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ACUTE TOXICITY TO FISH TEST SUBSTANCE Identity:Perfluorodecanesulfonatea,mmonium salt:may also be referred toas PFDS, FC-1 20, or 3M Sample J2905-1.(1-Decanesulfonic acid,1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,9,9,10,10,10heneicosafluoro-a,mmonium salt,CAS # 67906-42-7) Remarks: Test sample isa mixtureofthe testsubstance inwater and butylcellosolve(approximately25% testsubstance,37.5% butyl cellosolvaend 37.5% water).Allvaluesreportedrelatetothismixture. No calculationwsere made to adjustforthe actualconcentrationofthe testsubstance inthe testsample. Test sample was from 3M production Lot 164. METHOD Method: OECD 203 Type: Staticacute GLP: No Year completed: 1992 Species: Pimephalespromelas Supplier:AquatiRcesearch Organisms,Hampton, New Hampshire Analyticalmonitoring:pH, dissolvedoxygen and conductivitaynd temperatureonly.Test substance concentratiownas not measured. Exposure period:96-hours Statisticamlethods: Probitanalysis. Test fishage: Not noted otherthan "juvenile". Length and weight: Average length 26.5 mm. Average weight 0.14 g Loading: 0.09 g fish/ L Pretreatment: None Test conditions: Dilutionwater:Aerated,filterendaturalwellwater Dilutionwater chemistry (from a representativesample): hardness: 160 mg/L as CaC03 pH: 7.0 Conductivity: 640 pmhos/cm Stock and testsolutionpreparation:The testconcentrations were prepared by transferrinagppropriateweights of FC-120 directtloythetescthambersT.estconcentratiwoenrseprepared basedon totaslamplen,oton percenctoncentratoifotnhetest substancienthetesstample.AlltesvtesselcsontainiFnCg-120 wereiniticalelayarndremainecdleatrhroughotuhtestudy. ConcentratiodnossingrateO:nce StabiliotfythetestchemicaslolutionNso:tnoted Exposurevessels2:0 litgelrasasquaricaontaini1n5gLoftest solutiowna,terdepth- 18cm. Number ofreplicattewso: Number offishperreplicate:n Numberofconcentratiofnisv:pelusa negativceontrol Waterchemistrdyuringthestudy: Dissolveodxygenrange(0- 96 hours): 6.8-8.5mg/L(contreoxlposure) 5.7-8.6mg/L (15mg/Lexposure) pH range(0- 96 hours) 7.0-8.2(contreoxlposure) 7.0-8.0(15mg/Lexposure) TesttemperatureM:ean temperatusrteateads 23 + 20C RESULTS NominalconcentrationBsk:contro2l.,54,.0,6.0,10,15mg/L Elementvalue: 24-houLrC5o= > 15mg/L 48-houLrC5o= > 15mg/L 72-houLrC5o= 11(9.-7 12.5m)g/L 96-houLrC5o= 4.8(4.-0 5.7)mg/L Allelemenvtaluebsasedon nominacloncentrations StatistiEcvaalluationfMortality: Probiatnalysiwsas usedtocalculaLtCe50valueasndthecorresponding confidenlciemits. Cumulativpeercenmtortality: Nominad Test Conc., mg/L 24-hours 48-hours 72-hours 96-hours Neg. Control 0 0 0 0 2.5 1 0 0 0 10 4.0 0 0 0 25 6.0 0 0 0 85 10 0 1 0 40 85 1 15 1 0 20 85 100 CONCLUSIONS The testsample 96-hour LC5o forfathead minnow was determined to be 4.8 mg/L witha 95% confidenceintervaolf 4.0- 5.7 mg/L. Testingwas conducted on the "as-sold"mixtureofthe subjectmaterialin water (-25% subjectmaterial)T.he valuesreportedapplyto thatmixture -andnot directltyo the subjectmaterial. Submitter: 3M Company, Environmental Laboratory,P.O. Box 33331, St.Paul,Minnesota,55133 DATA QUALITY Reliability:Klimischranking2. This study,whilewellconducted, lacks analyticadlata fordeterminationofthe testsubstance concentrationinthe testsolutionsand determinationofthe sample purity.Because a mixture was tested,care must be takenwhen interpretintghe results.The solvent inthe mixturecould have enhanced or restrictebdiologicaulptake and/or toxicitoyfthe fluorochemicalcomponent. REFERENCES This study was conducted by EnviroSystems DivisionResource Analysts, IncorporatedH,ampton, New Hampshire on behalfofthe 3M Company. OTHER Last changed: 5/18/00 Study Title Static Acute Toxicity of FC-120 to the Fathead Minnow, Pimephales prome.Zas Authors Timothy J. Ward Robert L. Boeri Study Comipleted March 1992 Performina