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ACUTE TOXICITY TO FISH (BLUEGILL SUNFISH) TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonatweh,ich may alsobe referredtoas PFOS, FC-95, or as a component of FC-780B. (1Octanesulfoniaccid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was 788-20. The testsample is FC-780B. Currentinformationindicatesitisa mixtureof0.58% PFOS, 14% diethyleneglycolmonobutyl ether,76.7% water,5.5% urea,1.17% Hydroxy foamer, 1.15% sodium octylsulfate0,.8% polyoxyethylenemonooctylphenyl ether,0.05% sodium laurylsulfatea,nd 0.05% benzotriazole. The followingsummaty applies to a mixture with incompletely characterizedconcentrations of impurities.Data may not accurately reflecttoxicityof the fluorochemical component of the testsample. METHOD: Method: Methods forAcute ToxicittyestswithFish,Macroinvertebratesa,nd Amphibians, EPA-660/3-75-009,1975 Type: Acute static GLP: No Year completed: 1978 Species: Lepomis macrochinis Supplier:Dale FaffigFishFarm, Brady,NE. Analyticalmonitoring: Dissolvedoxygen,pH, and temperature. Exposure period:96-hours Statisticamlethods: LC50 valuesand 95% confidenceintervalwsere calculatedusingthe U.S. EPA (Duluth)Probitcomputer program. Test fishage: Juveniles Length and weight: Average length 45 mm Average weight 0.92g (wet) Loading: 0.57g/L Pretreatment:None. Test conditions: Dilutionwater: Carbon filterweedllwater,St.Paul,MN Dilutionwater chemistry: Hardness: 232 mg/L as CaC03 Alkalinity: 220 mg/L as CaC03 Conductivity: 405 pmhos/cm Lighting:A dailyphotoperiodof16 hourslighatnd 8 hoursdarkwitha 30 minute transitiopneriodwas maintainedthroughoutthe testingperiod. Stock and testsolutionpreparation: The testmaterialwas added directly tothe dilutiownater on a weight/volumebasis. Concentrationdsosingrate:Once StabiliotfythetestchemicalsolutionsN:otnoted Exposurevessels:Allglassaquari(a35x2Ox2cOm) containi1n6gL oftest solution. Number of replicates:2 Number of fishper replicate:10 Number of concentrations: fiveplusa blankcontrol Water chemistry during the study: pH range (0-96hours): 7.7- 8.0 (controlexposure) 7.6- 7.9 (1800 mg/L exposure)* Temperature range (0-96hours): 18-18OC Dissolved oxygen range (0-96hours): 5.2- 10 mg/L (controelxposure) 0.1- 10 mg/L (1800 mg/L exposure)* 1800 mg/L (intermediatceoncentratione)xposure valuesused because the highesttwo concentrationsresultedintotalmortalitbyy 48 hours. RESULTS Nominal concentrations: Blank control,1000, 1350, 1800,2400, and 3200 mg/L. Element values: 96-hour LC50-= 1500 (1@00 -1700) mg/L 96-hour LC50 = 1600 (1400 -1800) mg/L (replicate) Element valuesbased on nominalconcentrations Remarks: Testingwas conducted on the mixtureas describedintheTest Substance Remarks field.The valuesreportedapplytothatmixtureand not the fluorochemicalproportionalone. CONCLUSIONS The testsample 96-hourLC50 forbluegilslunfishwas determined to be 1500 mg/L witha 95% confidenceintervaolf 1300 to 1700 mg/L forone replicate, and 1600 mg/L witha 95% confidenceintervaolf 1400 to 1800 mg/L forthe otherreplicate. Remarks: Low DO levelsinthefirs4t8 hours (<60% saturationc)ouldhave initialilmypacted the organisms. Even though itdidincreaseslightliynthe next48 hours,itwas stil<l40% saturatedby 96 hours. