Document 8JxepYJjBVLLwOpkmkwxMXXe
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AR2IE- 0106
EFE-I3
TOXICITY TO AQUATIC PLANTS (E.G., ALGAE)
EST SUBSTANCE
Identity: Perfluorooctanesulfonate; may also be referred to as PFOS or FC-95. (1-Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,56,6,7,7,8,8,8-
heptadecafiuoro-, potassium salt, CAS # 2795-39-3)
Remarks field: "The test substance is a white powder (3M Lot 583) of
uncharacterized purity.
The following summary is abbreviateddueto the fact that this study has been superceded by a more recent test.
M -- EO--------------
Method: Test protocol utilized was modifiedafterthosedescribed by USEPA -- 600/9-78-018, 1978; ASTM-E-35.23, 1981; OECD 201, 1979. ASTM STP #667 Type: Semi-chronic GLP: No Year completed: 1981 Species: Selenastrum capricomutum
R -- ET ------
ia ECg Values, mg/L (Cl)
[a | 115(1885) | 82 (NoConf. Limits) | [0 18@e372)| o8(i643n) |
[14a 833350) |1 5(12485) |
Remarks: The statistical program used was questionable.
DATA QUALITY
Reliability: Klimisch ranking = 2. This study satisfied criteria for quality testingatthe time performed, but actual concentrations were not measured. Results were based on nominal concentrations. Additionally, sample purity was not properly characterized.
REFERENCES
001456
This study was conductedbythe 3M Company, Environmental Laboratory, 1981. OTHER Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 Last changed: 5/3/00
001457
| remourane
TECHNICAL REPORT SUMMARY
2/16/81
| omen sommmarmon awn sain | Suimmmpoarrtaynta-- t1iporreosmpnroirnmtatniobnoLisncoenoofp7a6p6a,(s3e6n5d4o)ws cle 0TCC)So"Guidlines forComplcionofTechnic Report
Ir
ntal Laboratory (EE & PC)
0535
= FC-!
Lot 583
9970030000
Huei
Algal Assay Test Method
007
Be
/A. welter
BAT,
46981
M-- TD--EcL oir aBo Gmes or | SEwe RED a
evRanse
Croemet 3S5RCTeIvVaE,luate the algal growth response of freshwater
green algae "Selenastrum capricornutums over several
gfelnueorraotcihoenmsicaals tFhC-e9y5,mayLotbe 5a8f3f.ected by exposure to the
2. Teoxpdoesvuerleoporanreaclogvaelrytepsetriopdrsototcoolthebasteedstoncovmaprouinadb.le
SE 4 PC Div. [REPORTAETRAG: temen emeior hereto TeilComerGr
HB
(Env. Lab)
Company RAD.|
Sr
In these triplicated four-phase exposure/recovery assays,
tehxeposiunrietsialtoefFfCe-c9t5s woenreoragsasneisssmesd ofovlelrowsienvgeravlarigeednerations. Taqelusgatanltirtepsaoutpliutlvsaetliygoanvisen.atnerAmlisgnadloifcabtciieoolmnla-sodsfrywatshweeiagsphsotessssi(ebndlge/l)effaencdts on
cell-count (No./ml). The following biomass indicators were measured and calculated for:
A. Verification of inhibitory effects
1. Mexepdoisaunregsr.owth response ECsg (mg/1) from varied
2. ECjg and ECgq (mg/l) for same exposure periods.
3. No-effect levels.
B. Verification of algicidal effects
.
1. RSeusbucmuplttiuornesorduraibnsgencvearoifed lopgerairoidtshmiocf grreocwotvheryi.n
pIrn:
9
001458
FraCg-e95T/wmoTE Decenber 16, 1981
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CONCLUSIONS
1) McaotnesriidaelrsedpotsosebessisnlgigahntlyECpshytinototxheicratnogealogfae1a0n-d100aqmuga/tlicare plant species.
