Document rxJ8nx7NRgXDj93ykZrrEVpEq
BIODEGRADATION (WARBURG)
TEST SUBSTANCE
Identity: rPeefrefrirueodrotoocatsanPeFsuOlSfAonAy,laFmCi-do1(2e8thoyrhaascetthaetem;ajmoarycoalmspoobneent of FC-127, FC-129, FC-109, or FC-109-X. (Glycine, N-ethyl-N[(heptadecafiuorooctylsulfonyl}-, potassium salt, CAS # 299151.7)
Remarks: The test sample is FC-128. Current information indicates itis a mixture of 86% test substance and 14% C.-C; fluoroalkyl carboxylate: compounds.
METHOD:
Method: Warburg Determination
Type: Aerobic
GLP: No
Year completed: 1976
Contact time (units): 7 hours
Inoculum: Activated sludge mixed liquor collected from the Metropolitan
Wastewater Treatment Plant, St. Paul, Minnesota. Liquor was washed,
suspended in a basal salts medium, and used at a concentration of 2000
mg of biological solids per iter.
Analytical monitoring: Oxygen uptake
Temperature: 30C
Test Concentrations: 1667 mg/L glucose control, Endogenous control,
20 mg test substance and 1667 mg/L glucose combined solution, 20,
100 and 500 mg/L test substance solutions, and 500 mg/L and 1667 mg/L.
glucose combined solution,
Test Solution Preparation: Dissolved 1.5 grams of FC-128 in 500 mL of
DI water to make a 3000 mg/L stock solution. A 5 mL aliquot of the stock
Solution was added to 20 mL of DI water to make a 600 mg/L sample. A 5
mL aliquote of the 600 mg/L sample was added to 20 mL of DI water to
make a 120 mg/L sample. One half mL aliquots of one of the above 3000,
06.050
maLndDI12w0atmegr/oLr
s0o.l5utmiLonsgliusctohseen
added control
to 2 mL solution
of to
the mixed make the
liquor plus final test
substance concentrations of 500, 100, and 20 mg/L.
Remarks: Described method lacks specificity on test procedures.
RESULTS
00297
wAiptphrionxtihmeat7elhyou7r0e%xpoefrithmeentthaelorpeetriciaold mifabxiiodmeugmraodaxtyigoennfuopltloawkeedotchceurred theoretical degradation route: CAFySON(CHICH,COO K + 4250; + H'> CAF SONH, + 4C0+; 3H0 + K' eRxepmearrikmse:ntOxygen uptake appeared to be continuing at the end of the CONCLUSIONS This testing indicated that the hydrocarbon moiety of the test substance potentially biodegraded in aerobic environments capable of supporting biodegradation Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY bReiloidabeiglritayd:atiKoini,mibsucthornalnykfionrg a3.theToersettiicnagl ipnadritciaalteddegarahidgahtiloenveolfotfhe compound. Sample purity was not properly characterized. Impurities may have significantly affected recorded values. Test method, raw data, and an accurate description of the procedure followed during testing are lacking specificity required to insure validity of the study REFERENCES This study was conducted by the 3M Company, Environmental Laboratory, 1976. OTHER Last changed: 5/16/00
ents
FormSivar
.
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T TechnicaE Corruncatons
3m TECHNICAL REPORT SUMMARY
pare. August 12, 1976
J -- LABORATORY, DEPT. NUMBER _ 0222
"licrororn copies: |.FriBeioondegradation Studies of- Fluorocarbons
Fo
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Fate of Pluorochentealsin the Environmental
wR. 1. Bohon
|[_Ee. nA. Reiner
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ox iIFnfSomUaMtMioAnRiYn RhiEsPOreRpTat
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Tem12 ei
bean coTreedbyether roprts (FGSTRRCT and Conclusions. (System can occommedare 00.250 wordt)
" patty
Biodegradation studies using a Warburg respirometer were conducted
Plses i7y7oCmomdplmoiroendion | FonM 3F4C2-29."5, NFoM b3i4o2d2,egrFaCd-a12b8i,litayndwahsydroobgseenrveadnaolnogsFC-o9f5,FC-a9l5thoaungdh an
reFignidvieuipuudneinmaimnrdieomirosas|| Faplpr3o4x22imaatned hFyCd-r1o2g8ebnotahnawleorgeodfeFmCn-s9t5rawtaesdretaodiulnydedreggorasdoambele.