Laboratory EnviroSysteme Division Resource Analysts, Incorporated One Lafayette Road Hampton, New Hampshire 03842 Page 1 of 14 I.SUMMARY The acute toxicity of FC-120 to the fathead minnow, Pinophalos promelas, is described in this final report. The test was conducted for 3M Company for 96 hours during February 27 to March 2, 1992, at the EnviroSystems Division of Resource Analysts, Inc. in Hampton, New Hampshire. It was conducted by Jeanne Magazu, Peter Kowalski, Ellen Stanford, Robert Soeri, and Timothy Ward. The test was performed under static conditions with five concentrations- of test substance and a dilution water control at a mean temperature of 23 t 20C. The dilution water was filtered natural well water collected at Hampton, New Hampshire. Aeration wai not employed to maintain dissolved oxygen concentrations above an acceptable level. Nominal concentrations of FC-120 were: 0 mg/L (control), 2.S mg/L, 4.0 mg/L, 6.0 mg/L, 10.0 mg/L, and 15.0-mg/L. Nominal concentrations were used for all calculations. Fish used in the test were purchased from a commercial supplier (Aquatic Research Organisms, Hampton, New Hampshire) and acclimated under test conditions for more than IS days. After 96 hours of exposure the control fish had an average total length of 26.S m and an average wet weight (blotted dry) of 0.14 g, resulting in a loading rate of approximately 0.09 g/L. All fish were in good condition at the beginning of the study. Exposure of fathead minnows to the test substance resulted in a 96 hour LCSO of 4.8 mg/L FC-120, with a 9S% confidence interval of 4.0 to 5.7 mg/L. One of the eight surviving fish exposed to 15.0 mg/L at 48 hours and one of the three surviving fish exposed to 15.0 mg/L at 72 hours exhibited erratic swimming. The three surviving fish exposed to 10.0 swimming. mg/L at 96 hours exhibited a loss of equilibrium and erratic No other sublethal effects we're observed in any test vessel during the 96 hour exposure. Page 2 of 14 XX. TABLZ OF COKTZ"S SECTION: PAGE I. Summary 2 ii. Table of Contents 3 III. Index of Tables 4 IV. Introduction 5 V. Methods and Materials 5 VX. Results VXI. References Appendix A. Water Quality Data from Toxicity Toot 12 Page 3 of 14 Ill. INDEX OF TABLES 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 lethal concentrations toxicity test (LCSOS) from Table A.l. Conductivity, pH, temperature, and dissolved oxygen concentration measured during toxicity test PAGE 6 9 10 13 Pag* 4 of 14 IV. INTRODUCTION This study was sponsored by 3H Company, St. Paul, Minnesota. The objective of the study was to determine the acute toxicity of FC-120 to the.-fathead minnow, a freshwater fish. 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. V. METHODS AND MATERIALS TEST SUBSTANCE: FC-120 (EnviroSystems sample Number 51033) was delivered to EnviroSystems on November 13, 1991. It was contained in a 250 ml plastic bottle that was labelled with the following information: "J2904-I, FC-120, Lot., 164, 2-Butoxy Ethanol". FC-120 (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 (approximately 1.3 grama) will be retained at EnviroSys@, s for a minimum of 10 years. DILUTION WATER: Water used for acclimation of test organisms and for all toxicity 'testing 'was collected from wells at EnviroSysteme 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 Table 1. TEST ORGANISM: Juvenile fathead minnows employed as test organi ma were from a single source and were identified using an appropriate taxonomic key. They were purchased from a commercial supplier (the Aquatic Research Organisms Division of Resource Analysts, Inc., Hampton, Now Hampshire) and acclimated at the EnviroSystems facility for more than 15 days. Control fish were weighed at the conclusion of the toxicity test. Prior to testing, fish were maintained in lbO% dilution water under flow