Document pzw6p5xvN6R225pdme2R03aw

ACUTE TOXICITY TO FISH (FATHEAD MINNOW) TEST SUBSTANCE Identity:A mixturecontainingperfluorcoctanesulfonatweh,ich may also be referredtoas PFOS, FC-95, or as a component ofFC-600. (1-Octanesulfoniaccid,1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-p,otassium salt,CAS # 2795-39-3) Remarks: The 3M productionlotnumber was Belgium 2367. The test sample isFC-600, referredtoby the laboratoryas "3M-1".Current informationindicatesitisa mixtureof 1.0% PFOS, 81.20% water,12.00% diethyleneglycolbutylether,1.00% sodium octylsulfate2,.00% propane Sultonefoamer, 1.00% sodium decylsulfate0,.85% xanthan gum, 0.1% N-(3-chloroallyhle)xaminium chloride0,.80% starch,and 0.05 % tolyltriazole. The following summaty applies toa mixture with incompletely characterized concentration of impurities.Data may not accurately reflectthe toxicityof the fluorochemical component of the test sample. METHOD: Method: EPA-660/3-75-009 and Std. Methods (APHA), 1980,15 th Ed. Test type: Acute flow-through GLP: No Year Completed: 1983 Species: :Pimephalespromelas Supplier:In-houseculturemsaintainedatABC LaboratoriesI,nc. Columbia, MO. Analyticalmonitoring: Temperature, pH, DO, and ammonia Exposure period:96-hours Statisticamlethods: LC50 valuesand 95% confidenceintervals calculatedusingbinomial,the moving average,orthe probittests. Test fishage: Juveniles. Average Length and weight: Length = 30 mm Weight = 0.52 g Loading: 0.35 g/L Pretreatment: None. Test conditions: Dilutionwater: Wellwater. Dilutionwater chemistry: pH: 7.8 Hardness (as CaC03): 255 mg/L Alkalinity(as CaC03): 368 mg/L Conductivity: 50 pmhos/cm LightingA:dailpyhotoperiofd16 hourslighatnd8 hoursdarkwas maintained. Stock and testsolutionpreparation: A 381,000 mg/L stocksolution was prepared by dilutin3g81 grams oftestsample to 1-Lwithdeionized water. Test solutionswere prepared by a dilutedreliverinogne nominal concentrationofthetestsample and controlwater. Flow-through rate:At least5 volume replacementsina 24 hour period. Stabilityof the testchemical solution:Not noted. Exposure vessels:Aquariawitha maintainedtestsolutionvolume of 15 liters. Number of replicates:two Number of fish per replicate:10 Number of concentrations: 1 plusa blank control,induplicate Water chemistry during the study: pH range (0-96 hours): 8.0 - 8.1 (controlexposure) 7.8- 8.2 (1,000mg/L exposure) Temperature range (0-96hours): 21OC (forallaquaria) Dissolved oxygen range (0-96hours): 7.6- 8.8 mg/L (controelxposure) 2.7- 9.2 mg/L (1,000mg/L exposure) Remarks: The dissolvedoxygen concentrationfsorthe 1,000mg/L exposure aquariafeltlo5.7 and 2.7 mg/L after96 hours. Since fathead minnows are generallymore tolerantoflow DO concentrationsi,twas notconsideredtobe a significanftactorin the resultsofthetest. RESULTS Nominal concentrations: Bk controland 1,000 mg/L Percent mortality(based on 20 fishper concentration) Nominal Concentration (mg/L) Control 1,000 24-hours 0 0 Percentag mortality 48-hours 72-hours 0 0 0 5 96-hours 0 35 Element value: 24-hour LC50: 48-hour LC50: 72-hour LC50: 96-hourLC50: >1,000 mg/L >1,000 mg/L >1,000 mg/L >1,000 mg/L Element values based on nominal concentrations. Remarks:Testinwgas conducteodn themixturaes describeidntheTest Substance Remarks field.The value reportedappliestothatmixtureand notthe fluorochemicalproportionalone. Behavior observationsduringthetestindicatedthatmortalitwyas preceded by surfacingand lossofequilibrium. A referencetestwas conducted usingAntimycinA. Test resultswere withinpublishedliteratuvraelues. CONCLUSIONS The FC-600 96-hour LC50 forPimephales promelas was determined to be >1,000 mg/L. