Document OJ1daaqLzvQg27NB4a9MmaEDQ
BIODEGRADATION
TEST SUBSTANCE
Identity: FPeCr-f9l5u.or(o1o-cOtacnteasnuelsfuolnfaotnei;c amcaiyd,a1l,s1o,2b.e2,r3e,f3e.r4r,e4d,5t,o5a.6s,6P,F7.O7S.8.o8r8-
heptadecafiuoro-, potassium salt, CAS# 2795-39-3)
Remarks field: The test substance is a white powder of uncharacterized purity.
Tbihoedefgorllaodwaitnigonissaturdeidesu.ceWdhiolevtheesr oe fspv trudei iveise ouhsaw lvyeccoomnpclleutdeedd that 1Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-heptadecafiuoro-, potassium sal is persistent, new testing is underway to determine if a mechanism exists for the biological degradation of this substance.
METHOD
tMheethSoodasp:anWdarDbeutregrgDeentteArsmsioncaitaitoino'n;'SshparkeesuCmuplttiuvree tsetsutdfyormothdeeled after Tdeytpeer:miAneartoiobniocf ABS/LAS biodegradability'. GLP: No
Year completed: 1976", 19787
Contact time (units): 3 hours', 2.5 months"
Innoculum: Activated Sludge
RESULTS No biodegradation was observed in either the Warburg study or the 2.5 month shake culture biodegradation study.
DATA QUALITY
Reliability: Kiimisch ranking = 2. This study meets all criteria for quality testing, but analytical methodology is questionable.
Kiimisch ranking = 2%. This study meets criteria for quality testing, but
analytical methodologyis questionable. No reference compounds or abiotic vessels were included in eitherof the
studies.
Footnotes:
"`SWharabkuerCgulDteutreermsitnuadtyiomnodeled after the Soap and Detergent Association's presumptive test for the determinationof ABS/LAS biodegradability
00228
REFERENCES 1. Fate of Fluorochemicals in the Environment, Project number 75-6398-
29, EA. Reiner, August 12, 1976, 3M Company, Environmental 2. LaboFraatteoroyf.Fluorochemicals in the Environment, Project number 9970612613, EA. Reiner, July 19, 1978, 3M Company, Environmental Laboratory.
Qi TUBRi , cc r------------eee
Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 Last changed: 5/3/00
000229
Faith)
Technical Communications. Carnes, BE
MICROFORM COPIES:
3m TECHNICAL REPORT SUMMARY
.
oare, August 12, 1976
eras it_e_Envizonpental LABORATORY, DEPT.NUMBER _0222 Biodegradation Studies of Fluorocarbons vest Fluorochemicals in the Environmental
LE
onE. A. Reiner
see cuRIe TY .
Thiectver
To determine selected 3M
ftlhueorboicoadrebgornadcaobmiploiutnydsof
Tevtore Fomber 47816
WowbReskGRuference a ath, 4150
_pectl AuhoriatonClossd
Wo IiFnfoSrUmaMtMioAnRnY RhEiPsOrReTpo.rt
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Fo Teens cud coven 12
beincovebyreethedr wperts [RBSTRACT and Conclusions. (System can accommodate 200-250 words)
Brean PnolracisnecClkuJedeydCeomImndpolfteohrrremlrayteipoons ||
giv pose burs ou|
--te--i--l:----------"| 3MCHEMICAL REGISTRY| "eEw rcheemicals rCelpovreterd?
Biodegradation studies using a on FC-95, FM 3422, FC-128, and
hWyadrrbougregnraensapliorgosmetoefrFwCe-r9e5
canodnducted
FapMpr34o2x2i."mateNohbyidordoeggernadaanbailloigtyofwaFsC-9o5bswearsverdeaodnilFyC-9d5e,graadlatbhloeu.gh-..an
FM 3422 and FC-128 both were
biodegradation. Attempts to
demonstrated to undergo sone
isolate degradation products
of
FslMud3g4e22,Stufdryomwetrhee Wwasrubcucregsssftuuld.ies, and from a subsequent activated
Saloct ge"neKrEsYl,WOpRacDiSf.ic, and
-
I oroducs rer ram
pa
EE PC - Div. Envron - Assess BaBicotleorgiya BFilousocrroecehneimnigcal Biodegradable
.
`SPECIFIC PROBLEMS remaining to reach objective.
Continued attempts will biodegradation products
be of
made to FM 3422
isolate and identify the and other 3M fluorocarbon.
Work with radioactivity labeled FM 3422 is being considered.
Information Scientin
00230
BIODEGRADATION STUDIES OF FLUOROCARBONS
SUMMARY AND RECOMMENDATION
OfNoFbCi9o5degirsadiamtpiroonbawbalse boebcsaeursveeditinisWarcboumrpglestteuldyiefsluoonrinFaCte9d5.. BThieodergersaidstaatnicoen hoafstnhoits yceotmpboeuennd dtoemboinsotdreagtreadd.ation by an acclimated microbial culture, however,'
Wocacrubrurregd.stuTdhieesproonduFcMts342o2f athnidsFbCio1d2e8grbaodtahtiionndiacraetendotthaktnowsno.me bSieomdiecgornatidmautoiuosn
activated findings.
sludge
studies
on
FM
3422
did
not
confirm
or
disprove
the
Warburg
wFouutludrebienvgersetaitlgyatifoanc'iolfittahteedbbiyodetgheraddaebvielloiptmyenotf otfheanflaunoarloyctaircbaoln pcroomcpeoduunrdss tfhoer pFrCes95e.nt Wfairndbiunrggs.studies using purified FC 128 should be made to confirm
SSotluudbiielsitoyn. theThbiisodpergorbaldeambicloiutlyd boef FoMver3c4o22meweurseinghinFdMer3e4d22byradiitsoacltoiwvwiattyer labeled on its hydrocarbon portion provided this material had a high specific oacftitvhietylab(e55lendcic/oampmooulned)coaunlddpbueritmye.asurBeidodbeygraddeatteicotninogf g14Cs0a;tuervaotluetdiosno.lution
INTRODUCTION
TshteudySusocfptthiebienlviitryontmoenmtiaclrobfiaatle moofdiafniycactliaosns oisf caonmpiomupnodrst.antItparisamettheermoisntthe important forn of degradation for organic compounds.
ASovavsatstairnrayfaoctf otrhgaatniict wcaosmpoounncdesbeclanievbeedcboympsloemteelythadteggriavdeend beynoumgihcrtoiomregaannidss.
Tthhiesprdoopcterricnoendoiftimoincsr,obimailcroionrfgaalnliisbmislitcyoulisd
degrade still a
any organic material. common misconception(1).
PAletrhfoluugohrincaomtpeoduncdosmpwoiutnhdssianregleextfrleumoerilnyesrehsaivsetabnetentosbhioowndegtroadraetlieoanse (2f)l.uoride nieonvserabseeanrsehsouwlnt toof buinoddeerggroadnaattiuorna,l pdeergfralduaotriionna.tedFocromptohuisndsrehaasvoen,rnaorelmyodiofrication moofditfhiecapteirofnluoorfoictsomhpyodnrenotcsarboofncocmopmopuonndesntsinsetheimsedstpousdsyibwlaes. anticipated. However,
Aennviurnodnemresnttandwiilnlg obef etxhpeopsaerdtitaol thdeesgerapdraotdiuocntsproinducatddsitiisoni'mptoortthaentusnidnecgeradtheed materials.
METHODS AND MATERIALS Chemicals The chemicals used in these experiments are shown in Table I.
000231
.
2
TABLE I. CHEMICALS USED IN BIODEGRADATION EXPERIMENTS
Cats
mn se22
CaF17SONC HOH
HydrogesnazAnalog of
Clty SORNCCo,ltHsy0H
Fc ss
>
CaF7s05K
`
Sipex-ols
Fc 128
Cglly70503Na
of
Cg,50zGH;C00K
TSThehepeyteFwmCbe.er9re5, oub1st9e7ad5i.wnaesdFMFfrro3o4nm22D1o0nt(N5R-8ei3tc.keFrosLeootfanltuchomehboCelro)smewwreacrsieailndoeCtnhtegimifiviceeandl afsDoirv7i8t8sheiCoCnFMi7n4354-222. hydrogen analog or the sipex-ols (RY 26442).
