Document 44qVqVad8vyqp2XxXqmpdjboV

M 3 2 6 _ 0X37 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-600. (1-Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-, potassium salt, CAS # 2795-39-3) Remarks: The 3M production lot number was Belgium 2367. The test sample is FC-600, referred to by the laboratory as "3M-I". Current information indicates it is a mixture of 1.0% PFOS, 81.20% water, 12.00% diethylene glycol butyl ether, 1 .00% sodium octyl sulfate, 2 .00% propane Sultone foamer, 1.00% sodium decyl sulfate, 0.85% xanthan gum, 0.1% N-(3-chloroallyl) hexaminium chloride, 0.80% starch, and 0.05 % tolyltriazole. The following summary applies to a mixture with incompletely characterized concentration of impurities. Data may not accurately reflect the toxicity of the fluorochemical component of the test sample. METHOD: Method: EPA-660/3-75-009 and Std. Methods (APHA), 1980, 15th Ed. Test type: Acute flow-through GLP: No Year Completed: 1983 Species: :Pimephales promelas Supplier: In-house cultures maintained at ABC Laboratories, Inc. Columbia, MO. Analytical monitoring: Temperature, pH, DO, and ammonia Exposure period: 96-hours Statistical methods: LC50values and 95% confidence intervals calculated using binomial, the moving average, or the probit tests. Test fish age: Juveniles. Average Length and weight: Length = 30 mm Weight = 0.52 g Loading: 0.35 g/L Pretreatment: None. Test conditions: Dilution water: Well water. Dilution water chemistry: pH: 7.8 Hardness (as CaCC>3): 255 mg/L Alkalinity (as CaCC>3): 368 mg/L Conductivity: 50 pmhos/cm 002040 Lighting: A daily photoperiod of 16 hours light and 8 hours dark was maintained. Stock and test solution preparation: A 381,000 mg/L stock solution was prepared by diluting 381 grams of test sample to 1-L with deionized water. Test solutions were prepared by a diluter delivering one nominal concentration of the test sample and control water. Flow-through rate: At least 5 volume replacements in a 24 hour period. Stability of the test chemical solution: Not noted. Exposure vessels: Aquaria with a maintained test solution volume of 15 liters. Number of replicates: two Number o ffis h per replicate: 10 Number of concentrations: 1 plus a blank control, in duplicate Water chemistry during the study: pH range (0-96 hours): 8 .0 - 8.1 (control exposure) 7.8 - 8.2 (1,000 mg/L exposure) Temperature range (0-96 hours): 21 C (for all aquaria) Dissolved oxygen range (0-96 hours): 7.6 - 8.8 mg/L (control exposure) 2.7 - 9.2 mg/L (1,000 mg/L exposure) Remarks: The dissolved oxygen concentrations for the 1,000 mg/L exposure aquaria fell to 5.7 and 2.7 mg/L after 96 hours. Since fathead minnows are generally more tolerant of low DO concentrations, it was not considered to be a significant factor in the results of the test. RESULTS______ :_____________________________________________ Nominal concentrations: Bk control and 1,000 mg/L Percent mortality (based on 20 fish per concentration) N om inal C oncentration (m g/L) C o n tro l 1,000 24-hours 0 0 Percentage m ortality 48-hours 72-hours 00 05 Element value: 24-hour LC5o 48-hour LC50 72-hour LC50 96-hour LC50 >1,000 mg/L >1,000 mg/L >1,000 mg/L >1,000 mg/L Element values based on nominal concentrations. 96-hours 0 35 002041 Remarks: Testing was conducted on the mixture as described in the Test Substance Remarks field. The value reported applies to that mixture and not the fluorochemical proportion alone. Behavior observations during the test indicated that mortality was preceded by surfacing and loss of equilibrium. A reference test was conducted using Antimycin A. Test results were within published literature values. CONCLUSIONS_______________________________________________ The FC-600 96-hour LC50for Pimephales promelas was determined to be >1,000 mg/L. 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 testingHowever, the sample purity was not properly characterized and it lacks analytical confirmation of the amount of fluorochemical proportion in solution. Additionally, DO concentrations fell significantly below acceptable levels at the 1,000 mg/L exposure. REFERENCES___________________________________________ _ This study was conducted by ABC Laboratories, Inc. of Columbia, MO at the request of the 3M Company, 3M Lab Request number 8875 and 9063, 1983. OTHER_____________________________________________________ Last changed: 6/28/00 002042 FC-Coo ANALYTICAL BIO-CHEMISTRY LABORATORIES, INC. P.O. Box 1097 Columbia, MO 65205 (314) 474-8579 LR SV?* LR ' Flow-Through Acute Toxicity Report #30175 Submitted T o : 3M Company Attn: M. T. Elnabarawy 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 002043 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 fc '-L .'fw u s ' David L. Kintner Biologist z /u h i Date 3Ae/r3 Date Approved By: Carl M. Thompsons Aquatic Supervisor Atfrn es A. Ault lity Assurance Supervisor /gsl Lyle. Johnson Laboratory Manager Date 3- 30-81 Date 301750 2 0 4 4 TABLE OF CONTENTS Page No. LIST OF TABLES......................................... . . . . 30175-iv SUMMARY ............................................... . . . . 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....................................... . . . . 30175-9 QUALITY ASSURANCE STATEMENT ........................... . . . . 30175-10 APPENDIX - RAW DATA ................................... . . . . 30175-11 30175-iip LIST OF TABLES Table No. 1 Page N o . The estimated LC50 values with their 95% confi dence limits for the 96-hour fathead minnow toxicity study of 3M-1............................. 30175-5 2 Chemical characteristics of well water at ABC's Aquatic Bioassay Laboratory ....................... 