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking2. The studymeets the criterifaorquality testinghowever,the sample puritywas notpropedy characterizedand it lacksanalyticaclonfirmatioonfthe amount offluorochemicalproportionin solution.AdditionallyD,O concentrationfsellsignificantbleylow acceptablelevelsatthe 1,000 mg/L exposure. REFERENCES This studywas conducted by ABC LaboratoriesI,nc.ofColumbia, MO at the requestofthe 3M Company, 3M Lab Request number 8875 and 9063, 1983. OTHER Last changed: 6/28/00 o ANALYTICAL 810-CHEMISTRY LABORATORIES, INC. P.O. Box 1097 o Columbia, MO 65205 o (314)474-W79 IA Flow-Through Acute Toxicity Report #30175 Submitted To: 3M Compaay Attn: M. T. Elnabaravy Environmental Laboratory 935 Bush - Building 2-3E St. Paul, Minnesota 55144 Dynamic Acute Toxicity of 3M-I to Fathead Minnows (Pimephales promelas) March 29, 1983 Submitted By: Analytical Bio-Chemistry Laboratories, Inc. 7200 East ABC Lane P. 0. Box 1097 Columbia, Missouri 65205 Prepared By: Alan D. Forbis Aquatic Biologist David L. Kintner Biologist 31?d Date Date Approved By: Carl M. Thompsonk,/ Aquatic Supervisor 04-pl@ut * Jae elsjstA A .A-ult Qu lity Assurance Supervisor /gsl Lysleeon@. JJoohnson i Laboratory Managbr Date 31FO Date 3-30-8-3 Date 30175-ii TABLE OF CONTENTS Page No. LIST OF TABLES . . . . . . . . . . . . . . . . . . . . . . . . . 30175-iv SUMMARY I . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30175-1 INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . . . . . 30175-2 METHODS AND MATERIALS . . . . . . . . . . . . . . . . . . . . . 30175-2 I. Test Fish . . . . . . . . . . . . . . . . . . . . . . 30175-2 II. Test System . . . . . . ... . . . . . . . . . . . . . 30175-3 III. Test Compound . . . . . . . . . . . . . . . . . . . . 30175-3 IV. Test Procedure - Biological . . . . . . . . . . . . . 30175-3 V. Test Procedure - Chemical and Physical . . . . . . . . 30175-3 VI. Statistical Analysis. . . . . . . . . . . . . . . . . 30175-3 RESULTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30175-4 LITERATURE CITED . . . . . . . . . . . . . . . . . . . . . . . . 3,0175-9 QUALITY ASSURANCE STATEMENT . . . . . . . . . . . . . . . . . . 30175-10 APPENDIX - RAW DATA . . . . . . . . . . . . . . . . . . . . . . 30175-11 30175-iii Table No. 1 2 3 4 LIST OF TABLES Page No. The estimated LCSO values with their 95% confidence limits for the 96-hour fathead minnow toxicity study of 3M-I. . . . . . . . . . . . . . . 30175-5 Chemical characteristics of well water at ABC's Aquatic Bioassay Laboratory . . . . . . . . . . . . 30175-6 Water quality measurements during the 96-hour toxicity study of 3M-I to fathead minnows . . . . . 30175-7 Percent mortality of fathead minnows exposed to five concentrations of 3M-I during a 96-hour toxicity study. . . . . . . . . . . . . . . . . . . 30175-8 30175-iv SUMMARY A dynamic 96-hour toxicity study was conducted to determine the _acute toxicity of 3M-I to fathead minnows (Pimephales promelas). The methods employed were those outlined by the Committee on Methods for Toxicity Tests with Aquatic Organisms. Water quality parameters of temperature, dissolved oxygen, pH and ammonia were measured throughout the test and were within acceptable limits. A flow-through proportional diluter system was used to maintain constant test concentrations. Nominal exposure concentrations of 3M-I were duplicate 1,000 mg/l and duplicate controls. As a quality check, the fathead minnows were challenged with a reference compound, Antimycin A. The observed 96-hour LCSO. and 95% confidence limits (C.I.) were withi'n the 95% confidence limits reported in the literature, indicating that the fish were in good condition. The observed mortality data for the study indicated that the LCso would be >1,000 mg/l. 30175-1 IWMODUCTION 3H Company contracted the Aquatic Toxicology