`Sa--twyr--cione--mc--aL--RE--GIS--TRY|| bsFil3oudd4eg2ger,aSdtauftdriyoomnw.etrheeAtuWtnaesrmubpcutcrsegsstsfotuuldi.iseosl,ateanddegfrraodnataiosnubpsreoqduuecnttsaocftivated
+Hg ichvaicals roevpeorrt
S SuetngrTere,vnwsopaoemsci, and
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EEEnvronPC- - AsDisve.ss BaBciotleorgiya BFilousocrorceheenmiincgal Biodegradable
. SrPEeCIiFnICePROBLEMS remaipningEta--rea_ch s--biec--tive--. ------------------------
bCoinotdiengureaddataitotnempptrsoduwcitlsl boef mFaide342t2o ainsdolaotteherand3Midfelnutoirfoycartbhoens. Work with radioactivity labeled FM 3422 is being considered.
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BIODEGRADATION STUDIES OF FLUOROCARBONS
SUMMARY AND RECOMMENDATION
oNfoFbCi9o5degirsadiamtpiroonbabwlaes boebcsaeursveeditinisWarcboumrpglestteuldyiefsluoorninFaCte95d.. TBhieodergersaidsattainocne hoafstnhiost yceotmpboeuennd tdoemboinsotdreagtreadd.ation by an acclimated microbial culture, however,
Woacrcbururregd.stuTdhieesproonduFcMts342o2f athnidsFbCio1d2e8grboatdhatiionndicaraetendotthkatnowsno.me bSieomdiecgornatidnautoiuosn afcitnidvinagtse.d sludge studies on FM 3422 did not confirm or disprove the Warburg
wFouutludrebeinvgerseattilgyatifoanciolfittahteedbbiyodetgheraddeavbeilloiptmyenotf tohfeanflaunoarloyctaircbaoln pcroomcpeoudnudrse
"
ftohre pFrCe9s5e.nt Wfairnbdiunrggs.studies using purified FC 128 should be made to confirm
Seutied.on the biodegradability of TH 5422 wore hidered by its low waeer
Sloalbueblielditoyn. itsThihsydprroocbalrebmoncopuolrdtiboen opvreorvciodmeed utshiinsgmaFtMer3i4a2l2 hraaddaioahcitgihvitsypecific
aocftitvhietylab(e5lemdcic/onmpmooulned) caonudldpbureitmye.asurBeidodbeygraddeatteicotningof14gC0s;atuervaotluetdiosno.lution
a) INTRODUCTION
Tshteudysusocfepttheibielnivtiyrontmoemnitcarlobfiaatle moofdiafnyicactliaosns iosf acnompiomupnodrst.antIptariasmettheermoisntthe
important for of degradation for organic compounds.
ASovavsatstarirnayfaoctf otrhgaatniict wcaosmpoounncdesbeclainebveedcboymplseotmeelythadtegrgaidveedn beynomuigchrotoirmgeanainsdms. Tthhiesprdoopcterrinceondoiftimoincsr,obmiialcroionrfgaalnliisbmislictoyulids dsteiglrladea acnoymmoonrgmainsicconmcateeprtiiaoln.(l).
PAelrtfhlouuogrhincaotmepdouncdosmpowuintdhssianrgeleextfrleumoerliynesrehsaivsetanbteentosbhioowdnegtroadraetlieoanse (2f)l.uoride nieonvserasbeeanresshuolwtn otof buinoddeerggroadnaattiuorna,l pdeergfrladuaotriionna.tedFocromptohuisndsrehaasovne,rnaorelmyodoirfication moofdithfeicpaetirofnluoorfoictsomhpyodnrenotcsarboofncocmopmopuonndesntsinsteheimsedsptousdsyibwlaes.anticipated. However,
Aennviurnodnemresnttanwdiilnlg obef tehxepopsaerdtitaol tdheesgerapdraotdiuocntsproidnucatdsditiisoni'mptoorttahnetusnidnecgeratdheed materials.
METHODS AND MATERIALS
Chemicals
\
The chemicals used in these experiments are shown in Table I.
000200
2
Ny
TABLE I. CHEMICALS USED IN BIODEGRADATION EXPERIMENTS
FM 3422 HydrMo3g4en22Analog of
Cas
CgF 1750,NCH Cay750,N(CaiHs O
FC 95
CgF17505K
H
Sipex-ols
Cait 7050 Na
"
3
FC 128
.
