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ACUTE TOXICITY TO AQUATIC INVERTEBRATES TEST SUBSTANCE (DAPHNIA MAGNA) Identity:A mixturecontainingperfluorooctanesulfonatmea;y also be referredto as PFOS, FC-95, or as a component of FC-206CF or FC-206CF-X. (1-Octanesulfoniacid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was 101. The testsample isFC206CF. Currentinformationindicatesitisa mixtureof0.5% PFOS, 10% diethyleneglycolmonobutyl ether,78.08% water,1.375% acrylicfoamer, 1.5% sodium octylsulfate1,% sulfonatedalkylethersalts,6.0% urea,0.5% triethanolaminea,nd 0.05% tolyltriazole. The followingsummary applies to a mixture with incompletely characterizedconcentrations of impurities.Data may not accurately reflectthe toxicityof the fluorochemical component of the testsample. METHOD: Method: OECD Guideline202 Type: Acute static GLP: No Year completed: 1991 Species: Daphnia magna Supplier: In-houseculturesmaintainedatEnviroSystemsDivisionR,esource Analysts,Inc.,Hampton, NH. Analyticalmonitoring: Temperature,pH, conductivitya,nd dissolved oxygen. Exposure period: 48-hours Test organism age: <24 hours old Statisticalmethods: No calculationpserformed, based on study results. Test conditions: Dilutionwater: Filterednaturalwellwater Dilutionwater chemistry: Conductivity: 840 pmhos/cm pH: 7.6 Lighting: Cool-whitefluorescentlightsat20 pEin/seC/M2. A daily photoperiodof 16 hours lightand 8 hours dark was maintainedthroughoutthe testingperiod. Stock and testsolution preparation: A primary 1000 mg/L stocksolution was prepared and added directltyo createtestsolutions. Exposure vessels: 250 mL beakers containing200 mL exposure solutionat a water depth ofapproximately6 cm. Number of replicates:4 Number of daphnids per replicate:5 Number ofconcentrationsf:iveplusa blankcontrol Water chemistryduringthestudy: pH range(0-48hours): 7.6- 8.3(contreoxlposure) 7.7- 8.3(1000 mg/L exposure) Temperature range (0-48hours): 19.0- 20.2 'C (controelxposure) 19.1- 20.3-C (1000 mg/L exposure) Conductivityrange (0-48hours): 840 - 880 pmhos/cm (controelxposure) 850 - 880 pmhos/cm (1000 mg/L exposure) Dissolved oxygen range (0-48hours): 7.6- 8.3 mg/L (controelxposure) 7.7 - 8.3 mg/L (1000 mg/L exposure) RESULTS Nominal concentrations:Blank control1,50,250,400, 600, and 1000 mg/L. Element values: 48-hourEC50 = >1000 mg/L Element values based on nominal concentrations. Remarks: Testingwas conducted on the mixtureas describedinthe Test Substance Remarks field.The values reportedapply tothatmixtureand not the fluorochemicalproportionalone. CONCLUSIONS The testsubstance 48-hour EC50 was determined to be >1000 mg/L. No mortalitoyr sublethaleffectswere observed. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking= 2.This study meets the criterifaorquality testing.However, the sample puritywas not properlycharacterizedand the study lacksanalyticaclonfirmationofthe amount offluorochemicalproportion inthe solution. REFERENCES This testwas conducted by EnviroSystems DivisionR,esource Analysts, Incorporatedo,f Hampton, NH atthe requestofthe 3M Company, St.Paul, MN, Lab Request number J1884-4,1991. OTHER Last changed: 6/28/00. Study Title Static Acute Toxicity of rC-206cr to the Daphnid, Dapbais magns Authors Timothy J. Ward Robert L. Boori Study Comploted June, 1991 P*rformi@a 16aborit@@ --ZnviroSystess Divisi Resource Inalysts,Incori*rated One Lafayette Road-'@' Hampton, Now Hampshire- 03842 Fag* :L of 14 1. SURMT The &cut* toxicity of FC-206CY to the daphnid, Daphnis magna, is described in this final report. The test was conducted for 3H Company for 48 hours during June 10 to 12, 1991, at the EaviroSystess Division of Resource Analysts, Inc. in Hampton, New Hampshire. it was conducted by Jeanne Nagazu, Peter Kowalski, Zllen Stanford, Robert Doeri, and Timothy Ward. The test was performed under static conditions with five concentrations of test substance and a dilution water control at a temperature of 20 t IOC. The dilution water was filtered natural well water collected at Hampton, New Hampshire. Aeration was not employed to maintain dissolved oxygen concentrations above an acceptable level. Nominal concentrations of FC-206CF were: 0 mg/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. After 48 hours of exposure the control dapbnids had an average wet weight (blotted dry) of 0.0006 g, resultingin a loading rate