Document DvmQO1qJ5eqk7aexYZ7BJK2ao
AR226 _ 02897
BIODEGRADATION (301E)
TEST SUBSTANCE
Identity: A mixture containing perfluorooctanesulfonate, which may also be
referred to as PFOS, FC-95, or as a component of FC-3017. Octanesulfonic acid) (CAS # 2795-39-3).
(1-
RFeCm-a30r1k7s:prodTuhceed3iMn Apnrotdwuecrtpi,onBellogtinuumm.beCrurwraenst2i3n3f0o.rmaTthieonteisntdiscaamtpesleFCis3017 is a mixture of 0.67% PFOS, 20% diethylene glycol butyl ether, 55.41% 0wa.t5e%r,p2ol1y%oxeytehtyhlyelneeneglmyoconlo,oc1t.y3l3p%heHnyydlroexthyerf,oa0m.e0r4,%1s%odsioudmiulmauorcyltyslulsfualtfea,te, and 0.05% tolyltraizole.
cThhaerafcotlelroiwziendg scounmcmeantrryataipoplnisoesftiompaurmiitxiteusr.e Dwaittha imnacyomnpoltetaeclcyurately
reflect the biodegradation potentialofthe thetest sample.
fluorochemical component of
METHOD
Method: Modified 1979 version
OECD
Screening
Test,
OECD
301E
with
DOC
Analysis,
Test type: Aerobic Ready Biodegradation
GLP: No
Year Completed: 1980
Analytical Statistical
monitoring: Dissolved organic carbon methods: Results were determined by
(DOC) calculationofthe
%
DOC
removed as compared to the blank controls.
TTehsetsoercgoanndairsym esfofulurecnet:waAs 5f0r:o5m0 amnixacotfivsoaitledexstlraucdtgeanadersateicoonndbaasriyneafftlutehnet.
sMoeitlrforoWmasttheewtaotwenrofTrWehaittmeenBteaPlranLta,keS,t.RPaamuls,eMyNC,ouwnhtiyl,e tMhNe.soil was garden
Test condition: Dilution water: Deionized water
-
Mineral (1979).
Nutrient Initial pH
Medium: 7.0.
Nutrient
medium
per
OECD
301E
method
Test and prepared
reference solution preparation: by dissolving 208 mg in on liter of
The test material was mineral nutrient medium.
Trheifserseonlcuetisounbgsitvaenscea,fhinyadlrtoeqsuticnoonncee,ntwraastipornoepfa4r0edmbgy DdiOsCs/oLl.vinTgh3e0.6
mg in one liter of mineral nutrient medium. test concentration of 20 mg DOC/L
This solution gives a final
~
Tsoelsuttivoensasneldsc:a5p0p0edmLwitEhrlfeonammeypleurgsf.lasks containing 250 mL test
Incubation conditions:
Lighting:
Constant dark conditions.
002747
Temperature: 23.5-24C
Agitation:
Continuously
sNuubmsbtaenrdo)facnodncbelanntkr,aatliloinnsd:upl1icpaltues. hydroquinone (reference
Iarneocrueqluuimrecdopnedrittihoenroefnerteesntceidnimteiatthioodn):.Not given (No measurements
Total suspended solids and Element Basis: Decrease in
pH on dayoftesting: Not given. dissolved organic carbon compared
to
Ttheestblsaunkbss.tance flask conditions: Not given.
|
RESULTS
Nanodmianta~l2c0omncgenDtOrCaItLiopnlsu:s HBglCakn,k c(ionnhtirboilt,edh)y,tdersotqumaitneorniealata~t2~04m0gmDgODCO/CLIL and at ~40 mg DOCIL plus HgCh, all in duplicate.
Element values: and 94.6% DOC
28-Day removal in
Degradation duplicate 2.
= 96.2% DOC Mean value =
removal 95%.
in
duplicate
1
Remarks: Testing was conducted on the mixture as described in the Test SthuebsftluaonrcoechReemmicaarlkpsrofipeoldr.tiTonhealvoanleu.es reported apply to that mixture and not
CONCLUSIONS
Percent biodegradation of FC-3017 value was 95 after 28 days.
was
based
on
the
mean
DOC
removal
Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St.
Paul, Minnesota, 55133
DATA QUALITY
tReesltiianbgi.lityH:owKeivmeirs,cthhreasnakimnpgle= p2u.riTthyiwsasstundoyt mperoeptesrltyhechcarritaecrtiearfiozreqduaalnidtythe isntusodlyutliaocnk.s analytical confirmationofthe amountoffluorochemical proportion
REFERENCES
The studies were conducted by the 3M Company, EnvironmentalLaboratory,
St. Paul, MN, Lab Request number 5541S, 1980.
oo
OTHER
Last changed: 6/26/00
002748
iimmena
TECHNICAL REPORT SUMMARY
3 TECHNICAL COMMUNICATIONS CENTER -- 201.208
portan--t Ifrepisporinrtedton both siodfpaepesr,send te copes to TCC.)