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY Reliability:Klimischranking= 2. The study meets the criterifaorquality testing.However, the sample puritwyas not properlycharacterizedand the study lacksanalyticaclonfirmationoftheamount offluorochemicalproportion inthe solution. REFERENCES This studywas conducted by the 3M Company, EnvironmentalLaboratory, St.Paul,MN, 1978. OTHER Last changed: 6/28/00 .;T,@ 6747 11 A TECHNICAL REPORT TO: TECHNICAL COMMUNICATIONS CENTER - 201-2CN SUMMARY E('vj:e:me5:r=b,I 197a riportan-tIfreporitsprinteodn boO sFaWosfp*w, swd tm copiestoTCC.) Division Prolw-t Ropcwt Title -fo- EnviroTmental Laboratory (EE & PC) Comercial Chemicals CFC 780B 7.8 -20): Results of Aquatic 96__Hr.LCr!kn Tests ep*- um OS3S ProWt 9970012600 Report Num-Sir- 0 04 D. L. Bacon Au thor(s) Notebook ftforem* "I' /t@ A. N. @r@. Welter -employ" Numb-or7s7-@ - 09362 Na6 of Papo Including CZ;;rowet' 6 SECURITY 10, rClOpen (Company CoMkIentW) 11 Closed (SpeeW Aijtborizeftni 3M CHEMICAL REGISTRY Now Chank0s Reported 0 Ys$ (3.NO KEVWORDS: Isola"torrmfrom3M ThomruL Suggm other *Ok$Wo tormIl CURRENT OBMCTIVE: To determine the acute LC value for the bluegill sunfish EE 4 PC - Div. (Environmental- Vepomis macrochirus) e;;82od to varying concentrations of PC-790B. Lab), Toxicity ,(Aquatic) REPMT ABSTFTACT1:200-25w0ordsT)hisdwrectintomutiiosndistribubtyed TechnicCaolrnnwni=tioCnssntetro aiwt3Wws toCompanyR&D.ItisCompanyconfidmtwmaterw. The acute static 96 h LC for bluagill sunfish (Leponds mwzwlirus) exposed to @grying concentrationsof FC-7808 ranged from 1.5 to 1.6 g/l. Information Liaison Initialc$-: 3PACONFIDENTIAL 2 CONCLUSIONS In a replicated static 96-hour IX aquatic test, the calculated iz so values for bluegill sunfish ILepomis macrochirue) exposed to FC 780 B 788-20 were 1.5 and 1.6 g/l, respectively. Materials possessing an LC of this order of magnitude are considered to present an insiikificant hazard to the aquatic environment. INTRODUCTION The test substance FC 780B (788-20) is a yellow minimally viscous fluid. Tests performed on this material evaluated its aquatic toxicity and the data obtained form the basis of this report. METHODS A. Aquatic Testing The test protocol utilized(fyr this study was modeled after that described by USEPA (197S). ACCLIMATION PROCEDURE The bluegill sunfish (Lepomis macrochirue) were obtained from a. private hatchery.* Stock fish were held in a fiberglass holding tank filled with carbon-filtered well water at 14-15 C. A 4aily photoperiod of 16 hours light and 8 hours dark with a 30 minu' te transition period was maintained throughout the acclimation and testing period. The fish were fed Tetra-Min'daily, food being withheld 48 hours pribr to and throughout the test period. Fish were acclimated for 14 days prior to testing. An acute short-term (96-hour) static bioassay was performed on FC780B. Carbon-filtered well water of known composition was used as the diluent (Attachment 1). All glass aquaria, (3S x 20 x 20 ca), containing 16 1 of'diluent or diluent plus toxicant comprised a. study chamber. Ten bluagills, uniform in size were tested at each chemical concentration. Test fish were randomly allocated to@each 8 test chamber within 30 minutes f llowing toxicant addition. Bluegill experiments were conducted at IS C. Mortality, temperature, dissolved oxygen level and pH of all test solutions were measured at 24-hour intervals or until total mortality had occurred. General observations were recorded relative to swimming pattern and behavioral changes induced by exposure to t.hetest chemical. Dale Fattig Fish Farm, Brady, Nebraska Cownercial Fish Food of known composition 3 These studies were performed in replicate using the following FC 780 8 (lot 788-20) concentrations: 1.0, 1.3S, 1.8, 2.4 and 3.2 g/l. LC values with 95% confidence limits were calculated using the US/EPAso (Duluth) Probit Computer program. Organisms used in this study were considered to be generally healthy and free of disease. RESULTS The bluegill sunfish (Lepomia macrochirus) used in this study had an average weight