Document zQdEvQNJxRbVEXLNyV6Eg4zqm

AfLza.G _ 0274 ACUTE TOXICITY TO FISH (FATHEAD MINNOW) TEST SUBSTANCE_________________________________________ Identity: A mixture containing perfluorooctanesulfonate, which may also be referred to as PFOS, FC-95, or as a component of FC-780B. (1Octanesulfonic acid) (CAS # 2795-39-3). Remarks: The 3M production lot number was 2. The test sample is FC780B, identified by the laboratory as "3M Sample 1-3". Current information indicates it is a mixture of 0.58% PFOS, 14% diethylene glycol monobutyl ether, 76.7% water, 5.5% urea, 1.17% Hydroxy foamer, 1.15% sodium octyl sulfate, 0.8% polyoxyethylene monooctylphenyl ether, 0.05% sodium lauryl sulfate, and 0.05% benzotriazole. The following summary applies to a mixture with incompletely characterized concentrations of impurities. Data may not accurately reflect toxicity of the fluorochemical component of the test sample. METHOD:____________________________________________________ Method: Methods for Acute Toxicity Tests with Fish, Macroinvertebrates and Amphibians, EPA-660/3-75-009, 1975. Type: Acute flow-through GLP: No Year completed: 1980 Species: Pimephales promelas Supplier: Fattig Fish Hatchery, Brady, NE. Analytical monitoring: Dissolved oxygen, pH, ammonia, and temperature. Exposure period: 96-hours Statistical methods: LC50 values and 95% confidence intervals calculated using a computer program by Stephan, et al. 1978. Test fish age: Juveniles Length and weight: Average length = 27 mm Average weight = 0.34 g (wet) Loading: Not given. Pretreatment: None. Test conditions: Dilution water: Filtered well water Dilution water chemistry: Hardness: 255 mg/L as CaCC>3 Alkalinity: 368 mg/L as CaC03 pH: 7.8 Conductivity: 50 pmhos/cm Lighting: A daily photoperiod of 16 hours light and 8 hours dark was maintained throughout the testing period. 002569 Stock and test solution preparation: The stock solution was prepared in deionized water on a volume/volume basis. Flow-through rate: Approximately seven volume exchanges of water every 24 hours. Stability o f the test chemical solutions: Not noted Exposure vessels: Glass aquaria containing 30 L of test solution. Number of replicates: 1 Number of fish per replicate: 20 Number of concentrations: five plus a blank control Water chemistry during the study: pH range (0-96 hours): 7.9 - 8.2 (control exposure) 8.0 - 8.1 (1500 mg/L exposure) Temperature range (0-96 hours): 21 - 22 C (control exposure) 21 - 22 C (1500 mg/L exposure) Dissolved oxygen range (0-96 hours): 7.9 - 9.1 mg/L (control exposure) 7.9 - 8.6 mg/L (1500 mg/L exposure) Nominal concentrations: Blank control, 90, 200, 380, 770, and 1500 mg/L. Element values: 96-hour LC50 = >1500 mg/L 96-hour NOEC = 380 mg/L Element values based on nominal concentrations Remarks: Testing was conducted on the mixture as described in the Test Substance Remarks field. The values reported apply to that mixture and not the fluorochemical proportion alone. CONCLUSIONS____________________________________________ The test sample 96-hour LC50 for fathead minnow was determined to be >1500 mg/L. The 96-hour No Observed Effect Concentration (NOEC) was 380 mg/L. The 1500 mg/L concentration resulted in a 15% mortality. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY________________________________________________ Reliability: Klimisch ranking = 2. The study meets the criteria for quality testing. However, the sample purity was not properly characterized and the 002570 study lacks analytical confirmation of the amount of fluorochemical proportion in the solution. REFERENCES This study was conducted by Analytical Biochemistry Laboratories, Inc., Columbia, MO at the request of the 3M Company, St. Paul, MN, 1980. OTHER_______________________________________________ _ Last changed: 6/28/00 002571 ANALYTICAL BIO CHEMISTRY LABORATORIES, INC. P.O. Box 1097 Columbia, MO 65205 (314) 474-8579 Dynamic Acute Bioassay Report #26241 Submitted To: 3M Company Attn: Mr. Moe Elnabarawy Environmental Lab 935 Bush - Bldg. 2-3E St. Paul, Minnesota 55144 Dynamic 96-Hour Acute Toxicity of 3M Sample 1-3 to Eathead Minnows (Pimephales promelas) September 29, 1980 002572 best copy available Submitted By: Analytical Biochemistry Laboratories, Inc. 7200 East ABC Lane P. 0. Box 1097 Columbia, Missouri 65205 (314) 474-8579 ; Prepared By: Carl M. Thompsol Aquatic Supervisor ---- V- o /g o Date Alan D. Forbis Aquatic Biologist Paa unli BPoAuiidHrreoaa iui Biologist ' Date &Q Date Approved By: - Cl it James A. Ault Quaaity^lSssjLirance Officer Lyle Jo. Johnson Laboratory Manage' a? Date 7 -J o - a Date 002573 TABLE OF CONTENTS LIST OF TABLES.......................... SUMMARY.... ............................. INTRODUCTION............................ METHODS AND MATERIALS................... I. Test Fish......... ....... .......... II. Test System....... .......... III. Test Compound...... ............... . IV. Test Procedure-Biological....... . .. V. Test Procedure-Chemical and Physical RESULTS.................................. LITERATURE CITED..... ................... QUALITY ASSURANCE STATEMENT............. APPENDIX I RAW DATA Page No. ii .1 .2 .2 .2 .2 .3 .3 .3 .4 .8 .9 002574 LIST OF TABLES Table No. Page No. 1 The estimated LC5o values with their 95% confidence limits for the 96-hour fathead minnow toxicity study of 3M Sample 1-3......................................... ............ 1 and 4 2 Chemical characterics of well water at the Aquatic Toxi cology Laboratory of Analytical Biochemistry Laboratories, Inc. (All analyses performed by ABC Labs)..................... 5 3 Water quality measurements during the 96-hour toxicity study of 311 Sample 1-3 to fathead minnows (Pimephales promelas)..... ..................... ' . . . .... 6 4 Percent mortality of fathead minnows exposed to five con centration of 3M Sample 1-3 during a 96-hour toxicity study.......................................................... 7 ii 002575 SUMMARY A dynamic 96-hour toxicity study was conducted to determine the acute toxicity of 3M Sample 1-3 to fathead minnows (Pimephales promelas). The methods employed were those outlined by the Committee on Methods for Toxicity Tests with Aquatic Organisms (2). Water quality parameters of temperature, dissolved oxygen, pH and ammonia were meas ured throughout the test and were within acceptable limits. A flow-through proportional diluter system was used to maintain constant test concentrations. Nominal exposure concentrations were: 90, 200, 380, 770 and 1,500 mg/1. As a quality check, the fathead minnows were challenged with a reference compound, Antimycin A. The observed 96-:hour LC50 and 95% confidence limits (C.I.) were within the 95% confidence limits reported in the literature (5), indicating that the fish were in good condition. The observed mortality data for the study gave the following esti mated IC50 values. TABLE 1: The estimated LC5<> values with their 95% confidence limits for the 96-hour fathead minnow toxicity study of 3M Sample 1-3. Hours of Exposure 24 48 72 96 3M Sample 1-3 (mg/1) 95% Confidence LCrq Limits >1500 >1500 >1500 >1500 In addition to the above LC50 values, the results indicated a 96-hour, no observed effect concentration of 380 mg/1. 002576 INTRODUCTION 3M Company contracted the Aquatic Toxicology Division of Analytical Biochemistry Laboratories, Inc., to conduct a five-concentration plus control, 96-hour, flow-through bioassay to determine the toxicity of 3M Sample 1-3 to fathead minnows (Pimephales promelas). The primary objec tive of the study was to define the acute toxicity of 3M Sample 1-3. by determining the median lethal concentrations (LC50's) during the course of the test. The study was authorized by Mr. Moe Elnabarawy, 3M Company, in a protocol dated September 15, 1980 (Appendix I) and was conducted from September 25 to September 29, 1980. The term LC5o as used in this report is that concentration of 3M Sample 1-3 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 Examina tion of Water and Wastewater (1) and Methods of Acute Toxicity Tests with Fish, Macro invertebrates and Amphibians (2). The study was con ducted following the procedures outlined in ABC Protocol No. 8007 (Ap pendix I). I. Test Fish The fathead minnows used in the test were obtained from Fattig Fish Hatchery in Brady, Nebraska. 