2) Tvhaerieadlgaplerigordoswthofrreescpoonvseeryiinnmdoiscattesdubmciunlitmuarlesalfgoilcliodwailng
aelfgfaelctscel(ldseatrhecoofveraeldgalandcelrless)umeadt cloognacreintthrmaitciongsrowtteshtewdh.'en The
rteesstuspmeantdereidali.n fTrheeshmenduitarniengtromwethdiurmespionnstehe(aECbssge)ncevasof the
tihnehriebfiotroery ienftfeercptreatsedopapsosbeadsitcoalalny ailngdiicciadtailveefoffecatn. In the
afroermreerduccaesde,whpehroetaossynatnheasligsi,cidcaelll-egfrfoewctth,cauasneds cdeilrle-cdtivision,
>
cell-destruction and cell-death.
3) Riensduulctesd ninodigcraetaetderthianthibeixtteinodnedbuetxpsolsouwreersre(co.gv.e,ry>7radtaeyss)in the subcultures were noted.
SUMMARY
Aevaflouuart-pehaaslegalexpgorsouwrteh/rreecsopvoenrsye aoflgatlheasfsraeyshwwaastedrevuenliocpeeldlutloar agsreetnheaylgmaaey b"eSealfefneacstterdumbycaperxipcoosrunruetutno tohveerflsueovreorcahlemigceanle,ratFiCo-n9s5. Tdhuirsingmulftoiuprhacsoensealcguatliveasseaxypoesvuarleu/arteecdovaelrgyalcogmrboiwntahtiorness:pons4e/s10, e7/x7p,osu1r0e/4,pearnidods14 idnacyrse.asedA,s iwshilcelearrelsypecitnidviceatreedc,ovethrey dpeurriaotdison of . dweacsrefaisxeedd. atTh1e4 cdaoymsb.inedAlgdaulratgiroonwtohfreeascphonseexpovassurmee/arseucroevderyinphase cteerlmls-coofunatn (iNno.c/rmela)s.e in~Drbyi-owmeaisgshtasandcelcle-ldlr-ycouwnetighrtesu(lmtgs/l)useadndfor stehetscaoflcutleasttiocnulotfureECsfgl'assks(.mg/lS)eewetreestavsecrhaegmeastico:f triplicated
Phase Exposure (Day) Recovery (Day) Total No. (Day)
11
74
107
1144
1w
1u0
None4
144
cIanlctuhleasteedtriEpClsicavtaelduesmul(tmigp/lh)aseformulftriegsehnweartaertiognreeanlgaallgaaessays, the v"aSleuleesnarsetprruensecnatprithceornmuetduiman agrreowtphresreenstpeodnsein (TEaCbslgemg1/.l) Thfeoslelowing exposure to the test material for 4, 7, 10, and 14 days.
001459
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FPaCg-e95T/hMrTeEe
i
December 16, 1981
TpheeriovdasluelsisotfedEeCa1grliaenrd wEerCe c(amlg/clu)latfeodr eianchteromfs tohfe cexeplols-ucroeunt and are presented in Table 2.
No test concentrations were defined as a no-effect level.
gTorowvtehrifiyn atlhegicaibdsaelnceeffoefcttsh,e atnedsttomaatsesreisasl psousbsciubllteurersesuwmeprteion of ecsotmabbilniesdheadt fthreomeenadchoftreaicphliceaxtpeodsurseetpeofriotde.st cAluglatlurerecfolvaesrkys r1e0,spo7,nseandwas4 edavyasl.uated following respective recovery periods of
~
INTRODUCTION
aIqnuatteisctinegnvithreonmpeonsts,ibluenicefeflelcutlsarofalgchaeemiacrael rseucbosmtmaenncdeesd oasn athemodel s(yasqtueamticforpreivmaalruyatpirnogductehres)i,nfalqueunacteicofplacnhtesm,icaalnsd opnhytaolgpallankgtroonw.th
Tcoexlilc-deifvfiescitosn adruerintgesttehde otnestg.rowTihnegsealggarlowtchultruersepsonstehsatmauyndbeer:goa) stimulatory, b) inhibitory and/or algicidal.
MATERIAL AND EXPERIMENTAL DESIGN
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Tshoelubtleestinsuwbasttaenrceat (aFmCb-9i5e,ntLrotoom583t)empiesraatufrien.e whTihtee bpioowadsesra,ys apnedrfotrhmeeddaotna tgheinsermaatteedrifaolrmevthaeluabtaesdisitosf ptohtisentrieaplorta.lgal toxicity
TthheosetesdtescprriobteodcolbyutUiSlEiPzAed- 6fo0r0/t9h-i7s8-0s1tRu;dyASwTasH-Em-o3d5i.f2i3edDarfatfetrNo. . 2; OECD; A. G. Payne. (1), (2), (3), (4).