[=oom 6/30/80
="Environmental Laboratory ( & pC)
0535
Commercial Chemicals Division
9e97e00r12600
Biodegradation of "LIGHT WATER" Products in OECD Test-6/80 | 040
John A. Pignato - Commercial Chemicals Division - 236-24 (01)
Eric A. Reiner - Environmental Lab (EE&PC) - 21-BW (63) 47816
None - See 3M ----
Environmental 0cows
Lab
Request No. 55418 ERS
31 meen
ESwoensas:rmomas CURRENT oRIEGTIVE:
a athe
bvoirraotnomreyntal
TpTeorsotudsucewtistthheaDrOeiCnhtiaegnrhanllayytsiibosinoadltelogyrsahrdoeawcbolgtenh.iatzed"LOIEGCHDT WSAcTrEeRe"ning
E & BC)
FFF
aLEPOS RT ABSTCRAoCT,m200250 word) Tis snvact formations deed oytoTemi Commansioms Som
oc
sAT"hnoLianIslcGylHsuTsistsiWu'vAdeTyElaRynu"dsdedeBsmruoaptnnhpdeslterm"aeMtnoetdailftiheepdaerxOatlEelCneDlsiSvcserteeebrniiiolndegegcrToaendstatrboilwlsiitthcoy.DoOfC
lFCe-v3e0l1s7.of FInC-21046dadyesg,rAaFdFteFhedpdrbyiodsus9co0t8lsv:eadndoFrFCgC-a-2n3003i1,c7 c0aw-ra2bso0n6'9,45(DaOnCg)
nTdeohgetradpduaeerda.ltloelcECh-es2mt0ie3craillsehoorwceodpnhtyr9so3il3csaDlOp.Cr-opdrveoegcdreastdshaeatstiostnuhcihsinaDgO2C1 dlaoysss. was
maadtseorripatli.on, volatilization, or precipitation of the parent
HE SRa0
002749
nls
r
BG
am
TECHNICAL . REPORT SUMMARY
TO: TECHNICAL COMMUNICATIONS CENTER -- 201.20
pane sorsn bo ss ofapr nd cio 76)
FL FC) =
6/30/80
"Environmental Laboratory (EE & pC)
0535
Commercial Chemicals Division
9970012600
Biodegradation of "LIGHT WATER" Products in OECD Test-6/80
040
John A. Pignato - Commercial Chemicals Division - 236-24 (01)
emEoriRac A. Reiner - Environmental Lab (EE&PC) - 21-BW (63 SP47816 Ce
None
ecomvy
-
See 3M
8 oom
Environmental
T coma
Lab
Request No. 5541S
mcnameALy
31
on re rs
Ke eroroS : romau CURRENT GRIECTIVE:
ost To
To use the internationally recognized OECD Screening
Environmental
Tpersotducwtisth aDreOChiagnhallyysbiisodteogrsahdoawblteh.at "LIGHT WATER"
L(EaEbo&ratPCo)r]y
SEE Rc-203
rE
REPOSRTNABESTRnACTe: (2a0s0.:250word) Tos somvectinfGoviromteadtoyioneashen Sommers Some
TOC
gonclusively demon TAhniaslyssitsu"dy aundsedsuptphelem"eMnotdailfiepdaraOlElCeDlScsrteeerniilneg cToensttrolwsithtoDOC
strate the extensive biodegradanitity of "LIGHT WATER" Brand AFFF products:
FC-3017. In 14 days, the dissolved
FC-203, organic
FC-206, and carbon (DOC)
levels of degraded.
FC-206 degraded by FC-203 showed 93%
90% DOC
and FC-3017 degradation
was 94% in 21 days.
The not
dpuaeraltloelchesmtiecraille orconphtyrsoilcsalProPrvoecdestsheast-stuhcihs
DOC as
loss
was
material. adsorption, volatilization, or precipitation of the parent
ste: --Fe-2,e MFP eo Sin AplyeaShreds 0
w[ahSt Eover ST
J
oe
002750
.
-2-
3
BIODEGRADATIONIOnF02"CLhIGTHETSWTATER" PRODUCTS
INTRODUCTION
hbieode3g4raEdnavbiirolnimteyntoafl "LLaIGbHoTraWtAoTrEyR"chopsreodutcotsattuesmipntg ttohedeOmEoCnDstsrcarteeenitnhge nEentvhiordonmbeenctaaulse Ltahbehatsesthadhasexwpiedreienicneterunsaintgiontahle aOcEcCeDpttaenscte,asanad the Cpaormtpilceimpeanntt thien eanxisitnitnegrnaBtOiDondaaltaritnhgattesshto.w tThheesseeprroedsuuclttss wtiollhave a high degree of biodegradability.
TchaerboOnE.CD Fteosrt tihseserunwataetr-2m0i-s2c5i0bCleand"LIitGHmTeaWsATuErRe"s opnrloyducdtiss,solwvheidchoragraenic dntoeetgsrtaiendxgap,teicottneh.de OtTEohCeDvoTltOaeCtsitlainizasel,yasnipsraepcpmierptoihptoraditaete,evenomretmhbeeoadsuardfesosorrbmteehdaesudprueirrinfnglguoritnoattaeld portion of these products. METHODS AND MATERIALS
FHetohodls lantodheCwSoneidpittneimogbnesr-19W,ade197A9., ScvheerislionperoffortmheedMotdhiisfietdestOiEnCgD aSctrteaecnhiendg (TAepsptenwdiixth1)D.OC Analysis{l). A copy of this method is
) Raevdauicltaibolnesshiankitnhge esqcuailpemeOnft.theFitveesthwuenrderedn-enclessEarrlyenmtoeyaedrapftlasitksto rceeppllaacceedd 920-0limtle,r aflnadsktsh,e s25a0mplmle osfizeinwoacsula2t5,ednontutr30ienmtl. solAsutiionnthe pErielstcrraitebedforproDcOeCduarnea,lystihse.analyst used only the last 10 ml of sample
wAitchailnibrtaheted20c-h2a5r0tC rraencgoerdedrurimnognittohreed28-thdeaytetmespte,rateuxrceepwthifcohr sata3y-4edhr. pheerairodthwehehnighitenrdeaocfhedthe25.r5a9nCg.e (T2h3e.5-t2e4m9pCe)r.ature was most frequently
Chemicals - All chemicals were reagent grade unless otherwise noted.
oTfheAn3tuwe"rLpIGmHaTnuWfAaTcERt"urep.roduTchtesirusleadbelweriedenstaimfpileeds otfhemcoamsmerFcCi-a2l03maNFtPe,rial FC-3017 Lot 2303, and FC-206 (FM 3824) Lot 2330 10/79.
gThreadehydhryodqruoiqnuoinneoneuse9d8.5a%s, thLeotca1l0i0b14r7a.tionFocuormpcohuenmdicawlassAulpdprliychhourseeasgent pWerreescrciobnetdactbeydthien ManodiaftiteedmptOECtDo oSbctraeiennintgheTe9s9t.,5%bupturethihsydrpouqruiitnyonies SZpepdaurceendtypurnoittyrweialdliyhaavvaeilnaoblseignifnictahentUSeAf.fecWteonexpreecsutlttsh.at Tthheis ) tfhuepiurritpireessenhcaevecnoouldappmaarkeentonliynhiabittroirviyaleffcehcatngeonimnicTrOCoormgeaansiusremmsenatnsd.