TFhleusoeroccahrebmoincalcsonwsetrietuesnetleocfteda floarrgea mummbbeerr ooff Teasfalnuso.rocaFrCbo9n5 icsompeosusnednst.ially the
FMFi3n4i9s2h15flaunoroicnatrebromncdpiraotdeucitn.theSippreoxd-uocltsioanndotfheIHyfldurborgoecnarAonoanlso,gaondf FFCH 1343282is
Co
wheyrderogseenlecatmeadlogfoorfcFoCmp9a5r.isonTh1e0sethhyedrfolgueonrocaanarlboognss.werSeipteexs-toelsd biescaaunsespbpiroolxoigmiactaelly
.
bTyibeinlezymfelsuorwohciacrhbonnosrmhaalvley rfermeoqvueentalhyydbreoegnenf.ounTdhutso;beitgrsaeetmueidtopursolbyabdleeflutohratinated
cmiocurlodbimaolregcroowttphletoenlhyyddreoggreadneanfalluoogroscacrobuelnds.select populations of organisms which
MARBURG DETERMINATION
W(aArtbtuarcghmesnttu)dieMsicwreoroegracnoinsduusctweedreacccoorldliencgtetdoftrhoem tahtetamcihxeeddstlainqduaorrdopfrotcheeduPriegss. Eayteatcroonactennetnrtatsiyosnteomfva2s0h0e0d,Bganodf bsiuoslpoegnidceadlinsolaidbsasaplersallittser.medium and used
WtuahletterWraasroibnnuisrcoglpurbfollbeaess,kusb.bsatsreEamtu1el/ss2wieoinrnsechw,eemruaeltsiB1fa0id0ee%dpuosiwnienrwg.ataerBLlopagrcaikrosirtcohnateoicamoddddieilltuitoiEnoPn-s2to
iSnidweataerrm Solution
owefrethemaWdearobufrgtheFlatseksst. or deionized water in
SubCsotnrtartoelss, coanndtai1n/2edul1/v2asml this side am. The second
ploafce1d0 gi/nl tghelucfoisrest side ara contained
Cither glucose or deionized water.
000232
oh
-3-
.
.
>
IS 4)
JW", \ x
Owixtyhgenreaudpitnagkse
waats 10f-i1r5stmionb.serivnetdervinalseactho
flash fora period up to establish the endogenous
1.5 hus. activity.
mTohniistowraisngfolfloorweadppbryoxiamdadtietliyon2ohfrst.he fAidrdsittiosnideofartmheasnedcocnodntsiindueedaromxygen
containing glucose, a readily degradable material, allowed a further
evaluation of the toxicity of the previously added material.
Semicontinuous Activated Sludge Studies
A week-long semicontinuous activated sludge (SCAS) study was conducted
TorneaFtMme3n4t22.PlanTth.e mOincerooHrugnadnriesdmsFiufsteydmlweroef oabcttaiivnaetde,d asslubdegfeorwea,s fardodmedPitgos Eye 3 SCAS reactors and tap water as a control to a fourth. FM 3422 was added taoddi3tiroenactionrcsrebaesleodwthteheFwMat3e4r22sucrofnacceentrianti1o/n2 bmly o3f3 magb/slo.lutePuraelcoehtohla.nolEwaacsh asedmdiecdonttoinounoeusslurdegaec--tocrosntaisinisnhgownreianctFoirguasrea1.control. The operation of the
Tthhee ScCoAnSternetsactoofrseawcehrereaacetroartewderfeors2ti3rhrresd.wiwtihtha50m0agmnle/tmiicn.stoifrrearirtwohiplreevent settling. After the aeration period, the sludge was settled for an hour and oEnyee Plliatnetr.of supernatant was replaced with primary effluent from the Pigs
wFeMre342t2akweansataddtheed asttarttheabnedgiennndinogf eoafchtheteTsetstcycClyecslesand1, f2r,omantdhe4.supSearmnpalteasnt after settling.
T1h3e" haiegrhatiwointhchaas4b"erisntuersneadlindiathmeeteSrC.AS sAtusdiideesawremrealplloewxeidgldarsasinacgyelinodfertshe Supernatant leaving the 500 ml with the settled sludge undisturbed.
Analytical
wSearmpeleesvatlaukaetnedatbythtehinterlmaiynearticohnroomfatotgherapfhiyrst(CWeanrtbruarlgRsetsuedayrcohnanFaMly3t4i2c2al work r0eq.a Nsom.allAS9v4o1l2u)m.e, Tahned ssaemppalreastewderoen eWxoterlanctesdiliicntaopldaitcehsl.oromTehtehadneev,eldorpiiendg wseorlevenvtissuyalsitzeemdwabsy 1t0h:e90ioedtihnaenoslt,arcchhlotreocfhonrimque(Va:nVd).C.oTmhpeareddevetolopkendowpnlates standards with a detection limit of one yg of FM 3422.
SgaasmplcehsromfaotrogtrheaphSCyAwSitshtudayn weelreectreoxntraccatpetdureintdeotne-ctoocrt.anolExtarnadctsieopnasrawteerdeby bpyerfcoerntmreidfuingincgappaetd2560,7m0l0XpGolfyoprro1p0ymliennuetecse.ntrifuge tubes and phases separated
RESULTS AND DISCUSSION
Warburg - FM 3422
ReexspuelrtismenftronwasthpeerWfaorbrumregd sbtyudfyirosnt FsMoni3c42a2tinargeFMsum3m4a22rizaendd iintsFiagnuarleog2.in This wga/tleroftotmhaekeFMem3u4l22sioannsaloogf. apSprionxc.e 2F4M,0304022mga/nld oiftsthheydFrMoge3n422anaanldog11a,re00n0ot very soluble in water, it was felt that forming an emulsion would put more of these compounds in contact with the microorganisms in the Warburg study.
00233
Lo
~4-
-
STEP 1: mAedddiat,estancdompound, |
microorganisms fo
KC i |
~
7
" [ster 2 aad SR
i
aerase ana mix
for 23 hours,
vw
aa
Lad
7 SARL b~
StTeEsPt 5c:omRpeo-unadd/d stmiargrnientgicbar air sparger
aRnedpCeycamlteed.ia./ 7
es
7
pa
a
ee]
rei Det
i
supernatant.
i]
ii
--
;i
. n==}F] N drain
STEP 4: Drain supernatant.
stutge~ . = aC rdi r
STEP 3: SLteotpsaleurdagteiosneccalned.mixing.
FIGUE 1: Tost cycle for semicontinuous activated sludge reactor, 00000
.
-
!
:
FM 3422
5
-
!
830FM003422mg
Ci
(16 11 mol.
4|
i
idsl
=
EE
2:
:
5EO
0
2
5{
ol
!
S 14
2 i.
~ ;
& Js
#
A Tr
/
&
&
.
--7QF zz Adgjtion
5
rS
Analog
EN
"
\\
FM 3422 4300 ng/1
(1.6 a mol.
