Document 6b1Lv6R7nKZwan0zDyEzo3B2m

ACUTE TOXICITY TO AQUATIC INVERTEBRATES (DAPHNIA MAGNA) TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonatweh,ich may also be referredto as PFOS, FC-95, or as a component ofFC-203. (1Octanesulfonicacid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was notnoted. The testsample isFC-203. Currentinformationindicatesitisa mixtureof 1.34% PFOS, 35% diethyleneglycolbutylether,37.85% water,20% ethyleneglycol,2.66 % Sultone foamer, 3% sodium octylsulfate0,.1% sodium laurylsulfate,and 0.05% tolyltriazole. The followingsummary applies to a mixture with incompletely characterized concentrations of impurities.Data may not accurately reflecttoxicityof the fluorochemical component of the testsample. METHOD: Method: OECD Guideline202 Type: Acute static GLP: No Year completed: 1991 Species: Daphnia magna Supplier: In-houseculturesmaintainedatEnviroSystemsDivisionH,ampton, NH. Analytical monitoring: Temperature, pH, conductivitya,nd dissolved oxygen. Exposure period: 48-hours Test organism age: <24 hrs.old Statisticamlethod: LC50 values calculatedusinga computer program from Stephan, 1983. Test conditions: Dilutionwater: Filterednaturalwellwater. Dilutionwater chemistry: Conductivity: 700 pmhos/cm pH: 7.9 Lighting: Cool-whitefluorescentlightsat 12 pEin/seC/M2. A daily photoperiodof 16 hours lightand 8 hours dark was maintainedthroughoutthe testingperiod. Stock and testsolution preparation: A 1000 mg/L primarystock solution was prepared by dissolving2.0 g oftestsubstance in2000 mL ofdilution water. Exposure concentrationwsere prepared by additionofthe appropriate volume of stocksolution. Exposure vessels: 250 mL beakers with200 mL exposure solutionand a water depth of approximately6 cm. Number ofreplicate4s: Number ofdaphnidsperreplicat5e: Numberofconcentrationfsi:vpelusa blanckontrol Waterchemistrdyuringthestudy: pH range(0-48hours): 7.9- 8.3(contreoxlposure) 8.0- 8.3(1000mg/L exposure) Temperature range (0-48hours): 19.7- 20.6OC (controelxposure) 19.4- 20.6-C (1000mg/L exposure) Conductivityrange:(0-48hours) 640 - 700 pmhos/cm (controelxposure) 640 -660 pmhos/cm (1000 mg/L exposure) Dissolved oxygen range (0-48hours): 8.3- 8.8 mg/L (controelxposure) 8.0- 9.9 mg/L (1000mg/L exposure) RESULTS Nominal concentrations:Blankcontrol1,50,250,400,600,and 1000 mg/L. Element values: 48-hourEC50 = >1000mg/L Element valuesbased on nominalconcentrations Remarks: Testingwas conductedon themixtureas desc(ibedinthe Test Substance Remarks field.The valuesreportedapplytothatmixtureand not the fluorochemicaplroportionalone. CONCLUSIONS The testsubstance48-hourEC50was determinedtobe >1000 mg/L, the highestconcentratiotnested.No mortalitoyrsublethaelffectswere observed. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking= 2.Thisstudy meets thecdteriaforquality testing.However, thesample puritywas notproperlycharacterizeadnd the study lacksanalyticaclonfirmatioonftheamount offluorochemicaplroportion inthe solution. REFERENCES This testwas conducted by Envirosystems Division,Resource Analysts,Inc., of Hampton, NH atthe requestofthe 3M Company, Lab Request number H2864-9,1991. OTHER Last changed: 6/27/00 Amended 8/9/91 study Title Static Acute Toxicity of FC 203 to the Daphnid. Daphnia mauna Authors Timothy J. Ward Robert L. Boeri Study Coapleted July 29, 1991 Report Amended lugwt 9. 1991 Perfo@ni@ na Laboratory Envirop3ys teas Division Resource knalysts'.Incorporated One Lafaye@te,Road Hajipton,'Now Han@sbire 03842 Envirosysteas Study Number 90174-3 P age --Iof 17 !Y E G= LAMUTORT PRIMCZ STITCONR This study was conducted according to OEM Good Laboratory Practice Regulations. Neither the Study Director nor the sponsor are aware of any circumstances that would affect the integrity of this study. 