002751
oa
wwaasterSt.usePdaulincittnye wparteeprarpaatsisoendofthraollughsolautciaornbso,n beexdc,epmtactrhoepoirnooucsulum,
)
atnhiroonuigch ares0i.n2-ubend,filatnder.2 miTxheids wbaetderdewiaosnizaelrsso, usaendd ttohenmakfeilutpereldosses
etxopeerviampeonrtatieonns.uredStcoocknssiosftenwtatewratecrolqlueacltietdy.at the start of the
Itnhepprreepasrcirnigbednut3r3.i4entg OsFoluNIt,iUoInPsO,4-22161.340, g3o6f.3agnhOyfdrCoauCslyN-a3p1l1i5P00, rerpelpalcaceedd 2i7d.e5ntigcaClaCly5. tho`sheesereqrueiprleadcempeynttsheyOiEeClDdedMetshooldu.tioTnhseaonpdticoonnaclentyreaatsitons e(x1t.r6a.c1t.2(so)l)u.tion`Thweas100usemdg oifn pFlea-cceheloaftetheusevditaimninthesolturtaiceonelement sanodlut70ionmgcoEnDsTAi)s.ted of equimolar amounts of FeCl and EDTA (30 mg FeCly
TSeesrtveCdoancsetnthreabtaisoinss =foDrOCadmjeuasstuirnegmeinntistioafl staemsptlecosntcoecnktrdaitliuotnisontso
acpopnrcoexnitmraatteiloyn 4i0n mtgheofracngaerbornecpoemrmenlditeedr.by TthhiesOEisCDthteesth.igheUsste of this
mheiagshurienmietnitasl. coTnhceentlroawtitoonxiciintcyreoafsed"LIthGeHTsWeAnTsEiRt"iviptryodoufcts& DtOoC removal
microorganisms unlikely.
made
significant
inhibition
at
this
concentration
Acdajrubsotnnepnetr olfitheyrdrwoagsuidnoonnee,bythweeicghatlibbraastediononcoimtpsouknndo,wntocar20bonmg of ) concentration of 65.4%.
Tcwoorbrelcatnikoncso.ntrols, instead of 1, were utilized in making blank DOC
1I.no6c.u2l.u1nan-dTh1e.6.i2n.o2cuOlfumtwhaesOE5C0D-50temsitxtuusriengprferpeasrhedsoialccofrrdoimngE. tAo. steps Rseliundegre'saegraartdieonn banadsintheatsutpheernSta.tanPtaulofMaetrsoampPlleantf.romTahne afcitlitveartepdaper cusheldorwianse-fWrheaetmawenlkl54w,atearn.d the water for- soil extraction was
aSsterilecCoosterilennttrorlosl-sRefpolriceaatceh vteessstelmsatceroinatlainainndg th4e00cmagl/ilbraigtCiloyn served choamnpdoluinndg. ofiltahnedlvinigabOlfe ctuhletsueresst.erilTeheyconhtardoltshewassameideinntnioccaullumtoantdhe s0-ampalnidng28f-rdeaqyuesnacwyp,lesb.ut the analyst only made DOC measurements on the
Sample Preservation - The sample preservation method used was that dpsraromeppslceorfibtecodon<pcbieylnt2tr,haetaenTddOC HsCmtLaonruitfnoagctthtuehree1r0s(a4mm)lp.lefsiTlhtueenrdemederthsroaedmfprliiegnsev,roalvtbeirsoinngaidindnignvtgihael1s pwirtehseravlautmiinounmmet0hiold-.lineTdhecampasn.ufaTchteuremrethionddicisataens etfhfaetcttihveemethod can J pshttahbaillaitzee, caalirberaadtiiloynbsioolduetgiroandsablmeadematferroimalp,otafsosriusmevehryadlrogmeonnths.
002752
Soa
in
0
Since the
necessary
acidification in this preservation
step in the TOC analysis protocol,
method was also
its use was the
a
most
dpmeorsnactcrhtiibcoeafdl thaienndptrhaeelslOeoErwCevDdatimeoeltnihmooidfn.attihoAeLnLsoafTmOplCtehsea.naglCylsyespwreerseervmaadteionwittheicnhniaque
DIonnsrtmraunmenDtCs-52aAn.d per liter.
eqIutsipmseenntsi-tiTvhietyorlgiamniitc iscarbbeotnweeannal1 yzaendr
2usmedg
wOafs caarbon
Tpthlieungesss.taonpdGpeerlssmtaonrfeodr0.2tinheudmerimeoeanmcibtzrieaodnnewfalftaieslrkt,esrsws,eerrev(ePdcalrettaonH,of.iplot6re4o8ru1s4s,)ampbploleiaslsetdifcor3 foDaOCm analysis.
G(lCahsrsowmaerregeRc)leafnoilnlgoweidnvobylve6ddesiooankiinzgedinwatcehrromFiicnseasc.id cleaning solution RESULTS AND DISCUSSION
Tcaobnlteain1 stuhemmaarcitzueasl function of time.
dtahtearesshuelettss oafndtphlisotsstuodfy.theAdpepegnrdaidcaetsion1 aasnda 2 All 3 "LIGHT WATER" AFFF products degraded nearly
completely.
3
Percent
Degradation
of
HydroquiTnAoBnLeE
1 and
"LIGHT
WATER"
APFF
Products
FC-203,ModFiCf-i2e0d6,OEanCdD FSCc-r3e0e1n7ingWitThestTime in
* Degradation at Day:
Product 1 14 21 27
28
FC-203 FC-206
13 23
85 90
9943
9931_
9923
FHCy-d3r0o1-7 8397 992a
9976
996
9935
quinone
ovIrenrpytrheeucniclapisiketealtyofiotnhtahectauhpsihegydhsliyctahelwatmleeorasnsssOoflsuubcslhoeluabs"lLeIaGdTHsOoTCrpWtA(iTDoEOnRC,"). pvroTolhdauitcsitlsii,sz.atiitoni,s
steusbtsstaonftiattheed 3Mby EnthveirornemleanttiavlelyLabhoirgahtoorxyyge(nFCd-e20m3andBODob5s/eCrOvDed= FC-206 BOD5/COD = 0.5).
0i.n5,BOD-
J "e"TvLhiIedGeHsnTtceeWrAiTltEehRa"ctonpttrhroeodluscotdlsautblaies inTnOoCtTabdlluoeess2tofprarodomsvoitrdhpeetiftouenrs,tthevsroalmapstluieblssitzacanottniitoaanli,ninogF psraemcpilpeistaetxicoenp."t tThhaetsethesyamcpolnestaiwneerde hHgaCnldpledto ipdreenvteinctalmliycrtoobitahle gtreoswtth.