Test Species:
Aalbgaaecte(rCihai-ofrroepehycceualst)u,re Soeflenthaestrfurmeschowpartiecroprnluatnukmtonoficthgereoecnder c(hFleobrroucaorcyca1l8,es 1w9a8s1).obtTahienedtesftroamlgUSaEePAare-- non-+motiColrev., Oregon ualngiaclelcluulltaurrecewlalss,sthoarveidnginthetheappdearakranacte4o9Cf. a new moon. The
Inoculum:
gAilvgeaea fsrtoamrtain7g-doapyt-iomludmstioncokcucluumltulreevelwerofe
used as inoculum green algae
to
T S5eleO nastT ru0m18capriSEcocokrnGuutlutmie1 xins1u0r4edceltlhse/mplr.esenTchee uofseaof a 7
sgruofwftihciephnatsen.umbeThreofinviitaiabllealaglaglalceclelllscouinntthine ethxeponsetonctkial
cceullltsu/rmel)w.as determined using a hemocytometer (300,000
001460
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DFPaCeg-ce9e5n/bFMeouTrrE16, 1981
Algal Nutrient Medium:
aMnidnertaelsti(nignoarlggaaniec)wasstapnrdeapradrednuatsrieonuttlimneeddiumin fAotrtaccuhlmteunrting#1.
Tfhoirsalnguatlriegnrtowtmhedianudm aplrsoovidseedrveadllasminteheraldilnuuetnrtienftosr ealslsenatligaall
oPpHerOfatitohniss siynnctlhuedtiincg atlhgealprmeepdairuamtiwoansofadjsutsotcekd stoolut7i.o5n5s..1 The
prior to use in assays.
=
Culture Flasks:
cEoamcphri2s5e0dmla tEersltenmfelaysekr. coCnutlatiunriengfl5a0skmsl oafnd taelslt ostohleurtion
g42l.asswAaurtoeclvaevreedspfeocaimalplluygspruespeadredasasfladseksccrliobseudreisn, Atptearcmhimtteendt
free gaseous exchange to occur.
.
Range-Finding Test (Exploratory):
5
bTihoemaesxsploinrattorriyplitceasttedconfsliasstksedcoofntadientienrgminsitnagndatrhde n4u-tdraiyenatlgal mseevdeiruamlploursdertsheoftesmtagnmiattuedrei:al a1,t c5,onc10e,ntr5a0,tio1n0s0, coanvderi5n0g0 mg/l.
Full-Scale Test (Definitive):
Aulslingdef2i5n0itmliveErlalegnamleyearssafylsaskwsereconctaarirniiendgou5t0 miln otfritpelsitcate ssiomluulttiaonn.eousTheexpdoesfuirnei/trievceovaesrsyaytecsotnssis(t4e/d10,of7/f7o,ur10/4, and t1o4/0verdaiyfsy. inThhiebitpourryposeeffeocftst.he Tehxeposruerceove(rcyont(ascutb)culsttuargee) was vsteargiefieedvalaulagtiecdidatlheefvfieacbtisl.ity of growth-inhibited algae and
T4h0,e f61o,llo9w3,ing145F,C-9a5n,d L2o2t5 m5g8/3llvoegraerituhsmeidctoconicneinttiraatteiotnhse: 26,
ebrxapcokseutrethsetag4e-.dayThEiCssgdvosaeluersangperedofictdeidlutfiroonmsprwaeslimsienlaercyted to
exploratory testing. algal nutiient medium
pTlhurseealfglaalskscelclosntacionmipnrgise1d00%thefresh
snuobnsttraenacteedvacsonptrroelpsa.redAinfrethseh asltgoaclk nsuotlruiteinotn mofedituhem test
siomlmuetdiioantelwyaspr7.i3o.r toProtceesdtuirnge.andThesteipnsitifaolr pplrepofaratthiiosnsotfock
algal test flasks are outlined in Attachment #3.