through conditions in an all glass aquarium. During acclimation fish were not treated for disease and they were free of apparent sickness, injuries, and abnormalities at the beginning of the test. During the acclimation period 14 days prior to the test initiation the temperature ranged from 21.10 to 22.SOC, and the dissolved oxygen concentration was maintained above 8.0 mg/L. Fish were fed a commercial fish food (EnviroSysteme lot number TH03) once or twice daily before the test. Page 5 of 14 Table 1. Chemical characterization of a representative sample of natural well water used as dilution water for toxicity test Parameter pH Conductivity Hardness Organochlorine pesticides Organophosphorus pesticides. Polychlorinated biphenyle Unit of Measurement pH units Reporting Limit umhoo/cm mg/L as CaC03 ug/L O.S ug/L O.S ug/L O.S Measured Value 7.0 640 160 ND ND ND Notes: 1. ND - not detected at or above the reporting limit. 2. pH, hardness, and conductivity were measured in dilution water collected from the control test vessel prior to introduction of organisms and pesticide and PCB data is collected during routine biann-jalwater quality testing. Page 6 of 14 TOXICITY TESTING: A screening test with the test substance was conducted during January 13 to 17, 1992. - The test was conducted at 1, 10, 100, and 1,000 mg/L. After 96 hours there was Ot survival at 100 and 1,000 mg/L and 100% survival at all lower tested concentrations. A definitive toxicity test was attempted during February 20 to 24, 1992. Nominal concentrations of the test material were: 0 mg/L, 15 mg/L, 25 mg/L, 40 mg/L 60 mg/L, and 100 mg/L. The test was repeated because less than 50% survival occurred at all 5 concentrations after 96 hours. The definitive toxicity test was performed during February 27 to March 2, 1992 according to procedures of the OECD (1984). The test was conducted at a target temperature of 23 20C with five concentrations of test substance and a dilution water control. No stock solution was prepared and test material was added directly to dilution water contained in the test vessels without the use of a solvent. Nominal concentrations of the test material were: 0 mg/L (control), 2.5 mg/L, 4.0 mg/L, 6.0 mg/L, 10.0 mg/L, and 15.0 mg/L. Twenty fish were equally distributed among two replicates of each treatment. The test was performed in 20 liter glass aquaria (approximately 20 cm in width, 40 cm in length, and 25 cm in height) that contained 15 liters of test solution (water depth was approximately 18 cm). Test vessels were randomly arranged in an incubator during the 96 hour test (a random numbers table was used to select the location of each vessel). A 16 hour light and 8 hour dark phoioperiod was automatically maintained with cool-white fluorescent lights. Aeration was not employed to maintain dissolved oxygen concentrations above acceptable levels. Fish were not fed during the test. The number of surviving organisms and the occurrence of sublethal effects (loss of equilibrium, erratic swimming, loss of reflex, excitability, discoloration, or change in behavior) were determined visually and recorded initially and after 24, 48, 72, and 96 hours. Dead test organisms were removed when first observed. Dissolved oxygen (YSI Model 57 meter), pH (Beckman model pHl 12 meter), conductivity (Cole Parmer model number 1481-60), and temperature (ASTH mercury thermometer) were measured and recorded daily in each test chamber that contained live animals. STATISTICAL METHODS: Results of the toxicity test were interpreted by standard statistical techniques (Stephan, 1983). The 24, 48, 72, and 96 hour LCSO values were calculated by the binomial, moving average, or probit method, when warranted. All c&lculations were performed by the author using nominal concentrations of the test substance. Page 7 of 14 VI. RESULTS All