002753
SoA
s-
) AnttohetthiaenmieatnijdaolrofDfOaCtc.hteor,T2h8u-astdaylloesatssetstoffpoersroilnouodbn,lmeettTahObeCoylibszyteidltlhe"scLeoInGtpHhaTyisnWieAcdTaElRn"eamordleysawllas cnootmporneemnotvse. "LTIhGiHsT cWoAnTtErRo"lmdeoteasbno'ltitepsrovien tthheattpehsytsircuanls,prsoicnceessenso did mfeotrambaotlioinsmofoccluersrsedwatienr tsheosleublsetermialteericaolntsroilss,unbluitkeltyhe bceactauasbeolic* catabolic products are usually more polar and smaller.
TABLE 2
DOC Loss from Sterile Controls
Product
% atDODCayRem2a8 ining
Cc--220063 iicy-a3r0o1q7uinone
9914..32
8787..15
-
Adsnalyiogf0hft-lpywrhiiobtrreowtnpoirsefhci,ilptieatsraitnwegasffotorhremeDdsOColinuatniatolheynsihisyn.drtoheqTuhiehnyodpnrreoeqcusiitpneiortinaleteescwtaoesnritlreol on
) OcatEoCntDlreoaelsxtpearfiitnemretnhte2.1prdeaTyshsei.snceaTphpoifasrelinsgtuCgLgyt,eesntdtsehnrcotyuhgahtofouhthyyddrthroeoqqucuioinunoronsneee iostfonotthest2a8b-ldea,y
sipnoanptparnoeporuisaltye precipitation
cfoonrtmroilnsocloumbploeundhumfuors-ltihkeeOmEaCtDertieaslt.makIetss piatrtainal with HgCly also casts some doubts about the
maeptphroodp.riateness of using the HgCl, preservation procedure in the OECD
concLusToN
Tpihrneod2up1crtedsas,yesntFuC-ns2dt0eu3rd,ytchFeoCn-2cc0lo6un,sdiivtaienoldnysFdCe-om3f0o1nt7sh,teramatoredesifniteehadartlyOE"cCLoDIGmHpSTlcerWteAeeTnlEiyRn"gdegTresatd.ed REFERENCES
(1) To1er97sg9ta,innigTzeastPtiroonGguriafdmoerlEiExncpeoenrotfmoirGcrotCuheop-MooopndeirDfaeitgeirdoandOaaEtnCidDon/SDAcecrveceeulnmoiupnlmgaetnitToensC,themwDiiectcah.lsDOC
Analysis (Level I) (H. G. Nosler) Revision of Sept. 19, 1979.
(2) Dohrnan DC-52A Operating Manual, 4th Ed., 1978, p. 3-1.
.
Appendices: 12 -- ODaEtCaD TSheesettsGuidelines (Sept. 19, 1979)
h)
3 - Graphs
* Cnoalteacbuolleiss.m is biologically facilitated breakdown to less complex
002754
3.
APPENDIX 1
:
| Tokyo, December 1st, 1979
D
C 118/79/1nt. OECD - Chemicals Testing Programme Expert Group C, Degradation/Accumulation
Modified OECD Screening Test with DOC Analysis
002755
Bes
Test Guideline C 118/79/Int. Tweistth DGOuCideAlnianleysifsor the Modifid OECD Screening Test Date of last revision: Sept. 19,1979 Levl I test for ready biodegradability
1. Prerequisites sItoluhabsletoinbetheknocwonncwehnettrhaetriontherantgeestemmaptleoryieadl inisthe test (corresponding to 5 - 40 mg DOC/1).
2. Guidance Information
Kpnroowpleerdtgiecsofof thtehebatcetsterimaaltertioaxliciitsymootr uinmehqiubiitvoorcyally
)
creoqnudiurcctdionbutofctohnestittesutt.es useful information for the
3. Qualifying Statements
Tuohrlegtaimnmeiatctheocdobmipoiosduensgdursia.tdeadbItilfioirtsyutnhoesfuiwmetaaetsdeurrfesomorelnuttbhleeo,fbiotnhdoeeng-rvaaoedlroaabtbiiiclleity
cvaluation of mixtures.
_
002756
Bar
Ds.
Groa/s
Sept. 19. 1979
Modified OECD Screening Test with DOC Analysis Preamble
It has to be realized that the following procedure
for the modified OECD Screening Test has to be
)
regarded in some points as provisional since se-
veral important features such as the calibration
compound and the new test duration of 28 days are completely untried yet. However, since the test
constitutes a modification the official, widely practised and well accepted OECD Screening Test for the biodegradability evaluation of surfactants there should exist a good prospect for success.
J
002757
D4
D
Septemder 19., 1979
OECD Chemicals Testing Programme
Test Guideline for the Modified OECD Screening Test with DOC Analysis
0. The test procedure constitutes a modification of the OECD Screening Test (OECD Environment Directorate,
Proposed Method for the Determination of the Biodegradadbility of Surfactants Used in Synthetic Detergents, Paris 1976, and council directive of Nov. 22,
1973, on the approximation of the laws of the member
states relating to methods of testing the biodegrada-
)
bility of anionic surfactagts (73/405/EEC), Offigial
Journal of the European Communities No. 4 347/53 of
Dec. 17, 1973) for the application of the dissolved
organic carbon DOC analysis.
1. Method
1.1 Introduction: Purpose, scope, relevance, and application of test and explanation of-limits.
The purpose of the method is the measurement of the
ultimate biodegradability of water soluble, non =
volatile organic compounds in an aerobic, aqueous
medium at a starting test concentration correspondirg
to 5 - 40mg DOC/1 ( In order to avoid inhibitory
effects it is in the investigator's own interest
-
-2-
002758
oy
d
Re
to choose as low a starting concentration as his
analytical capability permits )
1.2 Definitions and units
1.2.1 Definition of biodegradability
oe | LL oom] % | 1-
L
c, = , |i x 100
bl,
where
Dy = degradation in.percent DOC-removal
)
at time t
Cy = starting DOC concentration of the culture medium (mg DOC/1)
Cy = DOC' concentration of the culture medium at time t (mg DOC/1)
Cy,= starting DOC concentration of the blank (mg DOC/1) -
Cpy= DOC concentration of the blank at
time t (mg DOC/1)
1.2.2 Units
The degradation is stated as the percentage
DOC-removal within 28 days with respect to the test material ( % DOC-removal )
2
002759
LS
@
)
-3-
1.3 Reference compounds
1.3.1
Calibration compound The calibration compound used in this test is
hydroquinone at a concentration corresponding
to 20 mg DOC/1. Hydroquinone has to exhibit a
DOC-removal of 260 % within 28 days, otherwise the test is regarded as invalid.
