
which cODt .. illate ibis aatural .. eao_ce. III rotltiae u ••• f tbe
4 ••• for ox"e. eaalyeia t.e •• mea obeeryed a precisio. of within
two per ce.i wbea toaometric.S6.', Wiakler iitratioa,60 oc
colort.etric a.thoda61 were ••• 4 to e.tablish the collceatratloD of
ox, ••• in solutioa. with solid eleetrod •• or in tbe pr ••• Dce of
contaaiaallta, parti_1arl, tbOM of OI'cuie ori,ln. tile aceurae,
was betw •• D two &ad five per cent of tbe value obtained by Wiakler
tl tratloD. 62-67 In aoat c ..... the iavestigatoz •• e1:e lllclilled to
attribute the errors to tile Wiakler titratioD, whicb is extremel,
II
I
I
42
sensitiye to imparities present during titration. Tbis contrasts
with the five-tentha per cent error wbicb the chelllica! &oa1y.t ill
able to obtain13 and emphasize. the 1.~taac. of the cootrol of
the contents in aolvtioD ill order to ... s._ aaxi ... acc .. acy.14
Tile pol"oarapbic _ihod h ... au.4e available to the en_Deer
&ad to the biologist aa i •• truaeatal method for the d.termiaatioa
of dis.olYed oxy,en to replace a slower aDd le •• precise metbod.
In addition it baa made it po.aib1e to replace diaeontiDuooa
aam,lia. p!'oeed_.a witll a conti_ .... cordinl OJ: 1lO.1ltorinl
..,ste. pd _de deflait. techaolopca1 Wyuc.a pouib1. in tbeM
fielda.
TIle are.. ia 1IIbich pol .... epapllic _tbod ha. coae clo.eat to
aCbievi., it. axi_ .. potatial valae ia ill the applic:atio.D to the
stud, of respiratioD i.D .. lutiou. Oae of tae 1i1litationa of the
eORftniiou.l I.'eapiccmeiua ia tbe t4 .. required for equill_i_ to "
be eaiabliab.4 between tbe plsu •• la "the .. uuri .. .,.ata, inter-
ior of cell, cell Mmb.tue, aolutioD, ad pa ,ba... In Warbar,
aD4 IM'croft type &ppuatua this ia baateoed by _akill, the rea
piro.eter. In the reapirometer. used for ainBle cella or small
phe.a of tl.a.e the YOIWlea must be Ude quite ama1l to make it
poulb1e to meUUl'e rates of the order of 10-6 liter per !lour. 68
The poluoar aphie ... thocl e lWaa tea tile aece •• i ty of .hakia, aad
baa the .... aeaaitiylty aa haye the aingle eell respiro •• ters,
altboulh it introduce. the reactive error. of tbe po.aible tox
ici iy of mercury69 aud tbe coanaptioD of ox,sell by the
43
electrode. 70
80th qualitative &ad quantitative .eaaarementa have been used
in the application of polarosxapbie measurements to the deter miaa
tion of respiration and photosynthesiS. The maximum accuracy for
the droppiul electrode waa claimed by BaWllberger71 to be four
tbouaaAdths per cent, cu~p.ria. favora.bly with the three tbousaa4.
the per cent accuracy obtaiaed in the volumetric measurements.
The sreatest accuracy obtained with solid electrodes i$ reported
by 01408. 16 Usinlthe 110dified circuit he was able to obtain an
accuracy of within one per cent. Darie. and Brink report &J1
accuracy of within three per cent21 for the reces.ed electrode,
while cOD.lleII,.'10 fro. 'the aue labOratory rapor'ts tha.t bue wire
electrodes were accurate to between five aDd ten per cent.
Knowled,e of respiratory proceasea &ad photosynthesis in
uscellal" orpni ... baa beell exp~.d in aUf &reU by applica-"
tioll of tlae poluosraphic method. A critical oxrlen ten.loll of a
few millimeters of mercary at which re.piratioD become. oxy,en
dependeat .&$ verified for bacteria by LOD,auir.7l and for reaat
cella b1 Wiaal.e,7a who alao democatrated the depeudeace of the
absolute ftlue of crt tical p02 upon teapel'atw:e. GoodkiDd aad
Harfty?3 were able to relate 'cU.uilll' of photoluaineacellce to
critical ,02. Similar atuaiea 08 photoayathetic prodU(:tioa of
oxygen revealed the initial burst'. of OXJ'.en prOduction 01: re
leue at tbe be.i .... lIl. of illwaJ.aatioD, the inElueace of tbe
leDltk of d .. " iatervala, aDd tbe quantum efficiency of photo-
44
.yntbeala.?S,?6 Ad •• ntase baa al80 beea takea of tbe ~apldii1 of
the reaponae of tbe polaroaraphic measure of oxy,e. coaceatration
to map tbe photoaynthetic spectra?? aad tbe intenaity effects of
ligbt UPOD oXYI.a production.?a IauJIlbe~ ... '1.80 haa abowo tbe
usefulnea. of the d.e ... a rapid meana of .atlaatiDI the
oxidation-reductioD poteatial of a auapenaioa and the relationship
of OXJ,en con .. .,tl08 to eytochro.. reductioa.
Siailu atudlea on uiaal t1 ..... llaft not bee. as allCe ... 'ul
in quaati. tative deteniDatlou, althoap a ~.at _y Mv_cae la
mowl •• ,. haft aeened fna the polan paphle studi.a. Duacllerl
81,83 ... able to show that ase Iua4 relatlftl, little lafl11enee Oil
lIOuae ape... • •• pll'atIOl1 in !! vi tro atudi.s. ..4 W •• 8D83 t.sted
Barcroftta tbeol'Y .1 el'ytbropoeiaia abowl., a critical pOa of .ix
.illimetera of mercul', &ad a •• ciprocal retatio.ship between
reapiratioD ... ,1ycol,al8. D. I. Hl1184 .. «4 "6, .. ac~e !! "
vitro aDd ... pled tbe 801.'io. fl'oa abo •• tbe ... cl. d~l .. ex-
po ... e of the lower aurface to nr.,ID, OX".. teadOlla. He ...
able to demo.strate a critical ,02 for fro, ... cle ia the ....
range .. that found tor Ilicroft'aai.... Tile JObUOD 'oundatioR
laY.atl,atora have utll1_.4 a flow reapirometer for the atudy of
respiration in i801ate4 ae'Ye aDd .yapathetlc ,&aelia.8S •• 3 Tbei%
findin •• aot oal, ideatify tbe respirator, rates of fro, axona,
tbe lncre as.a aaaociate. wi. tit coDdtlc tioD of lap.l ... , lAd tbe
kiaetica of re.,lratloll, bUt alao tbe probable mechani .. by whiCh
oxy,ea re.plration ia tact.ased. I '
45
Other s'bldiea OD ... e.pl.l'at108 in cell. aad atttrpea.lon. haft
rel1ect upoa tbe qualltatl .. polu08l'aphic 4etel'aillatioa of oxy, ••
eo.centratioll. Thi. aethod utilia.s th. rate of ell_,. 01' I'ela.
tiY. catea of ell..,. fzoa .teady atat. n.1u •• as a _u. of 4e
teclI.inia, tb. Dature of nspuato~ peGc...... co .. ell,10 baa
desc ... ibecS tb. principl.. of the .. thod la detail. Tile .s.em lal
upect. ae j)I' .... ted ia ,1,.e 10. III 'i",,8 lOa tbe two coa-. IT .
A.
PISUlE 1'0.
t B.
N2 GAS PHASE STARTED
o I 2 3 4 5 6 SEC. I
APPLICATION O' SOLID ~BS 10 1HB STUDY OF.
CHAM'CIiJ INDU!l!D IN 1ll •• I.RATION
atant levela of IT .... p" .... t atead, atat.. of oxy.e. coaceat ... _.
tloa. Tbe i"J:'e .. larlti •• at th ... level. aad oa tke a10pe r.p .....
aeat l .. tabl11t, of the 81.ct.-octe, aDd It i. appue.t tbat til ...
will DOt l.flue.c. tbe filldiala. If the atead, ata •• leftl. ue
kaow •• tbe "ate CaD be Ii ..... I.. absolute ... l .. a. Otbemae.
relad. ve .a'. cbaa ... or tbe "el*-iioaatU, betweell .. ate of oil ......
aDd ainlt_eou. _tabolie activit, 01' "hel' chan,. is "epol'ted.
ID 'I,... lOA tbe atead, atate system ia diaBlamed. The altea of
46
oxygen eODaumpt!OD are-ladicated .. X ae4 XI. Po~ purpoa.a of
aimpliflcati •• , X. the cellular co.s .. ,tloD. la a.aured to be uDi
fo'. thl'ouSbout the cell. X'. the electrOde cOD_aptlo •• i.
(iepen4 •••• poll tile ox, .. a ift tbe .. e,ioa of the electrOde. Aa la
dlcated ia the fi,. .. e. thi8 coRce" .. atioa for a ateady atate ca.
be represented by ... "04S of slailar c.Bceat~atlo. (dotted 11 .. a).
Alteratloa of tbe ea.ir ..... tal ~xYlea oGDeeatratioa or 1. tbe
,ate of reaplJ!'atioa r ... lta lD traaaltioR to a aew ateady atate.
In the t,uaitio ... epre.eated la 'i ..... 108 the oxy,.a aY&ilabll-
It., ia defte&HCl by .. ohan,. la eafiro ... atal ox, ... f~OII 156 _
HI to ca. 0 _ H,_ ftia ob .... la reflected by .. fall ill iT to •
DCW atead, atate.
Thia metbod .... x, .. alYel1 ap,lie. to fl'o, .erYe by iayeat.
l .. t0l:8 ia the Jo_8On ,euadatl.a LabOl'atori •• at tile tIB1.era. ty
of .e_yl ... 1&. It ... fO.1ld 'lbat:: heat pcoductJ.o. at&d alteratloa ..
of ox".a coa .... *io .... oolated wita ... iYity follow differeDt
time cour .... ,. Studl •• of tbe ex,,.. 0 ••• umpll08 .. elatiye to a
l.'est181 .. _.ane aDd ... ad.ctlDI ........ IDdlcaiM that utl
flo1al pol&1."ls.tloD9. of tbe _.ue 4Jd not lac...... ."eD COD-
... ptloa aad tIlat ttle claaB .. 1 ... , .... 00 .... ,t1 •• w .. aot duect ..
1, related to frequenc, of Coad .. tl ... ,.,9? The addl110D of
phoaphate to preparation •• u oae aetbod of iocreaain, tbe rate of
oxy, •• uptake, aad It ... 8GBcladed tbat oa. poaalble repl.tioD
of tbe I.'&te of ox,,.a .,take could be tbe •• a1labllit, of
phoaphate. TO
41
arittoll Chuce9S baa used tbe .... t. of utlllaadOD of -ftea
b1 a •• pealoa of .itrochOadl'l .. ill coaJuction witll spectro
photometric .. aaU%eme.t of tbe oxidatioa-reductioD atates of Yal'-
10U8 c.llular co •• tit ... ts ia elucidatiD, tbe blochemical baa'. of
respiratloa &ad formatioD of bilk •• er., pboapbate. Tbrou,b t ..
use of a riftatla, _lid electrOde ia a apectl'opllotoMtl:ie eel1,
the ... peasl •• uel .. added. aubatrate cou14 be abe4 ad thea
all0we4 to ... ad. uadistulMd f .... tbe tille I'eqall'ed to reduce tbe
0XJ, •• la solution. '1_ vibratia, ele._ode in one "POB of tile
cuYette followed the rate 01 OXJ ••• l'ellOYal _i1e tbe other po .. -
tiOD waa uaed t.... epectropbotGaetrlc d.termiDatloa. Cbaace
fouad98 by ........ Ia. ' .. .rate pcoductlo. Ie a ... cieic oxida ••
a.,st ••• tbat luau .. te ,,04uctlo. be •• &ad eDcMd with -Jlen
availabilit.,. ChaaCe _d W1111 ... 99-101 studied the respiratory
cbaia in latact dt"houdria aad foulld that a.u1ahI1!ty of AUf "
I1ml teet the UIOUIIt of ox.,,.. uti lbeef. 'ftey -ueeS thl. Imowledp
to cletehiae othel' z:ate lWttll, futora &ad tbe "que •• of
enz,... ift the eeapil'atocy cbaia. Cbaace aad other.102 utili8ad
these flDdia,. for the ia ••• tigatlon of Ike .ite. ID901 •• d in
oJridative pboaphol'ylaUoa. The rapidl ty aDd ae.altinty of 'lb.
polta'~srapbic _ ...... at. eoatributed s'plfleytl,. to theM
atUdi •••
III .tudies Oil blOOd aa4 U .... t a.dd! tional probl._ .".e
eDc.unt_ed wtdch I ...... tbe c1iff:l.culty of obtaiail'll quantita.
tive ~ •• ult.. The det~.iD&tiOD of ox,. •• in blood baa been
· evaluated by 8er'll'ea. 103 8e c01lCludes t "--, til. alnl.aeo ..
preseDce of both chemically aad physiCall, COabiDed oxy,ell Jive.
rise to such complicated conditions ia polaro,raphic ..... r .... '.
that it seems impo.sible to arrive at a quantitatiYe deteraiDatioa
of either tbe oxygen coDteDt or t .. slon directly in bloOd." Slaee
the poluogr.phle a.thod .. uu.s te •• loa, la ••• ti,.tors haft
801ved this difficult, by r •• oving the !nflueDc. of ~~emi~&lly
combined oxy, •• _1' haft coa •• rted it to dl •• 01 •• d OXYIt. by ox!
diziD' hellO,lobi. to 801M fel'l'ic compo.... Aa ludleated in the
fol10wla, ,.ap'apu, aD ace.a., of withla two and fi ... percent
w.a obtd.eel witll the polaro_apble method ba.aed o. the ataa4aH
obtained by .... Qalyala of tile e .. l11H_tin, .... _ til. ,as ex-
tracted uDder YU1IUJIt. PUt of tbe •• 1att •• ia r ... lt ... , be
attributed to failure .f .... l .... ti,atora to correct for tbe
yuiati ... iD t •• ioa ... ociated wi ~ tbe co .... tio. of ox"ea "
by blood or pi ...... 1Id ... lllfl •• ce ,., oxyllello,loblD diaaoci ...
tlol1.104 ... well .. YU'!abillt, i8 tlte polU'o,raphlc ..... c ••• t.
It i. po.alble to aake the polarographic __ relleDt in whole
blood depeade.t _poa onl, the pIl.,alcally dl •• olvecl .,.pn in
seYel'al 1 •• taacea.1U .are.103 aad C1&l'k104A pol.. .p tile fact
that if tbe OX".. tenaiea reBin. abo... tile 1 • .,el of o.,lleao
,lobia di •• ociation. tbe.... 1. DO coatrlbatloa of cheal call, bOaRd
OX" •• to the iT- It is alec po .. ible to displac. th. che.ically
bound oxy, •• with carboa aoDOXi4e. 103 la ... ti,atora t10l ,10S,106
howevel'. haft Ibown that .h.,drop. peroxide dee_pod tioD ia
49
catalyzed by tbe ,,"esence of he.o,lobi., resul ti., ill iac&'eaaed
pelaro,rapbic current. This CaR be peeveated by the additioa of
cyanide ion101 01' by the use of ... applied po tea t1 a1 wbich .r:educea
ox,sen duectly to h,droxyl 10as. 103 "Vq thea, the .e ....... at
of oxygen in wbo1e bloOd la le •• than 10 p1aaaa equilibrated with
the sue gaa pbaM, a facto. atai_ted by JJer'gI".ul to the alowe ..
rate of diff.aloa of exnell ia reel blood «e11 .. 103
It ia also possible to aae a meua.re4 b1004 _,18 &ad to
.atl_te lta oxy,en coateat tllzougb the co.veraioa of the eIleld
cally cU.Mo1ved. ox"e. into pltyaleall}' dissolved _,pn la a
kDOWR volume of a red.cia, "1~tl0 •• 108.109 Bauaberaer used acid
to rel.ase the bo.ad .. ,,.. and cGB.ert the oxyh .... 1obi. to .aid
be ... tia. Tyauowaki110 cOllver ted the oxyheao.l.obia to cyaaohe.
slob1a, .. la, pot ... iua fe.r:.r:lc,aaide ia a aanae. reported earllel'
by wei .. l.,v.111 T,auowald. repo.c~ed DO valu.a for ,I'edai", ..
but BaUlBber,.1' c1aiaed .. accuuy slliil" to tbat obtaiaed with
van Slyte dete.rlldaat1ou, and W.l .. i.,el' ... ported aD &eo.", of
withia two pez ceBt.
A modificati •• of the polUoJl'.,ld.c PEIDc.i,1e _ac.a it
posaible to 46tel'm1 •• the oxy, •• In aolutioa b, poteati ••• t.-!c.
tech.lque. 112 Slace veE, little ox"ea ia used by the eleet ... 4e
dUiD, me ... ,.e_at t it 'a eq.ally effecti.e ill bl .. ct or 110lI0-
,.aeo.. .olatioa.
In other i .... tl,atio •• tbe .eaa._ ... t. of ."ea 'e .. loa
1. bloOd haye beea di .... ct.d towud _aa_ .... t la pi ....... ed
so
eithe¥ 00 physical .. paratlea o. upoa physical lactora .bien
.arkedl, redace the contribution of ehellica1l, bouDd oxy,ea to the
polUOp.pllic .. asu_at. leecher aDd Ilia colleape.lll were the
ftrat io ... cell-free bioloaical fluidS as a meaD. of eatimat! ••
.."ea cODte.t or ienaOA ill bl004 and otbal: body fluids. Ia
tbell' atudie. DO epeeial cue .aa takeD to ute the aeparatioD at
the tcape ... tIl •• of tile o.rlaiaal sample. In latel: atti4!Q6 by
1Mi •• ill ... 114 aad by Wilaoa _4 coworke .. s 113 apecial __ f_
Ulaeroblc aepaz •• loa auck .... aN by the pre",iou poup wa.s
aceo.pll.hed at bOdy t .. ,el'ature. Ia addltloa. Wi1aoa'. ~k
•• pllaslaecl the cO_lIPtioa .f ."e. by red bloOd cell. aad ,1 ...
daria, tile iate •• &1 betweea ... ,11a" aael •• pa.:ratiOll ud a.u •• -
aeai. witli adequate cue and col'J:ectioa fo. oxv.ea coaauapnoD,
thea. _thod. •• eault ia u .atPation of the -".a teasioll witb.
ia five to tell m11Ii_tel'a of "1:"1',.0£ the oxyg •• putial ,rea
.. re ill tbe .quilib.cati,.. ,.a pbaM.IJ6
Aa it ia iaGOAYetieat Mel diffinlt to Obtain ace.ate ...
ti.t.e of tbe blOOd oxypa teaaioa b, ... ueMata Md. on pl_
M,uated by oe • .-i_f._alioa, otbe~ _thode haft beea ",uabt which
do DOt •• quire .. pac.tiOll bat baD the advutap of be.taa lade
peD4eat of Chemically beaad OX,18" Dreackbara111 iDtcoduced the
ua. of a collodt_ stocki. ov .... the 80lid platilla electrode, aad
later iacladed ibe ... of 0180B'S clccuit. 42 'articularl, -'th
the ... ad lIOdifieatioa the _at ... iINtioa of Gbealcally bouDd oxy_
lea is aiaiaal. The accU'acy obt ... 4 is within e. per cat of
51
tbe .quilibcatiDI saa partial pr....... C1a~k118 aad hi. co-
1fOS'kera iDtl.'od1lced a eovel"ed e1ectl.'ode and later the i.alated
electrolysi8 cel1119 whick were discu.sed earlier. Theae elec
trodes lIake 1t po.aible to .... uretealioB, but unlea. 110_ form
of coaveetlon ia praseDtl20,121 the teaaioD i. le •• tbaa that of
the equilibrating gas, becau.e of the diffusioA .,..adiellt to the
suface of the eleetl.'ode lIe.brue. DepeDdeut upon coDtrol of
cODvective effect. and teaperatv .... the accuracy of th ... aodlfled
electrOdea ia withill two pel' e.eat of th. pa&'tial pre •• ueof ox.,
pD 11l aa .quilibcatia, ,aa uxturel22 or the pre •• ue •• tiMted
froa ... ext.actioD aoalyaia. 120 When a ~. platin .. el.ct~04e
ia &'ap1dl, rotated in blood tbe diffusion la,er &sQuD4the elec
trOde atabilla •• to ... ell free layer of pl...... "klan aDd
Nahaa123 baYe applied the rotated electrode to .. ole blOOd aad
obt .. i.e4 a lilM'" I'.spona. f •• «e.8£o .. of f ift, to two baadred "
flft, ail11_te ... Ilel'CUJ.'J .itl1 1e •• fbaa cae to lift pel" cea ..
deviation f.-OIl tile curve. Tile .. aoctU J.ed metboda fol' blood la
creue tile conyenienee 01 ... uria, tenaloa of .'Iea, bat tile"
are special problema with ea.ch method, as well u the contut
problem .f teape .... tur. aDd time laflueace. OD t.e .&aple Cb~
aeteriatic ••
Meas ___ t. of .,,.11 tendoD ia blOOd ladieate tlaat aiail&&'
factora .y be expected ia ti ••••• ".VesteD'S. The bu. wise
electrOde mu.t be uaed in ti.... meaauremeata and is subject to
the SUte contudutlollS found in bloOdstudies. 21 ,.S ODe findlDS
.5a
ia blood wIllcb iDdlcatea clec~e ... ed seaal tl.i tf 111 tl Mue ....... e
_Dta ia the 4e"e ... e ill IT PCOPQtiODU to tbe red blOOd cell
coacelluatloa.103,123 Tlda wou14 pve expel'iaeatal •• pport to
tile .. tbematl •• 11, derived. illf .... ce of slaw .... diff .. l .. i.