Ne
3 pt Te ega aGdditEion E
2s ss 10 12D Ls 20 tine hours
FIGURE 2: Warburg study of FM 3422 and its hydrogen analog)
liydrogen Analog
002>.35 mL
:
-6-
-
eW7mh5ui%llseoifftihteehde,FFMtMh3e452422F2Mai3n4na2tl2oogweawmsauslsnioroten.l,atAaipnvpderlotyxhiiesmaastmiealtlyeyreimoanulelsphiorfuoirecdeweaadsneddrtesoqtuasibrlleeodwloytncoeput
come back out of emulsion. In about two to three hours, excess FM 3422 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
5a,t btahesehdigohnesthte cofnoclleonwtirnagtioenqua(t1i6on:u moles) would require 87 u moles of
CaF 7508(CHIC HO +5.50, - gF)750M, + aco, + 4H 0
Microbial oxidation rarely exceeds 60% of the chemical oxidation. In this
experiment, only 2-3 micro moles of oxygen uptake was observed. However, oxygen uptakewas.continuing at the end of this experiment. Addition of
Cglouncfoisrenintog tthhaetFMthe342F2M c34u2l2tuermeulaslisoonpwroadsucneodt iinnchriebaisteodryoxtyogetnheupmtiackreo,bial culture.
OUnpontheaddoitthieornhaonfd,thethmeoshtydrcoognecnentarnaatleodg eomfulFsMio3n422ofsthhoeveadnaslioggn,ifiecnadnotgentoouxsicity.
oxygen uptake ceased and was not restored even after the addition of glucose to, the culture. `The negative slope ofthe hydrogen enalog's oxygen uptake. curve
(Figure 2) is due to the endogenous correction and not oxygen evolution. Similar results were obtained in a second Warburg experiment with FM 3422 and its hydrogen analog.
How1#7o_bysAnearlyvseids woafsFgMre3a4t22erhatshanshwoowunlditbetoebxepecqutietde fpruorme.impTuhrietioexsygeinn tuhpetakceompound.
It is `conceivable that sonication produced degradation products that were `biodegradable, but not detectable by thin layer chromatography. It is also
padonesygsrmiaadbteldee.ritahlHasotwesfvooemrrem,edoufsaistnheg4 hrtyehdsirunolctlaaryobefornthcehcormbopimooandtesonggtrrsaadpahotyfiowFneM wo3ef4r22eFMmuon3l4a2eb2cl.u"eletoIwtedreiestencott kcnhorwonmatiofgrtahpehyhydsrhoogweend tahniasloBgatoefriFaMl3t4o22beitismeplufrei.s toGxaisc.chroThmiantoglraaypehrasnhdovgaesd. the analog to be 0% pure with two major contaminants. The contaminants may
have been the cause of the observed toxicity.
SCAS - FM 3422
The semicontinuous activate sludge (SCAS) study was a second attempt to isolate the hypothesized degradation products of FM 3422. This study was
ecnodndoufcteeadchove2r.-har.perciycolde.of T1h'eweeFkM w3i4t2h2 ssaammpplleess atdadkeedn iant atnheetihnaintoilatisoonlutainodn
rapidly separated from the liquid phase, and as a result may have had too
small a surface area to allow significant microbial degradation.
n-octanol extractsof the samples were analyzed by gas chromatography.
No new leaks were formed as a result of exposure of the FM 3422 to the
microorganisms. If some of the FM 3422 had been degraded to the sulfonic
acid, it would not have been detected. The sulfonic acid is not sufficiently
volatile to pass through the gas chromatography colum.
* performed by Commercial Chemicals Division
00236
:
a
-
bTSehtceialunls-eoecxtoiasfntotlhaetexlttohrwiascvtodslaattceioluailntddyncoootfulbdtehbiessespeoaxlrtvaretanectdt.ebdyFirtnhotizonenalnoaoyrmeeerxtvcrohalracottmiealdteosgasrmoaplplvheeysnt for thin layer analysis. Teihxnrpeeterhiemaedrndetia.tcitooTnrshewhoefFnMFMt3h4e23242s2sueptietnmlae3td3anpwtiptmwhaisnthcwerietmsheodnlrtiasdwsnw.earnedThmfeaodreftidhnauelrimnocgsotnctpheaenrttSrCaAtrSieomnained h(oamlotgheoungehounsolty idnisstorliubtuitoend) awnadsaacpcpurmouxliamtaetdeloyn t1h0e0 msgi/dle.s oTfhitshemarteearcitoarls.was not
WARBURG FC-95 The results of Warburg studies with FC-95 are graphed in Figure 3. Nmoateorxiyaglenaulpstoakceauwsaesd noobsetrovxeidc aesffeactrse.sulStipoefl-tohle, adanditaipopnrooxifmaFtCe-95h.ydrTohgiesn aefnfaelcotgs.of FC-95, was shown to be readily biodegradable and to have no toxic TFhCe-95Siwpaesl-oilnscowmapsletseollyublseoluabtleallatco4n0c0e0ntmrga/tl,ionbsuttweasstedcom(apslehtieglhy iasn s1o7l00utmigo/n1)a.t 4c0o0mpmogu/nld.s aTrheechlaarcakcotefridsetgircaadlaltyionnownibtiohdeFgCr-a3d5sbwlaes.expected since perfluorinate
WARBURG FC-128
OTbxhiyeogsdeeengrrauedpsatualtktiesonciunorcdvcieucsrastefrasothmaiWstarFsbChu-or1wg2n8bsetilsuodwir,eesadaioplnpyrFobCxi-io1md2ae8tgerlaasydas7bh0lo%ew.nofinAtshFsiugmtuihrneegorde.tical
moeaxnxydigemonufmutophxetaykgeeexnpeisruipmtgearnketea.toecrcTuhtrehrsaeenderwexisptuehlcittnsedtahraeend7s-oahmprep.wehaearxtepdeirtniomqebuneetsatclioonpnteirnsiuioindnc.ge
aTthitshe this
FC-128 is known to be an impure chemical.
CgF 1SONGH,CO0K + 4.250, + H' + CF, SO, + 40+ 34,0 +
00237
a:e . `
7]
"- . :
172 mg
Sipex-ols
4
.
:
.
83 5
.
3.
Hi3
FE
2Bosg{
S:
3
i.
&
i
]
i
24
|
i
1d
| i
0 RC
:
1ii
.
.
substrate
+ addition
Y
Do 1 H
time hours
glucose
addition
},
'
00F0C n9g5/1
p 00 m-- g
:-
FC 95
:
~
;
74
CtCh eed \
ee
2 RgiluvTceosnes
.
FIGURE 3: Warburg study of FC 95 and Sipex-ols.
0238
ae ate
!!
2|
.
:
54 sd
i
!pe
a
1} 2 2a
3
.
;
.
=n
Boz
.
.
70
}
.
ean
-
6
500 mg/1
-
50
i} ]:
|*
:
eTe220]
al
100 mg/1
:
10]
|0k;
|!;
"1 baa
Suaubdsitzriaotne
pear
PLN tt
1
2
3
time hours
FIGURE 4: Warburg study of FC 128.
dh
4
5
|Fi
i|;
`Yen
6
7
00239
>
-10-
REFERENCES: (1) AMliecxraonbdiearl,FMa.l;libBiiloidteyg.radaAtdivo.n:ApplP.roMbliecmrsobioaflMo7:lec3u5l-a80r,Re1c96a5l.citrance and (2) CSth.apmPaanu,l,P.MinJn.e;soDteap,arPtemresnotnaolf CBioomcmhmeimciasttiroyn,sU2n/i2v4e/r7s6i.ty of Minnesota,
00240
F . h .
. ATTAGRENT 1
7E/.10A/.75Reiner .
STANDARD PROCEDURE FOR WARBURG DETERMINATIONS
1. Design experiment and calculate concentrations of materials to add. 2. Fill vater bath (DI water if left in bath). 3. Adjust temp. of bath (sveral hrs. or overnight). 4. Place manometers in desired order. 5. Prepare themobarometer. Add about 3ulof H,0 to 1 flask. 6. Stehteyouwetreglacsaslwiabrraetdedn).desired order (to match the manometer with which 7. wLiitghhtlsyolvgerneta.se cAedndte0r.-2wemlll 1t0%opKwOHi.th stopcock grease that can be remoAved 8. PKreeepparreefrsiamgpelreastedinunDItilwatuesred.(or according to Tequest) to add to side arms. 9. Prepare cells (keep cells cold at all times but avoid freezing).