1.4 The orinciple of the method
.
A predetermined amount of the compound is dissol-
ved in an inorganic medium (mineral nutrient solu-
tion, fortified with a trace element and essential
)
vitamin solution), providing a concentration corres-
ponding to 5 - 40 mg DOC/1. The solution is inocu=
lated with a small number of microorganisms from a
mixed population and aerated at 293 - 208 (20 - 25C)
in the dark or at least in diffuse light only. The
degradation is followed by DOC analysis over a 28
day period. The procedure is checked by means of a
standard (hydroquinone).
=
A control with inoculation but without either test
material or standard is run parallel for the deter-
mination of DOC blanks.
1.5 Quality criteria
1.5.1 Reproducibility
The reproducibility of the method is appropriate
oo
for a screening test which has solely an accep-
)
tance but no rejective function.
.
002760
97,
5 4-
1.5.2 Sensitivity
The sensitivity of the method is largely deter-
mined by the sensitivity limit of the organic
carbon analysis which is 0.5 mg C/1 at the present
state of the art.
-
1.5.3 SAppepcliifciacbiltey,forapptlheicabbiioldietgyradability evaluation
of water soluble, non - volatile organic compounds.
1.5.4 Possibility of standardization
)
The test version with specific analyses for anionic and nonionic surfactants is standardized
as "OECD Screening Test".
1.5.5 Possibility of automation Parts of the test, e.g., the analysis,can be automated, although hardly the total procedure. The procedure is, though, well suited for being operated with whole series of test materials.
1.5.6 Costs ( in 1978 Swiss Francs )
1.5.6.1 Equipment
Glassware
2000,-
shaking machine
6000 = 20000,-. _
Carbon analyzer
42000,-
Miscellaneous (PH meter, balance,
J
provision for air conditioning)
5000,~
"gw
!
002761
@r
) -5-
1.5.6.2 Person - hours
10
1.5.6.3 A(pipnrvoexsitmmaetnets taostaslumceodsttopebre test 750, = 1000,amortized)
1.6 Description of the method
1.6.1 Reagents and materials
-
1.6.1.1 Deionized water
Deionized or distilled water free of
substances (copper in particular), for
use as a solvent. Water which has been
zed by distillation or ion exchange is
toxic general ceionisuitable.
)
A high purity of this test water is necessary
in view of the DOC analyses in the concentra-
tion range of 0 - 40 mg/l. The contaminations
result from inherent impurities but also from
the ion exchange resins and microbial devel-
opments (bacteria, algae under the influence
of light ete). Only one water charge must be
used for one test series which is to be controlled
beforehand by DOC analysis. If necessary,
suitable water may be gained by UV irradiation
or other means.
1.6.1.2 Nutrient solution Mix 1 ml each of the following solutions (a) o (f) and make up to a volume of 1 1 with water 1.6.1.1
J -6-
002762
3 -6-
(a) KH, PO,
AR.
8.58
K HPO,
AR.
21.758
NaHPO, . 2 HO AR. 33.4
NH C1
AR.
2040 &
in 1000 ml of water 1.6.1.1
the pi value should be 7.2
(5) 22.5 g of MgSOy . 7 H0 A.R. dissolved in 1000 ml
of water (3.3.1)
(3.3.1) )
(e) 27.5 g of CaCl, A.R. dissolved in 1000 ml of water
(d) 0.25 g of FeCl, . 6H,0 AJR. dissolved in 10CO ml
of water (3.3.1)
This solution is prepared freshly immediately
before use.
(e) Trace element solution
MnSO, . 4 H0
H380,
nS0, . 7 H0
(NE) ghog0,,
39.9 mg (30.23.mg MnSO, . Hy0)
57,2 mg
42.8 mg
34.7 mg (36.85 mg (NgioK0,,,.4) 0)
Fe = chelate (FeCly,EDTA)
100 mg
water 1.6.1.1
1000 ml
B
Sterilisation of the trace element stock solution
J
a3 393 (X) (220C), 2 atm., 20 min.
tT
002763
`a .
@n
) -7-
(f) Vitamin solution
Biotin
0.2 mg
Nicotinic acid
Thiamine p-Aminobenzoic acid
Pantothenic acid
Pyridoxamine
2.0 mg
1.0 mg
1.0 mg
1.0 ng
5.0 mg
Cyanocobalamine
Folic acid
2.0 mg
5.0 mg
water 1.6.1.1
100 ml
The solution is filtered sterile (0.2 um). Instead
)
of solution 1.6.1.2 (f) 15 mg of yeast extraxt
may be used per 100 ml of water 1.6.1.1.
1.6.1.3 Biodegradability standard
Hydroquinone 99,5 DAB Erg. B. 6
1.6.1.4 Mercuric chloride solution
1 per cent ofHell, in water
:
1.6.1.5
Shaking machine accomodating 2 ltr. Erlenmeyer flasks either with automatic temperature control or used in a conrtant temperature
room at 293 - 298(K) (20 - 25%)
1.6.1.6 Narrow neck - 2 ltr. Erlenmeyer flasks. (Creased fluted _ flasks are recommended) The flasks must be carefully cleaned with, e.g., alcoholic hydrochloric
2
-8-
002764
y
-a-
poe -e use, rinsed and dried in order to avoid
1
.nation with residues from previous tests.
~t .lasks
ist use
also
since
have
they
mtaoybebeclceoannteadminbaetfeodr.e
their
1.6.1.7 Membrane filtration apparatus
1.6.1.8 Membrane filters 0,2 um
-
1.6.1.9 Carbon analyzer
1.6.2
Incculation EIther of the following three alternatives may be used as inoculum or a composite sample thereof.