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 prophylactic and therapeutic disease treatments, is included in Appendix I. During the holding period, the fish received a standard commercial fish food (Rangen's) daily in an amount equiv alent 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 0.34 g and a mean standard length of 27 mm. Weight and length measurements were made on the control group of fish at termination of the test and are included in Appendix I. As a quality check, the fathead minnows were challenged with a reference compound, Antimycin A. The ob served 96 hour LC50 and 95% confidence limits were within the 95% confidence limits reported in the literature (5), indicating thajt the fish were in good condition. Results of this test are included in Appendix I. II. Test System A proportional diluter system described by Mount and Brungs (6), with the modifications of McAllister et. al. (7), was used for 2 002577 the intermittent introduction of 3M Sample 1-3 and diluent water into the test aquaria. Aerated well water (Table 2) was delivered to the glass aquaria at a rate of 150 ml/minute/aquarium, an amount which was sufficient to replace the 30 liter test volume at least 7 times in a 24 hour period. The test aquaria were immersed in a circulating water bath held at 22C (1) by submerged heating elements. III. Test Compound The 3M Sample 1-3 standard was received on September 10, 1980, in good condition. The sample upon receipt was observed to be an amber liquid and was stored at room temperature (22C). The stock solution was prepared in deionized water on a volume/volume basis and corrected for specific gravity to a weight/volume basis (mg/1). Standard weights and dilution volumes can be found in Appendix I . IV. Test Procedure-Biological The test was initiated on September 25, 1980, by random as signment of 20 fathead minnows to each of the 30 liter test aquaria after test solutions had been flowing through the aquaria for 8 hours. The fish were exposed to the following nominal concentra tions of 3M Sample 1-3: 90 mg/1, 200 mg/1, 380 mg/1, 770 mg/1 and 1,500 mg/1. The fish were observed for mortality and abnormal be havior initially and once every 24 hours for the 96-hour test period. Dead individuals were removed at each observation.- A computerized LCS0 program developed by Stephan et. al. (8) was used to calculate the LC50 values and their 95% confidence limits. 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 concentrations of 3M Sample 1-3 were determined by measuring predetermined dilutions of a stock Rhodamine B dye. A standard calibration curve was constructed to measure the absor bance (Coleman Spectrophotometer Model 55) of known dilutions of the dye that bracketed the absorbance value obtained from the proportional diluter. The dilution factor was determined by inter polation from the standard curve (Appendix I). The stock 3M Sample 1-3 concentration divided by the dilution factor determined the highest nominal concentration. The remaining concentrations were calculated by multiplying the highest concentration by the percent dilution (% difference) obtained from the calibration curve. For example: _ 631.5 g/1 (stock concentration) y 421 (dilution factor) % dilution for aquaria #4 , aquaria #4 380 mg/1 nominal concentration 3 002578 The nominal concentrations of 3M Sample 1-3 calculated in this maimer were: 90, 200, 380, 770 and 1,500 mg/1. The maximum test concentration was set at 1,500 mg/1, as requested by the study sponsor. RESULTS Table 1 presents the predicted LC50 values and 95% confidence intervals for 3M Sample 1-3. These values were obtained by employing a computerized LC5q program developed by Stephan et. al. (8). TABLE 1: The estimated LC50 values with their 95% confidence limits for the 96-hour fathead minnow toxicity study of 3M Sample 1-3. Hours of Exposure 24 48 72 96 3M Sample 1-2 (mg/1) 95% Confidence I<Cso Limits >1500 >1500 >1500 >1500 -------- -- -- a . In addition to the above LC50 values, the results indicated 96-hour, no observed effect concentration of 380 mg/1. Mortality data is presented in Table A. Water quality parameters of temperature, dissolved oxygen, pH and ammonia were measured in the control, low concentration and high concen tration throughout the test (Table 3). The dissolved oxygen concentra tions which stayed between 85 and 100 percent saturation were considered adequate for testing (2) and ammonia concentrations were below the toxic level (9). 