AB.. CWeansthriwfiutgheco0ldC.BSM - centrifuge.
C 5.
RADedestjueusrsmtpiennredemaicinonndcceoelnrdtrtaBoStNid.oensiorfedanconacl.iCquobtaswiicthsatlhetssmpeedcieurma.nicR2e0fraitge6r0a0teni.
E.
until use. Take sample
of
final
adjusted
sludge
for
standard
= MLSS
analysis.
10. Add samples to side arms (usually 1 ml if one side arm, k ml to each side arn if
4
2 side arms).
11. Add 2 ml of washed cells to flasks. 12. Add filter paper strip to alkali in center cup.
15. Actach flasks to the correct manometer. 14. Retighten flasks after about S min. shaking in bath. 15. Leave stopcock open to atmosphere, and let temp. adjust for an additional
10 min. 16. Adjust level in manometer to 150 with stopcock open (close stopcock). 17. Begin readings (always adjust closed arm of manoneter to 150 mm before reading). 18. Add contents of side arms according to experiment design requirements.
00241
.
2
19. Take readings periodically (on open arms) throughout course of experiment.
20. Disconnect and clean flasks.
:
AB.. RWianssheowfifthgrweaatseer.with acetone.
CDo. ARciindsewawsiht.h DI Water.
-
.
.
00242
Ford 6747-1 4
TECHNICAL REPORT SUMMARY
TO: TECHNICAL COMMUNICATIONS CENTER -- 201.208
ips pr oost mtpe mncoi 629
7/19/78
For Environmental Laboratory (EE & PC) Fate of Fluorochemicals in the Environment
Biodegradation Studies of Fluorocarbons - IIT
D. L. Bacon
0a 535 9970612613
5
Homes BE cova Eres | MED framers T5e.4!4445R77e20i73n,,erppo. 326--1343,;21. 4,543050-2,7, 2p. 011,-12. 35, 30-43, 457 [Meme 471781n6eseen
KEvwoonosn: an CURRENT oBsEcTIVE:
ei ao (Biodegradation)
Ttoo emivcarloubaitaelthdeecosmupsocseipttiiobni.lities of FC-95 and FC-143
EEnEvi&r.BC-ADsisve.ss.
F"leuroardoacthieomnical
| [Semen Ama: oe as Tr oration net Fe Som
Es
Aetovbailaouedaretoigboirncadaomfitcirtoohnbeisastluusdycdeegp1rtsaidbdaietlsiicotrnyi.boefdThwFeCh-i5cc5hulta`aulnrldionwFgso-p1teahoge-
MfcaionecsddtruoDrbeefitsoaerlrugtseheinedntodicenAutslestarohmciwisienaratsetitiounod'nysoapfrreeAsBmuSom/dLpeAtlSievdebiaof(dtseehgrarkaetdhsecbuilBiyoiuursee.)
TcLLroCel,al.tepcgtnCet-dspcalitanntCtihsle.lmaoltiiAtonena,lyctoDoiebuccntaataltiiunnrgperdoaacnnfeddrdoMumarenetaasrlcoytsiiwinvsacacltsueeddeds1uGLdCe,y
tfhleuorsiudiet.abilDietgyradoaftitohne boifodreegfreardeantcieoncotmepsotundcsonfdoderimtorinemslmeeeeassreodd
Re
00243
.
i.
y suuARY
FrelsuiosrtoacnhtemitcoalbsiodFeCg-r9a5daatnidonFC-i1n43a w2e4r-emonshtohwnshatkoebeculctoumrpelebtieloydegradadteiroinvesdtudfyr.om aTchteivmaitxeedd smliucdrgoebiailnoctuelsat ocbutlatiunreeds fursoemd tihnretehiwsasstteudytrweearte mCounlttusryesstewmesre(Cmhaeimnotlaiitnee,d Diencadtiulru,te&ytehaestTweixntrCaictti-ebsasMaeltrsoaltplsanmte)d.iaThe Siucaplpsl.emeTnetsetd wciutlhturtehse ahlysdorogceonntaainnaeldogFoC-f95theorrFeCs-p1e4c3t.ivePhfelnuoolrocahnedm1rdeofedreecnecnee-cdoemrpiovunedds.lineTahreiraldkeylgrasdualtfioonnatedem(oLnASs)trwaetreed uthsaetd absiodegtrraadnastfieornredcou15ldtoicmecsurovuenrderthethe2i-tmesotnthconpdeirtiioodn,s.andAltlempceurlattuurrees wwearse controlled at 25 C. during the latter half of the experiment.
Ifnluotrheochfeinnailcalgsrowwtehrepearsisoady,ddefgorradbyatitohnin-plraodyuecrtschorfoma"t4ocg-rlaapbheyled(TLC) waenrdegavsisulailqiuzieddcbhyroTmLaCt-oagurtaoprhaydi(oGgLrCa)p.h. CheMmeitchayllsatesdepaanrdatneodnbmyethTyLlCated NcoultduergeradeaxttiroanctsprosdeupcatrsatweedrebydGeLteCctweedr.e dSectiencttieldlabtyioenlecctoruonntincgapture. schheomwiecdaltshatremalalinerdadiinoactthievictuyltuarsesomceidaituemd. with the labeled fluoro-
In all but the final growth period, fluorocarbon biodegradation was ` ) cmoonncietnotrreadtisoinmpliyn btyhememaedsiuar.ingNotheincirneiatsiealinandflufoirnaildefclounocreidnetration
wnaost orebssuelrtvedinintdheicarteilenagsethaotf fifluobriioddee.gradCaotnitornoldcidultocuoruers, suitppldeidmented wtihtehexfpleuroirmiednetaslhowceodnditthiaotnsfluusoerdi.de is mot lost from - the media under
Wchoiulled btehisfosutnuddythactanwnootuldrulaelloouwtthtehebipoodsesgirbaidlaittyionthaotf ctohnedseiticoonmspounds,
ptehresirsetsulitnstohfe etnhvisirosntmudeyntsufgogresetxtetnhdatedtpheerseiodcshemuincaalltsereadrebylikely to
microbial catabolism.
.
J
C0244
3
3)
INTRODUCTION
Thahveefpleurorfolcuhoermiincaatlesd scealrebcotnedchafionrsthainsd satruedyc,hemFiCc-a1l4l3yasntdabFlCe-.95,
Tthoebepersfulsuceoprtiinbalteedtpoobritoiloongiocfalfludoergorcaadrabtoinosn
have (1).
not been found Therefore,
bfioordetghreadsaatkieonof stcuodmipelsetweneerses.condWuicthtoeudt osnucthhesteestcionmgp,ounidts cporuilmdarniotly
bmeodisfaiicdatwiiotnh. certainty that these compounds would rsist microbial
Since biodegradation was unlikely, the best feasible test condi-
tfiroonmsarfeoarsbicoodnesgirdaedraetdiolnikweelryetsoelceocntteadi.n acIcnloicmualatewdermeicroobotragianneidsms.
Ldeovnegloapccploipmualtaiotnionpseroifodsmiwcerroebesusecdapainbleanofatdteemgprtaditongsetlheecste acnodmpounds,
and hydrogen analogs of the select organisms that might
fgrlautouriotcoaursbloyns "wceormeetaabdodleidzet'o
ttrhye
to
.
luorocarbons .