J
1.6.2.1 Ino:ulum from secondary effluent
The inoculum is gained preferentially from a
secondary effluent of good quality collected from
adortersetaitcmenstewapglea.ntThdeealeifnfgluewnitthmuastprebedomkienptantulnyder
aerobic 21d use.
conditions To prepare
in the period the inoculum
between sampling the sample is
filtered through being ciscarded.
a coarse filter, the The filtrate is kept
first 200 ml aerobic until
ised. The inoculum must be used on the day of collec=
tion.
1.6.2.2 Inoculum from soil
100 g of soil (fertile, not sterile) are suspended
_
in 1000 ml of chlorine-free drinking water (soils
with an extremely large content of clay, sand or
"
7
soursgpaneincsiocnarbisonalarleoweudnsutiotesde)t.tleAftfoerr
stirring the
30 minutes.
-9-
002765
.
@r
9.
D
The supernatant is filtered through a coarse
filter paper, the first 200 ml being discarded.
The
The
filtrate
inoculum
is aerated immediately and
must be used on the day of
until use.
collection.
1.6.2.3
Inoculum from a surface water
An
The
inoculum
sample
is
is
drawn from a suitable surface water.
filtered through a coarse paper,
the first 200 ml being discarded. The filtrate is
kept aerobic until used. The
on the day of collection.
inoculum
must
be
used
1.6.2.4 Composite inoculum
Equal volumes of the 3 inoculum samples are united,
mixed well, and the final inoculum drawn ffom this
mixture.
tT
)
Tohfe thseuitsatbanidlairtdy ohfydrtohqeuiinnoonceu.lum is checked by mears
1.6.3 Conduction of the test
1.6.3.1 Procedure
The test materials are evaluated simultaneously in cuplicates together with the standard (1.6.1.3) and a control test with inoculation but without either
test or standard material for the determination of DOC blanks.
The standard
moval within
material has
28 days at a
to attain 60 % DOC re-
starting concentration
corresponding to 20 mg DOC/1. If (60 % DOC removal
7TRISTIRIt1sbased on preventexperienceWTR are achieved the whole series has to be discarded +)
tthheis 19lidniaty ovrerseivoenn ooff tthhee stetsatn.darAdrmeivgihstionhavoef
D)
to be considered after the accumulation of experience with the new 28 day version of the test.
-10-
002766
on. .
:
) -10 -
A stock solution of the test material in water
(1.6.1.1) is prepared. So much of this stock
solution is added to the nutrient solution
(1.6.1.2) that a carbon concentration of 5 = 40
mg DOC/1 is attained. The starting concentration Doc/1. of the standard hydroquinone is, though, 20 mg
Two reaction vessels (1.6.1.5) are each filled
with 900 ml of the nutrient solution and inocula=
ted with 0,5 ml/1 of the inoculum (1.6.2). The
opening of the vessel is covered with, e.g., alumi-
)
num foil in such a way that the exchange of dir
between the flask and the surrounding atmosphere
is not unduly impeded (Cotton wool is unsuited
because of the DOC analysis). The vessels are
then inserted in the shaking machine. The tempe-
rature of 293 -298K ( 20 -25C) must be maintained
unchanged during the test, and the vessels should
be shielded from light. The air should be free of
pollutants and
vents ete).
toxic
materials
(chlorinated
sol-
In the course of the biodegradation test the DOC
concentrations are determined in duplicate (1.6.4.2)
at the beginning (day 0), and on the 27. and 28.
day. Three additional analyses have to be performed
or
J a1 -
002767
.
=
)
-11-
in rather regular time intervals (~7., ~21. day)
~1i., and
The
and
analyses are
evaluated.
registered
in
the
attached
form
sheet
Only the necessary volumes of culture medium may be
drawn for each determination; however, they have to
be large
fugation
enough for
preceeding
the
the
mcaermbbornandeetfeirlmtirnaattiioonn.orThceentlrait-ter
requires differing volumes for the different instruments.
Evaporation losses
up by adding water
o(f1.6t.h1e.1c)ultiunrethemedrieuqmuiraerde
to be made
amounts.
) The culture medium is to be mixed well before withdrawing
2 sample. Material adhering to the wall of the vessel has
ftoiltberatdiisosnolovredcenortrisfuusgpaetnidoend
before
has to
bseamdpolnieng.imTmheediamteemlbyr.ane
The filtered or centrifuged samples.have to be analyzed
on the same day, otherwise they must be preserved with
0,05 ml of
of nutrient
the
me
HgCly
dium or
solution (1.6.1.4)
by storing them at
for
2
=
each
4C;
10ml
The
bexihoidbeigtrsadaabdielgirtaydatteisotn isratvealiwdithpirnovitdheed sptheecifsiteadndraarndge.
Tmhienertaelstizactainonbe isfiancicsohmepdlibsehfeodr.e the 28. day if complete
All steps require
pipettes etc. but
great care and
not sterility.
cleanliness
of
the
vessels,
-
J
-12-
002768
Ds
J -12 -
1.6.3.2
Calculation of results
The degradation at the time t is calculated
from the determinations of the DOC concentrations at the beginning (Cy) and the time t
(C,) according to
Dr |I 1m em Co = C2oyM, | lxi100 .
|
% "Ca, |
where D, = , =
Cy =
Cpy =
degradation in per cent at time tT measured starting DOC concentration of
the inoculated culture medium (mg DOC/1)
DOC concentration of the culture medium at time t (mg DCC/1)
starting DOC blank of the mineral nutrient
solution with inoculation but without test
material (mg DOC/1)
SY = DOC blank of the mineral nutrient solution
with inoculation but without test material
at the time t (mg DOC/1)
The degradation rates are calculated to the nearest
0,1 %. The means of theDy values are calculated ard
J
reported to the nearest full per cent. Results
tis.
002769
@-1
)
"13 -
ending in 0,5 are rounded up to the nearest whole
nfuomlbleorw.edThgerapchouircsaellyof inthea
degradation test
diagram as shown
is
in
the
attached example. The results are reported on the
attached data sheet.
The results of the degradation test are valid condition is met that in the same test series
standard yields = 60 % DOC-removal.
if the the
1.6.4 Analytical means
1.6.4.1
Membrane
paration
filter
of the
0f:i2ltpemrs,:
25 mm (1.6.1.6).
membrane filters
Pre-
are
impregnated with surfactants for hydrophilization.