CCMHs
Cg,7S0pNCH,C00K
3
TTShheeepyteFwmCbeer9re5, oub1st9e7ad5i.nveads FffMrroo3mm422Dloont(NR5-i8e3ct.keFrosLeootfantlhucemobhoeClro)smewwraecsrieailndoetCnhtegimifiviceeandl afsDoirv7i8ts8hieCoCnFii7n4354-222.
hydrogen analog or the sipex-ols (RM 26442).
Tfhleusoeroccahrebmoincalcsonwsetrietuesnetleocfteda floarrgea nuummbbeerr ooff Teasfolnuso.rocaFrCbo9n5 cisompeosusnednst.ially the, FM 3422 15 an intermediate in the production of SM fluorocarbons, and FC 128 is
|
vaerfeinissehlecfltueodrofcoarrbcoonmpparroidsuocnt.to Stihpeexf-loulosrocaanrdbotnhse.HydSriopgeexn-oAlsnaliosgaonfaFpMpro34x2i2mate
hlyadbriolegenflauonraolcoagrboofnsFCh9a5v.e fTrheeqsueenhtyldyrobgeeenn afnoaulnodgstowbereegtreatsutietdobuesclayusdeefbliuoolroigniactaeldly
bmyicreonbziyamlesgwrhoiwcthh noonrhmayldlryogernemosvnealoaghsydcroougledn.selTehcuts,popiutlasteieomnesd porfoboarbgalneistmhsatwhich
`could more codpletely degrade fluorocarbons.
WARBURG DETERMINATION
W{aArtbtuarcghnesnttu)dieMsicwreoroegracnoinsdmusctwedereacccoorldliencgtedtoftrhoem tahtetamcihxeeddstlainqduaorrdopfrotcheeduPriegss. Eatyeatrceoantcmeennttratsiyostnemo,fwa2s0h0e0d,mganodf bsiuoslpoegnidceadlinsolaibdassaplersallittser.medium and used
WtahteeWraribnusorlgubflleasskusb,strEamtuelsswieornes weemruelsmiafdieeduisninwgataerBlparcikosrtonteo maodddeiltioEnP-2to
ultrasonic probe, base 1/2 inch, at 100% power. Logarithmic dilutions
A
iSnidewataerrm Solution
wooefrredtehmieaodnWeiazrebodufrwgtahetfelrtaseksistn. stuhbiCssotnrtsaritodeless,acmao.nndtaTi1hn/2eedmsle1c/w2oansdmlpsloiafdcee1d0argimn/lcotghneltuacifonisreesdt
either glucose or deionized water.
0c0o291
2
3
.
Owxiytghenreaudpitnagkse waats10f-i1r5stmionb.serivnetdervinalseactho felsatsahblifsohr athpeerenidoodgewnpoutso a1c.t5ihvist.y.
)
mTohniistowraisngfolflorowaepdpbryoxaidmdaitteiloyn2ohfrst.he Afdidristtiosnideofartmheansdecocnodntisnidueedaorxmygen
ceovnatlauiantiinogn golfuctohsee,toaxirceiatdyiolyf dtheegrpardeavbiloeusmlatyeraidadle,d maaltleorwieadl.a further
Semicontinuous Activated Sludge Studies
AonweFeMk-3l4o2n2.g seThmeicomnitcirnouoorugsaniascmtsivuasteedd wselruedgeobt(aSiCnASe)d,stasudbyefwoarse,confdroumctPeidgs Eve T3reSaCtAmSenrteaPcltaonrts. anOdnetaHpuwnadtreerd Fasiftaycmolntrooflatcotiavatfoeudrtshl.udgeFMwa3s422adwdaesd taodded taoddi3tiroenactionrcsrebaesleodwthteheFwMat3e4r22sucrofnacceentrianti1o/2n mbly o3f3 magb/slo.lutePuraelcoehtohla.nolEawcahs saedmdiecdonttoinounoeussluredagcet--ocrosntaisinisnhgownreainctoFriguasrea1.control. The operation of the
"
Tthhee ScCoAnStenrtesactoofrseawcehrereaacetroartewderfeorst2i3rhrresd.wwitihtha50m0agnmel/tmiicn.stoifrraeirrtwohiplreevent
:
soentetlIitneg.r oAfftseurperthneataaenrtatwiaosnrpeeprliaocde,d wtihtehslpurdigmearwyaseffsleutetnltedfrfoomr atnhehoPuirgsand
"
Eye Plant.
wFeMre342t2akweansataddtehde asttarttheabnedgiennndinogf oeafchtheteTsetstcyCcylcelsesand1, f2r,omanthde4.supeSramnpalteasnt after settling.