J
000245
ae
"METHODS AND MATERIALS
|
Chemicals
F(Ct-h9e5,hydFrCo-1g4e3n, atnhaeloghydofrogFeCn-14a3n)a,locgarofbonF-C-1945,laabmemleodniFuCm-1o4c3t,anoaantde Dciavribsoino-n1.4 lTahbeesleedcFhCe-m9i5calwserweeroebtauisneeddasfrroemcCeoimvmeedrcuinallessChedmeisciaglnsated otherwise (Arthur Mendel-Report in Progress). cSotmapnoduanrdd floirnebairodaelgkryaldaatteiosnulsftoundaiteespwraespaorbetdaifnoredusferoamsthaerUeSfe/rEePnAce LcaobmoproautnodrsyweirneCirnecaignennattig,radOeh.io, Except where noted, all other
Culture Media >
Tinhe_TcAoBnLtEro1.l medium used in these studies had the composition shown
TABLE 1
CONTROL MEDIUM COMPOSITION
1) Basal salts solutions:
>
1.0 g/1 - NE,CL
:
2.0 g/1 - KHPO,
0.25 g/1 -MgSO, -7TH,0
0.002 g/1 -FeSO, 7H,0
2) Well water - 25 ml/1
3) Yeast extract - 0.3 g/1
4) HoyrdrFoCg-e1n43a-nal20ogsmgo/fl either FC-95
bMreoduigahtwerteo pvroelpuameredjusftropmrisotrocktoseoalcuhtiocnusltwurheichtrwanesrfeerc.ombAinefdreashnd
sinolmuetidoina porfepFaerSa0t,i7oHn,0atwaesachpretpraanrsefdera.nd dTrhye ypeHasotf eaxltlramcetdiwaawsasused
a1.d0juNstNeadOHt.o 7T.5hewwietlhl1w.a0teNrHwCaIsanadddeidf otvoerisnhsoutreaadnjuasdteeqduabtaeckswuiptphly of
pterracieodelpermieonrtst.o tAhenaliynsietsiatoifonthoefwetlhliswasttuedrymasdheowdeudriintgs cthaelci1u2-mmonth
hiatradtenesrsesutoltirnagngefrofmromthe92adtdoit1i4o4nmgo/flweelxlprweastseerd filtration through a #54 Whatman filter.
CaCO. Any was femoved
precipby
00246
>
-5-
D TbDhiaeoodleupggureradidfaiitneidoanhnydtaretostgteemnmptedanitaaol-oasgnesdleoccftontFarCo-ml9is5c.roabnidTahleFsCe-p1o4pc3uolmapwtoeiurnoednsulwsieekrdeelyin to Gnooaagetrbiaoodnne-hytrdheeraocgtfeilonunosrboocnaardrebcolfnersae.vqaugeenEtnlz(yy1)m.eisdeAcnadtipdcaiabtllieontoaoflencczaoytmmapeloysnzeinintngsvodolefvfeldoutohirne-r Specific media are listed in TABLE 2.
TABLE 2 GROWTH MEDIA FORMULATIONS
Media
~
Components
FC-95
FC-95 Control Medium + 50 mg/l FC-95
:
FC-143 Test
Phenol Controls
FC-143 Control Medium + 50 mg/l FC-143
FC-95 or FC-143 Control Medium +
30 mg/1 Phenol
LAS Controls
F3C0-9m5g/lorSFtCa-n1d4a3rdCoLnitnreoalr MAlekdyilu-m +
)
Fluoride Controls
benzenesulfonate (LAS) FC-95 or FC-143 Control Medium +
-
33.2 mg/1 NaF (15.0 mg/l F)
4c pc-05 Test
FC-95 Control Medium + 50 mg/l
4c_rc-95
14c_Fc-143 Test FC-95 + LAS
FC-143 + LAS
FC-143 Control Medium + 50 mg/l 1c pe-143 F+C5-095mCgo/n1trFoCl-95Medium + 30 mg/l LAS F+C5-0143mgC/1ontFrCo-l14M3edium + 30 mg/l LAS
Culturing Procedures
Tmheediuinmiwtiiatlh Sludge used
g1rmolwthofpearcitoidvawtaesd sstlaurdgteedsbuyperinnaotcaunlta.tingThe49amcltiovfateeadch was a mixture of two sludges collected on the day of
CCSohnneotmcrouollliattieCoonmW.maisstseTihoTenr'essaltumMdeegntetrowPaplslaanontbttianiinnCeSodatitfnartgoemPaGturhloe,veM,MeitnrMnioenpsnooeltsiaot,taan.anWdastthee
00247
.
6
y Following inoculation, the cultures in polypropylene Erlenmeyer flasks were shaken at 200 rpm on rotary shakers at room temperature (4).
Aitdetnhteicaelndfrofesheamchedigarowutshinpgeraio1d%, ienaocchulucumltfurroem wtahse tprraencsefdeirnrgedcutloture (i.e., 0.5 ml of existing culture to 49.5 ml of identical new medium).
TAhe10gmrlowtshamppleeriwoadsbettawkeeenn ftrroamnsefaecrhscvualrtiuerde aats i1s0 mniontuetdesinaTfAtBeLrE 3. iSanmopcluelsatiwoenreorcencturlitufruegedtrafnosrfe1r0 mainnd. atatth1e7,e0n00d oxfgeapcrhiorgrotwothanpaelryisoids. nofotethdeinceTnAtrBiLfEug3a.te. Deviations from this culturing procedure are
TphreepafrienadlwigtrhowtCharpbeorn-i1o4d dliafbfeelerdedFCf-r9o5m panrdecFeCd-i1n4g3.periOondes.hunMderdeidamwlere ccuolntturroelslewderaet 2g5rownC. in+ f1.laskTswenotnyamlrotsaarmyplesshawkeerreintakaengroatwth10cmhianm.b,er 2 days and at 7 days.
Chemical Analysis
.
Fluoride ion concentrations were measured using a fluoride ion
ealnedctarosdteand(aOrrdioncurivoen darnaawlnyzefrromflutoheridreeseulletcstroofdememaosduerelme9n6t-s09)o,f
) athcecsueratfelluyorpidreepasrteadndafrldusoribdreacksetatneddartdhse. conTcheentcroantcieonntsratpiroenssentofin
:
sthaempelxipnegripmeerinotda,l seaxmcpelpets.forFlturoarnisdfeercu1.rvesFowrerteheseatnalupyseast efaoclhlowing
tthheisitnrsatnrsufmeern,t wait1h.0thpepmafslsuuomrpitdieonsttahantdartdhewasslopuesedofttohecaplriebvriaotues
fluoride curve remained constant.
PEhxeanmoilnatainoanlysoifsWwaatserdoanned aWacsctoerwdaitnegr,to1E S4ttahd ndai rd t Meti h1o9d7so5.fn or t,hLienear `alkylbenzenesul(fLoASn)atweas anaiyzed for by the methylene blue, ScthalnodraorfdorMmetehxotdrsact(i3)o,n meextchepotd diensctrriabnesdferisn 8th-e14,14tLhASewdaistioannaolfyzed bsyampalemsodwiafsicadtiilountedoftothi1s00memtlhoidn. a sIenpatrhaitsormyodiffuinenedl.methAolds,o tahdeded tboluethesolsuetpiaornatoarnyd 1f0u0nnemll woefrech2l5oromfloromf. StTahnidsardmixMteutrheodswasmetshhyalkeenne dfroarwn30ofsfecotnhdrso,ughallgolawsesd twooolsetitnlteo, asw2i.r5lecdm, diaanmdettehre, cshpleocro2f0ormcurvette. Pceurrcveentpretpraarnesdmiwtittahncesurwfasactraenatd satamp6l5e2snmofaknndowcnompcaornecdenttroataiosntandard treated in the same manner.
J
0243
Fi
i.
TABLE 3
hl
IN TSHUEMMASRHYAKEOFFCLUALSTKUBRIIONDGEGPRRAODCEADTUIROENSSUTUSDEYD OF
FC-95 AND FC-143
Transfer # CPuelrtiuorde G(rdoawytsh) Notes
3
MUesterdoaacntdivCahteemdolsiltued.ge in. oculum from
1
3
cFuCl-t1u4r3e-shydarnodgecnontarnoallso.g added to 143
2
a
3
3
A1tmlthoeftDiemceatoufrcsulltuudrgee struapnesrfneart,ant
added to cultures.
a
3
LcAoSmporuenpdl.aceLdASphemneodliaaswasa rienfoecruelnacteed
waintdhChaemmioxltiutreeanodf Dcoenctartoulr scullutdugree
supernatant.