7 hird A@rbyaotTrnani Study Direct 2AI1L1@14@t R6beri L: B&ri Coauthor J6&nne P. ma L@@tic Toxi=t govalsld Aoatic To3dcolt)9L1st INV. %7. Stanford, 7 Aquatic,,Toxicologait -k ftvirosysta'asStudy Number 90174 -3 Pa 2 of 17 k@@- 9,JADAOWdkyd U. QUI.TffLssn"a ST=ONT Subaitted by: EnviroSystess Division Resource Analysts, Incorporated One Lafayette Road Rampton. low Rampsbire 03842 Certification: Data presented in this report were derived by methods and with materials identified in the section of the report entitled -Methods and Materials." The test was performed in accordance with EnviroSystess Protocol 90174-3 and the Product Registration Aquatic TomicolM Laboratory Standard Operating Procedures Mmual. The toxicity test was performed by Z3LlenStanford, Jeanne Magazu, Peter Kowalski, Robert B"ri, and Timothy Vard. The original raw data and final report will be archived at Resource Analysts, Inc. for at .least 10 years. all data transcribed froa the raw data to the report were checked for accuracy, and all data were verified by Quality Assurance Auditors. Quality assurance audits were performed on Audit Date 11/14190 113LO/91 2/ 7/91 7/30/91 8@i fit Reported to Study Director ]Ll/lS/90@-' -1/10 2/ 7/91'* 7/30/91 Reported to Management 7/30/92 7/30/91@@: 7/30/91, ty Assurance tepr@santative N Inv rosystess Study Wmkber 90174-3 Page 3'of ''17,,,:- RemurceNO^ kv.@SAWclay d L III. TABLZ OT COCRMS SECTION: 1. Good Laboratory Practice Statement II- Quality Assurance Statement 111. Table of Contents IV. Index of Tables V. summary VI. Introduction', VII. Methodi and Naterials VIII. Results-@, . Ix e erences @4, Appendix Qu ality Data from Toxicity Tes t. Page 2 3 4 5 6 7 7 14 15 At, 1., in EnviroSy teiiii@@S d y N Page@4 of er 90174 j,"A' e@;@ e,4 IV. INDKI OF TkBtaES Table 1. Table 2. Chemical characterization of a representative sample of natural well water used as dilution water for toxicity test Survival data from toxicity test Table 3. Median effective concentrations (EC50s) from toxicity test Table A.l. Conductivity, pH. temperature, and dissolved oxygen concentration measured during toxicity test Page 8 12 13 16 -,-,Envirbsistess,StudyNumber 90174-3 'Page 5' c ResotxmAr krc d Amended 8/9/91 V. SUMMY The acute toxicity of PC 203 to the daphnid, Daphnia magma, is described, in this final report. The test was conducted for 3H Company for 48 hours during, March 19 to 21, 1991, at the EnviroSysteas Division of Resource Analysts, Inc. in Hampton, New Hampshire. It was conducted by Ellen Stanford, Jeanne magazu, Peter Kowalski, Robert Boeri, and Timothy Ward according to the protocol. developed for EnviroSystems Study Number 90174-3. The test was performed under static conditions with five concentrations of test substance and a dilution water control at a temperature of 20 t 10C. The dilution water was filtered natural well water collected frou wells at Hampton, New Hampshire. Aeration was not employed to maintain dissolved oxygen concentrations above an acceptable level. Nominal concentrations of test substance were: 0 ag/L (control), 150 mg/L, 250 mg/L, 400 mg/L. 600 mg/L, and 1,000 mg/L. Nominal concentrations were used for all calculations. Daphnids used in the test were produced from an in-house culture and were less than 24 hours old at the start of the test. kfter 48 hours of exposure the control dapbuids had an average wet weight (blotted dry) of 0.0017 g, resulting in a loading rate of 0.004 g/L. All dapbnids were in good