0811a124 aad would wrlf., l'.I'o_,.125 obae ... vatiou on ti .......
'helm.loglcal ad.aacea _4 Mttu .... l'.t .. d~ of pbyaio-
logic fu.ctiou as •• 11 .. ad ... uce. la polUOlrapll, bave re .. l ted
fro. iaft.ti,&tio.8 of bloo4 ox, ... t.ui... TIle iIIs-oYed polU'
opapblc electrGde42-44 baa pr.dd .. tile .u. fol' coati .....
.. ldtor!., of blOOd .. "eutioD ud eff •• ti ....... of corporal pu ..
fuaioll d_ia, tbol'ule ... ,..,.44.12 .. 27 Wbkh reqail' •• atilb ...
tloa 01 the "titleial be~t-lull' app ... taa.
Th. aimplified ........... t of .,.,e .... ceatratioa with tbe
,.luo_.pIlio aethod baa Md. addltlo.a1 tecUuli.... a ... llable f. ,
blood .. 1,..ia. III til. a'.'.4 1 ..... ato,.i •• it baa beea de .... ..
• 'rated that ox,beae,lobia 4i ... c1atlea • .rYealaS &ad tbe .. tur.
of bloOd. "'_l1ob1.129 ... be obt.hed ia a few lliaute. wi tato.t
I'ea .. tia, to tbe ... or 81wel: &ad Hre cube...... .. ........ ic
toCbal. .....
l.api4it, of tbe poluop.,pbic .eclud.qu .. ea It '.uibl. to
.tudy 4,....1.0 &apect. of _loed ox' .... tiOD. 1'e fifty to one
hund ... ed bloOd ... gal, ... l'.q.l~e4 III tile c ..... of • w,le
expe ... iment oould bazdly be bandied .tth ... aD Slyke or aicJ:o,&ao-
aetric .. tbOds .f OX" ..... ly.i.. Aa a re .. lt of multiple blood
oa, ...... 1, ... the abuatin, of blOOd fro. aonYeDtilated alYe.li
.53
in noraallOJ ,130 and ,atholo,i8 131 atate. of the re.plratory a
tem ba. become eYidelilt. Silliluly. Imowledp of ,.lao."y reM ..
Gapaclt, to .aintaiD oxy,en 8&t8l'atl08,132 tbe lucr ... ed OXYfeD
affinit, of foetal blood,ll3 and tbe role of co.nterf1ow betwee •
•• ternal aad foetal eircalatioBa in oxy,.. tran.fer to the foet ••
baa benefited froa tbe reduced time required for oxyp. analyds.
polaropapiliC "UIIl'UeDt of tl •• ue .,seD t ••• iOD lacks a
readil, aY&11al»le _tlto4 f_ •• tiaaiilll tile acc."., of the ......
... e ... ta wIlicb ue Md •• * la tlt.e pre..,I._ ,u&lI'apll.8 application
of polU'oll'apllic .. &8ur .. eata wu IUd. to .yate.. in whleh a ... t
probable val .. eould be •• tlu:ted or ....... 41 for actual _,po teasioa. la ti •••• the ,&E ... ter8 of ozr, •• te .. l0 •• di£f •• iyit,.
blo04 flow. blood oxy,.a capacity, OXY __ Ilobin diaaociatioa,
'li ...... tOI' ..... aetabolic rat •• teapel'atun and pre •• ure, ... d
ti ••• e di ... WOd have yaryi., <kll' __ ea of iaflu.llce in diff.re.t -.
atate. of the ti .... aDd uka it difficult t. predict a re .... ab1.
value. 'urtber eoaplicatioo ~i ... ia the bet.rolenelt, of tb.
ox.,,.1l teuloa patteI'll of tbe ti..... UDeleI' auc:b coaditlons the
tel'll "ox, .. o ayail.bill.,.. ia .st deHl:i,tl .. of the ••• of the
iaflaeaces &ad tbe ten ladleat .... be u ... rtalat, ... to wbat i.
bela, .. aaued. po1U'oll'apblca11y I.a tbe tl .....
la sPlte 01 tbe difficulti ..... tioaed abo ... quantitative
-*fbe metbod .f calcu1atioD 18. other available •• thoda of
meaaurin, tt ... e ox,,.a te •• loD .t11 be diacu.aed i. the Dext Met!o. of tile paper.
I I
tissue oxygen ten.lou have beea obtained 1. a few Instaac... la
v.sti,atioDs usia, the recessed el.ctrode134 report accurate me.s
ueaeats of tbe teaaloa at the tip of the electrOde, aad ,01&1'0-
graphiC Jleaauremeats can be .ad. OD fluidS which baft beea ia con
tact with tbe tl •• ue .. a ••• _ of calculatiD, the ttssue O%.,,.n
tensloD. 13,S,136
OX".8 utili •• tioa i •• ti.sue bas been qa.litat! •• l, .ati
aated polaro,raphica11y. ~rest of the circulatloe in the r.sion
of tbe electrode aakea it posalble f. lavesti,at.s to co.,..r.
tbe rate of fall of iT uDder different coeditl0.8. This teohaiqUt
w .. firat applied to the braiD cortex b., Broatll1 and later b,
Opitz aDd Kreuaer. 118 Urbach uaed a .iail~ proce.u.e in .... ur.·
.ent of l'elatiYe rates of OX" •• utilization 01 Doraal t inflaaaed.
irritated aad patholotical at.tea of the skia.139-14l Aa the
iacbemia .as obtained by co.pr ••• ioa or tensiOD of the ti ..... , "
the experlmeatal cODditioDS lack co.plete zoepcodacibillty. La ..
variatloa ia ••• oaat.red ie 18 •• atl,.tlou ia wbicb tbe circul ....
tiGD Ie drested by oCClUaiOD of the ..... la aupplyiq the repoa
under 1 ..... tiptiO'. 0'1' by c •••• t108 O'f perf •• IGIl tI.tll tile ."eD
teaslGo fall. to a.rG. This tec.tmlqae haa also been applied to'
the bcal. cortexl42-46 aD4 la a,-patbetic .an,lia. 81- 93 ,147-49
In a few ill .... atiptiou in whicb .. taboll_ waa made tbe irade-
pendeDt variable, tbe interrelationship of oxy,en availability ~
_tabolia. bas been a'u4i.<1. Davie.1SO observed tile rate of de
eliae In IT durin, mu.cle coatr~tioD, aDd .everal love.tisatora
II
I I: I
I!I I
:1' ,'I
I',
I,
i'l
$5
.. ave iadaced iaueased braiD .. t&boll_ while makin. polarop'aphic
.... .r.sent •• IS1-S3
ouria, the cour.e of inveati,atioaa, chaD, •• ia iT Ila.,.. been
obServed to be depeadent .poa Y&riatlon8 in blood flow, arterial
oSYIen •• tlll'atioa ad physical f act.rs. ft ••• ob •• Yationa la
ellcate tile direetloBal chaa,e to be expected la the poluosraphic
.asw:em •• t of oxygen tea8ioD, but offer Ilttle toward q.otit ....
tl Vit calibration. of the obeerYed diff.lon cureata. De"e ... _
ill the po1uopaphic e_ .. e.t &l'e ualfom1,. ob .... Yed dur:i., putial
IDd total obatructioD of retioD&1 blOOd flow154-59 and duria,
lower.cl uterial blood ,.. •••• e. 160-63 Wbea tbe circulation ...
altered by the UM of dru,. _lell affect Ilow,. pre •• urea, aad
"aaoaoto. to.16l-63 or tty ai.:.llar reflex cbUlg.s,163-... the
poluO ..... pbic oxygen teaaioa ill a few caMS did Dot change, but
•• uall., waa altered in tb.e predieted::d.irectlon. T1l1a Y_iability ..
la reapoas. may be due to interrelationship. between blood flow
tlu:oulba tisaue, which determifu~. the meaD cyil,larl oxygen
tension, aad the ratio of tbe capillary radius to the radiu of
the tis ... 8upplied by that capl11&., beia, modified and conae
".ntly chaasiD, tbe tiaaue oxY88n tenalon. l &5
Directlollal chaatp. in pola.r:08l'aphica11y m.aaured ox." ••
availability are obaeryed in atudies in wbleb arterial 0"1"'. aaturation 11 ... beea varied. In two stud! •• tbe arterial oxygen
a.turation va. cpaanti tated by oxiMter meuaemeDt, 166-61 &I1d i.
others the cbaD,. .as induced by alteratioD of the compositioD of
.5,
inspired ,as.s.134,138, 168.12 ID eitber cas. tbe ret.tio.ship ie
bOuDd to be DOIl-lia.u becaua. of the pre .. ace of be_,lollia.
Several other expel'iaental deaips haft iDtroduced factors
wbich would modify the ox,,.. aYailabillt,. The oll .... ed polaro
,raphic cur •• t ia tile .. experiaeata had tbe .... qualitative
characteristic. aa did tbo.. obSel'ved in otbe. tia.ue .... ur.
.e.t.. SO ••• "heia'a experJ.aeatl'l placed the aDiaa1 In aD eo.
virooe.t GbudMr aDd v_ied the _ ...... fto. 0 •• to aix atao ..
,here.. QUantltati .. I'e .. lte .ou14 be difficult to prediot fro.
tile pr ....... c1aaap. becau.e of aaaooiated alt ..... tl0. i. tte
ph,sicall, 41_1 v.'" .,.... &ad the pecoeat... of the .. tabollo
reqai.e .. nt. it .upp1i •• , aa. because of tbe Circulator, aDd
metabOlic ob ..... ill41aoed by the pre ...... ia additloD to the al
tel'atl08 of tbe othu , ... _t •• of ., .. aanilabill .,. Gl'avi.
tatioaal 10I'c •• ,113-74 beat. l ?' aad GOld1'6-?? ha .. be •• iade--.
peDde.t n.riabl.. ia in ••• ti,ati... iJl _1Gb. oxy,.. a.ailabiU t.,
was ...... d poluo ..... hically. A,aiD it would be difficult to
quatitat. tbe poluo,raph!c .a •• e.Dt. bec .... of alteratioD
of tbe par ... t.l'. of OXf,en availability aDd tbe iat.raetl0. of
tb ••• witb tbe phy.ically iapoae4 .tl .. 11.
BveD tho1tp polU'oJl'apbic ti ........... at. of OXJp.
a.a11abilit, bave be.n ... litat1 •• in aat ... tbe cODt~ibatlo .. of
thi ... tbod to ltD.wled, •• f •• taboli •• circulation aad. clinical
medici .. bave be ••• xteui'ft. SOIte.f th. GODtl'lbati ... ".Ye a
d.il'ect beuia •• R the ' ...... ti.-tio .. "'.,o.l't.4 lat." ia tbe the.ia.
I , !
These eontributloas ba." been __ arized below. Otber aspects CaD
be foulld In reriews by M08t,om_,118 _d Paaduzll79 aad a • .,..
posi_ oa OX,,.8 ten.toll ill tluue. 165
The fact baa beeD lulOWI'1 for Dum, ,eus tha.t Pl'ot080a &ad
ttuua in vi iro tespire at a rate iadepelKleat of ox.,,.. teasiol1 -abOve a 'ftt, low u_i.a1 '9'al.e.87,180-81 but this tact waa Bet
established for ti ••••• in riyO prior to the applicatioD of polar --op:aphle .... ~ement to the ,roblem. A. V. Hil1 t a124 _peculation
that there ... t be a muted diffel'eace between oxy,ea conauaptioD
of different relloa. of a cell 01' tlasue ha.e been aubataatiated
by polarop:aphic uYeatiaationa of l'espiratioD in various repolls
of the aerVOd .y.t ••.• 137 tl82-84
The fact that oXYHlIO,lobin dia.oeiation in ti.aues is the
aame .. diaaociation in vitro h .. beea eatabliabed16J.166.l36 to - .. SOil. de",ee. Otbu atadi •• of ox,seil & ... ail&bi1i i, in tiaaue
emphaa!ze the vuiabili ty of tile .iruetva! aDd functional char
acte~lsti •• of ci~culatioa 18 differeat ti.suea. Some ti •• u.s
such .. tbe brain152.182-83 and beartlS6-S9 have little col1atera
eirea1atioD, while other ilea •• s auch .. 1"B,l03,130 appear to
have exceptional eapacit, to abUllt bloOd. In atudi.a .ade of kid
ney eireulatloalaS it appears tbat the blood paa.ina throup the
capillari •• doe. DOt bay. the sue red blOOd cell composiiioD as
bloOd in tile veia. _d uterie ••
Two faceta of poluolI'apbic M.aurel1Kulta of oxy,ea U'e ~.
vealed ia as 0¥era11 perspeeti.e of the I1teratare on this
i
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r---------I ~
I
I
L
subject. The encouraging aspect of this new type of measurement
for oxygen is the veraatill ty and usefulness of application. The
challenging aspect of polaro~r&phic measurement of oxy,en is that
while the measurements have an accuracy of within two per cent
In vitro. it is difficult to obtai. measurements in tissue that _ J
remain witbin fifteen to twenty-five per cent of the averale
steady state value.
, I'
I,i
II "I "I
~ I , ~ !
CHAPTliR II
liXP.D.IM.J1NTAL PROC.lDlllBS
Tbe expe~i .. nta1 procedures uae4 in COBJUDctioD with the
doctorate prosea. iaeluded two experimental deallDs. Tbe firat
dealp conai.ted of a _tbod of ........ Dt of diffy.lvit, of
OXl,e. 18 s01"tlO8. Ttlls desi,. is d.acribed (a) as a aetllod of
.......... t aDd calc.latio. of tb. coefflcieDt of diff.aioD of
ox, ... i8 •• li •• , (b) .. the val .. obtai.ed for tbe coeffie!eDt,
aa4 (0) aa a 41 ..... 10. of the reliability of tbe co.fflel.at aDd
tb. illplicailoa of tile rel1abilit, for the ace_ac,. of the polu
oll'apbic appuatua aDd techlliq.. beia, ... 4 iath ... ex,.&,l_ata.
The MCOad expert ... atal d •• l,. waa .. _tlaN of t.s'iol tile ueu
ac, of polU'op'aphic ••• a •••• nts of oXYle. t ... ion i. vivo. Ia -till. cllapt.&, tbe hypoth •• ls aad tbe up.rl..-.t.l aapecta of tbe
Meoad "aip .. e pr.M.ted.
"
l'OLAROCIlAPHIC INSTlUMJUft'ATION
'8 tile .. exp.~be.t. tile pelar:opqb!c cll'ctalt. o~i,ioa11,
d •• l_4 ." 01 ... ,16 ... aodlfied to 611_ tbe use of .. ltlp18
electrodes. Tbe modified eireait, In add1tl08to havi., the
f .atar.s of tbe aillp1e polU'op'apbic elrcai ta 4 •• cri bed !a tbe
pr.do .. chapt •• , proyid •• for.
1. Application of alttrnatel, poslt!T. aDd Dlfatiye pot.atial. to the analySis electrode .tth atersparsed periods of abortia, to tbe refereaM electrode.
-59-
, I i ! I
rr r
2.
3.
4.
aececdin, the current flow of a ain,le analysla electrode oaly dari., the teralaa! portion of the period of .. ,atiYe applied poteatial.
Stabili&IB, all analyais electrOdes by cOBtlauous cyclin, eftn when tbe electrode current is 80t beiD, recorded.
Autoattic recocdla, of c-v cur ....
60
This circuit differs froa .q.~e wave aDd alter.atl., curr •• t
polaro",a,ll1c circuit a in that the referace electrode potential
i. Dot a04ulated dari .. poiuizatiOll. wbile square wave ud al
ternatiD, current tecbDique • .adulat. tbe polarized electrode and
..... 1'. tbe IIOd1alated _I'r •• t flow •
• i ... e 11 111.atratea tbe .ajor feat ••• of tbe cueult. A
block 41a",_ at tbe top of tbe 111 •• tl'at10B ladleatea tJut eireui
fucile.a. Tile circuit ,. the ce.tel' of the fipre ahow. ~e
location of yar1... electrical COat0D.ata aDd the .lcre .. ltebe ••
Proyisions OM tUGU,b foul' ... tioae.;a abOve .... obtailled by a aet "
of prop'aain, Stdtclae. wld.ch complete "one cyCle pHalDute aad
iaclflde .ix tea-Meoad aub-cycles. Tbe atd tchia, prop_ i. COD
trolled by ucroawitcb ••• ctiYated il'oll a fou-left1 a .. driveD
a .yacMoDOIl. olle-rpli _tor. * AD illuttl\t!oll of pro_wD,
sequence aay be fouDd i. the lowea: left COI'D.r of tbe fi,... ••
The oae .. "",t. cycle conal at. of aix tea-aecoD4 periOd. ill
wbich each electrOde, iaplaced in the .... co ... dio' cia:cuit by ita
*80diDe a. c. .yn~ODOtl. capaci tor motor, lYC-22RC. Bodine Electric CO.PaDY, Chicaso 12, 111iGoia.
POWER a
VOLTAGE REGULATOR
RI
POLAROGRAPH SCHEMATIC
IPOLAROGRAPHIC CELL I IIIIII
~MMETER ELECTRODE a SHUNT SELECTOR 1
INSTRUMENT
°2E
TIMING CYCLE I MINUTE 8=MERCURY DRY CELL
MS- MICROSWITCH V-VACUUM TUBE
M S I T06 I I I 2 I 3 I 4 I 5 I • I MS 7 (+) MS 8 (-) MS 9 MS':':IO-
------- - - ------_. ~~~~ -.- -..-" ----. .-~- -.
E - ELECT"ODE R- REFERENCE ot-4NALYIIS
SPST- SWITCH R.- 35 OHM
VOLTMETER A-NANOAMMETER
Rf 1000 OHM
R2- 100 OHM
RJ-R 201 ooot4M
.-~.~-----.- -- -- --------
PIGURB 11. • ...