)
5
3
6
3
dThiescounsteinoufedf.luoride control was
7
6
pSohsaskiebrlywasforin5addvaeyrst,entdluyrintgurntehdisoff,
growth period.
8
3
w4asmlaodfded Mteotraoll cuslltuudrgees.supernatant
6
"
10
4
:
1:
4
cInulttuhriessawnedresubgsroewqnuenitn garorwetchippreorciaotdisn,g
msihna,keran-dwa2t5eg bC.ath at 100 strokes per
12 13 14
2D
-
15
6 6 8
+6 (2)
7 78 days = Total Enrichment Period
000249
s-
d Carbon14CountingTechniques
SCceinnttriiltluagtaitoen acdoduendtitnog AqwuaassopetptfhgTrmeadndoncou1ntmledswaimtphlessnofinceualreteusre wSatsandcaormdparqeudenctho kcnoorwrnectwieoing.ht Tshaemplreasdioofacgj'vCi-tFyC-o9f5 otrhegq1isCa-mFCp-l1e4s3 added directly to Aquasol.
Sfoillitderssamcpolmepsosweedreofccoelllelcutleodsediarceecttaltye aonndtocemlillulliopsoerenHitAra0t.e4.5 yTmhe
cfoimlbtuesrtsiownerecontghgeyn wweatshewidthwitChomdbeuisotnaiizdg\d)w,ataenrdgaonmdbupsltaecdedininAtogrpia-per
chen's Packard")
combustion was
tracpopmedbusitnioan secqiunitpimlelnatt.ion'Tfhle uid14Cc0,8ntraeisnuilntgingan trom
ocrougnatneirc. amSinaemplaends cwoeurneterdecoiunntAegdricwhiethm'sanPaicnktaerrndalscisnttainldlaradtiofnor
quench correction.
Thin-Layer Chromatography (TLC)
.
wTmeehrtieanb-olclaoiylteleresctceohdfrogangd"t-oCgi-rFmCam-pe9hd5yiaawtneadlsy,p1ef9rrCof-zoFeCr-nm1.e4d3. TthoeTsedmeetsmelacmtpclurelastduirwoeearcestaimsvpteloersed R) caffertnoetzrerniftufhgoarewdinagabtowuti1t7h,100m10o0ntmxhl.goftToheetehnyssluarmepalceetsthaetwees.reepaTrehaxettirosanacmtpeolfdestihmwemeeredetihayttlehlesy
oafcettahtee,thwaytlera,cetaantdespohlaisdes wphearseese.vapTohreatweadtetro pdhraysneessandunpdoerrtiNoo.ns msTihpxeottutdrireni.gedwes(raSemopmlereesssuawsmepprleeensderdwehsiuicnshpmeenetdvheaadpnoorilan.t)ead9T:th1oehderrxeyansneuess:speetnihdnyeldairsetahmbepeffloerse wwreeerrseeidusreepfowetertreeendceoadnlsEao.gaaipMnpeslrtcikedasmidilixirtceaucrtlegyetlofoGF,Rc1,Cs.-FeC-pS1lm4aa3tlelasn.dspo1t4s11i1Co-fFsCa-mS6po8.lleisds Tvfhoiersupoanlleaitzeweesdekbw.yereexpdoesvienlgopeKdodawkithno-1s0c%reeetnhanxo-lrayin f.eitlmhylon actehteatpelataensd TTrhLeeCsiwdsauoselverrneetpseuwasatpseenddaeolndlowtinehedmerttoehmaeanvioanlpi.onrgatpTehoerstteioonpdlrayotfneesstshweeirnSeolaivsrep,notttaensddampmtloheerse. h2eawveielkys,. developed as before, and visualized with x-ray film for
00250
--o5
Gas-Liquid Chromatography (GLC)
Eltahyyelr acchertoamtgejogerxatprhayctsmewthegrdes.preCpoanrterdolassdoelustciroinbsedweirnethmeadethbiyn-
dtihessoelthvyilngacet-*aCt-eFC-e8x5traancdt sa~mCp-lFCe-s14a3ndintehtehyelthyalcetaacteet.atePocrotnitornosl of
nsoolnumteitohnySlwaetreed eatlhsyolmeatcheytlaatteede.xtraAcltisquoatnsd cofonttrhoelsmewtehryelatiendjeacntded
conatpoturtheede5t7e1c3toHre.wletMtetPhayclkaatredd sgaamsplecshrwoemraetogirnajpehctweidthwitehlienctr3onhrs.
o1f/8"th0e.iDr. msetthayilnaltisosn.steeTlhepacchkreodmawtiotghrap2h0%icDCco2l0u0mn(w1a2s,50102 CfSt.) oxn
10% Bentone `temperature
34 and 70% 80/90 was 250 C., and
mtehsehdAenteacktroorm
Pt.eAm.perTahteureinj30e0cgCi.on
pThoert
tcoolruimsetteompe18r0agurC.e waats8groC.grapmemredmint.o, hoalndd tfoorh4oldmina,t a18t075C. C.,The
flow rate was adjusted to 35 ml/min. of Argon/methane, 95/5.
MCeotFhCylOa0tHionssoluwteiroen,perafsoramerdefbeyrenacdedincgompaou20nd ulto aelaicqhuotsaomfplea. 1 ug/ml
Dsbalmapzloemsetwhearnee wtahsenthleonosealdydedcapupnetdi,l aswiyerllleodwacnodloarllpoewresdisttoeds.tanTdhefor
)
15 minutes. Nitrogen was blown over the samples until the yellow
-
vcoollourmedwiistaphpeeatrheydl, acaentdatteh.e sample was returned to its original
RESULTS AND DISCUSSION
A
Fluoride Release
In all FC-143
but was
the final monitored
ognrloywthby pearniaoldy,sisdeogfradfaltuioornidoefcFoCn-c9e5ntraandtion
atthattheifbetghiennfilnugoraoncdheemincdalofpoeracthioncsultofurethpeesreimoodl,ecuIlteswasweraessumed
dtehgartadfeldu,orifdleuorwiadsenoitonlwoosutldfroamccutmheulactueltuirnetbhye amebdsioar.ptioTno, epnrsuerceip-
oiftatfilounoriodre.volTahtiislizfaltuioorni,decocnotnrcoelntrcautlitounresiswearpeprogxriowmnatewliythwh1a5tmg/l
wpoeurlcedntresfullutoriidfeFCr-e9le5asoer. FCT-h1e43ruesnudletrswenotf dtheegrafdlautoiroindewiatnhaly50ses
wcaosndudcuetedtoonthdeifvfaerrieanbtledaaynsd svehroywedslucgognissihderraebslpeonsvearioaftitohne. flTuhoirside
SThAeBsLiEro4tbe.s,hoJwAsBLtEhe4aresshuolwtss tohbetarienseudltwsheonbtiahieneSdamset seanmphletsr,anwshfiecrh.
haafdtebreetnhesttoerremdinaitniopnolyofethtyhleeneexpecroinmteanitn.ers,Dewsepriteeantahelyzveadriatboigleitthyer
J
diuferteoleathseedantaolytthiecamleditaechtnhirqouueg,h btihoedergersaudlattsioinn,diwcoatueldtmhoatt bfeluolroisdte,
from the media.
The results of the fluoride analysis on fluorocarbon-containing
cultures and controls are shown in TABLE 5. The results show that
no biodegradation with fluoride release occurred.
C0023!
To
-10-
)
TABLE 4a
(ImNgI/T1I)ALOAFNDFLUFOIRNIADLEFSLUUOPRPILDEEMENCTOENDCENCTORNATTRIOOLNS
MEASURED BTHYESTPIEMCEIFOIFC TIRONANESLFEECRTRODE AT
FluoriFdCe-95Control
FluoriFdee-1C4o3ntrol
Transfer # Initial Fimal
Tait--iFianall
0
21
23
20
21
1
23
22
21
20
2
20
22
1.
