Document YDb3Gj1EzEgqDVBG9g39eBqz8

4R - 0 2 7 & ACUTE TO XIC ITY TO FISH (BLUEGILL SUNFISH) TEST SUBSTANCE________________________________________________ Identity: A mixture containing perfluorooctanesulfonate, which may also be referred to as PFOS, FC-95, or as a component of FC-783. (1Octanesulfonic acid) (CAS # 2795-39-3). Remarks: The 3M production lot was not noted. The test sample is FC783 (cc784-1). Current information indicates it is a mixture of 1.0% PFOS, 2.0% Sultone foamer, 2.3% sodium octyl sulfate, 1.6% polyoxyethylene monooctylphenol ether, 0.1% sodium lauryl sulfate, 25.0% diethylene glycol monobutyl ether, 0.05% tolytriazole, and 67.95% water. The following summary applies to a mixture with incompletely characterized concentrations o f impurities. Data may not accurately reflect the toxicity o f the fluorochemical component o f the test sample. METHOD:________________________________________________________ Method: Standard Methods, 1975, and USEPA Methods for Acute Toxicity Tests with Fish, Macroinvertebrates and Amphibians, 1975. Type: Acute static GLP: No Year completed: 1979 Species: Lepomis macrochirus Supplier: Osage Catfisheries, Inc., Osage Beach, MO. Analytical monitoring: Dissolved oxygen, pH, ammonia, and temperature. Exposure period: 96-hours Statistical methods: LC50 values calculated using the statistical methods of Litchfield and Wilcoxon, 1949. Test fish age: Juveniles Length and weight: Average length = 28.6 mm Average weight = 0.60 g (wet) Loading: 0.2 g/L. Pretreatment: None. Test conditions: Dilution water: W ell water Dilution water chemistry: Hardness: 255 mg/L as C aC 03 Alkalinity: 368 mg/L as C aC 03 pH: 8.2 Lighting: 16 hour daylight photoperiod. Stock and test solution preparation: The test material was added directly to the dilution water on a weight/volume basis. Concentrations dosing rate: Once 002594 S tability o f the test chem ical solutions: Not noted Exposure vessels: 5 gallon glass vessels containing 15 L of test solution. Num ber o f replicates: 2 Num ber o f fish per replicate: 5 Num ber o f concentrations: five plus a blank control W ater chem istry during the study: pH range (0-96 hours): 8.2 - 8.3 (control exposure) 8.2 - 8.3 (1000 mg/L exposure) Tem perature range (0-96 hours): 22-22 C Total am m onia range (0-96 hours): 0.68 - 1 .1 mg/L (control exposure) 0.60 - 1 .1 mg/L (1000 mg/L exposure) Dissolved oxygen range (0-96 hours): 5.0 - 8.7 mg/L (control exposure) 3.8 - 8.7 mg/L (1000 mg/L exposure) RESULTS Nom inal concentrations: Bk control, 100,180, 320, 560, and 1000 mg/L. Elem ent values: 96-hour LC50 = >1000 mg/L Element values based on nominal concentrations Rem arks: Testing was conducted on a mixture as described in the Test Substance Remarks field. The values reported apply to that mixture and not the fluorochemical proportion alone. CONCLUSIONS ________________________________________________ The FC-783 96-hour LC50 for bluegill sunfish was determined to be >1000 mg/L, the highest concentration tested. A 20% mortality was observed. Subm itter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY___________________________________________________ Reliability: Klimisch ranking 2. Testing meets the criteria for quality testing. However, sample purity was not properly characterized and the study lacks analytical confirmation of the amount of fluorochemical proportion in the solution. REFERENCES____________________________________________________ 002595 This study was conducted by Analytical Biochemistry Laboratories, Inc., Columbia, MO at the request of the 3M Company, 3M Lab Request number 4896S, 1979. O T H E R ____________ Last changed: 6/26/00 002596 ANALYTICAL BIO CHEMISTRY LABORATORIES, INC. P.O. Box 1097 Columbia, MO 65205 (314) 474-8579 Static Acute Bioassay Report #23897 Submitted To: 3M Company Attn: Dale L. Bacon P. 0. Box 33331 St. Paul, MN 55133 " Acute Toxicity of 3M Sample No. 6 (F ^ 7 P-0 to Bluegill Sunfish (Lepomis macrochirus) June 19, 1979 002597 Submitted By: Analytical Biochemistry Laboratories, Inc. 7200 East ABC Lane P. 0. Box 1097 Columbia, Missouri 65205 Prepared By: Aquatic Biologist ft Alan D. Forbis Aquatic Supervisor Date Approved By: Ct. d ujjt____tiiaba Oamps A. Ault Date ility Assurance^Officer Tlfe^D. Johnson Laboratory Mimager Date 002598 BEST COPY AVAILABLE SUMMARY The acute toxicity of 3M Sample No. 6 to bluegill sunfish (Lepomis macrochirus) was assessed usiug the methods outlined by the Committee on Methods for Toxicity Tests with Aquatic Organisms (1). Water quality parameters of temperature, dissolved oxygen, pH and ammonia were measured throughout the test and were within acceptable limits. As a quality check, the bluegill sunfish 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 (2), indicating that the fish were