62
tespective microa.itch (1-6). All t ..... coRd cycl •• are identi
cal. In the l.itia1 third of a t.u-second cycle, Rlcro.-!tcb 1 1.
activated. Darin, the following oae sixth 01 a cycle no voltage
is applied., ad the aaal.,sis electrodes are aborted to tbe refer
ence electrode. MicroswitCb 8 is activated daria, tbe aext ODe
third of a cycle, ~icb is a,ain followed by a oae-sixtb cycle
period in wld.eb the &IIalysia electrodes are ataorted to the refer
ence el.ctrode. WIlli •• iero_itches 7 <d 8 ue operatiul, the
potential from the batteri ••• placed ia the circuit with oppoaing
pol ..... l t., is applied to the poluopaph!c cell. The recordla.
circuit Is activat.d for ODe .1ectrOd. " .lcroawitch 9 d~ln,
the t ••• iDal partioa of *be applied volt.,e at the analya'. elec
trode.. The electrOde. ue the. aesaU •• with re.pect to tbe
refereaee el.ctrode potential. Two variable potentiometers t wired
in parallel witll tbe pot .. tial aouc •• , replate the applied YOlt-"
..... A lI&Dually .et I'Uae potellti_t.r" replatea the applied
volta,. .pan. A .aDaal1y or 801enoid operated ateppin, potentio.
meter." allOWS tke cboice of & pr.-set and CO •• tlDt applied .olt
ap or of aD •• tOllatic at.pwl .. i.cr .... fro. _.1"0 to full .paa
volta,e in five per cent increments. Microawitch 9 actuatea tbe
·Mercury batteries, Mallory RM1R: 2 in aeries, 2.7 volta.
**2000 ohm radio potentiometer.
***l~ precision 10 obm realstors wired 1n aeriea to the CODtacta of a 881tiawitch to form a 2000 o~. pote.tloReter.
63
solenoid* durin, the last five <i_sree. of each one-aiaute
36o-cse ...... ,&le. A vacua ... tube 'f'Oltaetel\** i& parallel with tile
polaroaraphlc cell Ilonltora the applied poteatial ••
The polaroBraphic cell is Composed of a aaturated calomel
relerence el,aetl'ode connected to the aaalyala portion of the cell
ria & aatarated potaasium chloride ..,ar ... .,u Wid,.. Mercury and
mercuric chloride purified for use 10 electrolytic cells ~e COD
taiDed in the calomel half cell. The croas.a.ctioaal area ia
app~oxl.ate1y tweaiy square eeati .. t.r. for the calomel cell and
approximately ODe square centimeter for the bridle. The brid,..
are fou to aix iocua 18 1.8,tb. "ftle .. al,_i. electrode.
(Pisure 12) are coaatracted from platinum-iridium wire which . ,
vuie. from 10 to 65 .iol'oH ie diameter ud ia approxillatel,
four mil1iaetera 10ft,.
Electrode cu •• at .... e 1'8.01'4.4: by ___ of • pbotokymO.
grapb*.* from a reflect!D8 ,alvanomet.i**** which ia introduced
*24 volt power supply to 801eRoid ia regulated by aD off-en .. itcll. SPOT. I. off ,.litl08 of the SPur .. ito tile manually set posi tioll of the steppillg potentiometer is DOt I'". '
cbaRB.d.' I
I
input l=:c~~,32v!~ r=ti-::-I.':!eO::;~·~~:::-:o:·c. 11
ceDtel'i., polDtel' to aU-.cal.. Ii'
·* ... tel'a-coale, IS-laGb pbotok,.oaraph •
• *~ipp _d ZoneR. 4-23, Micro, Noll.type .alvaaoute%, 0.25 sec. undamped period, 2.2S to 3.8 x 1O-S-amp/ __ deflectlo. at 1 l1etel'.
I - INSULATION
W - ELECTRODE WIRE
J - HARD SOLDER JOINT
P
C - WATERPROOF CEMENT
G - SOFT GLASS BEAD
P - PLATINUM E"LECTRODE
fIGURE 12.
POLAROGRAPHIC ANALYSIS BLBCTRODIS
into tbe ci~c.it witb Ita critical dampia, reslataDe •• by micro
..,1 tell 9. The crt tical dupin, I'.alatane. also MIrYeS as a YUl
able "Daitlvit, abuDt.
DIPPUSlON
MBTHOD OP MEASlI'UUlWfl:. OP DX PPUSI YlTI
Both tbe laPPuat.a ad the _thad u.sed to obtaift UI iate .... &1
coeffleieDt fo~ dill .. l08 of oxysen in saline depended u.POD the
.... ure •• t of ox, ... ia a dlffuaion chamber with tbepoluo
_&phi. apparatu. which ba. beea <i.scribed. The diffu.eioD eIlaber
w .. composed of two cel18 separated by a cellophane me.braae. Oae
cell coat.bed two polU'op'apbic _a1ysis electrodes _lell were
used to .as ... e tbe coaeeatratlon chua •• la ox,.e. as difl ..... o.
.a. W. Bor,. Mod 205 .icropot, a teL~tUl'. precialoa poteatiometer with 0.1$ linearity.
II I
II I
'Ii I',
I' "
65
occurred.
Pipre 13 ia a diagr&lll of the diffusion eh8llber 1 which til" constructed. The diffusion path, apprOXimately ODe centimeter in
cro •• sectioDal area, tIIas drilled on the uia of the ch.Ulber after
the two cell alta bad been fUhioned and fastened together. A
new 1/16-1ncll drill t _Jcb tie uured 0.43' inches in diameter, was
used to drill the bore of the chamber. The diffusion cell was
cut to .lAure two ceatu.ters (0.188 inches by mieroJDeter meas
uremeDt). After the chamber had been drilled, the cells were
sealed at tbe outer eDds with plastic discs.
Dotb cells had opeainl'8 tor flushing and filling, tbe refer
ence cell had an acceaa opening to reee! ve the agu-asar: bridle
froa the caloMl referaees electl'ode. An .stiaated cross
S.ctioDal area of oae per cent of the cros. aeetioDal area of the
diffusion path WiI.' op.a to the &tlllO.~bere. PlaciD, theM opu
in,. at a aaxla_ diataace fl"O. tile c:I.lff.sioll illtutaee .iniaiaed
their influence.
Blectrodes in the diffusion cell were positioned accurately.
The elect .. ode neareat tile ._.ane. El t was f.atened in pla.ce witb
a set-screw while it was beld firal" againa. a teaplate tbat ex
tended foar milliaete .. s (0.1',0 iaches by micrometer measuremeat)
into tbe dil'.aioA cbamber. Sleet.Ode.2 waa flusb with the end
of the dll1.aloa •• 11. Alter the electrodes had been positioned,
they were cemented into place. Duriog tbe experiments the cement
a.al was checked to in8u. that the 81eClOro<1.. bad aot shifted
I
II'
F
DIFFUSION CHAMBER
c
P ~I , , E'; J"E(I,
h\\\J G ·\\\\1 t , , ",}" ) v-
S PT
E
P
v s
"160 POLYETHLENE TU BI N G, 0 062"0. D.
VARNISH :EMENT HARr SOL=EFI
~ Lr'<iINUM WIRE
A - REFERENCE CE LL (I CM2DIAME TER, 2 crv ~ IF-L U:.)I ',--,N DI STAN C [ )
B - DIFFUSION CELL
E' a E - A N A L Y SIS E L E C T ROD E S (20 G AUG E P L A TIN U M WI R E)
c- CALOMEL CONNECTION M - MEM!::H-<ANE
D-OVERFLOW TUBE FaG-FILLING INLET
PIOUiUi 13. :
61
pOsitiou.
OUla, calibratioD aad ..... eMat of dltt.aioa the eIl_ber
was .uppo~ted ia aD 011 bath froa a pl .. tic float ~cb alao sup
porte4 the cal ... l ~efe ... ace electrode. The float was centered 1m
th4 oil batb by .eale helical _pd...,a to .... 1a1a. t ..... i .. lo. of
~bratloaa to tbe a,ate ••
A dial,.,. _b ......... t.eulled &Cn._ tbe f.oat of tile
dilf .. loD •• 11 t. pz'e,are tbe ell_ber for 41ff ....... _aauneat.
The two .el18 were thea ..... led aDd placed la tbe 011 bath
a •• pe""8d 'rOIl the pl ... tlc fl •• t.
~ el •• t.od •• we.e calibrated prior to dilf.aio ...... re
aent ..,aia_t a_qal1A_at.. .aliae. The.... _eel iaeluded
tuk Illtrop •• ro_ ail', aad 'uk ox".a. A.aIl Sl,_ appuat1l8
was e.,10ye4 to .... ~r. the ox,,.n coat.at of the aaliDe of fi ••
ailll11te ... ..,le.. The callbratl~ ,coce4ue ia.lude4& (1)
fluni .. til. cellI' with the , .. colltu.ed I. tbe sallae Pd..~ to
filii.,. tile ell ..... (2) fl111., tile GhaabeJ' with 84alll_ate"
salla. aad ... 8 .. 41 ... tile poluopapblc cu" ... at f.1: at le .. t one
ai •• te, (3) •• 001'41. ...... c •• C.ft clvia, the ca1ibl'atloD wi til
aali ••• ,.i11b1'&t.4 &lda.t 1:0" d_. All poluopaplllo •••• e
meata, ex.ludl., the C-Y CUI'", .... e cOIapuative tteUUeMDta _ctl
at aD applied .olt&,_ of .. 0.59 YOlta l:'e1ati .. to tbe ealael
refer.ace e1ectl'ode.
To ....... dlffaalo., a ahup cOBe.ataatioD diffe •• ac. at tbe
cllalyala .................. , .. &t118bed at a Iea.a tlae ... 4 tb. cll_ ••
ia ox,.,e8 CODGe.trAtio. at tile electrode. la tile diffuioa cell
were observed for the aest bou.c. _til cella .ere drd.aecl aad
flushed with nitroll..... The ... fluabiaa 10 tile .cefe ••• ce
t:uJlbeI' wu coaUned while the dilfuaJ.OllC.U was filled wi til
aitl'opa-qullibl'ated. 011.e, ... all '" _bbl •• wen c •• ful1y
.... h.d out of tIlat alde ., tile cell. Tile ai'"." .... "as dis
t:oatl ... d. aa4 •• .c.fe.e.c. cell .... cllately filled with aali_
equillNated witll. t&llk ex"e.. Witld •• aeccnd afte.c the cell
... filled. the pola&'opaphio eircait va. tuned oa to ladieate
the atut of the e .. riMa. t ad tJae firat pip 08 ille "ace
l'epate"e4 the .ad of tile f1l'a' tea .. 00114.. After the .cefe .. eace
el.ctrode had beea i ... rt.d la •• cloreuit by ...... of tile a.u
a," bl'idP. the .,.t.s was ull41atwbed for tbe l'ellaill4er of tbe
boul'.
T • .,.ratu.ce " .. nCOI:dad with , peedaloa .. .ceal'J Dec __
.. t... ,.caduated ia t •• tlla of a de,.. •• 'c •• tipad.. Gas .aal.,.l •
... lIade GIl flft aillilite.c • ..,1 •• try the "u 81yo ,&8 ui.cac.
tioa ••• 04 deSCl'l1M4 ia a fol1 .. t., ... tloa. The toe .aa
M .. ued. 00 the basl. of the tea-aecoact cycle cOBtrolled by a
• .,BCU .... _tOI.". with the iait1&1 ti ... deteraiaecl as deaaibe4
aboft.
MBASUIlBMlNT OP 1'HB DI'FUSION COEP'lCIBNT
I. thl. esperi ... t the critlcal dlae .. io .. aad parameter. of
the .. aau.ceme.t of the latell'&1 diffusioa coefficient for oxygen
Iii ,! I
Ii
are time, distanc. aDd cODeeatratioa diaenai •• s and temperature
arad CODvect! Ye infl.aces. DietaDc8 was ooau011e4 .. "eported ill
the pl'eCecU.D, .. otiol1, bowever. tbere were DO available checks f.
accuracy 01 til. diataace between the two elec1:xod •• Bl _d l!2-
Tbe coacentratioa me .. ~e ... t. by ,asometric aDa1yaLa have aD
&ccilracy of witbia two per c •• t. Althoup the tirae was aot
checked, the ayacUODOU _tor atopa if it ia thrown out of ph ....
by a realstias Iftce, beace it Roald ltaft aa accuacf of bette ...
thaD O.l~. The .o1.tl0 •• aDd bath were at 1'008 temperature,
which did Dot yuy <I .... ia' the two-hour period of obHrvatioD. No
eyideace .f coD'ftcti ... e .ffects ... ONez .. 4 in tile ,.l.o"aphic
record, a .,s'" wblela la ext ... a.l.,. .... iti .... to COllyecUn
eff.cta.
!be ..abraD. ie the .yste. off.ra aD uakaowa •• aiat_c. to
dilt •• ioa. It CaD be r.pr .... ted 1t'Y .. equiva1 •• t DOHt of I'e-"
aiat ... ill tbe aallae .,lutioa. III order to .01"'41 tile dlff .. loa
equation .. laltiU loeatioa of tile beaduy ... t be •• tablished.
Jiace tbe tiae ~equixed for dltta.loD of a alYea eoaceatratioa in
a ,iftD • .,.te. to reach polata withi. that .yste. is a lia.u
lUIlCtloD of tbft a"uue of tbe diatuee fn. tbe illit1al boUJlduy,
the ~elatlo"blp betweeA tl.. aDd diataace ma, bt U&~d tu eatab
lish the laitia1 boaDdary 1. tee .. of eq.1Yalent distance in a
colwan of s&11... Tn .. 1 ... IOll for the equlyal •• t d:l.taD~ 111
this puticulac experiMat ia ,iven in Table 11.
'0 TAa..B II
ESTIMATION 0. Xo PItOM DIPPUSION 'nMB
11 .12
X 0.4 2.0 '* fro .... brane
t a8* 4.s0 Seeoada to appearaace of o.os Co at Ji
*Batlaated from o~aer .. tloD at 30 seeoads.
XO • .0.15 eM fro. Xo
Tabl. III coat.la. the tabu1atioR of tbe unlt8 of tJae
diffualoD OO8lll01 •• t .... ere ... t. Cal •• latioas u. baaed OD
_tbeds dl8CdaH 18 JOat181 &ad BIber .188 TIle eq •• tioa
¥o = [I -.fir ~q--ql dca] i (1">
"
i. applicable to ay ...... hleb have", laltlal .up boundary,
and 4iff.alOft doe. aot ,reacb a boUJJ4uy dviD, the _HUl'eMnta.
The ....... t lo~ tbe errol' fUDetiou. q, ia related to difl.aioB
coefficient, t1me &ad distance by the equ&tiOD
Co - cODeeatretiol1 differeace at ti .... ro C - CODCel1tratioo difference at tilte 'B' q - tb. U' ..... t for the .""01' IneUo"
- the error tUDction x - eli.toee fr01l tbe iai tial boulldu, t • ti_ D - integl'a1 dilt .. !o. coetflcieat
I
: (15)
-71
sinee the m.abrane provides a Be .. a for eatabll.hiOS .. initiall,
sbarp beuadary and the _uuelleata were concluded wben tbe
initial cban,e .in oxy,e. concentration oecarl'ed at the .econd
electrode Ba • .8quatioaa 13 and 14 CaD be .Md to obtai. tile yalae
of D ..
Tables of the tkror hactloD, AMS 41,189 were aMd to obtain
the aolatlo. to equation. a aDd b. Tbe int~gra1 ia tabalated fo«
upHata of q, and tlle tabulated values correapolld to the value
e 1 -2C/Co).. Tbe yalta. of q thus obtained waa llNd to acl"
equatloD b for tbe value of , for ox"en in aaline.
1ft Table III the calculatlona of D baaed on the aboy. aola
tion are tabalated. Tbe y.lU.8 reported for cOReentrat!o. are for
an initial ,alYaftometer deflection of 56 aillia.ters, whlcb
eorl'e.poD4a to the _ .. ured ox"e. coaceatration of 0.029 co of
oxypn per co of aali.e. M ... ure.uta were Mde to tile Mueat .'
0.25 .illi •• tera, and the ratio or .ach .. aaoreaent to tbe .alvaD-
o .. t.r deflection for initial oxysen concentratioa ia recorded.
The .... yarl.tioD ill lJ ia calcula.ted fro .. the SWi of aquue. of
variation. The fiadie,. were !k • 7.70 x 10-4 cm2.ec-l for a
putial pre .sure ,r&di •• t of 0.95 ata.o.phet:e. per cea tiaaeter at
230 C in 0.9, aaline.
DISCUSSION OP OIPPUSlON OOBPPICIBNT
The diffusion coefficient, ~ • 7.70 x 10-4 cm2aec-l ,
corrected to standard coaditioaa baa a value of 1 • 2.36 x 10-'
72
TA&a III
CALCULATIONS POR DIFFUSION COBPFICIENT OF OXYGEN IN SALINB
DBTBRMINATIOH PROM INITIAL JlETBCTIOH AT BlAND 82
t cleo 1-2C/co q ca2 .ec~ x 10-4 sec:
28 o.os 0.9' 1.822 6.82 450 0.05 0.99 1.822 8.40
O.B"lBRMIHATION PIlOM 02 CHANGES AT li1 AT TIME '.'
t cleo 1-2C/C. q 15K dDx: (~)2 Me
I ca2 sec·1 lit 10"
30 0.01 0.99 1.645 7.41 29 841 40 0.015 0.91 1.S1S 8.00 ;)0 900 .50 0.023 0.95 1.386 7.87 11 289 60 0.035 0.93 1.282 1.61 3 9 10 0.045 0.91 1.199 7",53 17 289 80 0.06 0.88 1.100 7.82 12 144 90 0.065 0.81 .9.4 7.35 35 12a5
"
100 0.08 0.84 .921 ,.
1.66 4 16 110 0.095 0.81 .906 8.00 30 900 120 0.10 0.80 .595 1.68 2 4 280 0.20 0.60 .582 7.63 7 49 290 0.20' 0.59 • .510 7.10 0 0 300 0.210 0.58 .559 1.76 6 36
Dr.. •• DID • 100.°1/13 • 7.10 %*10-4 cm2 .. c·1
(dD)/(n-l) • (4102/12) • 0.20
Measured ,aa cODceDtration., 1'42 pb... • 0 ox, ... 02 ph.a •• 0.02' 81/a1 oxygen
n • 0.029 lit 1.7Q x 10-4 • 2.23 x 10.5 c.aeec-1(2JOc, 0.91 Hac1)
13
cm2aec-1 for a cradient of ODe milliliter of oxy,en per canti.eter
distance at 250 C. The corrected value compar.a fa .. rably with
the val ... reported in the literature, aummarized in Table IV.
2.38
2.03
2.42
2.38
1.84-1.98
1.1'-1.94
2.36
TA ... .u IV
VALUES POI tHE DIffUSION COBf'ICIENT
0' OXYC'JEH IN WADa
Ori,iaal .ea ••• e Author
1.62 cm2/da,-16OC HufDer
1.72 ea2/day-18OC Carlaon
Il'idta and Wi.a.ee
Experiaea ta aad Method
(1) - 1J Stephan's Table.
(3) - 'D Sua of triJOaometric seriea
(1) - Id (1) 'ol arosrapbic equation ..... to1thoff (4) - Id (D)
and Lin,.. Po1arosraphic equation
a ...
a ..
3.23 x 10-.5 ca2/aec-2JOc
.Muller and 'ircher
b"euer
Kelao 8Ild Peiaa
(4) - (15) LaPlace Tranaform
(9) - (11) Lallace Tranafora
(1) - (11) SubstitutloD in i8tegral
aufuer l90 deterained the value by the classical .ethOd of
layer asal,sis USiDI Stephan's tabl •• to interpret the data ob
tained. Carlacol 'l .11181',192 Pucher193 aDd '1.'8"'1'194 used a
I'
'I': ',I !I 11'. i
1
'1',
:11.1
.',
I .
74
,ourier-type aDalysia to obtain a conat_t for their .rst.... If
tbe total oxyteD diffasin, i. diYided by the constant. the data
can be interpreted ia tena of diffuairity. ID Culsoa t a experi
_nta tbe a,st •• did not naeh eqllilibrl_ aDd tile ayate •• u
,,"I'd da,a in rescllia, tile atate upo. whicb the aDal, •• was
pu:foraed. I. tile 'ribou ... , X.atitute uperi_ats (Muller, Pirche ....