of 0.015 g/L. All daphnids were in good condition at the beginning of the study. The test substance was not toxic to daphnids. Exposure of dapbaids to the test substance resulted in a 48 hour IC50 greater than 1,000 mg/L rC-206CF. page. 2 of -14 II. TABLB Or CONTEM SECTION: PAGI 1. Summary 2 ii. Table of contents 3 III. Index of Tables 4 IV. Introduction 5 V. Methods and Materials 5 Vi. Results 8 VIZ. References Appendix A. Water Quaiii bita from Toxicity Test 12 "t 7:t Page 3 ixr.nmimorTuLzs nD rrcnzs Table 1. Chemical characterization of a representative sample of natural well water used as dilution water for toxicity test Table 2. Survival data from toxicity test Table 3. Median effective concentrations (EcSos) from toxicity test Table A.l. Conductivity, pff,temperature, and dissolved oxygen concentration measured during toxicity test PAGE 6 9 10 13 Page. of 14 KIIFORE nmommon This study was sponsored by 3H Company, St. Paul, Minnesota. The objective of the study was to determine the acute toxicity of FC-206Cr to the dapbnid, a freshwater invertebrate. The report contains sections that describe the methods and materials esploy*d 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. TEST SUBSTANCE: V. METHODS AND MATERIALS FC-206er (EnviroSystess Sample Number 29382) was delivered to EnviroSystess on Kay 23, 1991. It was contained in a 4 oz. Plastic bottle that was labelled with the following Information:IJ1884-4, FC206CF". rC-206CF (a light brown liquid) was supplied by 3M Company, 935 Bush Avenue, St. Paul, Minnesota. Prior to use the test material was stored in the dark at room temperature. A reserve sample (approximately1.0 grans) will be retained at EnviroSystess for a minimum of 10 years. DILUTION WATER: Water used for acclimationof test organismsand for all toxicity testing was collected from wells at EaviroSystem in Hampton, New Hampshire. , Water was stored in 500-gallon polyethylene tanks where it was aerated'. Results of chemical analysis of a representative sample of water are.presented in Tible@l. TEST ORGMSN: -Juvenile daphaids employed as test organisms were from a single source -'andL@w@ere .identified using an appropriate taxonomic key. Daphnids 'used'@ tliei-e7st'iere' produced from an in-houie culture and were ess than 24-; tha' start of the test'."Control dapbuids were weighed at@z' the toxicity test. -,Priorto testing, daphnids were iaintained-..:.1i0n0.% dilution.water under.static conditions in 4 liter glass D@iini'.,,.,.icclizatdiaopnhaids were @not:,treated for disease and they. of apparent,@sickness, injarlei,' and abnormalities'a b begin'n g'::'.-"of7"t@els?t.b,e During the.acclimation period 24 hours"prior to'---@, the-,.t-e,st'--i@nitiation the temperature was 19.5*C. Daphnids were fed yeast trout" and/or the freshwater alga'.S-o@lonastrus capricoroutuwonce or twice'dailybefore the test. k'@ NOU=Ar&^W-&bd*qd Olt Table 1. Chemical characteriz@lion of representative sample of natural well water used as'dilutionwater for toxicity test Parameter PH Conductivity Organochlorine pesticides Organophosphorus pesticides Polychlorinated biphenyis Unit of Measurement PH units unhos/cm ug/L Reporting Limit 0.5 ug/L 0.5 ug/L 0.5 Measured Value 7.6 840 I(D ND ND Notes: I. None detected at or above the reportino limit. 2. Ph and conductivity were measured in dilution water collected from a control test,vesselprior to introduction of collected organizas'arid,-Pesticidaend PCB d ata is during.routine biannual.water quality testing. 7,@ 7 P e 6 of 14 TOXICITY TESTING: The definitivetoxicity test was performed during June 10 to 12, 1991. It is based on procedures of the OECD (1984). The test was conducted at a target temperatureof 20 t 10C with five concentrations of test substance and a dilution water control. A 1,000 mg/L stock solution was formulated without the use of a solvent. Nominal concentrations of the test material were: 0 mg/L (control),150 mg/L, 250 mg/L, 400 mg/L, 600 mg/L, and 1,000 mtr/L. Twenty daphuids were randomly and equally distributed among four replicates of each treatment. The test was performed in 250 al glass beakers that contained 200 ol of test solution (water depth was approximately 6 ca). Test vessels were randomly arranged in an incubator during the 48 hour test (a random numbers table was used to select the location of each vessel). A 16 bour