)
T1h5e" haiegrhatwiiotnhchaam4b"erisntuesmeadlindiatmheeteSrC.AS sAtusdiideesawremrealplloewxeidgldarsasinacgyeliondfertshe
Supemnatant leaving the S00 ml with the.settled sludge undisturbed.
Analytical
x
WSearmpeleesvaltuaaktenedabtythtehinterlmaiynearticohnroomfattohgerapfhiyrst(CWeanrtbrualrgResstuedayrcohnanFMaly3t4i22cal work rteoq.a Nsom.allAS9v4o1l2u)m.e, Tahned ssaemppalreastewderoen eWxotorlanctesdiliicnatopldaitcehsl.oromTehtehadneev,eldorpiiendg wSeorlevenvtisusaylsitzeedwabsy 1t0h:e90ioedtihnaenoslt,arcchhlotreocfhnoirqmue(V:aVn)d.coTmhpeardeedvetolopkendowpnlates Standards with a detection limit of one ug of FM 3422.
SgaasmplcehsromfaotrogthreapShCyASwitshtudayn weelreectreoxntraccatpetdureintdeotne-cotcotra.nolExtarnadctsieopnasrawteerdeby pbeyrfcoernmterdifuignincgappated2560,7m0l0XpGolfyoprro1p0ylmeinnuetecse.ntrifuge tubes and phases separated
RESULTS AND DISCUSSION
Warburg - FM 3422
Results fron the Warburg study on FM 3422 are summarized in Figure 2. This
`J.
weaxtpeerrimteontmawkaesepmeurlfsoiornmsedobfyapfpirrosxt.so2n4i,c00a0tinmgg/lFMof342t2heanFdM i3t4s22anaandlog11,in000
mVge/rLy osfoltuhbeleFMin342w2atearn,aloitg.wasSinfceeltFMtha3t422foarnndingitsanhyedmruolgsieonnawnoaullodg paurte nmootre
of these compounds in contact with the microorganisms in the Warburg study.
00292
.
a
5
~-
STEP 1: mAedddiat,estandcompound, microorganisas |
" STEP 2: Afeorrat2e3 haonudrsmix
|
"A
dress) mpd ow i
k[
Co
Jha
2
7 SA
tanedstmecdoimap.oun/d' stir.ring bar
RepCeyactle. /
.
y
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~~
/
=&
.
Pu
ie
= iof
1
Ey
"
supernatant
i
;a
|i
i
| FRE drain
=Fi;
"
A
sludge
dl
7 =r
STEP 4: Drain supematant.
STEP 3: SLtetopslauedrgaetiosnettalned. mixing.
4
FIGURE 1: Test cycle for semicontinuous activated sludge reactor.
0en293
.
se
i4_ sz
Pi3: .
5 saz
.
7
,
A er
000 FM 3422
AEE
Towonsan22gs
(1.6 a mo)
8 SVo a
,&
/
.
:g 0 a _ i ~ S py F 3422 Mljtion
7
5
"xe
H~ E
wAinalllog \
.
:
N
2.
hN
w
:
-- Sadirion
.
-3
3
+4:
1 11 |
1
' ' ' Se '
25s as lo um
Be aims -- zoCL --. wt
time hours
FIGURE 2: Warburg study of Ni o54c22oamnnd eits hydrogen analog)
RFR i
~{21 au moles)
ge
Wehmiullesiftiheed,FM th3e422FM a3n4a2l2ogwawsas norte.latAipvperloyxiemaastiellyyemounelsihfoiuerdwaasnd resqtuabilreed ontcoeput
3
7c5o%neobfactkheouFtM o3f422emuilnstioone.mulsIinona,bouatndtwtohistomattherreiealhouprrso,ceeedxceedsstoFsMlo3w4l2y2
emulsion, which had not been used in the experiment, turned into a semi-
solid gel.