,reuael') tbiD lay ••• of beJlO,l.c,lJia-cOIltaiain, aolutioll, .50 to sao .lcrona thick. were placed over a ,botocoloriaetric detector.
whicb followed tbe aaturatioa of the bellO,lobiD. ....id1l:a aDd
.eisner19S aDd Colt.off aD4 Lia,..e 196 baye ... aured id and
applied tbe formula
i4 - diffualoD carreat
• - faraday./aole
f - coulombs/Paraday
(16)
A - .ve.
-,
C ·'-molu concentration
It ia obvious that tbe deflaitiy. yala. for diffuaiylt, of ox, ..
remains to be deteraiDed. Altboulb the re.ulta of the Pribour,
group of workers ia indicative of 11.ear .ature of 0, the iadica
tiOD was Dot a critiCal teat of tbe ... umptio.. Of all the .. tb
Ods applied to tbe proble. up to the preseat time, the Metbod '1'41-
aented in tbis diaaertatioD offera tbe moat promia. aa a diverd
fied meane of atad.,i •• diff.alv!t, of oxygen. Modification of the
Chamber not on 1, will provide a method for a critical te.t of the
linear nature of oxYaell dlff •• iritv. but will Derait the Iii Ii'
75
evaluation of the influence of Yiaeosity, temperature, and salin
i t1 on diffusion of oxygen. Incorporation of a lIlembrane into the
system will allow other aspects of tissue permeability to be
studied aDd permit the verilicatioa of Krogh'. observationa12S on
oxygen diflualon in ti ••••••
The standard deviation of ~ observed in these experiments
indicates a precision of about fift per cent for the measurement.
of oxygen concentration. In all probability, the asreement of tbe
y.lae of n with otber values reported in the literature ia a
better indication of the precisioD than is the variability iDdi
cated by the stllldard dtrriation. In either case, the use of the
standard denation or tbe use of values from the literature, the
results indicate that reliable measurement of oxygea coneentratioa
is obtained through the us. of tbe _:tiled. deacri bed in this
chapter.
The reliability whiGh W&I observed when a diffusion coeffi
cient was obtained for oxygen is similar to that obaerved by
others for measurementa made in aolution. In vitro polarographic --...... -meaaurements u8ually bave an acctll'&Cy of wi tbin OBe per cent;
rarely are tbey more than five per ceDt from tbe predicted CODcen
tration in solution. On the other hand, biologist., of necesaity
have accepted qualitative reault., since it is difficult to obtai.
reproducible results with solid electrodes in biological tiaa .. s.
76
In human tiasues it baa Dot been uDuaual to obaerve fluctuationa
of fifteen to twenty flYe per cent from tbe avera,. ateady state
.&1ue.21 ,156-59,1'8
It la ,I"opoeed that R!laro.r!phic _ .. ure_ats 01 tias ..
,2?'11!! t.l1alon have tbe sue inherent "cur &eZ ... tbe accul"&CI
obtained in Don-tiNue .uurea.ata. If the variability of oxy,. _ I r
CODceatratloD in tia.ue were oootl"011e4, the tisaue then would
cOl"reapoD<: to .. !! .... ''' .... It ... r .... o .,.ate. in _leb the .sol"te compoaitioll
varied aDd cODc •• tratioD aDd coDvectloa •• r. not u.iform duriag a
period of _ ..... aent. VadeI' the coadl tioaa Jut deacribed !! .ltl'o, aD accuracy of wltbin live to ten per ceat would be u
pected. If aa acoUl'ao, of withla five to tea per ceat is obaern
vtlder coatnl1ed coKitio .. ill tl ... , such aD accuracy would
pronde .videa.. to .upport tbe Yalid! t, of the abo.. hypotheais.
ODe difficult, in t •• tla. _ell ~ahJ'POtbe.ia la that ... aatl • . '
factory metbOd i. aYd.labl. for .... ula. oxn •• t ... loa ia the
tia.... Ther.lon it ia Dec.s.uy to •• tl •• te tb • .oat probable
valu. of tbe ox,.e. t._l0D i8 the cepoD of the electrode ill
order to teat the bypotb.els. The available _tbOds for obtaialll
the eati.te include _pectJ:Opbotoaetclc .as .. eMat of tile conceD
tratio. of oxypn compounda of ti •• ue pi,8.ta, pa asalysis 01
the ,U ph .... 1. equilibrium with tbe U ••• , ad calCUlatiOD of
the ox, .... te._loa froll 8e .. ".:.4 ,&I: ... t._ of tissue OXf,e.
17
tea8ioD.* The aetbod of caleal&ti08 baa been ChoaeD froa tbe
tIlra. lIethods .. tbe .oat a"itable IN prov.ldia, aD approxhlatloll
of tile .'180 teasiOD ill tbe repoa of tile electrOd ••
Oal, _yoalobi. &pIMa. to haft the propel' chuaotuiatlca
for estlaatlD, local ti .... ox, ... t .... ou la tile 1' .... of zero
to oae buur.d ailll_tera of , •• alOft. .pectrapbotoHuie .....
ueua t. of tbe ,. •• e.t ... of OX,,,1l a.tuatioll of 1M_l1ob!. do
80t 4iff •••• tiate betweea tile bellO,lobl. of tit. 1al',. blOOd
.. s"la &Dd tbe ha_.1obi. i. blOOd ia the U .... oaplliari.s,
•• kia, it '-Po .. ible to determine th. il .... oxy,.. ooD.88tl'atl08.
Spectl'opbotoaetrlc .... ar ... at of 1' • .,ll'atory pi .. e.t oxidatioa199
ladle.... tbat ..,..0 teadOD abo.... a tent alll1Mt.l'a of MI'01ll')'
does_sot affect the de,xe. of oxJdatioD of tae ,l,.eata.
la pr.Yioua 1 .... tl •• tloa199 the .... or .... t. of .,0,10bi.
a.turatloa h ... ,i.lde4 oal, quaIit_lye I'e •• lt.. It would be
aeceaaar, to d ... lop teeba!qu.. for the ...... e .. at of tia ...
• yollobia coaMDtr.t!oD and oxyaJO,lobia di •• oc! atioll cu.z"Y.. iD
order to ... tbi ... tIlod quaatltatlye1y.
Qu anal,sl. of ox, ... iD equ11i1triull with the ,68 phas. of
tl ••• e eo1utl0 •• or ieJected air bubble. i. limited b, the
-'*Two technique. that bay. beea used to _u..... oxypn
coneeatration 10 IOlutiOO. bio ... a, aDd oxidatioD of dye., have DOt been co •• id •• ed. Bio ... a, teehaique_197 do Dot ,ield quanti tatift 1' •• u1t.. Tile ox.i.datioD 01 • .,.. h .. bee. ill 4is .. epute f~ .......... ot of oxr,.. la bioloaical _,.t... for the pa.t tbree d.Cad ••• 198
78
quantitlea of eolatloD obtalAable and by the time required for ...
bObbles to eq.i1i~&t. wltb the tl .... a. 198 18 the pcoposed teat
of the hypotbeaia it ia t.pGrtaat tbat the ti •• ue be relatively
undisturbed aDd that determiaatloaa be completed witbi. a miDute.
The aethOd 01 calc.1aU .. 01 tis.ue ..,,.0 '.uioa fro. the
_dua'e_at of blOOd flOW. oxyeea extractioD dul'ln, tl8aue per
fusion aDd ti .... dilleD .... wa. de_loped by the Daalah .athe
aatlciao BriaR, fol' AUcuatl%ogb.8 This equation baa beea ex
tended by Kety16.5 for the calculation of the aYeI'.,. tl.sue oxypa
tensioa. fte , ... to .. a ulled ,. Bl'lu,'. equatlo. ue 111 •• tl'&ted
in Pi,.. 14.
are:
" " - .,t-. - ,..--...... I " ...... / ,
. I I \ - - - - - - - - - - T - "1 - -,,",, \ _ _ _ _ _ _ ~_ _ _ ~ __ \ __ U .)
\ ' / .... ./ -..-,--~ ... ,
.....
• IOUD 14.
DIM.l!HSIOHS POI. CALCtlLATION 0. DI PPUSION IN TISSUE
1. Thl'.. co.centric cylladera are aasu.e4.
I.. TIle flrat w:t.tll radius I' whoa. s.face coincid.s with the capillary wall.
b. Tile HcoAd with radiu x. c. TIle third wi til radiua R haa a sufaee whieh
~.Pte"At. tbe 1iaita of the ti •• ue supplied by tbe capilluy r.
2. Darla, a steady atate) the ox1,e. clUfula, ac.ro •• the .... face of cy11Ddel' X i. jut equal to the OXYP. coaswaeti by the U •••• betweea the sUJ:fac •• of cylinders x aDd R.
79
3. The qUaDt!ty of ox,. •• Q at a siyen respiratory rate (ceOa Icc tl •••• /.ia) fo. this YOl ... of tl.... ias
,
4. POI' a diff .. loa .ratea defiDed by D the rate of chaD,. ia tbe By ••• t.asio., P. ,. de.cribed by.
5. AD. latell'at equ.,tloa ,. obtd. ••• froa Iq8atl ... 11 _d 12 vbea tbey ue eet equal to Q &lid eacb otber. ad tbe bOull4_, coDdltlOll •• f tile dilfuaioD ays ......... <lefIDed.
-1P'I. d P = -15 [R1t·lIt -{ • «a]
~ " 6. Inte".atloD of eqaatloa tbll'teea aD4 .,lfta,
dell... tbe OXJ,eD teaalon at XI
(11)
I .(18)
(19)
p. -P, -m.[~ln.L-~] .- 0 0 2" r 4 i (20)
, The aver.,. oxy •• a coecaat.atlo. of tbe capillar, Is calculated
IrOll either the a-v oxneR differ •• ce
Co= Co -[ Co - Cv] Lei (21)
,
or froa the OXJleD extraetloa
C - CODeeatratioD
- , ... Plow
L - Lenita
D - Diltuaios eoefliele.t, 02
80
(22)
a ... arterial
o ... "'x". uaiped value
• - oxyae. utilization
Usia, Erlan,'. equatioD (I •• alioa 20) Ket, baa obtained aD equa
tion lor the _an tissue oxyp. ten.ioD by intesratia, the equa
tion ~t.e •• tbe ltait. of l' aDd at
·(23)
Equation. 22 aad 33 provide the meana for •• tlaatia, tbe
tl .. ue oxygen teasion in the resioD of the electrode. Ie the ua.
ol the .. equatioDS, It Ie pce.ued t~t tbe elect.-ode eaua •• a
aiaiaal reactive error aad that eireulattoR .. d metabol! .. 1ft the
re,ioD of the electrode ace alaJla:r to thoae of other reJi.as of
the ti •• "e. TIM _aD cap!11uy oXf,ea COlICeatretioa, obtalaed
fro. Bquatloa 22, 18 .. e4 to deteral.e the related ox,sea tearioD
from Dill'. oxybemo,lobla diaaociatloa ~ve •• a Sub.tit"tloD of
the oxyten teD.loD for '0 In JquatioD a3, in addition to substitu
tiOn of the values obtaiDed for at r &ad a by aeaauc •• eat aDd with
sub.titutioa of the publisbed value of 0125 pcovidea the data for
the calculailOR of the U .a.e oXJ'.ell tea_ioD.
Tbe extebt to _iel1 ttle cal_l.ted t1ea .. ox,pn t ... ioll
81
"oYleSe. a c~ltlcal te.t of the bypothesia depead. In pazt on ike
COl'l'e.poaderu:e betweea tile tla.ue ayatea aad tile ... th •• atlcal
aD.lo, pcoYlded by Ket,'. equatloa. PactOI'. la ti88.8 cil'culatloa
aad .eta-.ol1 .. iD cal'taiD cues (a. will be abowa la tbe f011owlD8
chapte .. ) aa, lIake tbe _the_tical lIOdel a poOl' choice 101' .. plct.
lD' the tla.ue OX" •• teBai.R. ' .. tber...... a ate ad, atat .... t
.. aalatai.ed ~11. the I'equil'ed 4ata ls collected.
MI'J.'II)DS USED IN Tl SSUB MBASURBMBNTS
A cup ... l ... 01 polU.,J:ap''' .• aDd calcnl1ated oxyp. te.sloa
... lIade In the akia .. d in tile _cle of the I'abltl t 1. ordel' to
teat tile .. ypotua'.. Stla.u .. lected 10.. the orittlnal atady
bee .. ae it .... poull)18 to uk ......... t. with "1'1 little
dlatul'baace ill •• 1101'11&1 eoadltiou .f tbe _11181. .BYeatIla11,
.ac18 ....... , atllce the biNd ",d •• loa oaruteriatl" of "
.. scle we... Mre .tU18 til... tb... of'~ aklDI uel tile cll'q1a1~,.,
patte"D .. uld baft a bettel' 401'1' •• poDd ••• e to tbe ... ...,t10 ..
made in tbe deyelopaeDt of tbe :lI'IUlI - t .. op equatioa. Other
aspecta of oxygen amlability in ti ... e, .) ti.aue temperattll'e,
b) eoastaat -perf.sioD characterlstics and c) tl ...... taboll ••
were evaluated for the presence of a ateady atate in tbe tiasue
durio, ....... u.ta. 1ft ordaz te obtai. sevu&! diffe .... t ti .. ..
oxygen conceute.tlona at whicb tile polaro,rapblc aDd calculated
tissue oxy,.. tenaloa eoold be cOII,are4, tbe composition of tbe
inspired gaa .as altered to vary tbe oxy.en content of the
82
arter ial blood.
White rabbits weiJhina 1.4 to 3.8 kilogram were used as
experimental animals. Nembuta1* waa administered intravenously
in sufficient quantity to depress the respiratory rate to
ls.a4/minute. This degree of anesthesia induced a mild hypoxia in
the animal: An initial dosage of 30.50 mg/kilop-aa body veiFt
was followed by a supplementuy dose a few hours later. Heparin ••
was administered intraYenously in doses of l-amg/ki1ogram bOdy
wei.Jht to pzennt blood clotting.
In both skin aad auscle experiaents a tracheal caDBala was
, ' I
I,
inserted to ill8Ul'e a pate at airway and in some cases to perait !!'
malatenaace of respiration witb aD artificial respirator. In
rabbit skin experiments the cervical sympathetic nerve was cut to
denervate the blood vessels and prevent vasomotor chaages iaduced
by nervous activity. In the muscle ~xperiments the nerve to the "
muscle waa cut to prevent aervons sti.ulation of the auscle.
Cutting the nerve to the muscle, ia conjunction with stripping the
arterial sheath or cannulating the artery, also served to remove
vasomotor innervation of the muscle preparation.
Prior to tissue measurements the polarographic electrodes
*PentObarbital, sodium. Abbott Laboratories, North ChicaSO, Illinois.
**Reparin Sodium, 500 ml (50,000 U.S.P. units)/cc, diluted to 1 -alcc. Purnished for research purposes by The Upjobn Company, Kalamazoo, Ntcbl,aR.
" ! 'I
Ii I,
;: , I I
83
.ere calibrated In a.l1ae f"e.hl, equl1lbcate4 witb atmospheric
ail' and a e-y curve .d recorded. Aiter tbe coapletioD of tl ....
measurements the calibration and c-v ~ecord was repeated, aad aDY
difference 1. temperature of calibratioa was corrected OD the
basi. of • chaa,. of l.6~ Increase /desree centigrade.. Althou,b
this callbl'atlon lndlcated tbat tbe elactr04ea and circuit bad
been unaltered 4uia. an experiment, it save no ill4icatioa of the
magnitude of oxygen •• ' •• iOD in tbe tis ... a.
poluo,....phlc 8lectl'ode co_ctle .. co •• '.ted of ce.tact wit
tbe refer •••• eleetrOde and atabilized temporary iaplant. of the I '
anal,s'. eleet.ode. 'a elthe .. atin or aac:le. To obtain tbe nf
ereace elect&-Ode c •• tact, either tbe akaved and debrided rabbit
ear t Or the • .,ftd aad debt.'i4ed fro.t foot waa placed ia a beaker
of s&11... and the beaker was CODDecte. to 'tbe aaturated calomel
refere.ce electl'ode tty an aau-acu bri4,..
AIlalyai •• 1ect~ode. were placed ,. tbe sId •• of tbe tabbit ....
whicb bad bee. ... .. 4 aDd depilated _11_ a mild depilat • ., cre ..
• eft~a1 da,. prior to esperl .. at. The 4lectrod. were located i.
tile tip of the ear ill a npoa in ... iela. DO 1 • .t:. .. ... 1. were
vlalble. A small ball of cottoR baDeatll the abaft .f tbe elec~
trode eaaed the tenaioll 011 the aida, IUld the entire eu t1p, i __
cludin. electl'04ea ud cotto ••• u covered wi til seyer&! coats of
co110d10n. Thi. beld tbe electrode. in position and pre.ented tbe
*loltboff aad Li ..... 13 ladlcate a cor •• ctl08 of 1.4-1.6%/OC. rimeDt determinations indicate that with 01800'8 circuit the
influence of ataoephel'ic oxy,en* Oil the px.paratioD.
Anal,sis elect_odes aDd tbe levator scapulae aiaor ••• cle in
the Ilf!Clt of the ra.bbit were supported by .. T-sba.ped bar. Bach
electrode .u inserted pel'pendiculu to tbe axis of the muscle
libel'S and it penetrated the wid'tb of the auecl.. 'I'M 0_ to
four electrod •• placed ia the •• acle were aupported by Ciampa oa
tbe bar of tbe T which faced the a.aele. The T-bu alao contained
two plastic Jawed cl-.,a which beld each eDd of the eascle after
it had been f.eed from adjacent '1..... Tn ftpport bar .. d Ita
associated C1...,8 auft4 to ,..e .. nt the movel1tua't of the electrod.s
relative to tbe _sele &ad alao aiDiaized the Jllltcb.aaic:a1 dist_1>
anee of th. pceparatioD.
The aiRaPl. method. of tlad ... , tbe collectioD of oM-teath
mill1liie1 blOOd 5 .. ,l.s ia a sxaduated I .1 .e~o10.ica1 pipette
was aMd to ..... re blt'tOd flOW ill th~ expertea... Tbe IIU".81 "
ear win 1. the rabbit eca- WU #atbetezlaed with a pol.,."h,l~M
cathete!'.** tbe tip of _lell .... located at the point repreaentiDI
tbe ft •• _ .etUl'a fl'oa the %egioll of the polarographic analysis
electrodes. III the ..... 1. exp8rilleat. t the nin draining the
repo. betweea the plastic-jawed clampa wu ca.theteriaed with a
*,it ... rald200 baa estimated from experimental measurements that atmospheriC oxJ,ea Call aupply the OXYPB needs of human skin to a depth of 100 micro •••
**Intramedic POlrethylenetublag. #160 - i.d. 0.011, o.d. 0.024 lach.s. SCieat llc PrOducts Corp., Bv ... toa, I11i8018.
I", ' , ,
1
I
. I
1
, ,
1
,I,
l
ii.:1
I '! ,Ii
8.5
,iailar polyethylene catbeter. Tbe blood flow from tbe c.tbete~
tip was collected in tbe pipette. After approximately one-teath
.il1iliter bad collected In 'the tip of the pipette, tbea the
eatbeter tip was pushed lato tbe pipette to tbe deptb of thi •
• ..,le. Tbe ti.e required for eacb additional oDe-tentb ail11-
Ii ter collectl.a .... z(tcorded. A .. rie 8 of tiMet aampl.a ... made
until tbe flow rate 1=41cate4 a atable atate of perfuaioa.
1l00d ,.. .alyeia .... UCle os ODe alllll! ter ud ODe-balf
ail1iliter blood _.,lea witll tbe van Slyke-Neil appuatuaJ03 for
tbe firet few expel't._'. I'.ported for a.bblt au akia. ID all
other experla.ata the ,&8 analyeie WU .ade wi ttl a Xopp-Natelsoll
2M,IOS .ten' ...... ter. '* TIle appuatua ia slallu in prinCiple to
the ... !lyke-Neil ." ... at.8, but it require. OB11 0.03 all1111tel'
blood aamplea for tbe ,&8 anal,et8. All blOOd ... ,l.a were aD" l'Ped witbiD tea Illa.tea of tbe tlae:'tbd tb. blood waa ... ,led."*
"
Artel'lal • ..,1 •• froll tile CUGtld uteS', were uaed to deterlli.
arterial ox,,.a coDteat, aDd were drawa 'maediately tol1owia, the
eo.,l.tioD of Yea .. a flow me.aure ... t ••
TIle orr, ..... ft1 .... 4 Iv all&1,.le fao. the bl00ct a.ple by
alkali .... lutioD of •• aep root (a lak1.g apat). pot ... 1UII
re"l'i.cyaald(a .11d oxldizia, aceat). and .vacuatio.. Oxypa iD
trCopp-Natel80D Micro,a.OMt ... , Model 600. Scientific laduatrie., lae •• 1$ ,uk Itreet, Sprl.,£181d .5, M .....