light and 8 hour dark photoperiod was automatically maintained with cool-white fluorescent lights that provided a light intensity of 20 UES-la-2. Aeration was not required to maintain dissolved oxygen concentrationsabove acceptable levels. Dapbnids were not fed during the test. The number of surviving organisms and the occurrence of imobilization (inabilityto swim within 15 seconds after gentle agitation) or other sublethal effects (lossof equilibrium,erratic swimming, loss of reflex, excitability, discoloration,or change in behavior) were determinedvisually and recorded initiallyand after 24 and 48 hours. Dissolved oxygen (YSI Model 57 meter; instrument number PRL-3), pE (Beckman model pHl 12 meter; instrument number PRL-51), conductivity (Cole Parser model number 1491-60, instrument number PRL-50), and temperature (ASTM mercury thermometer; thermometer number 2265) were measured and recorded daily in each test chamber that contained live animals. STATISTICAL EMODS: Results of. the toxicity test could not be interpreted by standard statistical techniques (Stephan, 198.3)because 50% survival or greater occurred in all tested concentrations. 7 of 14 R ArooP^kV@Sut)WdkdNMyTLFM vr. RESULTS All tsst vessels containing FC-206CP were initially clear and remained clear throughout the test. Biological and water quality data generated by the acute toxicity test are presentedin Table 2 and Appendix A, respectively. One hundred percent survival occurred in the control exposure. dry) Control dapbnids bad an average wet weight (blotted of 0.0006 g at the end of the test. Loading rate during the toxicity test was approximately 0.015 g/L. The 24 and 48 hour EC50s for daphnids exposed to FC-206CF are presented in Table 3. The 48 hour ICSO was greater than 1,000 mg/L FC-206CF. Page 3 of 14' PosaumAnotvfWt- SbddbV d Table 2. Survival data from toxicitytest Nominal Concentration (mg/L) rep. 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 1 000 2 Number Alive 0 hr 24 hr 48 hr 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5, 5 5 5 5 5. 5 5 5 Number Affected Ohr 24hr 49hr 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 0 0 0 0 0 0 0 o 0 0 0 0 0 o 0 4t Table 3. Nsdi&u effOctivO concentrations (ICCSO,)from toxicity test Exposure period 95 percent EC50 confidence limits Calculation method 24 hours )1,000 mg/L 48 hours )1,000 mg/L 'V@7 )f 14 vir.zznmcics OECD. 1981. Decision of the Council Concerning the Mutual Acceptance of Data in the Assessment of Chemicals. Aznex 2. OECD Principles of Good LaboratoryPractice. C (81) 30 (Final). 030. 1984. Guidelines for Testing of Chemicals. Section 2: Effects On Biotic Systeas. Method 202, Dapbuid sp., Acute Iamobilization Test and Reproduction Test. Adopted April 4, 1984. -tIk @4- -1" d AppendiX A. WATER OUALITY DATT FRom ToxrciTy TEST tie r 12,nr -IL4 Table A.I. Conductivity, pn, tempo-sture, And dissolved oxygen concentration measured during toxicity test Nominal concentration (no/L) Rep. 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, 11000 1- 2 3 4 Conductivity (usho/cm) 0 24 48 hr hr hr 870 ego $40 870 880 840 870 880 840 870 880 840 860 900 840 860 900 840 860 900 840 860 900 840 860 880 $40 860 880 840 860 880 840 860 880 940 860 870 $40 860 870 840 860 870 840 860 870 850 860 870 850 860 870 850 860 870 850 860 870 850 860' 8801@, $50 860 880 $50 -860 -880,- 860. 880 0 hr 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.7 7.7 7.7 7.7 PH 24 hr 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7, 7.7 7.7 7.7 48 hr 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 @S. 3 83 .8.3 8.3 .3 of@ld, Table A.I. Continued. nominal concentration (mg/L) Rep. 0 control 1 2 3 4 150 1 2 3 4 250 1 2 3 4 400 1 2 3- 4 600 1 3 Temperature (OC) 0 24 48 hr hr hr 20.0 20.2 20.1 20.2 20.2 20.2 20.1 20.2 20.3 20.1 20.2 20.1 20.1 20.2 20.2 20.3 20.0 20.1 20.1 20 .2 2 19.0 19.0 19.1 19.1 @19.1 19.3 19.4 19.3 19.1 19.0 19.2 19.1 19.4 19.3 19.2 19.3 19.1 19.2 19.0 19.0 19.3 19.3 19.3 19.3 19.0 19.1 19.2 19.1 19.3, 19.4. 19.4 19.5 19 19.3,: 19.4 19.2 19 2,,-'i@'.19 19.1' 19-'319.'3i- Dissolved Oxygen (mg/L) 0 24 48 br hr hr 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7 .7 77 7.7. 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 77 7.7 7.7 7.7 7.7 7 7. 7:7 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 8.3 .3 @3 S.@ 3 8 31,?,;, , .1@ps -14@ li7