condition at the beginning of the study. Exposure of daphnid3 to the test substance resulted in a 48 hour median effective concentration (EC50) greater than 1,000 mg/L PC 203,, the highest tested concentration. .7' EnvirbSystems'@Study Number 90174 -3 Page 6 of 17 -,IKO A"AgiM Amended 8/9/91 VI. 33MODUCTION This report was amended to include data from the final definitive toxicity test that was conducted during March 19 to 21, 1991. The study was sponsored by 3K Company, St. Paul, Minnesota. The objective of the study v" to determine the acute toxicity of TC 203 to the dapbuid, a freshwater invertebrate. The report contains sections that describe the methods and materials employed in the study, and the results of the investigation. The report also contains an appendix that presents the water quality data collected during the test. VIZ. NL'MDS MM NILZ&IALS TEST SUBSTANCE: TC 203 (EaviroSysteRs Sample Number 2699E) was delivered to EnviroSystems on November 15, 1990. It was contained in a 250 al glass bottle that was labelled with the following information: "H 2864-9. TC 203". The test substance was supplied by 3H Company, St. Paul, Minnesota. Prior to use it was stored in the dark at room teaperature. k reserve sample will be archived at EnviroSystem for a aininum of 10 years. DILUTION WATER: Water used for acclimation of test organisms and for all toxicity testing was collected from wells at ZnviroSystess in Hampton, New Hampshire. Vater was stored in 500-gallon polyethylene tanks where it was aerated. Results of chemical ,analysis of a representative sample of water are presented in Table 1. TEST ORGANISM: Juvenile-dapbuids employed as.:test,organizas were from aLsingle sourci:'ind were identified using an appropriat*elt-i_i6noaic ke*y."'Dapbnids used -in th4't.est ,.were'produced,fron ,an in-house culture.-and were less -tbaln24_,,houLrsiioolndrlaiDtt,,@ttbhee start of the test.@@ Control diphnidi".*ere weighed 'at thei,concli toxicity test'.@,Prior to testing, daphrlidswere maintained in 100% dilution water under static conditions in 4 liter During acclimation daphnidi'ver4.,,, .@:,.r'ntorteated for disease and they were free of apparent sickness,".injuries,-'-and abnormalities at.the beginning of the -test'. During the acclimation, period 24@@',*":@ hours prior to the test initiation tb,e,-,tesperatuwraes 19.60CZDapWd 'cultures ted yeast/trout chow and/or the freshwater.alga Selenastriums capricornutul daily before,tbe,test InviroSy'st'e'U'-3"St'UNduys-ber Page:.7,of-. la .41,rl(:Itvfitrlr &"dicryof :7 kmended 8/9/91 Table 1: Chemical characterization of a representative sample of natural well water used as dilution water for toxicity test Parameter Unit of Measurement Reporting Limit Measured Value PH PH units 7.9 Conductivity ushos/cm 700 Organocblorine ug/L 2 ND pesticides Organophospborus ug/L 5.0 ND pesticides Polychlorinated ug/L 0.6 ND biphonyls Notes: 1. ND Not detected above the reporting limit. 2. The, pE and conductivity values were detemined in a sample of dilution water collected from a control'test vessel at the beginning of the toxicity test.* ]Ldditionalchemical parameters were measured in '-dilutiownater during October, 1990 as part of,routine biannua water,.qualityanalysis., *F7, E nvi'ros9 udy Number 90174 -3 ,Pave 8 of 17', ROMUFOOkldvft SibockIryd Amended 8/9191 TOXICITY TESTING: A screening test with the test substance was conducted durinc December !i to 7. 1990. Nominal concentrations of test substance were: 0.1, 1.0, 10, 100, and 1,000 ag/L. Ifter 48 hours of exposure at least 70% survival occurred at all. tested concentrations. I definitive test was attempted during January 9 to 11, 1991. Nominal concentrations of test substance were: 150. 