Einachtherecaobvseernycestofagethweastesitnimtaitaetredialb.y suAblcgaulltucreilnlgs f(r0o.m5/5e0acmhl) trreispulsipceantdeedd sienttooffrceuslhtunruetrfileanstksmedwieurem.comSbuibnceudltuarnde tmheetnhods are briefly described in Attachment #4.
001461
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c-9s/uze
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PDaegceembFievre16, 1981
Test ALCLondailtgailonso:perations were carried out under aseptic conditions, in order to avoid contamination with bacteria and other algae. Algal cultures were maintained in an environmental chamber under the following standard growth conditions: Temperature - 23 + 20C (70-779F) Fluorescent illumination - 400 .C. + 10%
SFhraekeingagg,1e00xc+han1g0erp-m.Continuous Platform
Algal Biomass Monitoring:
Tauclhsogeiuannlgtaslecgaui(lltctheueglrrrleoss/wamltlgha)af.ltreercsepBleoolant-cshdhergyrwofaoswwtethhiaegpmhpetreaaxsip(suomersgdue/rmlee)qnutaaasnnndtd/iwotarrareetciobvymveaerdlcyeyellwbiyth
stages. Both procedures for algal biomass measurements are briefly described in Attachment #5.
ECs values and 95% confidence limits were calculated aetxipteirziinnegnta3lM SdaItXaC.UR, a TRAC System for regression models of
Test mesurs
`SSTehelelenrfaess-tuarlutxmspocifaparbniiacoanomtardsnosuartrmeuyemadseuutsraeeidmleeidnntstinheosftehetshetautdtgiareceshenedaraedl.agtaae- sheets.
Calculated ECsg (mg/l) values indicating algal growth response to
FdaCy-s95,areLotpre5s83e,ntefdolilnowiTnagbleexp1o.surTehe pevrailoudess ooff E4,Cyg7, an1d0, ECagngd f1o4r etasaneudcrmnhmsarorefiosfzuemtdpcheteiliolenn-xcpTooaousfbnultreleoag3n.paderriiatcohedmsicprleigssretonewdttehdabionivnesTuawbbeclrueeltuc3.raelscuRleaacrteoevderyindata ace shown in Figures 1 and 2. Utilizing the ECso values, To illustrate the action of test material and to give an overview
of what has happened in the exposure experiment, toxicity curves
toxicity curves were constructed against time on semilog plots.
data sheets comprise original data. The data reported herein are based on studies developed and
performed by M. T. Elnabarawy and R. R. Robideau. Accompanying
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001462
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FPaCg-e95/SMiTxE
December 16, 1981
cIatlcsuhloautleddbeonnottheedbatshiast oafllthteheinrietpioarltedconEcCenvtarlauteisonvsereof FC-95, Lot 583, in test solutions at the Beginning of the bioassay.
REFERENCES (1) MSiellleenra,stWr.umE.C,apJr.icCo.rnGurteuemneP,riantnzd AT.lgaSlhiArosysaamya.Bott1l9e78.TestT:he
EUr.pS.etiEnnevnitroanmdeenstTalon,ProatppelcitciaotnioAng,encayn,d Cdoartvaalilnitse,cpOrreetgaotni.on. EPA-600/9-78-018. 125 p. (2) ACSoTnMd-uEc-t3i5n.g23T.ovxic1i9t8y1.TesPtrsopowsietdh FSrteansdhawradterPraacndticSealFtowrater Algae. Draft No. 2. (3) EOfEfCeDctGsuiodneliBnieosticforSyTsetesmtsi,ngTeosftCh2e0m1ic"aAllsga(e1,98G1r)owtShectIinohnibi2,tion Test,* Adopted May 12, 1981. (4) APlagyanel, TAo.xicG.ityandandR. IHt.s AHpalpll,ica1t9i79o.n tAo MtehtehoSdafeftoyr MAesasseusrsimnegnt of New Chemicals. ASTM STP #667, p. 171-180.