*"The ox,,.. CODS...,ti •• by blood at I"ooa ad body teapera. tUNa reduce. the oxne. coateat of a blOOd ... ple "rJ' l"apl<11,.104
86
the extracted sample was absorbed with a solution of sodium thio
.ulfite containing anthroquinone. The oxygen coatent was deter
.ined from the partial pressure of the gas which was absorbed.
Bither a stream of nltrosea or oxygen from tank aources was intro
duced into the tracheal caaauia or the intake of the re.~irator to
vary the arte"ial oxygea saturatioll.
To control the temperature of the tisaue preparatioDs the
rabbi t ear .u held over a water bath, or a water pack was applied
to the ... cle. The water bath for the rabbit ear was covered
wi ill a piece of braaa ahim which was· in contact with the eireulat-
in, water. The water pack conaisted of & heavy pl~e of dialysis i
tubia, throup which waier cirCuLated. A iel1perai~e replated
water bath waa maiatained a.Yeral teatha of a deBree above the
desired ',eaperature (3lOC. for robit eu, 370C. for rabbit
muscle). AlteriD, the rate of flow of water from the water bath
regulated the fiDal temperature at the' ti .. u ••
T .. peraturea of the ti •• ue were measured by ~a of tberao
couples •• platinum wire resiatance thermometers. ~d thermiator
probea... The majority of the akin experiments were measured wit.~
the platin .. resistance thermometer t while the maj~ity of the
muscle experiaenta were measured with the thermistor probe.
*Co~per-constaatan wire fused by an arc-weld and recorded a refleet1D, ,alvanolleter.
**rele-thermomeier. Model 43TA, Yellow Springs lastrument Company, Yellow Springs, Ohio.
b,
I.
:111
'I,
II I,II I:
II,' ! 'i ! Iii
87
Blood samples wel'e alao ualped fu IMUlo,lobia coatea. u.d
blood oxy,en ooacentratloD. a •• ollobie waa dete~.i.ed co101'1-
.. trieally by the method recOIIINo4ed by a ... itiea of tt.~ Medical
SCience 01vl8ioa of the National Acade.y of Scleacea.20l.202 The
bellOllobin COBCeDtl'ation 4etel'aiaaUo •• in stin expel'i .. a.a were
.ade with Bye_l. ch •• ical I'ealenta aDd ia ••• cle experiments the,
were made witb AClI,loballn** .1'8a,.ota. Pl'om a lambda Jda:o
pipette 0.-02 .111111 tel' of bloOd was added to .5.00 Jaillili tel's 01
ferr1cyaDide vehicle. The optical de.aity of tbe solution was
read OD a Klett pbetocolol'iaete .. oain, a cree. (540 mu) filter.
CalibratioD ell.art. prepared fl'o •• &aueMnta .ade with ataadard
101utl0 •• furnished witb th ••• reasents were uaed for a sraphic
interpretation of the optical danalty of the blOOd • ..,18 to ob
ta1n tbe bloo4 beM,lobin coDe'e.tration.
Otber procedures were i.stltute . .s in a few experiMats 'to
provide additional informatioD on tlUI" approxi.atioD of the tiNue
oX1&en tosioa WIlicb were obtained by calculatioa. Temperature
... varied iu .. fev experimeata. ID SOlie expel'i_eQta pot_iUII
cyanide aDd dinitnpheDol .ere .ed to alter _taboll... The
lubetallee. were intrOduced ill tile ..... t.rialauppl., to tbe lI .. cle
.yeel Chemical lteapnta, C,_.theao,lobin standard aDd rea,eDt. Byeel Hormone Cheaistry Laboratory, HoustoD, Texa ••
**Acu81obin standard and diluent pellets for cyaametheaoglobin method. Ol'tbo ,bullaeeutica! CarperatloD, Ru! tu, New Jersey.
II ,I
III ,I
,
!
I: .' , , ,
88
or ear in micro,r- quanti tie. over a period of MYel' a1 miautea.
The cyanide cau.ed an accoaapaayla, dULll,_ ia blood perfusloD, and
the result. with dinitrophenol were v.ciable. The directional
chans.s obaerved in tis ••• oX1.m tension with both dn,_ were
indicative but did not quantitatiftl, indicate the reliability of
the calculated oxy,.. tenaioa.
CHAPTBa I I I
U=~~~::o=$~:":o:m
Tbe .xperimental reaulta obtained lor ti •• ue ox"en tensioa
in akin aDd 1ItIaC1e of tile #ablJit ue ,fts.nted ill Tablea v. VI,
VII and VIII aad 'iprea 15 _4 16 at the ead of thia chapter.
The ld aDd calculated '02 for electrode. in wbich reliable pol~
o,raph!c _as.e.llta were obtaiM4 dlIrill, a at •• dy atate in tbe
tiaa.e are preseat eel ill 'll1Ue IS for tbe rabbi t ... ele aad ill
Pipre 16 for tbe rabbit skill. Ja Tablea VI u4 VIII ud Pi .. r ••
15 aact 16 the -al.,..lal repr ... ate selected data. the .. leeti.a
beia, baaed oa the crituia pr .... t •• below. Tabl. IX ._arb.a
the ualyala of tbe expel'iMatal data for factor. which iDelicate
• enaa,. i. the ayet .. for wbich oX1.e. teJl&ioll wu calculated Or
a ch .. ,. ill tbe polu:opapllic electrodea.
Jxperi .. atal r •• ulta were .val"~.d for tbe exiateace of
reliable poluo~ap1dc aeuu ... ata ucl for tb. pr.aeace of a re
producible ateady etate in tbe ti.... before co.pariao.a .ere made
of the iel of aD electrode awl tllecalculat.d ,02 of the U •• ue.
Polarocrapbic ..... r .... t. were acreeBed foe coDditlon8 ia which
lUluysla for id wo'"lel sift valid _ ... re. of oxy,en tealloll.
aain coaaideratlone were tbatt
1. A c-v cur.. be pre •• nt in tla.ue .... ure •• at. aDd ift calibratioJl' prec:edi III and following tiaaue meuare••• ta.
The I
II
11'1,
I'
III1 " !
I
:Iii
90
2. A con.tartt electrOde au.face exi.ted dUiDf experimenta. It ... pl'e ... d tbat if the electrode seaa tiyit, .a. .imilar in ca1i~ation before and after ti ••• e measure... t., .fI ell .... 111 tbe electrode a .. face occurred duriD, experimenta.
3. CoDditioDS ill the region of the ttlee'trode ftc. stable 4uia. tl ........ U' .... t.. ai.. iT ill vitJ:o .aa observed 'to vary le.. tban ~ over seyera! Soura, repeated fluctaatioRs of over S% in ti •• ue Me ... re.. nt. were regarded as iadieatisl ••• table eoB4!ttoas in the tis.e.
The l'eprod~clbility _d exi.te.ce of a ateuy atate were deter
'ained indiJ:ectly l%'Om thel
1. Presence of constant iemperature dtll'illl JIIea.vementa.
2. VU'iatioll.f leaa tltu teD per cent in thr •• COIUMCUtl .. col1ecti.a tl ... for blOOd flow .... 1 •••
3. VU'iatloa.f l •• a tIl_ ••• pel' c •• t between 14 or blood flow observed 18 a liRa! control and the i.i tid .at .. el oNe""9atio ••
TIl ... atio.al •• f tbe poluo8l'aplllc criteria .u en_rated il1 the
41a .... i08 of tbe polKOII" .... ic .tb9d. i .. tl" .... t&tl0. Ud 11 ter-"
aturel. tbe fi •• , cbapt... The .r ...... lor the "1 t.l"ia .. 184ted
for the U ........... l1te will beeOlle appu •• t claia, tlla dia-
cueaioa of the r ... 1t ••
TIle .al .. 1& ted ... ,,.11 te •• lou baft bee obtaiud b, two
aeUlocla, both baaed on Bquatioll 23. Tile data ue ftpoJ:t.d for
the Me.ad .. tlaOtt uaiD., the fo11_1a, e.atiol&l
(24)
I
11'1
i .. .eu ti.... OXfPD tea.iOD
'0. a¥erase capillary OX" •• t ... loa • - oxysen extraction A .. factor baaed on ratio of capillary aad ti •••• radii Jl .. ti.... .,11114." 1'a4i.a D - Kl'op'. Yalge for difl,,_ioa of ox,.11 in .... cle
91
Tbe Heoad aetbod baa tb.&d.aDt .... of 8.,lIaaiaia. tile coauiH
tioD of capill", ox"e. teasioa and of ti.au« diu.aio ... &ad
metabolia to tbe .stillated a"I'&,. p02 ill tb. t.la_.
Ia Tables V aDd VII tbe yal •• fol' A ... obtai •• d fl'oa Eat,'s
Table 1. 16' The val •• for D •• p~ .... ts .tl1Illt .... of _, ...
cdsslll, a c88*lu*el' s.a •• a.face pez u.ute tude .. a "adieat
of a ad.l1laet ... of ....... , p......... (te •• 'oa) .... c •• tl_t_ path
leDlth. ,!a obtai ... _.be_ tM calculated factor t a(Alt2/4D), i •
• ubtl'actetl Ina ave ...... capl11 .... , OXJ ... teadoll, Po- TId .•
factor, '0 "I'"ected 'or deft aU ... :18 t.ape ..... t •• frOil 31OC., "
... obtalaed froa Dill'. upe.,lobiD dia ... l .. tloD cul've.2 TIle
bloOd ... d.t.rmtaatioaa and ....... d b .... lobi. c •• c •• tratioD
•• I'e ... d to obtai .. the be_l1obia aatuatioa of the blood.
ft. data froa Tabl •• , aDd VII hay. been .. p_ated e. the
baais of aD&1.,.,. ,.to two .... .Ie.. The fu.t .... i •• o ... 'a.a of
thOM .aa ••• at •• de .ill ti .... cturl., .. ep.od .. lble ate.,
atate. wi tll eleet .. ode. brie, coastut cbuact ... i.tl ea. The
heolld ae .. lea •• pr ... ata .......... at. Mde ••• _I til ... tale ti ••••
atead,.atate •• electrode eb~act.rl.tl •• were Y&l'lable.
Tabulatl.u for co.,ar'aoa of .. leete. lc.t aad lOa are ,i •••
I:
-92
for rabbit lIuacle In Table VI _4 for Z'abbit .Id .• in Table VIII.
Tbis eo.,aziaoD J:epr.aeate al .. 1taneo ...... ~ .... t. of Id .tth
polUOp:apblc eleetzod •• tilat: laad coutaDt cluu::aeterieUu
.h&'ou.u"t aD experiMllt, aad Jo2 obtained dula, l'epZ'04ucib1e
s.ead, atat •• in tbe ti •• ue. The tabulated aaterial i. ,Z'e.e.ted
p'apblca1l, in 'I..-e 15 for rabbit .".1e and I.a Pllue 16 for
r&b1;,1 -:: atin. 1& the .... .,b. tile ioJ bu beea made the abaci.ea
aDd id the orella.'-e. la the .............. 11&4 ••• r..,.,lt ••• cle
Mvea ael'l ••• f ...... e •• t. oat of tbirty-two sel'l •• were lIlacse
when ti .... aad electrode chu_te .. l.Uc. r .... l .. d eoaatut. T.a
out of tJllZ't,....ipt M"if':& .f .u.e.ah ill Z'ab1>it sid .• bad
siailar con.taat coDditlou pz'e ... t la ti •••• aad el.ctrOde. 18
thea. ael'i.. of ......... t. .... 14 .. 4 Joa .... coaapCll' able, tbecf!
ia le •• tball 10 peZ' cellt varlatioll be ..... laitial and fi ... l COll-I
tro1 ........ t.. If the •• 1atl ••• f ladlyld ... l pol&l'oarapllic
lI18uareM.ta i. It .... " Oft the eleou04e aeuitlvlt., .. 4 calculat.d
pOa for .. expel'iaeat there ia le.. thaa 10 per eent vuiatioD foJ:'
any indivldttal .... u ••• t and tb. •• wrap variation la akia 1.
1.3 per cent. while. yalue of 5 pel' cent 18 obaerv.d for ... cle.
Tabl. IX ._ul ••• the analy.ls of the experi_atal data fo ..
polaroaraphic electrode cbaracteri.tics and rep.atable .t.ady
atatea in tbe tiesue. The coluaa headi •• iDdic&te tbe baai. for
exclu.ion of tile electrode compui ... wi til calculated values.
The aUbel' of each of tbe cat.SOl'i •• has BOt beea _ui.aed,
ainee. ia tl .... , ac ... , factor. could coatl'ibute to the
93
variability. ADy auaaary would tead to empbasia. either elect~od
or ti •• ue y_iabill ty .a the primary cauee f. yulatioDs whicb
are u8uallyobaerved ill poluop:aphic _uure_ats of tia ... OXY
,en tellsioD, aDd it i. likely tbat both factors are .Hall, ill
volved. The only cle.r-cut eyidence for electrode unreliability
is to be fouM in the ia ri. tro callbratloDs. _ ................. In ........ .,. the .. experiaeata ladleat. that.
1. . tIader cODtl'olled eODdl tiODS polarGpoapllle Muue..... ue d ..... trated to .. w1tbla t.a per ceat of the averace ·U .•• ue ox,"'. te&aion.
2. Due to variabill t, 1. tlea.. aDd el.clrode. polaro-........ l .......... t. ill a _Jol'it, of i. "iYO .ltuatlo •• will yield .aly qua11tati .. -..llmat.a of " .... ox,.e. availability.
3. oa1,. few .f tile c.... of vulabill i, la polue. p'apblcal,l, ..... ued tia.ue oxy,.a teasioa au be attri~t.4 directly to faulty .l •• trod •••
4. The 14 obaerYed pcobabl, repre ••• ta true tias .. ox, ... tellaioll, bat ,the relationahlp to ti.s. circulatloD, utaboliu, aDd"oq, •• availability requ!z.o.a additioaal iaforaatio. to coapletely cb .. acterlae local ti •• ue coDdltlona.
I
I' :.11
sYMJIlLS nun IN TA&BS V AND VII
S,..bo1. ~ari •• }Jader !5'!J:Uaeat,.
H • coRkol
+ - i.cr .... d arterial ,02
- - decreucd utuial ,02
o - circulatory ure.t
JeCN • pot ... ,.. cryuide
DJIP - diu trophe_l
S,.1to18 Appear! .. 1ft Table
a * tJ a
( ) - eatimated .&lue
• - Dot .. ~rected foe Ii .... weishl
C - DO oai!bratioa check
I' - more than 101 4ill.reDe. in control yalu.a
II - .atable
1
T.A&B Y
M1iAStIUUO!It1'S AND CAtCIJI..ATIOJIS OP nSSUB OltGlUf TDSlOJi I. RABBIT MUSeU ..
r r a A • ·0 .. 2 l! 14
~1~. radi .. a-a4iu ~ extcaet ,1 #2 113 #4 UPEIlIUNT " u cc/P/. .. He 4D _Be jim deflection
DOl 14 .129* 55·59 ~s
UG2 (35) .0045· 1:12 R .092· (3.5) l.16 sa
+ .081* .0042* 90+ 141 - .100· .0041* 0 0
RM'lTG 1 M .042* (3.5) (35) 3.16 .0062* 62 2 60 ~l + .041· " .0069· tOt> .2 (100+) 137 - .048* .0068* 0 0 0 0
_._- .2 lUll""'"
~;:; N .29S· (3. S) (35) 3.16 .0098* 60 4- S~ + .350* .0123* 19 5 74 ~9 - .392* .0071* 11 3 8 5
~".LG 3 12.60 19.3 ~7 N .333 3 22.5 .0230 45 36 23 8 78
+ .299 .0230 53 !I.l 43 ~7 30 9 15 -~ .353 .0225 33 P.I 24 ~l 11 S 12 N .. 341 " .0228 42 ~.2 33 ~2 19 11 62 0 0 - 0 0 0 0 0
~CIl .576 .0010 94 0 94 ~6 18 13 11 '0 IP II A J, If If II' \It
..
, TA&E Y (Coat.)
p r It A • ·0 8 2 Y #2"',13 PLOW radi_ radi .. 02 extract ....-- #1 #4
aUEltlMlNT -t/P/. • .. cc/P/. -ft. 40 _H, .. deflec:tioa
aM'l'TG 4 )I .309 J.' 40 3.40 .007' 36 4.1 J2 23 11)( 32 - .365 .0078 22 4.2 18 15 I1j( 18 • .278 .0083 39 4.5 35 26.5 1_ 16 ,. • ass .0072 33 3.8 32 21.5 16M ao 0 0 0 0 0 0 0 .= 0
KCN .7SO ~ 0 'Oto 0 90+ 72 52 A . A A A IP
JUrrfG 5 • .425 4.0 ., 3.36 .01a, 47 6.2 41 30 2211 s - .458 .0192 42 6.4 3.5 25 • .1 11 11 • .375 .0189 57 6.3 51 36.5 27; 9 .,
N .489 .0182 48 6.1 42 31 23 10 0 0 60 0 0 0 0 0
KCH .868 0 78 0 18 59 4011 24 A A A If
JUtfTG 6 N .245 4 4S 3.36 .0102 42 6.9 35 2S.5 [22 - .275 .0100 33 6.8 26 20 19 • .220 .010' 48 7.2 41 31 127 It • ass .0101 41 6.9 34 26 126 0 0 - 0 0 0 0 0
A ,. A A IfP .. •
, TA&B Y (COat.)
p r R A • p. ...2 ., nld#3 PLOW radial 1" .. 1 .. O~r.:r.ci #1 #4 UPJlRINBJrJ ai/PAl- • • - ·1 4D -iii - deflectioa
IOt1TG 7 2.3(J If .364 4 2.5 .0222 38 6.3 32 38 19 - .341 .0253 33 7.1 26 29 21 + .361 .0231 41 6., 40 40 8 II .34' .0225 39 6.3 13 23 13 0 0 0 0 0 0 0 011 DNP .322 .0225 3S 6.3 29 32 20
A A C C
IUlT'tG 8 N .211 3.5 22.S 2.35 .0199 41 2.8 44 11 15 - .237 .0190 33 2.1 12 8 22 + .179 ". .0191 12 2.9 69 41 10 If .223 .0180 42 2..6 39 23 36 0 0 0 0 0 0 0 0 DNP .235 .0325 36 5.1 31 19 29 A A IP IP
IUITlG c) N .396 3.5 35 3.16 .015S 26 60 20 43 32J/ - .437 .0151 21 5.8 IS 31 23 • .361 .015S 32 6.0 26 54 40)( If .384 .01S2 21 S.9 21 45 33)1 0 0 0 0 0 0 0 0 A ~ A A A
I '" ....
----i~ TAall v (Coat.)
, ~ a A • '0 .£
, 14 PLOW radi .. ~adl .. o~r~,:=t #1 112 #3 #4
.lxpBltumrr -t/p/. • " _H, 4D _H. _ deflectioD
III'r'lG 10
• .384- 3.' 3S 3.16 .0112 42 6~' 35 13 54 .. .48S .0112 21 6~6 20 47 3' • .4a2 .0110 41 6.6 41 91 61 It .422 .0117 40 6.8 33 10 19 0 0 • 0 0 0 0
A A A I.