250, 400, 600, and 1,000 ag/L. After 48 hours of exposure sublethal effects were noted at all tested concentrations (sublethal effects were not seen at 1.000 ag/L during the screening test) and the test was repeated. The definitive toxicity test was performed during March 19 to 21, 1991,, according to EaviroSysteas Test Protocol 90174-3 (Static. Acute Toxicity Test With FC 203 And The Daphnid, Daphnia aa(rna),which was signed by the Study Director on November 16, 1990. It is based on procedures of the OECD (1984). The test was conducted at a target teaperatimreof 20 1 1*C with five concentrationsof test substance and a dilution water control. A 1,000 ag/l. stock solution was prepared by combining 2.0 9 of test substance with dilution water and adjusting the final concentration to 2,000 al without the use of a solvent. Nominal concentrations of the test material were: 0 ag/L (control),, 150 ag/L, 250 ag/L, 400 ag/L, 600 ag/L. and 1,000 ag/L FC 203. Twenty daphnids were randouly and equally distributed among four replicates of each treataeut. The test was perforaed in 250 al glass beakers that contained 200 al of test solution (water depth v" approximately 6 ca). Test vessels were randomly arranged in an incubator during the 48 boar test (a random numbers table was used to select the location of each vessel). A 16 hour light and 8 hour dark pbotoperiod was automaticallymaintained with cool-white fluorescent lights that provided a light intensity of 12 uft"ta7l.A,eration was not reqaired to maintain dissolved oxygen concentrations above acceptable levels.'.Dapbnids axe not fed during theLteSt. The number of surviving organismi and the occurrence ai" bletba effects. and imobilization (inabilityto swin within 15 seconds aftei gentle-agitation) ;@or other,subletbal effects (loss of equilibrium, @e.r,ratic swimming,,, loss, iDf-7 reflex, excitability, discoloration. or change in' behavior) @w.ere 'determined,, visually and recorded initially7and after 24 and 48 hours.. Dead test'o i were .removed @when first' observed.',.D-i,ssolved oxygen ,(YSI',"-@nodel a a instrument number PRL-3), pH ,(Beckmannodel pEl 12 neter; irlstranut PRL-4) conductivity I (Labcoup'..SCT .,m,e,ter instrument nuidmr'@,P temperature (KSTH mercury tberaometer,@'theraoseternumber 2265) were a recorded daily in each,test chamber'that contained live animls.@l."-* ms Study Humber,90174 9.o f 17';@'-, Rexxxx A-mlysk k-- Sib*" d MLLPORE . Azended 8/9/91 STATISTICKL METHODS: Results of the toxicity test could not be interpreted ',y standard statistical techniques (Stephan, 1983) because less than 50% of the daphnids exposed to the highest concentration were killed or immobilized. Nosinal concentrations of test substance and the rumber of daphaids killed or immobilized were used to deteraine the EC50 value. 7" -..EnvroSyst"e@@,Study") -3 of Page 10 17 'I@ 4 L Amended 8/9/91 Vill. RESULTS All test vessels containing FC 203 appeared clear initially and remained free of insolublematerial throughout the test. Biological and water quality data generated by the acute toxicity test are presented in Table 2 and Appendix k, respectively. one hundred percent survival occurred in the control exposure. Control daphnids bad an average wet weight (blotted dry) of 0.0017 g at the end of the test. Loading rate during the toxicity test was approximately0.004 g/L. Exposure of daphnids to the reference toxicant sodium dodecyl sulfate resulted in a 48 hour median lethal concentration (LCSO) of 7-9 mg/L. The 24 and 48 hour