test vessels containing FC-120 were initially clear and remained clear throughout the test. Biological and water quality data generated-.by the acute toxicity test are presented in Table 2 and Appendix A, respectively. one hundred percent survival occurred in the control exposure. Control fish had an average total length of 26.5 mm, and an average wet weight (blotted dry) of 0.14 g at the end of the test. Loading rate during the toxicity test was approximately 0.09 g/L. The 24, 48, 72, and 96 hour LC509 for fish exposed to FC-120 are presented -in- Table 3. The 96 hour LCSO value (and associated 95% confidence limits) is 4.8 mg/L FC-120 (4.0 - 5.7 mg/L). one of the eight surviving fish exposed to 15.0 mg/L at 48 hours and one of the three surviving fish exposed to 15.0 mg/L at 72 hours exhibited erratic swimming. The three surviving fish exposed to 10.0 mg/L at 96 hours exhibited a loss of equilibrium and erratic swimming. No other sublethal effects were observed in any test vessel during the 96 hour exposure. Page 8 of 14 Teble 2. Survival data from toxicity test Nominal Concentration (mg/L) rep. Number Alive Ohr 24hr 48hr 72hr 96hr Number Affected ohr 24hr 48hr 72hr 96hr 0 (control) 1 2 2.5 1 2 4.0 1 2 6.0 1 2 10.0 1 2 15.0 1 2 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 a 10 10 10 10 5 10 10 10 10 10 10 10 10 10 3 10 10 10 10 0 10 10 10 6 2 10 10 10 6 1 10 10 8 2 0 10 10 a 1 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 2 0 0 0 0 1 0 0 0 1 - 0 0 1 0 - Note: Al@l affected fish exhibited erratic swimming and affected fish at 96 hours also exhibited a lose of equilibrium. Page 9 of 14 Table 3. Median lethal concentrations (LCSOB) from toxicity test Exposure period 24 hours 48 hours 72 hours 96 hours LC50 95 percent confidence limit LC50 calculation method >15.0 mg/L >15.0 mg/L 11.0 mg/L 4.8 mg/L -9.7 - 12.5 mg/L 4.0 - 5.7 mg/L Probit Probit Page 10 of 14 VII. REFERENCES OECD. 1984. Guidelines Biotic Systems. April 4, 1984. for Testing of Chemicals. Section 2: Effects on Method 203, Fish Acute Toxicity Test. Adopted Stephan, C.E. 1983. Computer Program for the Calculation U.S. EPA. Duluth, MN. Personal Communication. of LC50 Values. Page 11 of 14 I Aipipendix A. WATER OUALITY T)ATA FROM TOXICITY TLPST I t I I I I I I i 1 4 1 Page 12 of 14 Table A.l..Conductivity, pH, temperatures and dissolved oxygen concentration measurel during toxicity test Nominal concentration (mg/L) Rep. Conductivity (umho/cm) 0 24 48 72 96 hr hr hr hr hr 0 (control) 1 2 2.5 1 2 4.0 1 2 6.0 1 2 10.0 1 2 15.0 1 2 640 640 650 650 670 640 660 670 660 690 650 670 670 660 660 650 670 670 660 670 660 670 670 660 670 660 670 670 660 670 660 660 670 670 670 660 660 670 660 670 660 670 670 670 660 660 670 670 670 660 670 670 670 670 670 670 670 670 670 670 pH 0 24 48 72 96 hr hr hr hr hr 7.0 7.8 7.3 8.0 8.1 7.0 7.8 7.3 8.0 8.2 7.0 7.9 7.3 8.0 8.2 -7.0 7.9 7.3 8.'0 8.2 7.0 7.9 7.3 7.9 8.2 7.0 7.9 7.3 7.9 8.1 7.0 7.9 7.3 7.9 8.1 7.0 7.9 7.3 7.9 8.1 7.0 7.9 7.3 7.9 8.0 7.0 7.9 7.3 7.9 8.1 7.0 7.9 7.3 7.8 8.0 7.0 7.9 7.3 7.8 8.0 Page 13 of 14 ab w Table A.I. Continued. Nominal concentration (mg/L) Rep. Temperature (OC) 0 24 48 72 96 hr hr hr hr hr 0 (control) 1 2 2.5 1 2 4.0 1 2 6.0 1 2 10.0 1 2 15.0 1 2 21.5 23.0 21.5 21.5 22.5 21.6 21.0 21.8 21.4 22.9 21.5 22.0 21.4 21.4 21.5 21.5 22.9 21.5 21.3 22.2 21.4 22.5 22.6 21.5 22.5 21.5 23.0 21.5 21.3 22.6 21.5 22.3 21.7 21.2 21.8 21.5 22.3 21.4 21.5 22.3 21.5 22.4 21.5 21.2 22.3 21.5 22.3 21.7*21.4 22.1 21.5 22.7 21.6 21.2 22.2 21.5 22.9 21.8 21.3 22.1 Dissolved Oxygen (mg/L) 0 24 48 72 96 hr hr hr hr hr S.S 8.5 7.7 7.4 6.8 8.5 8.5 7.7 7.3 7.5 8.6 7.9 7.6 7.4 7.0 8.7 8.0 7.5 7.3 7.0 8.6 7.5 7.6 7.2 6.7 8.6 7.6 7.4 7.0 6.0 8.6 7.5 7.5 7.1 6.4 8.6 7.5 7.3 7.1 5.6 8.6 7.6 7.3 6.9 5.8 8.6 7.5 7.4 6.8 6.2 8.6 7.6 7.2 6.8 5.7 8.6 7.6 7.3 7.0 6.0 Page 24 of 14