2
3
26
17.5
25
16.5
4
16.5 19.2
16
17.3
:
TABLE 4b
I(NmgI/T1I)ALOFANFDLUFOIRNIADLEFLSUUOPRPILDEEMECNOTNECDENCTORNATTRIOOLNS
,)
MEMAESAUSRUERDECDOLBLYECSTPIEVCEILFYICATIONENEDLEOCFTRSOTDUEDY
Transfer # 0 1 2 3 4 5
FluorBiCd-e95Control FInitiail mal 16.4 16.2
15.6 16.2 15.6 16.2 16.2 16.2 15.6 15.7 19.3 17.0
FluoridCe-14C3ontrol Teit--iFiagall
15.7 17.0 15.7 15.0 14.5 16.4 16.4 15.6 15.7 17.0 15.0 16.4
H
.
>
00252
1 5
TABLE 5 IN(ImTg/I1A)L OANFDFFCI-N1A4L3 FANLDUORFICD-E95C-OCNOCNETNATIRNAITNIGON CULTURES A1N4D3 COOFNNTORONLSUPCPULLETMUREENSTED 95 AND
FC-95 Test _95 Control ~~ FC-143 Test 143 Control
Transfe#r Init. Final Init. Final Init. Final Init. Final
0.46, 0.51 0.31 0,38 <0.1 <0.1 <0.1 <0.1
1
0.50 0.46 0.36 0.36 <0.1 <0.1 <0.1 <0.1
2
0.42 0.66 0.3 0.56. <0.1 -<0.1 <0.1 <0.1
3(s) 1.75 1.6 1.75 1.5 8 1
81 1
4
0.73 0.71 0.68 0.60 <0.1 <0.1 <0.1 <0.1
5
0.72 0.78 0.61 0.68 <0.1 <0.1 0.1 0.56
J
6
0.73 0.8 0.63 0.70 <0.1 <0.1 0.1 <0.1
a
0.14 0.17 <0.1 <0.1 0.1 <0.1 0.1 <0.1
8
0.90 0.84 0.66 0.66 <0.1 <.1 0.1 0.1
9
0.84 0.73 0.72 0.60 <0.1 <0.1 <0.1 0.1
10
0.72 0.81 0.60 0.68 . 0.1 0.1 <0.1 <0.1
1
0.81 0.80 0.69 0.62 <0,1 <0.1 90.1 <0.1
12
0.74 0.73 0.66 0.62 @.1 0.1 <0.1 <0.1
13
0.73 0.84 0.64 0.66 <.1 <0.1 0.1 0.1
.
14
0.81 0.78 0.66 0.64 0.1 <0.1 0.1 <0.1
J
00253
-12-
hl
Reference Compounds
Rteesfterecnocnedictoimonpsounudssedwewreereussueidtatboledemtoonsdtergartadee tchaotmpotuhendsbiokdmeogwrnadtaotiboen somewhat resistant to degradation.
cInulttuherestiwrhsitchfowureregroiwdtehntpiecrailodtso, t3h0e mtge/slt pchuelntourlesw,asexacdedpetd ttohattwo mteheaysurleamceknetd offluoprhoecnaorlbocnosn.centArnaatliyotnicadlurpirnogbltehmes fpirrestventtherdeethgerowth p%eoriloedsss. thaInn 1th.e3 mfgo/ulr,thtgherowltihmitperoifods,enpshietniovlitywasoffotuhnedmettohodedgraasde afoprplitehde. biTohdiesgraddeamtoinosntroaftedphetnhoalt. the test conditions were suitable
sInulftohenatfeift(hLASt)hrwoausghusfeindalasgrtohwethrepfeerrieondcse, cormepfoeurnedn.ce lThiinsearcomaplkoyulnd
is a standard TAhsissocimaattieorni'asl
brieofdeergenrcaedamtaitoenritaelstusmeetdhoidn
the for
is considered to be relatively
SaonaiponiacndsDuertfearctgaenntts(6). easily degraded. In
ttheest,SoatpheanrdesDuelttsergaernet cAosnssoicdiearteidoni'nsvalsihdakeif ftlhaesk rbeimoodveaglraodfat1i-odnodecene-
derived LAS is not nearly complete.
)
TshueppldeamtaenstheodwicnognttrhoelsexatreentdepofictdeedgraidnatTiAoBnLEof6. LASTheindsautrafaschtoawnitng
tcohnetreoqlusivanloetntsupampoluenmtentofedmewtihtyhleLnAeS balrueedeapctiicvteedsuibnstTaABnLcEes7.in tLhiettle
LTAhSreedetgrraandsafteirosn woecrceurrreedquidruerdingbeftohreefitrhsetmafjeowriatdyaptoifvethetraLnAsSfebresg.an
ttorandsefgerrasdeweirnethreeqsuuirrfeadctfaonrt LsASuppdleegmreadnatteidonconitnrotlhe f1o4r3 FCc-o9n5t.rol.Five
TdheerrievfeodreL,AS iwt eraeppneoatredintihtaitalolrygapnriessemnst cainpabslueffiocfiednetgranduimnbgers1-dfoordecene-
LAS degradation. these organisms,
bTutheentresitchmceonntditoicocnurraeldlowatedafsorloewenrricrhamteentthoafn
had
bteoeninacnrteiacsiepattheed,rate Caonndselqiukeenltilhyo,ocdhaonfgeasccwleirmeatmiandgeoringantihsemsproccaepdaubrlee
o3ftdoe,g4-r6adidnaygs,theandflutoermopcehreamtiucraelsw.as Grraoiwstehd pferroimordosomwerteemepxetreantduerde from
d(<e2g0radatotio2n2Ci)n ttoheafcionanlstagnrtowtthemppeerraitoudreareofsh2o5wCn. inRTeAsBulLtEs 8.of LAS
wIansthdeonegrotwothdepteerrimoidnse iffollo5w0inmgg/ltraonfsfFeCr-s9511oranFdC-1124,3 ainnheixbpietreidment
tahpepedaresgratdoathiaovne oafn LiAnSh.ibiTthiensgeefrfeescutltsonartehesmhoiwcnrobiinalTABdLeEgr9a.datiFoCn-95
ooCffomLpFAaCSr-.i9s5onHionwwheiivtbehirt,TeAdBiLtLsEASSprd8eesagenrndacdea6twisaohsnowsnboytth1ac8to%mpatlhneedtep2lr3ye%sedinnuchreiibniogtfotrh5ye0.semg/tlwo
)
otfestouprermieotdhso.d, FOnC-1th4e3 doitdhernothanadp,pewairthtionhatvhee alimsiitgsnifoifcatnhte pefrfeeccitsion
on LAS degradation.
000254
:
-13-
d
In the final growth period, 50 mg/l of carbon 14-labeled FC-95 and FC-143 were used as test substrates in place of the nonlabeled
ftrliupolrioccahteem.icalTsh.e
Both FC-95 and FC-143 cultures were prepared in concentrations of the radioactive fluorocarbons
parreesesnhtownininthTeABaLqEueo1u0.s pThhaeseiansitidaelterFmCi-n9e5dcboyncesnctirnattiilolnatiiosnmuccohunting
slyoswterematthianc eexrpreocrteidn. theThicsollleocwtivoanlueofcotuheldihnaivteialresFCu-l9t5edsafmrpolmesa,
cItultiusreaslswohepnossthieblefirtshtatsFaCm-p9l5e hwaasd ntoatkenc,ompbultetetlhyisdsiesesmoslveudnliikneltyh,e
sciunlcteuretsheweirneitiaallmosvtaluiedsentfiocralFC-(9350.3c,onc2e0n,t8raatnidon30.f4rommg/all)l. 3 pNaervaelri-el
wtihtehlesFsC,-95theanrdeFmCa-i1n4i3ngredmaatiansehdowin tshaotluttihoen radduiroiangctitvhietyentaisrseociated
s7h-odawyeddseogmreadbatiinodnintgesotf preardiiooda.ctivAenalmaytseirsiaolf, tbhuetbitohleogviacstalmsajoolriidtsy
remained in the 1iquid phase.