of 0.92 + 0.36 g and length of 4.5 + O.S8 cm. The loading factor for these studies was approximately-O.S7 g/l, below the maximum resomended loading factor of 0.8 g/l (1). Temperature was maintained at 18 C. while pilranged from 7.6 to 8.0 pH units throughout this study. Dissolved oxygen values decreased sharply 48 hours post initiation of this study. Test aquaria solutions were originally clear, becoming cloudy on day 2, an observation which coincided with low D.O. levels ( 0.1-O.S mg/1). As the test solution cleared the D.O. levels increased. These abnormally low D.O. levels did not contribute to an increased mortality. This is considered to be presumptive evidence that the calculated LC values are accurate reflections of test chemical induced mortak2ty, per se. In this replicated study the 96-hour LC values were l.S, range 1.3-1.7 and 1.6, range 1.4-1.8 g/l respseptively. These values translate to LC values of 1500 and 1600 ppm. Chemicals possessing an LC value qn this range are coinidered to be an insignificant hazardso to the aquatic environment REFERENCES (1) US/EPA Methods for Acute Toxicity Test with Fish, Macroinvertebrates and Amphibians, Ecological Research Series 660/3-7S-009, 1975. (2) Registry of Toxic Effects of Chemical Substances, 176 Edition, Christensen, H. E. and E. J. Fairchild, Eds. p. XV and Ci, U. S. Department of Health, Education and Welfare; National institute for Occupational Safety and Health, Washington, b.C. June, 1976. I-AFK RIP-&AEC.r FOO SAt4PL-L l@ES&lk$PTIO14: Fit."tf$tUVXLL. RIOA= shmm Im @&-_ dige db m __mumm_ gi- 7ff 4 @,IM - -,To -- IL . 0. 41%9% (STAmt@i ..\ I ATTACIMNI' I z 0 #I a -9 . 4 JL97 '7.1 lp.,I-l five vto - 17.7 ireIF 4000 its2 40&Jr SALL 13 t7 .3 lype'I*t-st---.JII.L_@', @-st Oi-gani.,;m I-NVIRONMI.N'I'AL ENGINEERING LABOIZA'TORY A(IUA-1-ICTOXICITY DAI-A SHEE-1- Material Tested 7@t - i9 lot Date Recld<l/).@/@A FJjtch No Avg. llt,... grams Avg. Size q.,Jt, :Fg CM. 'rest Solution Volume I c- Liters/Tank No. Organisms per dose io Time Started Date Started- iCiliolIy Date Completed jo/iy/i; Dilution Water Carbon Invest igatorakA&ML-(Full Signatur#) 0 filtered well (Temp. c water LC SO (9S% Confidence D. 0. Limits) ppm, pk__@? t * c. Conc. mg/l. Total Vwunt Added 24-H ours Total ' * * B.O. No. Dead pil pps 48-Hours Total No.Dead IpH D.O. ppo 72-Hours Total D 0. N6.Uead ph-lp*pm 96-flours Total D.0 iNo.De ad @H -PPM Control V. A 0 .7.7 la- a 0 ?.7 liq 2.-L 0' 2.A_ id 'rest1'elill 1;5 -;it@iatureF, I)ate Comuients: otlit@it-han Mortality )ng/I SYMI)toms or Reactions (:ontrol 1&#biw-& lot 24-Hr. 0 Total No. Affected 48-tir. 72-tir. 96-Hr. L WAAU. 13 (141"6 Ali Vi 0-IIA ENV@RoNMI-'N'l'AL ENGINEERING LABOI,',ATORY AQT)KI'ICTOXICITY DAI'A SHEE'I' ,I,Ypc i*C.I;Otrganism Avg. Wt. , il9t Test Solution Volume 4C6 Material Tested f( Lot,., ic Vcndor-h',,,.t*,, Date Recld Batch No grams Avg. Size CM. #C- Liters/Tank No. Organisms' per dose Time Started_//.,,,@-4,k_l Date Started 1(;,Ifv Date Completed _jt,V/2i Dilution Water Carbon Investigato (Full Signature# filtered well (Temp. vs 0c ., D.O. water LC SO (9S% Confidence Limits) ppa!j,Pk-@i-Y-) ).3 Conc. Tot-aI kAwdudnetd Control 24-Hours Total D.O. No. Dead pH ppo 48-Hours Total No.Dead PH 'f2-Hours D.O. Total D.O. ppe N6.Uead pH ppe 96-liours Totil 0.0 No.Dead @H lpym S-7 (-,i 4 E.1 -4! 1 .5.1121 0 to odWt Test 1'emp.c i&ili@ure ftDate hi"P Comments: @8@--tdA"ft Lq I117 ff Gl:w64 i)uwxw JA.. toll-I)a . . ......... leil.Ull A VW Effects otlierthan Mortality ton-rci.) 0 n,-, syiitptumsor Reactions mgg//l, C.oonnttrol 24-Hr. 0 Total No. Affected 4B-Hr. 72-Hr. 10 7 10 96-Hr. @ikiiature4 1)@ite Cominents:------ left3 171