Division of Analytical Bio-Chemistry Laboratories, Inc., to conduct a duplicate 1,000 mg/l concentration plus duplicate control, 96-hour, flow-through bioassay to estimate the toxicity of 3M-I to fathead minnows (Pimephales promelas). The primary objective of the study was to define the acute toxicity of 3M-I by estimating the median lethal concentrations (LC50's) during the course of the test. The study was authorized by M. T. Elnabarawy, 3M Company, in a protocol dated February 21, 1983 (Appendix) and was conducted from March 10 to March 14, 1983. The term LCSO as used in this report is that concentration of 3M-I causing 50% mortality after a specified exposure period. This report presents the results of the toxicity study and describes the technical approach used to meet the project objectives. METHODS AND MATERIALS The biological methods used for the 96-hour flow-through toxicity test are basically those described in Standard Methods for the Examination of Water and Wastewater (1) and Methods of Acute Toxicity Tests with Fish, Macroinvertebrates and Amphibians (2). The study was conducted following the procedures outlined in ABC Protocol No. 8007 (Appendix). I. Test Fish The fathead minnows used in the test were obtained from ABC Laboratories in-house culture. The fish were identified to species using the taxonomic keys developed by Eddy (3). All test fish were held in culture tanks on a 16-hour daylight photoperiod and observed for at least 14 days prior to testing. Fish culture techniques were basically those described by Brauhn et al. (4). A daily record of fish observations during the holding period, along with any prophylactic and therapeutic disease treatments, is included in the Appendix. During the holding period, the fish received a standard commercial fish food (Rangen's daily in an amount equivalent to 3 percent of their body weight. The test fish were not fed during the acclimation and test periods. The fathead minnows used for this experiment had a mean weight of 0.52 g and a mean standard length of 30 mm. Weight and length measurements were made on the control fish at test termination and are included in the Appendix. As a quality check, the fathead minnows were challenged with a reference compound, Antimycin A. The observed 96 hour LC.50 and 95% confidence limits were within the 95% confidence limits reported in the literature (5), indicating that the fish were in good condition. Results of this test are included in the Appendix. 30175-2 II. Test System A proportional diluter system described by Mount and Brungs (6), with a Hamilton dual syringe dispensor system was used for the intermittent introduction of 3M-I and diluent water into the test aquaria. Aerated well water (Table 2) was delivered to the glass aquaria at a rate of 52 ml/minute/ aquarium, an amount which was sufficient to replace the 15 liter test volume at least 5 times in a 24 hour period. The test aquaria were immersed in a water bath held at 22*C (tl*) by submerged heating elements. III. Test Compound The 3M-I standard was received on February 24, .1983, in good condition. The sample upon receipt was observed to be a viscous amber liquid and was stored at room temperature. The stock solution was prepared in deionized water on a weight/ volume basis. Standard weights and dilution volumes can be found in the Appendix. IV. Test Procedure - Biological The test was initiated on March 10, 1983, by random assignment of 10 fathead minnows to each of the 15 liter test aquaria after test solutions had been flowing through the aquaria for 24 hours. The fish were exposed to the following nominal concentrations of 3M-1: duplicate 1,000 mg/l and duplicate controls. The fish were observed for mortality