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 Biochemistry Laboratories' Quality Assurance Unit. All original raw data was provided to 3M Company, with a copy retained at Analytical Biochemistry Laboratories. 4 002573 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 Zinc Measured organophosphorus pesticides Measured organochlorine pesticides plus PCB's Concentration 9.3 ppm 7.8 255 ppm 368 ppm 50 pmhos/cm <0.05 ppm 0.15 ppm 0.10 ppm <0.01 ppm <0.001 ppm <0.001 ppm 0.001 ppm <0.001 ppm <0.01 ppm 0.012 ppm 0.009 ppm <0.0001 ppm 0.0157 ppm <0.01 ppm a a aSee appendix for individual analyses. i-i o qa 002581 TABLE 3: Water quality measurements during the 96-hour toxicity study of 3M Sample 1-3 to fathead minnows (Pimephales promelas). Hours of Exposure 0 24 48 72 96 Temp. C 22 22 21 22 22 Control _______ D . 0 .b NHad m g /1 e s ! m g /1 8.5 8.2 0.56 9.1 8.1 1.6 8 .4 7.9 0.56 8,6 8.1 1.0 7.9 8.1 0.64 Low Concentration_____ Temp. C NHsd Eg ! mg /1 22 8.5 8 .0 1.0 22 9 .0 8.1 1.3 21 8 .3 8 .0 0.62 22 8.5 8.1 0.98 22 7.7 8 .0 0.58 3 High Concentration Temp. D . 0 .b C mg/1 em! NH3d m g /1 22 8 .4 8 .0 1.4 22 8 .6 8.1 1.6 21 8.5 8 .0 0.48 22 8 .3 8.0 0.86 22 7.9 8 .0 0.60 High concentration water quality was determined in Tank 6 (1500 mg/1 3M Sample 1-3) throughout the study. ^Dissolved oxygen concentrations - YSI Model 54 Dissolved Oxygen meter and probe. cpH - pH Probe (Coming Model 476022) used with a Corning Model 125 pH and mV meter. ^Ammonia concentrations - Ammonia Probe (Extech Model 8002-8) used With a Corning Model 125 pH and mV meter. 319VWAV AdOO 1S39 I TABLE 4: Percent mortality of fathead minnows exposed to five concentrations of 3M Sample 1-3 during a 96hour toxicity study. 3M Sample 1-3 Nominal concentration (mg/l) Control 90 200 380 770 1500 ____________ Hours of Exposure 24 48 72 000 00o 0 0 ; . ' # o ;00o 10 10 10 15 15 15 96 . 0 0 .0 0 10 15 BEST COPY AVAILABLE 7 003582 LITERATURE CITED (1) American Public Health Association. 1975. Standard Methods for the Examination of Water and Wastewater. 14th ed. Washington, D.C. 1193 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 Pro tection 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 Re search 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. Dpeartment of Interior, Investigations in Fish Control No. 26. 21 p. (6) Mount, D. I. and W. A. Brungs. 1967. A Simplified Dosing Appara tus for Fish Toxicological Studies. Water Res. 1:21-29. (7) McAllister, W. A., Jr., W. L. Mauck and F. L. Mayer, Jr. 1972. A Simplified Device for Metering Chemicals in Intermittent-Flow Bio assays. Trans. Amer. Fish. Soc. 101:555-557. (8) Stephan, C. E., K. A. Busch, R. Smith, J. Burke and R. W. Andrew. 1978. A computer program for calculating an LCS0. U. S. Environ mental Protection Agency, Duluth, Minnesota, pre-publication manu script, August, 1978. (9) National Academy of Sciences. 1972. Water Quality Criteria. 1972. U. S. Department of Commerce, PB-236 199. 592 p. (10) Food and Drug Administration. Regulations for Good Laboratory Practice. 21 CFR, Part 58. 8 002583 Quality Assurance Statement for final report #26241 entitled, "Dynamic 96-Hour Acute Toxicity of 3M Sample 1-3 to Fathead Minnows (Pimephales p r o m e l a s ) for Mr. Moe Elnabarawy, 3M Company, St. Paul, Minnesota. In accordance with ABC Laboratories intent that all studies con ducted 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 Sample 1-3 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 were in good health and should not bias the observed mortality in the study. An in-progress study inspection was conducted on September 26, 1980. A final inspection of all data and records on September 29, 1980, 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. mm?. (James A. Ault Quality Assurance Officer Date 9 002584