TABLE 6
CONCENTRACTOINOTNROOLFS LAANSD %(mLgA/Sl)REINMOSVUEDPPLEMENTED
)
95 - Surfactant Control 143 - Surfactant Control
us _ TT ----"LAS
Transfer# Init.Final %Removal(7) Init. Final %Removal
4
31.5 26.8 18.4
35.5 20.5 10.4
5
28.3 27.5 0.1
32.8 25.87 21.2
6
20.8 25.5 15.5
27.0 24.0 7.6
7
25.0 12.0 91.1
25.0 30.6 -2.0
8
31.2 3.75 89.8
37.0 , 35.8 11.1
9
33.0 3.17 95.1
38.9 13.7 94.6
10
32.7 2.33 95.9
42.7 12.8 89.7
1
3.0 2.0 95.0
3 13.7 93.8
12
31.3 2.33 96.5
41.3 19.7 77.9
13
31.7 2.5 93.5
41.3 18.0 88.3
1
31.3 3.0 92.8
40.3 13.7 90.7
J
00255
.
-14-
0
TABLE 7
CONCENT(RmAgT/I1)ONIONFNMOENTSHUYPLPELNEEMEBNLTUEED ACOCNTTIRVOELSSUBSTANCE
Transfer #
95 - Control Init. Final
143 - Control Init. Final
4
1.0 19
4.5 4.5
5
1.15 .38
4.5 3.5
6
To.s0 78
7.1 5.50
-
7
5.25 10.2
5.0 10.2
8
3.0 0.88
2.0 10.9
9
5.75 1.83 0.9 12.2
10
437 107
12.3 9.67
1
4.33 0.67 1.5 12.0
)
12
3.0 1.33
12.3 . 13.3
13
3.67 0.67 1.3 14.5
14
3.67 1.0
14.5 11.3
TABLE 8 CONCENTRATION OF MBAS (mg/1) IN SURFACTANT SUPPLEMEDNURTIENDGAFNIDNNAOLNSGURPOPWLTEHMPEENRTIEODDCONTROLS
----#F1CLA-S95#C2onLtArSolsRon ---7E1CL=A1S43C#o2ntLrASols NomTime Suppl. Suppl. Suppl. Suppl. Suppl. Suppl.
Initial 28.7 20.3 1.0
34.0 36.7 6.5
Day 2 13.0 26.0 1.0
8.0 22.3 53
0
Day 7 1.67 2.0 7
6.3 6.3 4.0
HReLmAoSval (7)96.5 95.4 -
91.6 92.3
-
00256
i
9
-15-
TABLE EFBFIEOCDTEGORFADFACT-I95ONAONFD FLACS-1A4N3AOLNYZTEHDE
FOR AS MBAS
FC-95 + LAS Culture FC-143 + LAS Culture
Transfe#r -- Ini% t. L_ Fii naa l s Rem_ ova_ l(S)~ Init.~ Final %HLemaosva_l_(s)
1
58.0 34.7 73.6 53.7 27.3. 97.8
12
64.3 40.3 78.9 53.7 34.0 71.4
.
TABLE 10
THECOCNECNETNRTIRFAUTIGOANTEOFOF4TcE_SFTc-C9U5LTUORRE1D4UcR-IFNc-G143 IN THE FINAL GROWTH PERIOD
--Ye-- _pc--- 95 CulSttuarnedsard Concentration Deviation
Init. 30.1mg/l 0.3 mg/l
Day 2 52.8
0.5
Day' 7 53.5
3.1 :
1c-rc-1+4--3 culSttuaressdird" Concentration Deviation
46.2 mg/1 18.0
0.9 ng/1 0.3
49.7
0.4
00257
Bnd
:
.
-16-
.
D aThcitni-vleamyeertabcohlriocmaptroogdruacpthsy odfideinottherrevFeCa-l143thoerprFeC-s0e5n.ce oLifkerwaidsieo,bgeasforleiqaunidd cahfrteormamteotghryalpahtyioonf, thsehoswaemdenoculptruodruectesxtrtahcatts,werbeothmot cionmibtiinaaltliyonpreosfentthesoer nroetsulatlss,o pirtesceanntbeincocnonctlruodlesd. thFatrommotbhieodegradation of these fluorochemicals occurred.
REFERENCES AND FOOTNOTES
(1) DGeogldrmaadna,tioPneteorf, `SEynnztyhmeotliocgyOrgoafniCcarMbooln-eHcaulloegseninBotnhdes.Biosphere, Nat Acad. ofSof., WashingDCt(o1n07,2). oo oenerS
(2) gTrhoewrtehwapseriaodsixd-udraiyngpwehriicohd btehfeorteestthecuolntsuertesofwerteheshfaikneanl at 25 C. in the presence of FC-95 or FC-143.
(3)TSdttahndardEdMiteiotnh, oAdmserfiocranthPeubElxiacmHienaalttihon ofA.ssWoactieartioannd Wastew(1975).ater, |
(4) D22agtiF.me Ntiegmhpteratteumrpeesratwuerreesowbesreervenodttomearsaunrgeedbedtuwreienng 2t0hatandpart
y
oefrattuhreest(usdeye TiAnBwLhEic3)h, cuHlotwuerveesr,wermeeassuhraekmeenntatmaadmebiennetarttehmep
ntiegrhmtitniamteionteomfperthaitsure23-fmroenqtuhentsltyudyd,ropisn Jtoanu17a0gy,C. showed that
(5) Aathitghhis ftlruaonrsifdeer,conDceecnattruartisolnu.dge was added which: contained
(6) SDuebtceormgmeinttteAessoocniaBtiioodne,graAdaPtrioocneduTreestaMnedthSotdasndaorfdstheforSoatpheand ~ SDueltfeornmaitneatiaonnd oLfinetahre BAlikoydleagtreadaSbuilflointayteo.f AlJ.kylofBetnhzeenAemerican 0il Chemists' Society, 42:986 (1966).
(7) Percent LAS removal was calculated as:
(MBASG; -MBASy;) - (MBASgp -MBAS,p)
% Removal = ----SL CI"
SF
CF y 44,
h
BaSg; ~ WBASy;
Where:
MBASG; = Tchoenceinnittriaatlionmetohfyltehneesbulrufeactaacnttivesupspulbesmteanntceeds (cMuBlAtSu)re.
BAS(; = Tchoentrionliti(aTlABLMEBAS7).concentration of the nonsupplemented
J
MBAS, = TshuepplfeinmaelntMeBdASculctounrcee.ntration of the surfactant
MBAS; = Tchoentrfoilnal(TMABBALSE 7c)o.ncentration of the nonsupplemented
0253
STE
D
: )
-17-
(7) The percent LAS Removal was calculated as: (MBASgqpy -MBAS) = (MBASgyp -MBAS.)
% Removal = ------------------------------ 1 (MBASgy -MBAS)
Where: '
MBASgy; = Tchoenceinnittriaatlionmetofhyltehneecbulluteureactsiuvpeplseumbesnttaendcbeys b(oMtBhAS) LAS surfactant and either FC-95 or FC-143,
MBAS = pThleepiennittailalconMBtArSolco(nTcAeBnLEtra7t).ion of the nonsup-
MBASgnp = The final MBAS concentration of cultures supplemented with surfactant and fluorocarbon.
MBAScp = The final MBAS concentration of the nonsupplemental
control (TABLE 7). MBASg, = The initial MBAS concentration of the surfactant
supplemental culture (TABLE 6).
It was assumed that MBAS concentration due to FC-95 or
- oFfC-1t4h3esewascomnpootunrdesd.uced by the biodegradation or other loss
ol
EAR/cen
J
00259