Complete chemical oxidation of the hydrocarbon component of the FM 3422 0a,t btahseehdiognhestthecofnoclelnotwriantgioenqua(tvi1o6n:j moles) would require 87 u moles of
CaF) 7SON(CHHIOHC + 5.50, + CF,SONH, + 4C0, + 4H,0
Meixcpreorbiimaelnt,oxiodnlaytio2n-3rmaircelryo emxocleeesdsof60o%xyogefnthueptackheemwicaaslobosxeirdvaetdi.on.HowIenvetrh,is ogxlyugceonseuptotaktehewaFsM c34o2n2tincuuilntgureataltsheo eprnodduOcfedthiisncerxepaesreidmeonxty.genAdudpittaikoen, of ccuolntfuirrem.ing that the FM 3422 emulsion was not inhibitory to the microbial
4
OUnponthjeadodtihteironhaonfd,thethemoshtydrcoognecnentanraaltoegd eomfulFsMio3n422ofshtohweedanaslioggn,ifiecnadnotgentoouxsicity.
.
otxo,ygetnheupctulatkuerec.eas`eTdheanndegwaatsivenotslorpeestoofretdheevheyndraofgteenr tahnealoagd'dsitioownygeonfugpltuackoesecurve
r(eFsiuglutrse w3e)re15obditeainteodthien eandsoegceonnodusWarcbourrregcteixopnerianmdentnowtitohxygFeMn3e4v2O2luatnidchi,ts Similar
hydrogen analog.
:
:
3
Aonbasleyrsviesd woafsFMgre34a2t2erhatshanshowwonulditbetoebxepeqcutietdefpruorme.impuTrhietioexsygeinn utphteakceompound.
bIitodiesgrcaodnacbeliev,ablbeuttnhoatt sdoentieccattaibolne bpyrodtuhicnedldaeygerradcahtrioomnatopgrroadpuhcyt.s thItatiwseraelso
pdeogsrsaidbelde.thaHtowesvoemre,oufsitnhge htyhdirnoclaarybeorncchormopmoanteongtrsaphoyf wFeM w3e4r2e2 umonlaebcullteeo.wedreetect "
aknnyownmatiefritahleshyfdorromgeednasanaalorgesuolftFoMf34t2h2e biitosdeelgfraidsattiooxnic.of FThMin342l2a.yerItandisgansot '
cthheronaantaologgrapthoybesh9o0w%edputrhieswimtahtertiwaolmatjoorbecoinmptuarmei.nantGsa.s cThhreomactoongtraampihn*antsshowmeady
have been the cause of the observed toxicity.
SCAS - FM 3422
Tihseolasteemictohenthiynupooutsheascitzievdatdeegrsalduadtgieon(SpCrAoS)ducsttsudyofwaFsM
a second attempt to 3422. This study was
ceonndduocfteeadchove2r4-har.perciycolde.of T1hweeeFkM w3i4t2h2 ssaammpplleess taadkdeend iant atnheetihnaintoilatisoonluatinodn
rsmaaplildlya ssuerpfaarcaetedarefaromtothaellolwiqusiidgnpihfaiscea,ntamnidcraosbiaalredseuglratdamtaiyonh.ave had too
n-octanol extractsof the No new leaks were formed
sasampalerseswuelrteofanaelxypzoesdureby ofgasthechrFoMma3t4o2g2ratpohyt.he
..
maiccidr,oorigtanwiosumlsd.notIfhasvoemebeoefnthdeeteFcMte3d4.22 hTahde bseuelnfondiecgraadceidd tios ntohte ssuulfffoinciicently
3
volatile to pass through the gas chromatography column.
* perforned by Commercial Themicals Division
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ou
0 bTehceanu-soectoafnotlheexltorwacvtoslactoiullidtynootf btehissespoalrvaetnetd.byFtrhoiznenlnaoynerextcrharcomtaetdogsraampphlyes sftoirlltheinxisltayaetr athniaslysdiast.e and could be extracted into a Bore volatile solvent
Tehxrpeeerimaedndti.tionTsheoFfMFM34232422seitntle3d3 pwiptmhintcheremseonltisdswearned fmaodrethdeurmionsgttphaertSCArSemained i(naltthheougrheancottoriwnhesnolutthieons)upewransataapnptrowxaismawtietlhydr1a0w0n.mg/lT.he Tfhiinsalmactoenrcieanltrwaatsionnot homogeneously distributed and accumulated on the sides of the reactors.
WARBURG FC-95
The results of Warburg studies with FC-95 are graphed in Figure 3.