001463
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TABLE 1
:
AlgalFCG-r9o5w,thLoRtesp5o8n3se to
ECs0's (mg/1) (1)
(ECxopnotascutr)e
Days
Cell-Dry Weight(2)
Cell-Cou(n3t)
4
115(18-65) (4)
7
122(41-366)
82(o Limits)
$9(19-398)
10
128(46-372)
98(16-431)
1
146(33-350)
95(12-455)
(1) MreetghroedssiofonFmCosdyelcsalcouflaetxipoenr:imen3tHalSIXdCaUtRa,. a TRAC system for
(2) Girnowttrhiplriecsaptoendse speatrsamoeftecru;ltcuerell-fdlrasyksv.eight (ng/1), measured
(3) Gmeraoswutrhedresipnontsreiplpiacraamteetders;etcselolf-ccouulntture(nof.lacseksl.ls/ml),
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(4) 95% Confidence Limits.
001464
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TABLE 2
!
AlgalFCG-r9o5w,thLRotesp5o8n3se to
;
BasedECo1no'sReadnudctiEoCngp'isn C(emlgl/l-)Count
Exposure
(Contact)
Days
ECig
Eceq
4
10
173
7
18
179
10
Le
179
14
16
174
001465
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TABLE 3
Algal CelRle-cCoovuenrty, StCaeglels/Dmalta (SMuEmANmaVrAyLUES)(1)
Treatment | Phase I 4/10 DaMyusl|tiPhPahsaeseIIExp7o/s7urDaey/sR|ecPohavseeryIII 10/4 pays
Aitia
wa
nit7al Fina
RItial Fina
Control Loa2d9i%ng (2) coun,t0(003) | Lo7a5d2i0n3g04,Cou0n0t0 Loa2d5i6n0g Co8u,n0t00
126 mg/L | 1,047 330,000 | 1,483 312,000| 1,380 144,000
2 40 ng/1 967 298,000 | 1,333 252,000 | 1,300 124,000
3 61 mg/L 935 266,000 | 1,060 222,000 | 1,310 110,000
493 ng/1 2713 248,000 503 210,000
513 78,000
5 145 mg/l 57 244,000 123 82,000
97 6,000
6 225 ng/1 47 190,000
67 56,000
50
(1) tBarsiepdliocnate0d.5semtls/50ofmlculstuubrceultfulraessksecsotmabbilniesdh.ed from each (2) Measured at the end of each exposure period. (3) Measured at the end of each recovery period.
001466
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NUTRIENTMEDIUM FOR FRESHWATER ALGAE
A. S PACKOWVTRIENTS STOT CKSSIOLUTIONS (CONCE(NTeRAsTE)D)
1) 25.500 gn NaNoy in | Titer DI water.
2) 1.044 gn K HPO, In 1 liter DI water.
3) 12.159 gn Hoc; 61,0 In 1 Titer OI water. 4) 14.700 gn HgSOy THO in 1 Titer DI water. 5) 4.410gnCaClz 240 In 1 liter OI water.
6) 15.000 gn NaHCO In Titer DI water.
1) 185.5 mg 3803 2) 415.6 ngMnCl lipo
3) 3.27 mg ZnCl,
4) 1.43 mg CoClyr640
5) 0.01 mg cut,
)
6) 7.26 mg NayMoOys2H;0
7) %6 mg Fecly
:
8) 300 mg NagEDTA*2H20
PREPAATILON GF SYNTHETIC ALGAL NUTRIENT MEDIUM Add Moincermolnutorfieenatcsh msatcorcoknmuitrxiepnetr s1talciktersoolfutdieoinonpilzuesdownaetemrl:
- sonicate and then filter through 0.22 sm membrane;
~ adjust pH to 7.5 0.1;
~ store In the dark at 4.
001467
ATTACHHENT 2
.
PREPARATION OF ALGAL CULTURE FLASKS
All fbloasrkosstlusiecdateIngmlaaisnsta(iKnIinNg).and testing algas were made of = flasks ware brushed Inside with a Stiff bristle brush; - washweidtwhitthapnwoant-eprh;osphate detergent (MICRO) and rinsed 3 times - rinsed with a 10% KC solution; = rinsed 3 times with tap water and 3 times with D.1. water; - dried in an oven at 70 for 2 hours (placed Inverted);
= and autoclaved with foam plugs Inserted at 124C for 20 minutes.
001468
ATTACHNENT 3
Feds \ors8s
|
AEPARAT ON OF ALGAL TEST FLASKS
ore AT Gare
ro rs
conTRoL
----
_ _cowws
a
a1) or (ng)
al
N85 + None +o
|
ol . 50
12onfllisngsod)
om 200 |
Sulndl(309) |
Bes |
SHS (129 |
HipB.