EC50s for daphnids exposed to FC 203 are presented in Table 3. The 48 hour EC50 was greater than 1,000 mg/L FC 203, the highest tested concentration. EnviroSysteas Study Number 90174-3, page 11 of 17 Amended 8/9/91 Table 2. Survival data fron toxicity test Nominal Concentration (ag/L) rep. Number Alive 0 hr 24 br 48 br Number Affected Obr 24hr 48hr 0 (control) 1 2 3 4 150 1 2 3 4 250 1 2 3 4 400 1 2 3 4 600 1 2 3 4 if606 2 3 4 5 5 5 5 5 5 5 5 5 5 5 5 5 5 4 5 5 5 5 5 4 5 5 5 5 5 4 5 5 4 5 5 5 5 5 4 5 4 3 5 5 4 5 4 1 5 5 5 5 5 5 5 2 2 5 4 4 5 2 2' 5 5 4 5 5 3 5 5 5 5 3 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 @o 0 0 0 0 0 0 0 00 0 0 0 0 0 0 0 0 0 0. o o @o Affected dapbuids were iwwbilized@(unable to swim witbin 15 seco7. after;.'gentleagitation) LPniiroSystems Studi-kii"er@ 90174 -3, Page 12: of 17.. Remotxv ArK*Vftkir-Subk" d L4LLPORE' X7;, Azended 8/t/91 Table 3. Median effective concentrations (F.C50s)from toxicity test Exposure period 95 percent Calculation EC50 confidence limits method 24 hours 48 hours >1,000 ag/L >1,000 mg/L Enviro3yollis Study Number 90174 -73 Pave 13 of 17 d N IIPORE IX. RErn=CES OECD. 1981. Decision of the Council Concerning the Mutual Acceptance of Data in the Assessment of Chemicals. knnex 2. OECD Principles of Good Laboratory Practice. C (81)30 (Final). OECD. 1984. Guidelines for Testing of Chemicals. Section 2: Kffects on Biotic Systems. Method 202, Daphuid sp., Acute I"obilization Test and Reproduction Test. Adopted April 4, 1984. Stephan, C.E. 1983. Computer program for calculation of LC50 values. Personal communication. EnviroSystess'Stu, y Number 90174-3 Page 14 of 17:@t,- 4f, RE$OUMO Anci r-S.tA)shjkxdytALiFORE Appendix A. WATER OUALITY DATR FROM TOXICITY TEST lr EnviroSystems'Study Number 9017,9-3 Page 15 of 17 Amended 8/9/91 Table A.l. Conductivity, pH, temperature, and dissolved oxygen concentration measured during toxicity test Nominal concentration (mg/L) Rep. Conductivity (unhos/cm) 0 24 48 hr hr hr 0 (control) 150 250 400 600, i,ooo 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 4 2 3 4,,, 700 700 700 700 660 660 660 660 660 660 660 660 660 660 660 660 660 .1 660 660 660 660 660 660 660 660 640 660 650 660 650 660 660 630 650 630 650 630 650 640 660 640 660 640 660 640 660 640 660 640 660 640 660 640 660 640 660 640 @660 640 670 640--,--, 660 640 670 640 660 640 6 60 640 660 640 660 PH 0 24 hr hr 7.9 8.3 7.9 8.3 7.9 8.3 7.9 8.3 8.0 8.2 8.0 8.2 8.0 8.2 9.0 8.2 8.0 8.2 8.0 8.2 8.0 8.2 8.0 8.2 8.0 8.2 8.0 8.2 8.0 8.2 8.0 8.2 8.0 8.3 8.0 8.3 8.0 8.3 8.0 8.3 . 80 8.3 8.0 8.3 8.0 -:1,@ 8.3 8.0 83 48 hr 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.01 EnviroSystems Study Number 90174-3@ Page 16 of 17. Amended 8/9/91 Table A.l. Continued. Nominal concentration (mg/L) Rep. Temperature (OC) 0 24 48 - bLr hr hr 0 (control) 150 250 400 600 l'ooo 1 20.3 2 20.3 3 20.2 4 20.4 1 20.4 2 20.3 3 20.3 4 20.3 1 20.3 2 20.2 3 20.1 4 20.3 20.4 .2 20.3 3 f 20.2 4 ' 20. 3 20.3 ',g,,@'-@.20.4 20.4 , 20 2: --20.3 2'* 20.3 20.3. 20.3, 19.8 19.9 19.9 19.8 19.7 20.1 20.5 20.6 19.7 19.8 19.7 19.3 19.4 19.3 19.2 19.3 19.3 19.3 19.4 19.7 19.3 19.3 19.3 19.3 19.8 19.1 19.9 19.8 19.4. -'19.6 19.4 19.5 19.6 19.7 19.2, -,@-29 $-- 19 7":@@ 19.4 20 2 19.4 1,9, 20 6, 8 19. 7. Dissolved Oxygen (mg/L) 0 24 48 hr hr hr 8.5 8.4 8-5 8.5 8.8 8.7 8.6 8.7 8.6 9.6 8.5 8.6 8.7 8.5 8.5 8.5 8.4 84 84 84 8.3 84 8.4 8 8.8 8.8 8.8 8.8 8.7 9.0 9.2 9.2 9.4 9.4 9.5 9.6 9.6 9.7 9.7 9.6 9.6 9..87 9. 6 9. 8 99 97 8.3 8.3 8.5 8.3 8.1 8.2 8.0 8.1 8.0 8.0 8.1 8.1 8.1 8.1 8.1 8.2 8.2 8.2 s.o 8.1 8.1 8.0 .8.1@ 8.0 Envirosyste s page 17 of Let! er',-90174-3 1-7 r-M