)
Thus each filter contains up to several mg of
soluble carbon which would interfere in the biode=
gradability determinations. Therefore the filters
aorreganpiucrifiinetderfferroemncseusrfbayctabnotislinagnd thoethmer3
soluble times 1
hr
each in deionized water. These filters may be stored
in water (1.6.1.1) for at least one week.
Other membrane filters are suitable if it is assured
that they neither release carbon nor adsorb the
compound in the filtration step.
If
at
the samples are centrifuged,
40 000 m sec ( 4000 g) for
t15hisminhuatsest,o
bperedfoenreably
in a refrigerated centrifuge, in any case <40C.
(Remark: the
tion at very
dliofwfceornecnetnitartaitoinonTsOC:dDoOeCs
by centrifuga=-
not seem to
=
)
work well
or carbon
assincpearteitofherthenotbacatlelribaalcteprliaasmaareisrermeodviesdsolved.
Astmalhlighienrocutlesattiocnontcheentrcaetnitornisf
ug(
a)t1i0o
n
mg
e
r
C/1)
ror
and
seems
the
to
same
be
comparatively small ).
002770
.
Form Sheet for the Modified OECD Screening Test
@-
- oo
A: Controls:
Tot 2 00
renal ueriens finociton
)
Te
:
7
- =het
- TT
Ei
T1
Pie
nye
2
Flask
of
hae.|]
Fr =
omen
for day x
HERE
00
@r
.
d -a
1.6.4.2
The
The
DOC measurement
sample withdrawn
frem
the
culture
medium
i(mamboeudtiat3e0lmyl)in isthecefnitlritfruagteidonoarppmaermabtruasne (f1i.6l.t1e.r6e)d
using the
"The first
membrane
20 ml of
filters prepared
the filtrate are
acc. to 1.6.4.1,
discarded,
:
`The DOC concentration is determined twice in the
remaining filtrate (about 10 ml) by means of the
TOC/DOC instrument (1.6.1.8). If the filtrate
cannot be analyzed on the same day it has to be
y
c(omEnCs/e1r)vedobatcaci.nedtoar1e.6r.e3.g1i,steTrheed DOoCn
measurements
the `attached
data sheet and the DOC concentrations of the
culture medium and of
each sampling time.
the
blanks
calculated
for
J
002772
*
:
APPENDIX 2
te
-
or
t
resul3tHsEnovfirtonnemeMnotdailfieLdabOor6a2torSyirDeastnainSghTeeetst
:
LsAonbealyRsootfg:ueSnuatrdtweo.o:T_ASsenHorST ] ES
pHIranieaooscruuarlMtauetimdoc:reTT0al0l!:s(o0lAo)Nf:STTaned sTnseosRtmmacPRthaeltrFaioraerTlFe:2le:0s(CATAL,AFoTcLN0on0ECd0sPrUy LASF sTluyentESTats SteTnrodneard
Material:
sunt
hydroquinone,
o
98.5%,
Lot 100147,
Aldrich
en Te So SEE ora Draeneimalcloslwestensier Cross slant
A. Determination of voc! of Yost Material;
WA hstocdkatonsiozleudtiwoanteorfs the test material was prepared by diluting _ 1170 mg to a final volume of 500 mi
TRI TT TE) This esi was ved co calculate che seconsary
on
| re
iin SF Saget! to prepare cular
5. cordon veterminations; %
BEST COPY AVAILABLE
nCFeloatnsockr.,iaonmfe/tftne"at||Co{TFonteFSuelatatastmeadteomrauital O90C)~ conTc. aW fter x ders E Incubagtion, mal]
)
onan mtestet
Inoculum
MSToiosnTtearcamtloanrnsuethrrioeenTut|]
but with
282
40.0
F pT eet eee
aT
mm alefooe [on fee 282 40.0 | Fare r RR ppnAmT ePTRr oLee fSreSsxET
rs
Fe A
TE fumeo [e77 []7 os [eerHi e Se rTorr eTee s oTeTra 11sg |-02 |04 [12 0.5| 00
[orm froculm
signe
TT
ve
C. gesluotioofn fev vata:
To Jo [isJoe Len ra rr > Herertertreteary =o.T Cones |14 |-00 [od |Z [06] 6
Fro
redatio=n%DOCgesoval after xdeyy Tresbetion FRI UA.
L' [esTenefens[Mf3a0ns| talmoienvesreoviiemd mInsthme csal.eutastioon,s.
[=[rz Jos[esJai[as] J
Pave ~ --p----
1. 2.
The test material 13 completely soluble In water. 00 anslyses performed more then tice on the sam
sTahmeprleefowreer,e trhuenTfOoCr=qu0s0lKl.ty
assurance purposes.
002773
Lob hosuenc vo, S541 8 rest30stoGuFtrhoemarnacee)16L8aboBreaestoreyraGsatiengSheTeotse$SaCataetyaoatf:SStVA aardsoao.E FTcnn EFrS y or 235 m/77_Standard Materlals_bydrogulnons, 98.5%, Lot 100147, Atdricn pCTehhcsrcoournreidMm:aTTycsseorrotfSaTnoedsnntoramatetarBaElSTsEASVRoratraonicrCatcanuas glgfon rilosLenientte.rSromemnt slant
A. Determination of oe! of Test Material;
A1tshtocGkatonstoileudtiwoanteorfs the test material wes prepared by Logo diluting 4060 mg to a final voluse of So 500 ml TSStoocThk sSolutTion I|stTocnkaTslelutiIon |TtestnEamaltes]rial | ihd miesni rtooosffusounmnm easlatvainepdteiseelsos oco0aclPocsruelPitreetnheenat!oceensss
2,120
93,900 BEST COPY AVAILABLE
0. Carbon Doterminstions:?
`StMoibnueecrialornnwuatirnient na
) erm leeter sa for or Inoculum
or | mTeiteesr,ian ngtt | [iSnF fSaratemsaecoroion
ig) TS Eg
20
Yo.0
[er1923130 E5156159 1350
B 5 Pp T t r ca eS cer --------------------
Y2 Yop BE [CefrrsEPHEer ErT IEs EAe a ATy SR rTsoe
rd
1
= lle oa eo HSionheorratlonnuwthrslhaanntt]]
CeI PA TTCa haa
[w | Too = oui[eel C. Evalustion of Rev date;
Er r r erTe ey 5
II
=
Colne =
10Taer e re e oe ET
14 [=o
v2 [oe |
PE |i eo ewaela [] m |97 | gmmpeseme [a ro[os[353 [92 | EAT vere aad In th cateut Tons
J
lave" 22
1. 2.