NS22,
HBL
4s
BM IS
2.0m
+ B0Bwd + Nw + ISK
Sw
+ 1M
+ 17 +a
1
soy
- 50
|
- 50
- 50
- 50
-
50
s2Suttizs)| 31,0 7
coMBwl .on
+
.
- 50 - 50
1) Determine initial algal cell count in the stock culture.
2) Adjust oH of algal nutrient medium to 7.5 + 0.1 ml.
.
.
3) Make up stock solutions with algal nutrient medium as desired.
A) Control flasks preparations:
1) Add inoculum (volume predeternined); initial cell loading of 1 x 10 cells/ml;
2) Add algal nutrient medium to bring volume to 50ml; swirl flasks, and place
8) Test flaskasu:toclaved foam plugs.
1) Add algal nutrient medium;
2) Add test material (dissolved In algal medium If required);
3) swirl flasks;
4) Add algal inoculum;
* Sto5)ckswisrlofmlasaks:, anSd pOlance aautioclav(edSfooammaplug1s0. 00 u2) EacIwhd = 0S wg
miau pH oF soe
M.3.
001469
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ATTACHKENT
OBUECTIVE: PROCEDURE:
.
SUpBCUeLTaURsE MpETHOaD Iu N ALGAL ASSAYS
To determine algal recovery response in absence of test substance.
- TransCfoerrni0n.gi5Smalfrosmteeraiclhe ocfenttrhiefutgreipltiubceast.e culture flasks into - Centrifuge at 10,000 rpm for 5 minutes. - Decant supernatant liquid and save the precipi tated algal cells. - Usinnguttrhieenstammeedcieunmtritofu8geFitxuebed,vorleusmues:pen1d5mla.lgal cells in algal - Mix atnhdoroturgahnlsfyerandthdeecraenmtainthiengtvooplu1m0em!(oSafl)algoaflalcgealll sceulslpension,
suspension Into 8 clean 250ml Erlenqeyer culture Flask. - Usinnguttrhieenstamemedcieunmtriifnutgoethteubeasb,ovetrEarnlsefenrmekysemrl coufltufrreeshflaaslkg,al
to bring total volume to 50 ml.
001470
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ATTACHHENT 5
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ALGALBIOMASSMONITORING
A. GRAVIMETRIC CELL ORYWEIGHT (1) The fpioltreersirzeec.ommended is Killipore type 80 with an 0.6 micrometer The method Is as follows: - Ory filters for two hours at 70C in an oven; = Cool lfeaislttertswoIhnouarsdebseifcocraetowreicgohnitnagi;ning desiccant, for at
- Filter a sultable measured aliquot of the culture under a vacuum or pressure not to exceed 8psi;
- Rinse the filter funnel with 0.1. water;
= Ory dtehseicfcialttoerr tfoorcotnwsotahnotursweaingdhtweaitg7h0.%, cool ina Basic Instrument used: Analytical Balance; Mettler KE 30.
B. SPECTROPHOTOMETRIC DETERMINATION OF CHLOROPHYLL a (2)
Basic Instrument used: Stectrophotometer; Bausch & Lomb, Spectronic 20.
.
C. RIMPROHVED ENEEUBAUECRore0d.1TmonreDtEEoPuracHdENOoCmYTeOMrEToEFR-SCTOoUaTNTING CHNerTpgsa AMBERMAoNDCOPPTTIICCAALL
0. Miller,caWp.ricE.o,rnuJ.tuCm.PGrrienetnzs,Algaanld AT.tsaSyhiBrootystmlae,Tes1t9:78. ExSpselreinnaasnttrauml BeETons AnpTcation, and Data Interpretation Protocol. Ecol.
Res. Series EPA-600/9-78-018. Corvallis, Oregon. pp. 27.
(2). APHA-AVWaWtAe-WrPCaFnd (W1a9s7t5e).wate"rS,t"and(aHr,dFrMaentshoond,s mfaonragt.heedE.x)a,minlautthioendiotfion, Pp. 1030-1031.
(3). Brite-Line (R), American Optical Corporation, Buffalo, New York.
001471
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