and abnormal behavior initially and once every 24 hours for the 96-hour test period. Dead individuals were removed at each observation. V. Test Procedure - Chemical and Physical Water quality parameters of temperature, dissolved oxygen, pH and ammonia were measured throughout the test and were within acceptable limits (Table 3). The nominal concentration of 1,000 mg/l was calculated by dividing the working stock concentration by the diluter dilution factor. The dilution factor was determined by dividing the toxicant mixing box volume by volume injected by toxicant delivery system. Calculations are in the Appendix. VI. Statistical Analysis Statistical analysis of the concentrationvs. effect data (generally mortality) was obtained by employing a computerized LCSO program developed by Stephan et al. (7). This program calculated the LCSO statistic and its 95 percent confidence limits using the binomial, the moving average, and the probit tests. Three different methods of analyzing the data were used since no one method of analysis is appropriate for all 30175-3 possible sets of data that may be obtained. The method of calculation selected for use in this report was that which gave the narrowest confidence limits for the LCSO (7, 8). RESULTS The estimated LCSO for 3M-I would be >1,000 mg/l after 96 hours. Table 1 presents the LC50 values and 95% confidence intervals for Antimycin A. The 96-hour LCSO for Antimycin A to fathead minnows was 5.6XIO 5 mg/l. Mortality data is presented in Table 4 and was 35% after 96 hours. Behavior observations throughout the test indicate3-that mortality was preceded by surfacing and loss of equilibrium. Water quality parameters of temperature, -dissolved oxygen, pH and ammonia were measured in the control and 1,000 mg/l concentration every 24 hours (Table 3). The dissolved oxygen concentrations in the control stayed between 86 and 100 percent saturation at 22*C and was considered adequate for testing (2). The dissolved oxygen concentrations for the exposure aquaria were lower than the control (5.7 and 2.7 mg/1) after 96 hours. This appeared to be compound related but would not have adversely effected the Lts. Ammon I -o-n-s-v-e'@ 70-X,-C-IVvel-- (9T.- The pH values were consistent with the control throughout the study. The study was conducted following the intent of the Good Laboratory Practice Regulations (10) and the final report was reviewed by Analytical Bio-Chemistry Laboratories' Quality Assurance Uait. All original raw data was provided to 3M Company, with a copy retained at Analytical Bio-Chemistry Laboratories. 30175-4 TABLE 1: The estimated LCSO values with their 95% confidence limits for the 96-hour fathead minnow toxicity study of Antimycin A. Hours of Exposure 24 48 72 96 3M-I (mg/1) 95% LCSO Confidence Limits 6.5XIO-5 6.2XlO-5 5.6XlO-5 4.ZXIO-5 - 1.4XIO-4 4.2XIO-5 - 1.4XIO-4 4.2XIO-S - 7.5XlO-s 30175-5 TABLE 2: Chemical characteristics of well water at ABC's Aquatic Bioassay Laboratory. Parameter Dissolved Oxygen pH Hardness (CaC03) Alkalinity (CaC03) Conductivity Total Ammonia (NH3) N03-N Ortho-Phosphate Aluminum Arsenic Cadmium Chromium Cobalt Copper Iron Lead Mercury Nickel Silver Zinc Measured organophosphorus pesticides Measured organochlorine pesticides plus PCB's Concentration 9.2 ppm 7.8 255 ppm 368 ppm 50 pmhos/cm <0.10 ppm 0.15 ppm 0.10 ppm <0.007 ppm <0.001 ppm <0.001 ppm <0.002 ppm <0.002 ppm <0.002 ppm <0.004 ppm <0.02 ppm <0.0001 ppm <0.004 ppm <0.002 ppm <0.001 ppm a a a See appendix for individual analyses. 