IMT'l'G 11 If .283 4.0 32.5 2.1S .0111 as 1.9 23 S1 "" - .a8 .010S 17 1.8 15 40 33 • .263 .0125 3l 2.1 31 80 51)(
" .311 .,
.. 0131 24- 2.3 J3 50 36j1 ". 0 0 - 0 0 0 0 0
DIQI .211 .0310 34 8.5 a6 , , A A C C
Drn'G 12 N .411 4.5 4.5 3.16 .0111 IS 7.0 8 22 31 12 - .436 .0109 7 [6.9 0 4 9 9 + .40S .. 011S 22 1.4 15 49 56 128 Pi .400 .0112 14 1'1.1 7 24 41 11 0 0 - 0 0 0 0 0 0
A A A Il' Il' ~ ..
GO
~ , TA&.I Y (COat.)
p r It A • Po 2 y 112
idll3 #4 PLOW radi.a radi .. Oa rt:~ac1 .AIl #1
EXPERIMEMT ral/p/. • • C:C Po II -III 4D .. He .. deflectioD
RMTrG 13 N .272 3.5 u.s 2 .. 35 0.0117 43 1.4 42 31 41 11 - .325 0.0122 27 1.4 26 19 23 7 ... .. 300 0.012S 46 1.5 45 33 37 (0) N .288 0.0123 32 1.6 31 23 32 (2) 0 0 - 0 0 0 0 0
A If IP IP IP
JUnTO 14 If .411 4.0 30 2.60 0.0136 S7 3.2 54 70 18 2S - .431 0.0124 34 2 .. 9 31 35 14 20 ... .389 0.0144 66 3.4 63 lOS 32 .50 N .396 0.0134 54 3.1 51 69 22 36 0 0 '.
0 0 0 0 0 0 -CCN .582 0 Jo 9 O+) 0 60+) 135 052 64 A A A IP IF
"
"-
:
"'" 100
TA8LI Y1
COMPARISON Of id and. pO. IN '1'HB RABBIT MUSCLE
AlTDIAL OXYGBN SATURATION EXPBRIMBNT kx>NTIlOL BLBVATBD DECaMUD CON"fR.OL ZERO
lu.nTG 4 ~2 32 35 18 32 0 23 36.' 15 21." 0
RM'lTG ~ ~2 41 51 35 42 0 30 36.5 25.5 31 0
lUfrra 6 ~2 35 41 26 34 0 as.s 31 20 26 0
JUt1TG 9 ~
20 26 1.5 21 0 43 54 31 45 0
lU41'TG 10 ~2 3.5 41 20 33 0 73 91 41 70 0
1UlftG12 ~2 8 15 0 1 0 22 49 4 .34 0
lorna 14 ~2 54 6J .,' 31 51 '. 0 10 lOS 3' 69 0
---------a,"
OXYGEN TENSION IN MUSCLE , 1001 AMP 10-7
, /
r- " /
Z /
W / /
0:: 80 / , 0::
,
:J ,/ /, .
/, ' U
,
U 60i , ,/.
' /.
I 04
0... + , ,
<t 0:: 40· <..9 0 ~ ... , / RIIITTG 4 _t_
o:: . / ,~ RIIITTG 5 ...L
j 20j + .....:/ / RIIITTG 6 0
,'/'/'// , ..
RIIITTG 9 L RIIITTG 10 ..L_ RIoITTG 12 ..L. RIoITTG 14 ..L
0 0... I /" /.,..,. .........
O· 10 20 30 40 50 60 70 MMHe
CALCULATED TENSION . -- ~-~. -- .-
.... PJGOU 15. 0 ....
COMPAR.ISON OF 141 AND p02 IN RABBIT MUSCLE
.~
TA&B VII
MBASJ..JilEMENTS AND CALCULATIONS OP TISSUE OXYGEN TENSION IN RABBIT EAR
T P J. It. • Po' la
M2 y 1d
-r PLOW ... 1 .. radius 02 ex.raet #1 a #3
BXPBIlIMENT co ml/P/. 1i 11 cc/p/a _H, 4D _B, _ defleetioll
HGI }( 1 .066· (3.5) (105) .s.~ .0040* 55 2.9 S3 21
D02 , If ., .07S· (3 • .5) (lOS) 5.:J .0011· so 5.1 45 21 • .083· .0015· 110 $.3 105 40 - .092* .0063· 10 4.6 S 3
• I.B!'fJ 1 tf 21 .063· (3.5) (105) S.:l .0031· 55 2.2 53 31 39 + 1/1 -.068· .0034* 110 2.4 161 lOS 101 - as .073* .-0030* S 2.1 3 0 7
UTG2 N 27 .. 092· (3 .. 5) (105) S.:l .0039* 48 2.9 48 38 41 + - 21 .091- .0037· 100 2.7 97 57 74 - 27 .101* .0030* 4 2.9 1 S 9
It .
IUlTG 3
• 29 .18S (3.5) (120) 5.6 .OO~ 42 3.0 39 11 24 • 29 .110 .0030 125 2.9 122 41 60 29 .220 .0032 as 3.1 22 9 6 -
UTG 4 N 31 .180 (3.5) ~ 13,0 5."J .0035 42 1.4 41 30 42 5 + 31 • 190 .0039 170 1.6 168 134 1M 9 ...
0 - 30 .211 .0038 10 1.6 8 1 7M 0 ..,
-.--.---.--........ ~ - ..
---=-~~ ~---~--- =-=--""""
',., TABUl VII (COat.)
...... p ~ a A • '0 .2 l' icl PlDW Ifacti .. .. acU.ua 02 auact
40 ~1 112 #3 DAr&aIlIINT CO &lIP/_ a a cc/PI· -lie -III ~ 4eflectioD
Qi'.IQ 1 34 .345 ~3.S) 106 3.4 .004.5 8.5 [3.3 82 58 ~1 as N ... 31 .263 .0040 46 ~, 43 30 ~6 11
+ 31 .333 .0036 150 tal' 6 141 109 81 51
• 31 .293 .0038 31 ta.S 28 20 21 IS 0 31 o - .. 0 0 0 0 0 0
I. I. IP I. I. IP
lIrrG 2 31 .283 ~3.S) 111 .5.4 .0080 90 1 83 33 29 15 II - 31 .263 .00.59 40 ~.2 35 10 14 18
• 31 .253 .0056 151 ~., 152 SO 32 9
• 31 .313 .0061 88 ~.9 82 30 l' 12 0 31 0 ". ... 0 0 0 0 0 0
A A A A C C
QrN 3 ~3.5) 103 .0065 ~.a N 31 .215 5.1 90 8' 43 7 7 - 31 .213 .0062 52 ~.O 48 30 ? 11
+ 31 .221 .0068 110 14.4 106 ? S2 31 R 31 .20.5 .0064 83 fit. 1 79 31 42 16 0 31 0 ... 0 0 0 0 0 0
A A A Cay !c-Y ~-y ~TI1:i 4
.319 ~3.5) ~ 96 5.2 N 34 .005S 46 ~~1 43 31 37 29 ... 34 .294 .0053 72- 3~0 69 52 65 27 ... • 34 .268 .0058 3.5 .3 32 25 30 11 0 w N 34 .3"'\,3 .0052 48 3.0 45 33 33 15 0 0 - 0 0 0 0 0 0
It. A A A A IP -
...,
TAILI VII (ceat~)
T pt.&. r R A • It Alt2
., ~1 #;d. #3 IXPJIlIMBNT CO JIl/P/.
cacti .. cadi .. ~icr~t ., ,. !
" • •• 118 -.... u. .. cle 1ectioll
~ .. IU, ... G .5 If 34 .2SS (3.5) 134 5.9 .0085 30 10.1 19 14 12 17 - 34 .363 .0069 5 8.7 0 3 0 0 + 34 .342 .0092 18 11.6 66 21 36 33 N 34 .431 .0081 28 10",2 18 9 1 6 0 34 0 .. 0 0 0 0 0 0
IP A A C C C
.B'lTG 6 I
H 31 .260 (3.5) 111 5.4 .0057 68 4.5 64 SO ? - 31 .240 .0055 55 '·4.3 SI 33 'I + 31 .231 .0061 97 4.8 92 66 ? If 31 .281 .00J2 63 4.1 .59 ., ? 0 31 0 ". .. 0 0 0 0 I
~ 31 .509 0 1. 0 14 65 1 I
A A A A ~-y
~.Iu'1:v 7
• 31 .216 (3.5) 120 5.6 .0111 45 10.6 34 27 29 21 - 31 .2.53 .0114 40 10.9 29 22 25 11 • 31 .345 .0138 68 13.2 55 41 S2 31 II 31 .2BS .0128 39 12.3 27 22 23 17 0 31 0 ... 0 0 0 0 0 0
tOf 31 .369 .0018 4' 1.1 43 36 39 29 IP I. IP If If If .
... 0 ....
. .., 'fA... 'fII (Coat.)
T p ~ I A • P' Ma y i PLOW ,"ad! ... radiu ~r,:,:ct
0, ,1 #24
#3 raP.tiPIMENT Ct) .1/pJ. • .. jul., 4D
_S_ ~ deflectioD
Jur.rrG 8 II 31 .210 (3.5) 140 s. C) .0055 58 7.6 SO 31 22 27 - 31 .321 .00$1 31 7.8 23 19 11 11 + 31 .220 .0055 72 1.6 64 48 30 26 If 31 .230 .0055 52 1.6 44 28 16 14 0 31 0 .. 0 0 0 0 0 0 .. 31 .220 .0060 5S 3.3 47 2S 11 8
A A It A C C
IKlf.lTG 9 If 31 .241 (3.5) 98 5.2 .007. 52 4.4 48 35 22 27 - 31 .267 .0015 11 4.5 7 6 10 0 + 31 .264 ,. .0082 100 4.9 95 11 48 0 K 31 .319 .0093 61 5.5 55 40 21 .2 0 31 0 .. 0 0 0 0 0 0
IDHP 31 .3S1 .0115 65 10.4 55 22 11 0 IP IP I. C C C
J!'1"'tG 10 )1 31 .150 (3.5) 112 5.' .0069 55 5.7 49 37 28 - 31 .159 .0071 26 5.8 20 10 14 + 31 .169 .0011 83 5.8 17 48 44 N 31 .171 .0075 57 6.9 SO 34 25 0 31 0 .. 0 0 0 0 0
ImiP 31 .166 .0075 54 6.9 47 30 26 A
~
It A A A ..... 0 \It
.-.-~ .---..... ---- -- - .. ---.. ~
."
TABLB 'fIt (COat.)
-T P r a .A • '0: .
., i Ptow radius l'a4iu 2 extract 1 #ad. 113
CO al/"';. • • "IPI. a8 40 H • deflection
31 .243 (3.5) 116 5.5 .0060 60 3.3 55 41 29 31 • .346 .0054 11 4.8 6 5 4 31 .328 .0050 124 4.4 120 42 61 31 • lOS .0061 51 5.4 46 27 22 31 0 - 0 0 0 0 0 31 .293 .0102 55 9.1 46 24 21
I. IP If I' 612
N 31 .318 (3.5) 122 5.6 .• 0051 67 5.1 61 47 33 - 31 .a19 .0057 25 5.7 19 18 12 + 31 .210 .0063 79 6.2 11 60 33 N 31 .318 .0050 61 3.0 56 42 30 0 31 0 - 0 0 0 0 0 eN 31 .306 0 65 0 65 46 14
A A A A A
.... 0 ..
101
TABLB VIII
OOMPARISON O. 1d and Joa IN 'BIB IlAI8IT SlCIN
AJlTElltAL OXYGfAN SATURATION SXPD.IMBNT CON'1'JtOL IL!VATBD IlBCiU!ASBD CONTROL URO
RBTG 2,pOa 48 9"1 1 0 id 2S 57 , 0
JUrlTG a P>2 88 lsa 35 82 0 1(1 33 50 10 30 0
UTtG 4 pOa 43 69 32 4.5 0 .11 Id 31 .52 3.5 33 0 12 id 37 65 20 33 0
lurrrG 6 p02 64 92 51 59 0 1d SO 66 34 41 0
IUrn'G 8 pOa .50 64 23 44 0 1d 31 48 19 38 0
"
"
UT'l'G 10 pO; 49 17 20 SO 0 .11 Id 31 48 10 34 0 aa id 28 44 14 as 0
ItJ!T1'G 12 ,02 61 71 19 " 0 11 1d 47 60 18 42 0 .12 1d 33 33 12 30 0
z z o (f)
z w ~
z W
<..9 >X
o
r:-o o ex> o I'-
\. • \
, \
, , o It) ,
• o 10
o q
~'4i~'1·H ·1 .;r:t
NN
"
,,"" I-~~ ............ a: a: a:
\ • o r<'l
w:tw
:t ... 2 "" III
1-1
-1-
1-1
-1-
............ a: a: a:
o C\I
~
III I-l-.... a:
o
JIHd\1~80tPjlO d
(!)
2 2 0 ~
0 N
o o o CI)
o V
o N
z o Cf)
z w
r-o w
r« --1 ~
U
--1 « U
10
8
I"
r 109
TAILS IX
ANALYSIS O' .lLI!C1I.ODB MlAStltDllHTS POll ULlAJIII..ITY AHD VARIABlLlTY
VAR1AJlIt.m ULlAllLITr Plow COlltrols id stability Calibration e-y curve
JXlIEI.IMINT
UftG 1 X X '1,12.'3 #1,#2,#3
RETTa 2 #3,#3
JUltTG 3 #1,#2,#3
UTTG 4 113
a:srrQ 3 X X
uno 6 la 112
UTTG 7 X. ,l,IIJ.'3
IlBTtG 8 la,'3 IlITTO 9 ;X X '1,'3,'3 UT'lQ 11 X #'.A'2
lUC'lT8 ,3 X X
JUG'TG 4 X '3 '2 lM'.lTG 6 .2
RM'tTG 1 '1.'2 JUtl'TG 8 '1,'2 ttm'G 10 '2 lUtt'l'G 11 '2 '1,#2
a.MnG 12 #2,#3
JUn"lQ 13 X 11,#2.'3
JitM'lTG 14 '2.,'3 , refers to electrOde
r CHAPTmt IV
DISCUSSION OP Tl SSUE OXYGBN DNSION MBASf.JIUiMSHTS
The description of tissue oxygen distribution depend. upon
.. al,Diog yalues to factora related to deli.ery of oxraen to tbe
tisauea aDd to stora,. aDd utili.atlon. Tranaient atatea &l'e
difficult to treatquaatitatively aDd, .tth tbe exceptioD of
Rouptoa t a20• treatment of the proble., baYe not been described
in aatbeaatical ter... DYen III atead, atate. olll, approxlutiou
of tbe diatl'lbutioa ~e available, aa4 tbeae "pl'o.l.atlo08 depead
upon ai.pl11,1a, ... .,tiOd. TIM abapllficatioD Is viauali •• d
thus. tbe ... , factors rel.ted to ox"eD ayailablU t, III tis ....
var, l&4ivi4aal1, .hile all other factors rea.i. COllatarat. M.the
•• tical treat_at depeada upoa this aiatplified approach and apon
sepuation of tbe factors. Tile initial a.puatioD ia to co.aider
delivery as Olle aspect aDd tla •• extract!oll a. the other .,aspeet. "
Deliver, ia <I.paDdeDt apoa rat. of blood flow aad the oxypn
a.turatioa of tbe bloOd. AD iacrease either In rate of flow 01' i •
•• tuatioD of the blood in tbe abaence of any clump in the blood
pattern will result in an iaCftued .tora,. in the ti.aue beeane
of tbe elevated diffusion potential &ad aD iocre.s. in YeDOUS ox,
,en s.turatioD in the new ateady state, aiDCe _taboll •• bas DOt
chaa,ed. AS the effects of elthe~ increased flow or iacreaaed
saturatioD are Similar, tbey can be !'apreMnted b, a 8in,le
-110-
r~------------~ 111
factor. ID tbe use of BrlaD,'. equatioD or of equatioas derived
oa a .l.llar baaia. tbe a.., ..... captl1 .. ., O",PIl coactatr ati 0" II eM be uMd to repl'e .. a" deli.el'Y. Ia 'ety' .165 equatiou aDd ill
tb. experime.ts .bleb ..... ..eporte", deliv81'1 i. rep"e ••• '." by
Co, ae a .... a,. OSJpa collAatrati08 of .,1004.
A furtbe.. complicatioD of the ti.aue descriptio., aDd tbe
pri • ...,. .t .... 1in. block In ..... h .... ti.... treat.eat, ia tbe » .. e_o. of ..... l.bl. aD4 the _c ... , t, of Cb_.l .. f 1'0. tbe ... of COIl-
ceat .. ati •• to the ••• 01 dil, •• loa lI'adie.t 1. order to • ..,lete
the de ..... "tl •• of tile QX,,.. dell ... .., to tb. ti..... 'b_ _It
filtl'atloD will 'el1 .... le .. til ... De per ceat of the ox,,.. ie-
q.il' .... ' .f tile ti ••• if it is calcu1&t •• Ic.a '..,. .... :lMr •• .ao' data, wIll.1I ..... tllat tbe oal, 1IOd. ., 41stI'I btltio.. to tile
tlH ••• of aD, a'pilie_.e i. difl .. l .... ,TIM cliff •• l.a p'a4ieat
fro. _1ood , •• pa.eat .... the di~.i.ti •• of OXypD fro.
bemo.lobin .tel'e. &ad the ph,.icall, dis .. l .. " ox,.... At ox, ...
t.ui .... abo ...... pM1 ••• tuatl0."h. ato:r ... ia 1:11004 :I.. dia.
".,uded, bu.. at oxy, .... aai ... ,. tIM .. epoa of ox,laeao.lobia
41aaeciatioa, tile difl.i.8 srad:l.eat MAO .... ,..<lent UPOD iIle
__ l1obia .. tuatloll. _iell ,. a co.,1" facti •• of ox,,..
t ... 10.. Tbe .eat 84 ... ,.,. ia .. ia, :.:0 ,. appu_t, .. ace •
aia,l .... al •• aa, be ••• i,..d. to ttl. diff.aioa ......... bead if Co
is c ...... te4 to .... ce.t .a, .... tlO8. Tide diff.ai.a ,....... i •
.... :r .... ed .. Po Is ibe expel'iaeata. la ... t expet .. i._,. the
sltsht hypoxia aad oal, .ode .. &t. .leYati08 01' depre •• ioa f:rom tbl.
112
.11Ibtl, hypoxic at.t. ,.&,.e4 to, teep tbe val... of Po to tb.
telatlYeI, 1laear port10n of the oxybeao,lobla 41 ... clatioD carve.
TIM low Po ahcMl14 alao ...... to aate tis. pc •• au. head fot 41ff ...
a10a al •• , til. oapt.l1u, ... e aeul, linea .. 4, to •••• tai.
ext •• t, j •• tl" tile al.,UI,ill, ... ...,t ..... JUde with I'e,ud to
ox, ... del1ye.,.
TIle otb.... ..,... of cseaol'l,t\08 .f ...,,.. ill th. tl... cI ••
pead.a upoD~"th. detd.la .f ...,,.. otl'utl... ox, .... xtl'actl_
'epa"a ••• e41ff .. ioa paella •• aad .. -JOI' 'actora widell
....... the uy ... alo., tbat sradie.t f .t ....... ud .etabOlla.
Oae , .. , .. ,. _ft .. tloa ia ,..aued to 1' ....... coutut. III -.It .. o ti .... _tabOl1 .. of .,sell 1. I_e,._ •• , .f _"e. teuio
abO .. a cJ'Itleal 1 ... 1 of a few .il1a.te .... of •• eu,. Jt i.
,. ..... ttlat a al_lu .It_tioa exiat. !!!&!!. altboDlIl ao
•• itleal t •• t of tile ... aption hu ..... 11 .ade. "' The otJau llaJ_ facto ... iD auactlo. 18 ato&'a,.. whicll yuie
diz.ctl,. wi til ..,p. t •• 810.. eoaaeq •• atl,. dUl'la, fluctuatiou
la tellsi.a thue la • tl'aDaleat aodtflcatioa of the ..... tated
.tOI'".. Ill. a'ead, .tat. tbe oal, factOl' par .... 4 to be ill
o ..... tio .. i. ox, .. _taboll .. , tIli. factor , ......... 4 to be
.... poulb1. for OM of tile cluu:actel'iatica of the alope .f tbe
diff.dOll .... &di •• t. ID Sq •• tie .. 11 ad 1Ta. _ I' ....... t. the
_taboli .. of _,pa. Hewe.el'. tile fact that ato ....... , be 1 ..
clu4e4 ahould be bOt. i. Iliad.