The 00K
atneasltysmeasteprairaflorImsadcosmoprleettehliyn
stoullucbeleoninthweatsearm.s
STaheeprleefovreer,e
trhuen
TfooCr
=qu0aXl.ity
assurance purposes.
002774
} co
en BE 3M Environmental Laboratory Data Sheet
Date of StartofTest: S-1-B0
pr AERARR eie m eye
A. Determination of voc! of Test Material;
FEITE Ny: TEoe | Di Ea ne cso A stock solution of the test material was prepared by diluting L245mg to a final volume of S00 m1
tite . 5 [EE Mineral nutrient.
ton movertat
and with
St [ers 20% _ 40,0
208 | geo or
_ BEST COPY AVAILABLE
Te coe shes dv amg ;_--Fede Bree Ter 7] Zo
po Isp Ae TET
eT rr
EA tree Re Lt tEo ELS BE ees r----------
EE arms [ell[eie s forle mmr Miners! nutrient] | _ 1emer le 177ToTTos[7s169| t6eres7
--
EEE a
[[ oct W[o ou[0a]ec]
C. Evaluation of fer vata:
Cliawe= | 19 [00] 0 | 72 | 06 | o
[Th oTpiRn E1mfte i T Om T |ss snea s.
J [[ = [s2oJ [Jaoasae[Joosa ss[aenfa]oa aesweesa|||toC TmEs=eSay 2rd1CsmT (0o1amwsiaessim [==157 [04 Jae[2| ar]
eT ere weere t5 meetets aetde vere r Toretoresr m2s,y sr
. to
os 3805
TE roR s SE enayTYE t
Dateof StartoE fTerr TB
BEST COPY AVAILABLE
ER a eT heoretical JOTTen etre
TAITE ee BATACTA CARBON ASSUMED 1D
a
a
BC
coSOLUBLE
anORGANIC
CARBON,
1 Asisetmctksstttuosnioan1o0f!toht sunssstaartrasassrsbyaa1aaitns ot Fovt of a1
Ty eheT [orn veton| conrae|| Tei nt cctut cn econtry
[TT evacuarion or causramoy 8. Carbon Determinations?
COMPOUND-- SEE STANDARD
MATERIAL ABOVE .
St eranTom | coe onan rei wr
Mineral nutrient
J mnrn groia
306 | 200 PlFeoT eEZ fEZ ey3sT l--e2 E1r0 R--1 t1 T1i1e7y5 Tr3 [77
| 306ac r1|2m27en00 |Pm [ r Ema Ee se EEfRe ar E[eE2E7e rtTieetEe[ie
mmr |mers bTut wo ith,e]
ani
[or foo fos foe foes C. Galuosfthoiecourre;
-- EEer LEregloz| 04 | 12 |os [0.4
1
HEIelLe Tard =o
Con=e Sc [1g |01 | 04 | 12 [06| 0
[TR] opens1 ontSyoyvee | sent seit in
[ses
[= [907
Joa
Jono
oon yar gro |
lerc|laonwa|
IsemmTtorevtmAedossLwwom seoe
Jf Tea fez[orfou[95 |
21. oTron nesatmsevarritalom1escnmpaeicetyatseetne28voeere, sTreteorn e1 +e5a.ti EES FE
002776
EEE re SSgE EsmT ns hptelreen irT oanr nn SH Eimna ds .
Ee ee
A
Ee
E er EE ecwLab Request NouA: m55e41n'ss peas wee a Soin oo)
-- = -- ET -- E --_-- TE T e
= ges "
Eee
ees
A
= mre a = iE
a 70x
heed
hm
EET
Eos
Foe]
Mf epee
a =}
Al I
e ge re e eee e e e ee e e EE ry e Led a-
fy
E
Cme mf e ee
sr epe aee e e |
----
Ep
e Ee e e e p eeoTe h ee
&g
iirs 6 ORYS IICUBATED
sasLhl
3
a
` ween
$
= Eek == EESTI
S ay cty rl ba oSrea--te--------------S teEmL stjt ws
ET re
A`OnEaClDysHto:diVfaldeed AS.crSecehneiln|g Test with 0OC Analysls
FS Fe
=oj-moy
+
ry Date of Start of Test: 4-11-1980
iAra hh
e F e eEe ES eLasbtRevqueerstteNA o.L:o5r5e41a'aSseT fesoozjeagpR o3gae S S+sote o ]|
ys -
Er
owTE E a ---- F = -- SyST Se e y
e H ee e 7 = eea ee
soy
--
w=
Re
=
EE
nLi
- = EEE
ee
-He er e m ee Ee S p a
pe
JJ
7
Wirst of BAYS CATED
EE
PWdS
>
.
Co
wp
E NTT N
TTT
shimaTl
= E a e C: r = e T icohA aatu fiey d SL cm reening U Tebt ith 00C Analysts h Ha Ea Eas
=
eee eee
ey
ee
jn FESeEEre
=S =S = E S t s : Se
i SE Ba
Tani Ee =
SEE
i aE
i Te E See =e SHe EE y ER
m E EE e e = e e e e
S f e ee e ee
ge
=
>3
NUFSER OF DAYS INCUBATED
mmemele ee
i wiashie
Ee
SE em em m e
eee
Mopaivete aScreesnineg n Test wi
ols
Eee hal
= ss SN Lab Request No: 5541'S
E Ee ee
. (ca.panvion_compounin)
= -_ ---- ---- ---- ---- ---- --
ee
--
E RSEE IETsE ani
ES EEem
E Ee E
.
----
:~
ed =
Er "l-- s ny A 8 --
= =
= = fs--_------
5 af ee
=
n
=r eee
TS
re
|
w Ef Ee ee ee ee e ee e ee
wel ef
e e e e e e
33Ro
T7
iNtUFSER OF DAYS 13CUBATED.
ie
=
----
)
par
ey Prwb hd