30175-6 TABLE 3: Water quality measurements during the 96-hour toxicity study of 3M-I to fi Hours of Exposure 0 24 48 72 96 Temp. oc 21, 21 21, 21 21, 21 21, 21 21, 21 b D.O. mg/l Controla c pH 8.8, 8.8 8.1, 8.1 8.4, 8.4 8.1, 8.0 7.6, 7.6 8.1, 8.1 8.3, 8:3 8.1, 8.1 7.9, 7.8 8.1, 8.1 d NH3 mg/l 0.18, 0.17 0.261 0.31 0.35, 0.30 0.22, 0.30 0.18) 0.26 Temp. oc 21, 21 21, 21 21, 21 21, 21 211 21 1,000 mg/l b D.O. mg/l pH 8.9, 9.2 8.1, 8.6, 8.5 8.1, 5.1, 4.3 8.0, 5.8, 3.9 8.0) 5.6, 2.7 7.9, aDuplicate aquaria. bDissolved oxygen concentrations - YSI Model 54 Dissolved Oxygen meter and probe. cpH - pH Probe (Corning Model 476182) used with a Corning Model 125 pH and mV me@er. dAmmonia concentrations - Ammonia Probe (Orion Model 95-10) used with a Corning Model 4 Ln TABLE 4: Percent mortality of fathead minnows exposed to 1,000 mg/l of 3M-I during a 96-hour toxicity study. 3M-I Nominal Hours of Exposure Concentration (mg/1) 24 48 72 96 Control a 0 0 0 0 Control b 0 0 0 0 1,000 a 0 0 0 10 1,000 b 0 0 10 60 30175-8 LITERATURE CITED (1) American Public Health Association. 1980. Standard Methods for the Examination of Water and Wastewater. 15th ed. Washington, DC. 1134 p. (2) Committee on Methods for Toxicity Tests with Aquatic Organisms (C. E. Stephan, Chairman). 1975. Methods for Acute Toxicity Tests with Fish, Macroinvertebrates and Amphibians. Environmental Protection Agency, Ecological Research Series EPA-660/3-75-009, April, 1975. 61 p. (3) Eddy, Samuel. 1969. The Freshwater Fishes. 2nd ed. W. C. Brown Company, Dubuque, IA. 286 p. (4) Brauhn, J. L. and R. A. Schoettger. 1975. Acquisition and Culture of Research Fish: Rainbow Trout, Fathead Minnows, Channel Catfish and Bluegills. Environmental Protection Agency, Ecological Research Series EPA-660/3-75-011, May, 1975. 45 p. (5) Berger, B. L. , R. E. Lennon and J. W. Hogan. 1969. Laboratory Studies on Antimycin A as a fish toxicant. U. S. Department of Interior, Investigations in Fish Control No. 26. 21 p. (6) Mount, D. I. and W. A. Brungs. 1967. A Simplified Dosing Apparatus for Fish Toxicological Studies. Water Res. 1:21-29. (7) Stephan, C. 1977. Methods for calculating an LC50, p. 65-84. In F. L. Mayer and J. L. Hamelick (eds.). Aquatic Toxicology a'nd Hazard Evaluation. ASTM Special Technical Publication 634. ASTM. Philadelphia. (8) Stephan, C. E., K. A. Busch, R. Smith, J. Burke and R. W. Andrew. 1978. A computer program for calculating an LCSO. U. S. Environmental Protection Agency, Duluth, Minnesota, pre-publication manuscript, August, 1978. (9) National Academy of Sciences. 1971. Water Quality Criteria, 1972. U. S. Department of Commerce, PB-236 199. 592 p. (10)Food and Drug Administration. Regulations for Good Laboratory Practice in Non-Clinical Animal Studies. 21 CFR Part 58. 30175-9 Quality Assurance Statement for final report #30175 entitled, "Dynamic Acute Toxicity of 3M-I to Fathead Minnows. (Pimephales promelas)," for Dr. M. T. Elnabarawy, 3M Company, St. Paul, Minnesota. In accordance with ABC Laboratories intent that all studies conducted at our facilities are designed and function in conformance with good laboratory practice regulations and the protocols for individual laboratory studies, an inspection of the final report for 3M-I was conducted and found to be in an acceptable form by a member of our Quality Assurance Unit. An inspection of the daily mortality rate of the test organisms prior to the initiation of the study indicated they wdre in good health and should not bias the observed mortality in the study. A final inspection of all data and records on March 25, 1983 indicated that the report submitted to you is an accurate reflection of the study as it was conducted by ABC Laboratories. Should you have any questions relating to the information provided in this statement or the function of our Quality Assurance Unit, please contact me at your convenience. I 4JJam@,sFstAA.@Ault Date Qufity Assssuuulrance Supervisor 30175-10