The ti .... 4iaeaalo .... 4 4itl •• loa ~&dl.Qt ..... tbe
rada!., fact.a ..... k.d to <leeel:!be the .,.t... .9.. I:/J., aa4
r aid ia deta •• laia, the 810pe of the diff •• lon ,radi.ai. Tbe
oaly .... riaeata1 value foe the coeffloi •• t of dil, •• ioa of oXV"
iIa ti .... i. xropta coaa.aat. 12S Tbe ... ipaa1 coa .... t, 0.13 c
of ox,,.. au ... a aqaue ce.tiM'eJ: a.face pel' al.ut. uale.. a
",adient of ODe ataoaplle.. ,.1' 1110.08 at 20. C. t .ol'rected f_
t .. ,. .... t ... aad ebaaaed to fit tIae coac1itiou .f the ."pel'1M"a,
ia 3.1 oX'10·8 ce of OXJse. uNaia, a catl_tel' aquue aufaee
per iii.... Ud... a _adJ. •• t of .. e ai11imeter of "l'e8Y pel:
ceatt..t... ai 370 c. By ...... ae capillary l'adJ.u, 1', aad tile tia.e o,liad'"
1'adJ. ••• a, to deeel'i" tile aya' •• , 1: • .., IlU al.,11fle4 tile calCUlii'
latioa of the ti.... dJaeaa1... fOJ: tb.h' infl.ence oa tb. chaz:.
&Gte .. ia.ie. of til. cliffuioa pa41eDt. This t ..... t ... t aimpllfi.s
IhJUtloa,p to
(24)
Tbe coutut 1./4 •• _ub.a a aUlbel' of ihe factora la l34,,&tl.8 23
which are related to the ratio of 1'/1. The ratio. I'll ia •• ed to
obtaiD the value of 4 fro. Eat,'. table. l6S The Y&1141t, of .aia,
1'/1. la ,..e.ented ia tbe 0l'ie:1Da1 uticl., wbel'e it baa beea de_a
.t~at.d that the poia. ia tlse tl ••• s 1Idticb baa &R OlCJ,ea t ... i ••
equal to the 'atearated ae_ ox".a .... ioa of the ti •• ue c:,lIDd.~
ia !adepeadent of the _pltUde of .t D &ad ll. depea4:L .. 0111, .poD
tile I'atio of r/R. Tlli. fact baa be •• stated i. Y&I'I ... wa,. by
114
Jacoba.208 Hill.U4 &ael aa8hevakJ.109 who bave DOt ,iveD exp1ic1t
matb.aatlcal I'el.tloaabi,. toe tbe CODcept.
fte factora 41 ....... abo •• <1M COt Po. a, A/4, Ita. ad D -
ue tbua .fflci •• t to &lve an _""ox!_tloa of tbe a ...... ,. ti ....
oxyaea tea.ion if tb. ti&aue ay.t •• haa a reaaoaable ai~l~lty
te tbe ai.,11'1ed lIfId..l .. epr .... ted by aquati_ 23 01' tile cleriftd
Sqaatloa 24. Co or Po ue ladepend.at ... iabl •• , - &ad D ue
co .. t ...... ' All tllat ia required to fb tile ayatea foc de.criptiol
i. tile ,..tl0 .f ria aad tile _pitude of It. Slae. a chuB. 1.
flow I'ate, .-eawda, ttlat tbe blOGd ,..a_e padl ••• hu I'.-ieed
co.atant. t.plle. tllat tbere baa beea a chan,e in the ratio of
r;a, two .f Ike .... tricti... ,. the .,plicati.. of aquatloa 13 or
24.e that .. /I be co .. t .. t 41_'., .. a •••• at •• ud 'bat r/R be
alail .. fo. cOl9UI .... f tw a'tuUOIIa i. a ti •••• 01' tbat it be
accarately 4eteraiD.d fo. each of th.att..tio.a. The ....... for "
laeludia, tile ••• alre .• at that flow 1' ...... relati .. ly co •• taat
4ul., me ....... '. a. crltel'l •• tor •• &1otla, tb •••• ult ....
e •• abl1a.. 'I'M tile cODal"l'atl... p.... I. tbla par .... a'h.
TIM ....... crl ..... lo •• tbat te.,. .. atu. be coutut, 1. also a
.ace ..... ,. ........ e... 1. tbe appl1 ... U .. of •• equat10ft to obtai •
.. •• tlute of ti__ • ." •• t •• at... 'fellPe ... t ••• Mdtfl •• til.
blOOd pel" •• io. cIluactel'iatln. _tabeli ... diff •• t ... Ity .f ...,..
.. a, &ad tbe ratio of 1'/1. C .... qu.atlY. each Y&ciatl •• I.
te.,.".., ....... i •• a a Dew aet of par_t .... to cseauib. tile
tl ..... ,a" •• , 0 .. altel'aati •• ly, tile ..... ll'e •• tof .. fbe.
I I
I
I
11$
expert.eDtal t • .,eratare.
oa. othel' aet of ,. ..... *i.D. ,. lap1l«1 t ia the expe .. beata.
It 1. pr.aue4 that the .1ectr04e ie ia COlltact with tle._ wllkb.
haa u a ........ ox,p. , ... loa descrlllte4 by til ..... OXJ,e. t.ul.1l
of the tl..... It ia al .. pc...... that tbe peef.aloll character
lstlcs 1. tile •• poe of til. 81ectrode ue .!all&&' to tbe eIl_act
erl.tl .. etteer'ftCl tOl' tbe tis... ...,le. Obvious iaGheala la the
.... po •• f til. el •• uod. ia4i.oat •• that theae pce • ..,tl... ...... ia-
ftli4. Til. bea' aftil.ble .ri4e.c. that tbe ,......,*1 ...... e
¥all' 1. that .. ben is • 11M. r.1.tlo .... J.' betwee. the ONel'ft.
14 &ad tile calculated 'f'a.t ... , ,.
fte n_~t .f tile ."li.atioD of 'let,'. equaU ••• to tll •
•• ,I •• dOD ., tle,.e ..., ••• taalOft 1. that, for the fir •• tl .. ,
polu ... a"lc ...... e ... '. of tia ... oxn- tau. !! !!!! haft
beell ...... l.ated •• tl'ae bUb of .., .. ~ 'e .. l .. at til. pol&rolfaphle "
... lyala ele.t.04.a. o.lle.. 1 .... tl .. *0... bay. .... the ox, ...
teHi.. ...... i. I'IloM. tale ... t •• f uy,. .tl1iaatioll, 01'
.... plratory ell..,.a 1. eY&1aatl., the ........... t obtained witb
po1uolI'a,ld.c electnd... Al til..... til... t.cllalqae. .. ... d tb.
ptII'fO ... f ftllclatl., tile fact tIl&t ox,,.. te •• loa ••• belll,
_uued, it ,. evide •• f .... tM dianss'o. ,...8._44 In tbe first
put of till. chapte .. tIlat tile, dO aot sly ..... titatiye .atbata
of the • .,,.. t ••• lOD at the .1ectro1e.
At ttao.p there are no .ulal' .... tltatl" eftltlatio88 of the
polU'OJl'apJdc .uuUNtata ia tb. 11 teratue. oft_ 10.atlptl0.8
116
of ti .... oXJ,eJ1 ten.lOA obtaiaed by tbe qualitative polaro,raphic
studi.s aa4 by clas.lcal .'041 •• IDd!cate tbe Yalidit, ., ,be
' ...... t10 •• wIllcll ha .... bee •• &4. ia tbe ... ,.ted 8xpel'lae8t ••
Ptartll_ ... _, the d.ta Z'8po .. ,.4 for flow aDd _t&boli ... e .Wlu
ia aapitu4e to tbe ftl ... .:.,.t •• In tbe lite .. atUl'e.
'italU'ald2OO baa a_ube. tlte eYideac. '0 .. ox,p. diffa.
aioD tIl .. Mp the 8ld... This p ..... ili.., of the stia to ilXfP.
baa al .. · .... r8,.t •• by ....... ,. .. aDd .Mfriead,l3. wIlO ....
a po1.opapllle .. tIlod. I. apl'. of tbe .Id .• ,. .... bl11ty ..
0JIf' .... stadi •• nail as Ul'bUllf.139...41 ladio.t. tbat ... i. UD
atialated sti ... ta1lo1i_ ".,.lIds ,..!aa.il, .,.. bleocl for it.
0..,,.. Apply. TbU .... ,. .... c •• po. MoM fl_ -.14 lapl, tlta'
itA .. _ to uy •• t.o • ..... ce. otllel' .... til. blood ",,'ui •• will
BOt 1IOd1fy tile fl .. U .....
• t.U.e. at tile aolliJ. •• tte PotIa4 .. ~loa of tile Uai ..... ait, of
" .. ,lnal. OIl tile poluopapldc ox,,. ..... i •• ill atia 1ll4J.cate
ttla .. tbe ,..i.-., iaf'1ue ... of ' ....... t ...... bU.. on .ki •• "e.
ta.aloD , .... '0 altUM ... ,..... ""10 ... , •• 164.176-'8.
210-212 ....... ,..t.l.3,161 .twl .... cl ... 1., 1' ..... &1 ..... ked la-
....... ia blOOd flow with 1 ....... 1., ' .... ratare aDd aD ... oci.
ated I'i .. in .ti. OSJIeD , ... iOll ".peed •• t _po_ tlae lut!&1 atate
of tJut akia.TIl. atUi •• '''11'.41,1, laduate tlla' rll Is de
........ by Aac ...... '''''''&' ..... it ... ute. tba, ... il1_i_
wi til &1 tel'e4 ",,,8&'1.. .f bleod " •• w.. 1... th.aa a .a.'e la
YUOdilated. .kla &ad .. loa, ......... ill ....... uict.d. _1e
I' I
I II
:1 ,
117
ad .ltill.
laplu _d Clark186 _d ""8,a163 1\&" ObM"Ye4 the !! DYO
oxyIMu.lobia cli.aociatioB e."., widell baa the .... cbU'ae •• "i ..
tica .. 'lao.. obael'Yed .!Iyl i£p. The poluo,c.pbic.tudie ...
aki .. Juat ... 'ioaed, .. well ...... cited ia ,be 1' .... 1 •• of tile
Ute"atun, ha ........... ied by .e. '0 .... ty 'i .. pel' ceat cSuiq
....... 1 ........... '1. 1UIde •• W1u co .. iti ... .
...... ·,polU'oP .. W.O ..... le. baYe bees Md •• f ..,,.. __ loa
1 ... _18 tllU ba ... f oar ..... ui •• ill sk.u. 0 ... 1 •• ,150 who e
.*de .a.f the .aU •• t .tuell.a, dad ".latl .... a .. p, ia icI
ct818, _rlC18 actiri., to 10110w tl .... _,.. •• Quaeterl.tic ••
.... ....,..,.'. lI'oapt 176-17.206 uable t ••• t ............ itl.it' fol'
tbeil' ttl~.,t,,"e • .I.a •••• ,... ".lied e.ticel, .... "el.tl .. -haac ••
'.ONuftd. iT to iate.peet .... Wlu .... of tile applied expel'l-.-'
... tal ... lOle.. W01Ie"tb161 !1. ti,- &1 ... elied .,.a tile I'at •
• 1 iac ....... i ..... 1 • ..,. to ladle.t. 'be aatue of pOa 1'" _ole
.. 'U Y •• ti1&teel .... aWe"I'.' coataW_, .FI ••••• yul.4.
la,111 884 Id. col1 ..... &,0I", ... wel1- .. Be .... et. al. ,169 - -ba .... bad 'lllilU expel'ie"_ 1a .ttapt. to ..... "1 ..... JlU.lCle
..,,.. ieau ..
Of tbe otlle. elude.1 .t11dlea oa ti.... .,... t ... l .. the
.al... ehtaiaed f~o. ... equilthe.tloa tec-'lquea198,314 will
Pl'0bul, ilave to be clf."elUded. Tbo ... f *,.Gtl'o,... .... tl'le
u.t.e215 01' ttl... depeD4 •• t upoe Calculatloaa31J- 18 .1_lu to
tale ....... ,..ted h til. last cta .. ,ter 184icate yal._ for ox1lea
•
118
teasiOD la 'lbe ............ tbo .... e,.ted III the thesi.. ID_
&.&ly.I. of the pa bUbble ,.cllalqu., a .... a14 reported tbat tbe
.quilibrate .... t ... iOD probably reflect ... DOua ... t ••• l08
"a.tbel' til .. tlu. au '.aiou. TIlle fact c_ld bave beea ••• -
pected , .... tlle .tudles of _ett aDd SC .. lbyatt.219 wIlo cle .... l,
.tated tIlat tbe, ObHrYed tile .pread. of the .... babbl •• &1oa, tile
m.jor blood ..... 1. elRiB, injeot10. of th_ ,u. I. • ... 1.. o. blOOd flow &ad ox,.e. • ....... i.. of ..-ole
wi til e. _ ..... d .... l.becI I.. tid.. tIl •• is, v ..... u 211 obaerftd
blood flows of 0.015 '0 0.6 ~";mi. aa4 .., ... c ..... 'tioa. of
0.0024 .e • OOIS "/Plua 'a tb. Iliad li1lb of a restl .. cto,_
Mll1ikaa20T 18 hia apectl'opbot ... tl'ic detel'alaatioaa .f 0XJ,ell
cOD.8ua,li!,ul ad blood fl_. • .... t.d blOOd flo. at 0.036 to 0.600
.t/p/mill .. 4 0.." •• GODSUllPtl ... f 0.0018,to 0.0310 co/Plala f.r
r •• tl.., cat .. 1 ....... 1e. Mll1ilda:, 41 •• ob ....... d aD laft.... ia ..
blood flo. wileD tile •• rve to JUa __ l. ....p .. atloa lUt.d bee •
..... red. Mott"Wlt .... • Ie ••• utic _pe .. i •• at&1 ,...ooe4 .. e,
.. etro_ade c.tlll ... .riaatl ... f tile .acle Yeia. aad obae .. ftd aa
oxy,.. eo .. uptua of 0.0034 .1/al/._. Tble i.o .. thud luger
til.. tltat repol't.d f. tllia .etl1041. e ... lie... l ...... tl,.ti ••••
320.23 .. ie. Mott~ .. att.ribut •• to 4 MO .. e .. e, •••• at.tl .. v.aou •
• ..,1. ia Ilia ..... iae.... Tile •• y&lues fo.",cle blood fl_
UId 01lyaeD eo ..... ti08 .~ well a. etbe ... a ,ubl1_ed ia tile Hudbook
of 11010".&1 Data,' ,ladieate that .be ... al .... btalaed ill tile
pc.Mat at1l4y ue witbia ......
119
Stuctie. OD ~abbit eat .tia .. taboli .. aDd blood flow baYe 80t
beea .ade by _thode 410.ely related to tile •• e"perilleDt ••
Bartoa224 haa obMI'Yed pr •••• e.flow curyes ia the elltue per
f.aet rabbit eu. Oae factor obserftd by _ton was also Doted ia
tbe present exper ..... t.. Oil. tile preparation (rabbit .ar) re-
lIain. ria ... le. ttl .... e ia .. padual ell.... iD tbe pes-f.aioD rate wi tb
tille, IndicatiD, a differeDtperf.aioD pattera. This .. , be
.... clatett-,witll _erou. a-......... tUlO ••• O"Ml'ved 111 the rabbit
8&1'.22'-28 'lbe '9'&1 .. a fouad for *in blood flow Uld ox, ... con
suptl •• la tile HaDct_oll are of tile ... orelar of a.pltude ...
tJaOM reported in the ,..eft ... GUptaI'.
0 •• factoI', tbe ratio of I'/a '01' m.aole, 'S aubataatlat.d ia
tbe lite .. at.e, blat tile ... l.e uaed f. atiD could DOt ":verified.
la his lldtl .. l .tadiea ... the capillariaatio .. of _cle. b'op23'
lDdicated tbat tbel'e i. • __ ut a 0.. to one r.1atloaabip be ..... "
Hacl. filM .... ad op ... capillarie.. Tbla .... alao obs.r .. d by
... tiD et. &1. 230 aDd ia the .... artB.d ~alue reported ia tbe - -UaadbOOt' for reatia, steletal .... 1.. Clark'a obs.rvatio ... a2S-21
.. a _11 as tho .. of 0*""1.'.38 OD the 1.cl te cluullMr .tudie a of
bl004 flow ln tbe rabbit ear iDelicate that the capillar:y ai&e used
for rabbit ..... (3.' .. l'acU .•• ) 1. appnxlaatel, I.'iabt. SillCe DO
01&1. ia aad. for tlleabaclata acc_~y of tbe bloo4 ft ... l elia
•• ter, _acle fiber cU._tel.', or tblckal!U of the .kia. it ia
aYideat that tbere la a poaalbilit, that tbe ratio I'll i. ib .I.'ro~
TIlta .111 baYe DO effect OD the liaearit, of the calculated ti ....
lao OX" •• tensioa8 .iaee it will oDl, IlOdif, the fiaal value by a
fl'&ctloD of tile coutant A which wa. uaed. let,163 haa .bOWD tIlat
aD 81'1'01' ia a. wIllch ia the" .. pel'l.eats should ha .... beea DO 1101'.
tluUl tweaty pel' ce.t f 1'0 a the tl'.. ....1... dou,b1e. the .al.. of til •
•• ti .. ted pOa fol' a 501 aDde"e.tt.&te of R aa4 hal ••• tae valu. of
tb ••• tiaat.d pOa fol' a 401 o.eS'e.tl_t •• wi til ..... t.el illc .......
fol' lu,." el'''ol'.. .eace. the UMiat_ yal ... of tl •••• IJ02 haye
DOt beea' •• ttlraiDe. by the _tIlocJ of calculatioD. At tbe a_
ti_ it i. evid.at that if tbe ratio r/R ia coutant daria, u
experi.eat, tile calculated pOa will bear a constant relation.hip
to the true ,oZ. AD approxiaatloft of the a.lllitude of error Which
mlibt exist can be obtai.ed from the limitatioD of altered blOOd
flow to witllia t.a pel' CeRt of tbe cOIltrol value and applieatiOll
01 POl ... ille t • L.w. Siace flow will vary d1rectl, with a4, it i •
• "id •• t tbat a ten per ce.t lDcr ..... i. flow ahould limit cbaa,e. "
in tbe capillariaatioa of ti •• uea to .ufficiently ... 11 value.
which will .ot appceciabl, modify the a1.tea ~ich was beia,
.twlied. The tea per cent 11.1 t chos •• for th •• e experimeats re
preM.t. a chaa,. of abOut one capillary per huadred already
acti.,. ••
la au_ary. l.t,'. eqaationa aDd the simp11fication. which
are .ec •• aary in order to gain an approximation of the t1 .... oxy
,en t ... lon iadlcate that, witbia the limitatioaa used to •• aluate
the experi_atal data for co.,ar18011 of polaro,raphic aDd calca.
lated tl ...... oXJ,e. teauOlla, the calculated tisaue oxy,ea t ... 101l I
I: Ii
121
should provide a reuonable appJ:oxiaa.iion of the OllJ' cen ienaion
exiateni at the electrode. Tbe polaro,raphic literature on tie •• e
oxy,en t ... ion indicatea that the presumptions .eed in these ex
peS'iaeats are luad_eatally aoUDd. &ad that tile quantitative eyal
uatioD of the pol&.op'apbiC ...... S'e of tieaue oxy,.. teDalOil ia
the flrat to ha.e beea reported. !be calculated tiasue oxy,e.
tenaloaa, Mt&bOlia rat •• &lid blood flowa obaerftd la tb ... exper
iaeata •• ·of tile a ... 0I'4er of u,Dl tude as th.a. r.por ted in tb.
lit .. atllZ ••
,I
1
CHAF1'JiIt V
CONCLUSIONS
Tbe illYeati,atlOl1 of tbe accuracy of poluosrapbic _aaue
menta of OX".. teaaioa ia 'Ii.... baa made cODt~ibutioa. to sever
al uea. of bowledp. Thea. uea. iacl.de 1) additional laform ....
tioa cOlleeraia, tbe beb.vior of oxy,e. In aolation, 3) evaluation
of two aetbods of .. aaurin, tis.ue ox, ... teasion. ..4 3) ladi
catle.s (or,the lut~e 4e .. lo,...". Deeded to .ake absolute deter
lliaati.u .f ti •••• oxy,ell tea.1011.