30175-6 3 Water quality measurements during the 96-hour toxicity study of 3M-I to fathead minnows ........ 30175-7 4 Percent mortality of fathead minnows exposed to five concentrations of 3M-I during a 96-hour toxicity study..................................... 30175-8 30175-iv 002046 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/ 1 and duplicate controls. As a quality check, the fathead minnows were challenged with a reference compound, Antimycin A. The observed 96-hour LC5 0 .and 95% confidence limits (C.I.) were within 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 LC50 would be >1,000 mg/1 . 30175-1 002047 INTRODUCTION 3M Company contracted the Aquatic Toxicology Division of Analytical Bio-Chemistry Laboratories, Inc., to conduct a duplicate 1,000 mg/1 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 (LCso'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 LC50 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 Examina tion of Water and Wastewater (1) and Methods of Acute Toxicity Tests with Fish, Macroinvertebrates and Amphibians (2). The study was con ducted 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 de scribed by Brauhn et al. (4). A daily record of fish observa tions during the holding period, along with any prophylactic and therapeutic disease treatments, is included in the Appendix. During the holding period, gthe fish received a standard commercial fish food (Rangen's j 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 LC50 and 95% confidence limits were within the 95% confidence limits reported in the literature (5), indicating that the fish were in good condi tion. Results of this test are included in the Appendix. 30175-2 002048 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 22C (1) 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 lie 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-I: duplicate 1,000 mg/1 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/1 was calculated by dividing the working stock concentration by the diluter dilu tion 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 concentration vs. effect data (generally mortality) was obtained by employing a computerized LC50 program developed by Stephan et al. (7). This program calculated the LC50 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 002049 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 LC50 (7, 8). RESULTS The estimated LCS0 for 3M-I would be >1,000 mg/1 after 96 hours. Table 1 presents the LC50 values and 95% confidence intervals for Antimycin A. The 96-hour LCS0 for Antimycin A to fathead minnows was 5.6X10" 5 mg/1. Mortality data is presented in Table 4 and was 35% after 96 hours. Behavior observations throughout the test indicated 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/ 1 concentration every 24 hours (Table 3). The dissolved oxygen concentrations in the control stayed between 86 and 100 percent saturation at 22C 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 results-! Ammonia concentraiions"~tfeirH' below the toxic'^rSVeT^' (9)7 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 Unit. All original raw data was provided to 3M Company, with a copy retained at Analytical Bio-Chemistry Laboratories. 30175-4 002050 TABLE 1: The estimated LC50 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)____________ L C Sq 95% Confidence Limits ---- 6.5X10"5 4.2X10" 5 - 1.4X10"4 6.2X10" 5 4.2X10"5 - 1.4X10"4 5.6X10"5 4.2X10" 5 - 7.5X10" 5 30175-5 002051 TABLE 2: Chemical characteristics of well water at ABC's Aquatic Bioassay Laboratory. _____ Parameter_____ Dissolved Oxygen PH Hardness (CaCOs) Alkalinity (CaCOs) Conductivity Total Ammonia (NH3) N O 3 -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 . 1 0 ppm 0.15 ppm 0 . 1 0 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 002052 002053 TABLE 3: Water quality measurements during the 96-hour toxicity study of 3M-T to fathead minnows. Hours of Exposure 0 24 48 72 96 Temp. C 2 1 , 21 2 1 , 21 2 1 , 21 2 1 , 21 2 1 , 21 Control3 D.0.b mg/ 1 pH'c 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 NH3d mg/ 1 0.18, 0.17 0.26, 0.31 0.35, 0.30 0.22, 0.30 0.18, 0.26 Temp. C 2 1 , 21 2 1 , 21 2 1 , 21 2 1 , 21 2 1 , 21 1,000 mg/1 3 D.0.b mg/ 1 PHC 8.9, 9.2 8 .6 , 8.5 8 .1 , 8.2 8 .1 , 8.0 5.1, 4.3 8.0, 7.9 5.8, 3.9 8.0, 7.8 5 .6 , 2.7 7.9, 7.8 NH3d mg/ 1 0.19, 0.18 0.41, 0.36 0.26, 0.20 0.18, 0.20 0.31, 0.36 0 Duplicate aquaria. ^Dissolved oxygen concentrations - YSI Model 54 Dissolved Oxygen meter and probe. Q pH - pH Probe (Corning Model 476182) used with a Corning Model 125 pH and mV meter. ^Ammonia concentrations - Ammonia Probe (Orion Model 95-10) used with a Corning Model 125 pH and mV meter. 30175- TABLE 4: Percent mortality of fathead minnows exposed to 1,000 mg/1 of 3M-I during a 96-hour toxicity study. 3M-I Nominal Concentration (mg/1) Control a Control b 1,000 1,000 a b Hours of Exposure 24 48 72 00 0 00 0 00 0 0 0 10 96 0 0 10 60 30175-8 002054 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 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. 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 LC5o, p. 65-84. In F. L. Mayer and J. L. Hamelick (eds.). Aquatic Toxicology and 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 LC5o- U. S. Environ mental Protection Agency, Duluth, Minnesota, pre-publication manu script, 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 002p55 Quality Assurance Statement for final report #30175 entitled, "Dynamic Acute Toxicity of 3M-I to Fathead Minnows (Pimephales promelas) , 11 for Dr. M. T. 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-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 were 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. 30175-10 002056