Nmoateorxiyaglenaulpstoakceauwsaesd noobsetrovxeidc aesffeactrse,sultSipoefl-tohle,adadnitaipopnrooxifmaFtCe-95h.ydroTgheins
.
analog of FC-95, was shown to be readily biodegradable and to have no toxic
1
effects.
4 The Sip3el-ols was soluble at all concentrations tested (as high as 1700 ng/1).
F40C0-95mg/w1a.s inThceompllaectkeloyf sdoelgurbaldeatiaotn4w0i0t0hmgF/C1-,95buwtaswaesxpeccotmepdletsienlcyeipnersfolluuotriionnataet
compounds are characteristically nonbiodegradsble.
3 'NARBURG FC-128
OTxhyegseenruepstulatkse ciunrdviecsatefrothmaWtarFbCu-r1g28stisudireesadiolnyFbCi-o1d2e8graasdasbhloew.n iAnssFuimguirneg 4.
bmiaoxdiemgrmadoaxtyigoenn oucpctuarkse oasccuisrresdhowwnitbheilnowt,heap7-phrro.ximexapteerliyme7n0t%alopfetrhieodt.h.eoTrheitsical
oexnydgeonf uthpetakeexpeirsimgerneta.terThtehasne reexspuelcttsedaraendsoampepwehaartedintoqbueestcioonntinsuiinncge tAth:isthe
:3
FC-128 is known to be an impure chemical.
Cy1SONCCMHt,yCO0K + 4.250, + H' >CgF) SONH, + 4C0, + 3,0 + K
3
:
oeoz296
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? S1i72permtenls
6
;
/
g gi s
.
TF oe
5
EE 1S3
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:-
dgaltuicoosne //
oY
7
pro00s0 myg/1
[/ aoTa.n
af
i J } i
//
o
pitied
Nw, v
Na |
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ah
Ea ry
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3
11 ---
i1
r
Po
d1
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FIGURE 3: Warburg studyof FC 95 and Sipex-ols.
000297
.
-
s-
-
5
Fc az
;
7-
7
6-
,s-
.
/|
| 60
500 mg/1
: so
I
I
5a
4
:
i
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Lfao o
:
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9 ea>
3
a
:
at --
100 ngr1 ;
1-
oe
.
10]
oid
Sauwdsittriaotne
Angee
1
2
3
tine hours
FIGURE 44: VarburgvarburstudyoofFC 126.
L
Cd|
s
s
6
7
000208
-10-
.
REFERENCES: (1) AMliecxraonbdiearl,FaM.l;libBiiloidteyg.radaAtdivo.n:ApplP.roMbilcermosbioaflMo7:lec3u5-l8a0r,R1e9c6a5l.citrance and (2) CSth.apmPaanu,l,P.MinJ.n;esoDteap,arPtemresnotnaolf CBoimomcuhneimicsattriyo,nsUn2/i2v4e/r7s6i.ty of Minnesota,
D
x
J
oee299
ATTACHMENT I
-
E7./1A0./75Reiner
\
STANDARD PROCEDURE FOR WARBURG DETERMINATIONS
J. Design experiment and calculate concentrations of materials to add.
2. Fill water bath (DI water if left in bath).
5. Adjust temp. of bath (several hrs. or overnight).
4. Place manometers in desired order.
5. Prepare thermobarometer. Add about3mof H)0 to 1 flask.
6. Set out glassware in desired order (to match the manometer with which they were calibrated).
"
7. wLiitghhtlsyolvgerneta.se Acdednte0.r2-wemlll 1t0o%p KwOHi.th stopcock grease that can be removed
: " 8. rKeeep prefrsaimgpelreastedinunDItilwatueserd.(or according to request) to add to side arms. 9. Prepure cells (keep cells cold at all times but avoid freezing).
5
AC8..] 5.
CWRaeesnshturswipifetunhgdecionlcdco.lAdSMBS-N.centrifuge. Determine concentration of an aliquot
with
the
spectronic
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:
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15.
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17. begin readings (always wdjust closed arm of manometer to 150 mm before reading).
15 A contents of side aras according to experisent design requirements.
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19. Take readings periodically (on open arms) throughout course of experiment.
20. Disconncet and clean Flasks. AB.. WRaisnhseowfifthgrweaatseer.with acetone. U. ARciindsevawsiht.h OI Water.
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