A method baa be •• de.eloped aDd lutrUMated for the detes-
ainatio. 01 the •• 1u. for tbe diffusioa coefficient of oay,e ••
Throup tile UM of this _tbod a val_ for the GOeffiei.at of 2.23
, I
!
x 10-' ca2.ec·1 baa beea deteral_d. It h •• beea iBdieated that I
furtber applic:atioa of the .etbod will re.~l 'I ill the <leteroaatioa I'
of the infl.eace of factors in tile .,atea upon dllf.aivit, of ox,.
,ell. Th. _thod will alao provi4e information OD tbe dependeace
of the "'fficient upoa tbe -.ture of the dilf.aioll ~adieat. IR
acldi tioD tbe .. 'iJa04 CaD be ateaded to the stud, of 41 flusioR ,.
meabraDe. of oxy,.. &a4 lona. DiffU8ioD coefficient. deterained
i ... polat'e)grapbie diffulo. cell should bav. tile sue acewao, u
diff.aioa coefficients studied i8 a refractoaetric or int •• fero.
metric cell, because the accuracy aDd priDciple. of tbe .etbads
ue aWlar.
,oluoarapbic a.uuremen t of OXY,fUl t~D.SiOD 11 ... be •• compared
-122-
r ~----------------------------------------------------------..,
123
with tension calculated from experimental measurementa. This
eo.parison supports the hypotheais that £2lar0l!apbic m~aa~e.eat
of tlasue omea tensloll baa the sue ~Ilherellt aceur a.cl aa the
accuracy obtained ill DOD-ttl •• ue .aaurePt'en;'.. SiDCe the compul
eoa of poluo'l'aphic and calculated OIty,en tension dur in. stand
ardized conditions io the tissue iDdicatea that there is a quan
titative relationShip between the two measurements, it ia reason
able to ...... that pol~o.raphic diffusion current indicates iru
oxygen tensioft of ti.aue '''0 durl., va.latloD in the par .. etel'.
of ti .aue oxypn availabill tf. The .. thod of eval~ation of polar
osraphlc meaa~ .... t. for ~iabillty and reliability baa identi
fied factors in both ti .. ue &ad th. polarop'aphic _thod which
will contribute to the DOn-qDaotitative nature of ti.sue aeasure
.enta which have bee. repo«ted in tbe literature. Analysis of the
abaolute valoea obtai.ad for the two .•• tlma tea of ti •• ue ox,.,en .,
tension iadleate tbat aeither metbOd caa give a defiaitlv. value
for avera,. ti .... OXf,en tea.ioa without further modiflcatioD of
the tecbaique of .... ur ... at. The fiDal oont~ibutioft i. to
au.p.t tbat absolute values for ti ... e oX1,eo teD.ion can be
obtai.ed _., _ electrecle whiell caa be calibrate4 aDd l'aaiu ia
depeadent of ita environment.
r U&IO<I\APHY
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3. Heade .. aoa, lAWl'ellce J. 8004. tale Vld,verat t, 'P .. e •• , New Raven, 1938.
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TU1. 241, ,. a60.Tl •• u. OXypa Co_ptloa. ADi.al .••
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Table aD, p. 210. " •• piz'atory CllM acta.i.ti.a. 11004.
Table 213. P. 283. Blood Plow. Ti •• ues ad Organs. MaJlllal ••
.' Table 214, p. 284. capil1a1'y ' ... cal .. lt" Skeletal
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-124-
'/ !
1,1,
125
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,
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I' " ,I 'I
I
136
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"
29. Ko1thoff, I. M. aDd Jocdaa, J. "EXaltatioD of the limit-1., curea •• f oxr ... by Ii.,..,e. peroxide at rotated 1014 81ectrode as a ..... for .... I' ... t~lc dete.ainat! •• a of traces of OXfle •• " ADal. ell ... !!.10Tl.?a. 19A.
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1
" I,
121
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48. Vitet, Y. ",oluo_apllie atudl •• with tile <1I'OPP'81 mercury c .. thode. Part LVII. Tbe eatimation of ox"ea coata ned i. g .... aDd "1"tiou," COllection CZech. eMil. COIla" .. 1,.53': t 193".
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129
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- -
61. , ... to A.M., ......... , P. At. &lid Nlldtia, D. "Po1Uo-paphlc aa4 pIlot_trlc 4.t .... Jd ... Uoa .f oxysea la water," I ... data, .. Lab. .!1.3' .. O,· 1939. ' ... 011 ehe_. Abat. .!!11S87, 1940.
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"
64. Ie_. W. aacI All... W. ".,.wo __ hie. 4e ... _l ..... 1 .. of cIl ... lft4 .,.,.. I. wat ... COD"aillill, lata ... lal efflu.at," sew. udlad ..... te 23.'12-17, 1950. -
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:i
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! 'Ii I
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r 133
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ll
i
CIl ... 21,,40'.428, 19.55. . -101. Cll_ca, .1tt08 aad Wl111 .... , G. a ..... e.'irator' ... p.. il
III oxlet •• 'ye pho .... ~'1atiOD. If_ Tile ••• pbatory chal •• " ,.J- Biol. I Cbea. ~".429-43at 1955. . " . II!
102. Cbaaoe, Bl'ittoa, willi ... , G. I.., Ho1ae., W. P. _4 Hl"l.. J.. "I.e.,u&t •• r e.,..a ia wel.ts. ve pbo.,b.o~,latio.. ,. i
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103. .. ...... , I. M. "Tbe oxr ... deficit of arterial blood c ... ed by D ••• e.t l.tiD, parta of the lua,.," ACta Pbyato1. acaRd. 4 auppl. XI.at.54, 1.42. -
104. ll'_le,. A. P., Stupfel. M. ud S •• eriapave, J. W. "Iffect of tape •• tu •• 011 Pco:a .. " Po2 of bleod ia yltl'O," J. Appl. ,h,alol. 1.201-05, 1956.
10.5. ....cu .• ta, I.. ..4 Tropp, C ...... laro_aph:1c atwll .. e o. blOOd pipeata &ad tbe r der1 ... t1 .. 8. I. ACtl •• tioa of hydro, •• pel'oxide by h_llobi ... 4 lIe_tl.," Bloche •• Z. a8,,301-12, 1937. -
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133
106. Dauber,_, J. Percl "Tbe ac~uate 4eterai_tioa .f he_,lobla, oxyllel1o,lobin and ~ubon IU)JlOxide by MU8 of the 4ro"in, mercury electrode," Am_ J. Physio1. !a!.308P, 1940.
101. BaWlbe".-. J. ,ercy "The ..,pa tell.loa of _ole blood ....... 84 polu:op'aphicaI1,," latera. Pby.lo1. Coa_ea. XVII t OXf~d, 1941. Abat. 113-14.
108. 8&uaberpl', J. 'erey "The reduc.tioll of OX" •• by _laO,lo.,ln cOIlpOunda ia acid and a _thOd of poluolI'apld.c ... al,.18," 'ed. 'roc. 6.16, 1.41. -
109. _bel'S-, {_ '.:re, "Reustatic chu,es in ltellOlloblll, blOOd YOl ... aa4 ba.. ..tabolic :rate 1a rata with aarcOll&," All. J. 'h,.iol~·' J.!!.30a-09P. 1940.
110. Ty.U'owald., w. "pOlu:o,raphic: eletectieaof hemollobill boult4 OX,pll,· Meel. Do .... ede.at .. 1 Miluobiol. 1tI31-40, 19.51. Pro. Cbea. Abat. ~.4602, 1 • .s3.
111. Wied.,er f E. "Ole Poluosraplli.ehe Me •• un, del' Saueratoff.,aaaua, 4ea l1.te .... Relv. ,hfSiol. et. ,~.. Acta Sa"l. Dl11-aO t 1 • .50.
112. .....el., R. u.d Laue, D. "TIle potad_etric ........ e .. nt of exypa pn .. ". 'a _ole bloOd,· "lu,_ .ucb ••••• ,by.l01. 354.101-2,5, 1954. ' -
113. lee.er, B. 1:., Pellu.bee, :,a.. MUpll,. A. J. aa4 Crai" p. w. tlDeteni .. '1!o. of the oxy,en coateat of .. all _ua •• of b04Y f11l148 by pol_oaraphi.: _al,si.,'" J. BiOi. CIl.a. 146.191. 20~, 1942. ---
114. ..l.i ..... , I:lau8 ttpola:rop:apbic deterlliaa1ion of oxr,ea teasioa 18 u.saturated blood," Bely. ,by.iol. et Phar.acol. 6=CI3-14, 1948. -
115. Wll ... , R. H., Ebert, R. Y. t IOrde., C. W., '.ars .. , R. T., JobaMD, a. S., Palt, A. aDd Dempse" M. B. "Tile deterain&tloD of blood flow thl'oupaoltftDtilated ,..tlolls of tile ftOl'ul. and .1 ..... 4 lu.,," Aa. R.y. Taberc. 68tl11.85, 1953. -
116. WilaoD, a ..... l B., .uce t B. Jay aa4 HapdorD, Max B. "Collput.o. of the aouslltoll-Scholuele ... a1C!af-, and pol_oarapbie _tbod. for ... urla, te •• lo. of .." •• ill P ..... Ud _ole blood.· J. Appl. 'bru01. 8,.513-23, 19.56.
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)1
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118.. Cluk. Lelaa4 C. J".. Wolf. R., Granp .. , D. a.ad TaylOJ.'. Z. "Coati.aoua I'ecol'di .. of blood oxygen t ... ioa by polaro~apb't" J. Appl. Physio1. !sIS9-196, 1953.
119. Clark, Lelaad C. JI'. "Mordt_ aad coatrol of blood. and tiaa.e OX" •• teD.loa.," Tl'aaa. Aa. loc. Actlfie!al Intel'8al 01' .... 2,41-46, 19S6. -
120. Sproule, Brig J. ,Nil1el', WII. P .. , C •• bl.C. Ina J. _d ella,..., C .... leto ... "All iapl'oYH poluo'l'apbie _tbod 101' ..... 1 .. ox,,.. t .. ai •• ia whole ttl004," J. Applie4 ,",.101. 11.365-fO, 19.51. -
121. ..n .. iacb .. s, J. W.".4 Br&41e" A. P. ttEleetrodes f. bloOd OJ ...... coa .. t ... lutio ••• • .lXld ... t. ,eelel'&tlo. of Allel'l_ SOCietiea fo .. .axpel'i .. atal tiolo.,., , .. iladel,llia, 1958.
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laT. CI_Il. I.. a •• Jr •• He1uwortb, J. A •• "apl ... I. t au ••••• T. &ad Taylo., Z. " .. lUOeapblC .. __ .eMat of ox,.e. t ... ioa In wbole blOO4 ad tl •••• dtU III total .. " ... • 1 beut," au"cal ...... 4,93.". 1"3. -w. _bUpl', J. P. "Det.l'aiaatl .. of •• oZJ'_ cll.sociatin ..... of 0xy"e_atobi.a by a .... ethod," All. J. R,.lol. 133. lOP , 1938. -
129. "' __ PI', J. Pel'o,_ .una, G. ad lUdVel1, ~. "Lidtatl .... fthe poluo .... .,hic _tbOtt of detentDa"101l of oxy,.. teaai •• 18 Whole blood." 'ed. koc. StS-f, 1949. -
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130. Dirken, "I. N. J. ud Heeutra, H. "Alveolu oxypa ~; t ••• iOB aDd IUD, circulation," Quart. J. Exptl. ,h,aiol. ~1193-211, 1949.
131. Di.kea, M. N. J. aDd Heemstra, H. "The adapt.tioD of the lUllI circul .. tloa to the "Btil.t1Oft," Quut J • .Exptl •• It,dol. 34*213-31.. 1949. -
132. Norpa, B. H. a&ld Kabu, Gakiel "StUdy of "elettOllshlp of .... t ... tal oX'P" t._i.a to .. 1 .. o1u oxy •• n pr ....... ill aau, uaia, poluOII.tric _thOCl for Whole blood." M. J. 'hyaiol. !!.!a136'. 1950.
133. Be"', It., IUtela t H. aDd ltocakeWald., H. A. "0" '_1'atoff diaaoaiati .. atu..e d •• fetalea 81ut •• and 4er a..au.tauche. del' aeaacblicbea PlaceDta," ,flulel". Arcb. gea ,hyaiol. 360.306-19, 1955. ---
134. lIOchi •• te t M. ".01 ..... 011'.'1110 _asureseat. of oxy sea coaceatratioa ia viVO," SODe a.,.arChe. iD BlO~~.iCI' Y01. a, Y • .baai, eellt_. I. ... ua !llrtuh 01 XPJ11e11lG "icity, Bokbaido vai.el'sit,. lappeto, Japan.
135. Reefta, R. L t ReDDie, D. W. and 'appnheiJDel', J. I.. "Ox,PD tea"'OD of ui... aad 1 ta .'plfle_c •• " Pe4. hoe. 16.6'S. 1.51. -
136. 8au11b.~I •• t J. r., _4 OOodil'ieacl, I.. B. "Det.I' ...... UOD of "terial oxrP. teadoa ia ... Ify equllibra.l.D •• oup latact .kin." Ped. troe. 10.11, 1951. -
137. ..oak, D. W., Davie •• P. W., Brill", P. JI'. aDd Larrabee. M. G. "Oaypa •• pply &n4 ....,tea ...... tio. 'a tile Bel' .. a ay ••• ," ftaaa. All. Neuol. MU. 141-144. 1944.
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139. Urbach.'. cd Pl ... c. t G. ·POlu., .. aplalc .. uue.at, of lb. oa" •• co ..... tloa of the ski_ 1a Y1vo," Scl •• ce 112,78.5-86, 1950. ---
140. Uz' .... Il, F. -,athop .. ,ai.lo" of Mali~,. I. Ti .... 0&7, •• , ... loa Of beni,. aact mali ..... "o~. of atill," Peoc. Soc. lX,.r. BiOi. Med. !!s644-47, 19S6.
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143. Davies P. W. aad Grenell, It. G. "Effect of bigh pot ...... lUll OIl I'eapkatioo Ul4 detlUcatioo pote .. tl .. l of a small locally pe:1'1."d U'ea of cel'''a1 COl'text" Pe4. 'rO'c. '#29. 1950. -
144. Daylea,'. W., Broall, D. W. and GreHll I.. G. "Tbe tl .. cou .. of ill ft." .. "eD $O .... ,tio .. of cel'e.al co:rtex fol1owla, electr:lc:a1 ".tlaalatlOJ1," ,ed. 'I'O'e. 1s25, 1948. -
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146. • ••• 11, I. "1· Daviea. P. w. &lid aronk, D. w. "Bfleeta of ".da aIlemia GIl COl't cal ox, .... CO.S.,tlOD 'a ft ••• " Ped. 'I'oc. 7.44, 1948. -
147. "oat, D. W., t.uJ:abee t M. G. _d Davie., P. lI. "The rate of ox'1". c __ ptloa ill local,Ued l'e,10uof the .U"1I8 .y.tea_ la pre.,...,t!c eacll .••• &ad cell bodles," 'e.d. 'roc. 1.11. 1946.
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151. Day:1_, B. W., McCullocll f W. S. aad aoe.m ... , E. "Rapid ch ...... in 02 ten.loa of cerebral. cortex 4urial induced C08ftl • • ioaa.·'Aa. J. ,.,.biat. 100,83'-3'. 1944. -
1$2. Davie a. P. W. aDd aOOlld f A. tt()x1.ea eonauapti 011 of the ceJ:ea.a1 COI.'te.x of the cat dulac lIetauol cOIlwlaion.," h'oc. A.soC •• e •• N ... 4 Meatal Dis. 26,305-11. 1.46. -
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1'6. .. ~. J. J. f Sheldoa, W. P. t Henri.s, 0., 1:-, P •• ,'eI'M, G. . Ii ..... JI'. f It. '. U<I Mea4 t J. JI'. "studl •• of c.o..,. 41Mue 1. tile exptl'b •• tal "bal. II. P.l"op~bi. 48t8--'8&t1.8 of 1 ... 1 oar ... a •• ilabillty ia the do.* left veatl'iel. dual., COl'oaary oeel •• ioa and pur. oxysea be.atbla.," J. Clia. I .... t. 30,931-40, 1951. -
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160. .. ...... L. B ••• Bud. W. L. "'0 1 uo • ..,hl 0 ox, ... at.siea wi ttl apecia1 ......... 011 .,.p ....... tabi1l t, iatlle do, brat.t " 'roc. •• D. Acad. SCi. 33.50-51. 1953.
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.... luopapldc •• di •• Oil tbe Cll'01I~atioll of tbe do,. a.1aU.aalLip of ..,. ... t ...... to blood flow in isol&ted d.eryated. I'e.tla, ateletal ... 1_," Sue,. Po.-.. 5t1.57.161. 1954. -
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163. Peuys. It. and MOil:'"."" H. noxyaea i.udoll of iln. by the polu0ll'&pJa!c .. etlled. V. TIle rate .f lIOYement of OXYPll froa the .... lph_"at art., to tbe ald.Il," J. Clla. laveat. 31t1042-S5, 1952. --
164. MODtpae.cy. fluSh aDd HoI"wlta, O. "OxYPJ1 teuiOD of tl ••••• byibe polarop'apbic _tIlOd. I. IRtl'OdacticlU OX, ... te.ai .. aDd blood flow of tile .kin of bUllUl ex"eatl.a." J. Clla lo ... t. 3.,1120-30, 1950. -
166. ~et" Ie,.., S. "Deterld ...... f ti ..... ,,.. t ... 108' Ped. Pnc. !!.66 ... "O, 19.57 ..
166. lAabel't".t c. J- 1 BuDce, 8. L., Drabld.D t D. L. t &ad Scblddt.C~P .... latl __ It .f os., ... ,.ulo. to 1le1lO,1 • .,!. ox,p •• &tU'&t:loa la the _t ... 1al "loed. of D.a&1 -," J. Appl. Pllya'.1. 4,873-78, 1952. . -
167. PeaDe'.t Ra)'llOad "OX" •• tendoD of the ti •• ues by the poluopaphlc ... bod. IV. Sttila _.,... tea.oe ••• uterial ox,p • •• tvatloaa relatloaa"l, to oxybe_,lobln di •• eclatio. CUI''",'' J Clia. laye.t. 311204-208, 19.52. -
168. Mollt,... .. y, B •• 1i.Daaez, H. P. t J •• Uld Hor:wita, o. "0&,,. ... oaloa of tl ..... by the polU'op'aphic _thoo. II. OetectloD .1 .ilbt to left ........ by ....... ill RiB oxrt_ai.~! ... 1tl., fna lallalatioa of oay,ell," Ci.culat 08 2,84.5-849, 19SO. -
169. ~,Y. G., arl ••• , W. it •• Malap., CL It. Md-, .---1 P. w. ~ul ... of uterial 02 sat_atioll witll tlaaue OJ 1 ...... ," Ped,. 'coo. 1.10, 1.50.
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APPROVAL SKBBT
The dissertation sub_itted by Albert P. Kelso has beea read
_d app#oved by five members of the faculty of tbe Graduate
ScMol.
Tbe final copi.s kaye been examined by the director of the
dia.rtatloD aDd tile alpatura whicb appear s below verif i.. the
fACt. that any neoe.au., ell.,.. kave been incorporated, and that
the di •• ertation 18 DOW pyen final approval with reference to
coatent. for., and Mchanica! accuaey.
The di.sertation i. therefore accepted ia partial fulfill
.ent of the reqdl'eaenta for the Depee of Doctor of Philosophy.
Sip.ture of Adviser
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