in good condition. The results of the four day static fish toxicity study using blue gill sunfish are summarized below. Compound Antimycin A 96-hour ICso (95% C.I.) >1,000 mg/1 0.000029 mg/1 . (0.000021-0.000040 mg/1) A 96 hour exposure of test fish to a logarithmic series of concentra tions from 100 to 1,000 mg/1 resulted in only 20 percent mortality at the highest concentration. Also, the results indicated a 96 hour, no observed effect concen tration of 560 mg/1. INTRODUCTION BEST COPY AVAILABLE This static bioassay was performed at the aquatic bioassay labora tory of Analytical Biochemistry Laboratories, Inc., Columbia, Missouri, for 3M Company, from June 14 to June 18, 1979, as authorized in a letter from 3M Company on June 6 , 1979 (Appendix I). The purpose of this test was to determine the 24, 48 and 96 hour LC50 levels for 3M Sample No. 6 to bluegill sunfish (Lepomis macrochirus). A preliminary range-finding study was conducted from June 12 to June 14, 1979, to determine the concentration range from the definitive bioassay. The study was per formed following the procedures outlined in ABC Protocol Number 7601 (Appendix I). METHODS AND MATERIALS The procedures for static bioassay, as described in Standard Meth ods for Examination of Water and Wastewater (3) and Methods of Acute Toxicity Tests with Fish, Macroinvertebrates and Amphibians (1), were used in this experiment. The bluegill sunfish used in the test were obtained from Osage Catfisheries, Inc. in Osage Beach, Missouri. The fish were identified to species using the taxonomic keys developed by Eddy (4). All test fish were held in culture tanks on a 16 hour day light photoperiod and observed for at .least fourteen days prior to testing. Fish culture techniques used were basically those described by Brauhn et. al. (5). A daily record of fish observations during the holding period, along with prophylactic and therapeutic disease treat ments, is included in Appendix I. During this period, the fish received a standard commercial fish food (Rangen's) daily until 48 hours prior to testing at which time feeding was discontinued. The bluegill sunfish used for this experiment had a mean weight of 0.60 g and a mean standard length of 28.6 mm. Weight and length measurements were made on a repre sentative group of fish prior to the test and are included in Appendix I . The static fish bioassay was conducted in five gallon glass vessels containing 15 liters of laboratory well water. Characteristics of this dilution water are shown in Table 1. These vessels were kept in a water bath at 22C (1.0). The test fish were acclimated to the dilution water and test temperature and held without food 48 hours prior to testing. A 48 hour range-finding test was conducted to determine the concen tration range for the definitive study. The preliminary test concentra tions were set at 500 and 1,000 mg/1. Based on the results of this preliminary testing and results of toxicity tests with fathead minnows (Pimephales promelas) by 3M Conqjany, five concentrations of the test compound, ranging in a logarithmic series from 100 to 1,000 mg/1, with ten fish per concentration were selected for definitive bioassay. The--test was conducted with duplicate five gallon glass vessels containing five fish each as authorized by 3M Company (Appendix I). The fish were added to the test chambers by stratified random assignment within 30 minutes after addition of toxicant aliquots. 2 002600 The 3M Sample No. 6 standard was received on June 8, 1979, in good condition. The sample upon receipt was observed to be a yellow liquid and was stored at 22C. Sample purity was not provided; therefore, the test concentrations were prepared based on the total compound. The test concentrations were prepared by transferring appropriate weights di rectly to the test chamber. All standard weights and dilution values are listed in Appendix I. RESULTS Table 2 presents the predicted LC5o values and 95% confidence in tervals for 3M Sample No. 6 and the reference test against Antimycin A, a piscicide. These values were obtained by employing the statistical method described by Litchfield and Wilcoxon (6). Mortality rates, test concentrations, and water quality data are presented in Table 3. The dissolved oxygen concentration which stayed between 40 and 100% saturation was considered adequate for testing. The pH values remained consistent with the control throughout the test. The ammonia concen trations were below the toxic limit (7). The study was conducted following the intent of the Good Laboratory Practice Regulations (8) and the final report was reviewed by Analytical Biochemistry Laboratories' Quality Assurance Unit. Ali original raw data was provided to 3M Company, with a copy retained at Analytical Biochemistry Laboratories. 3 002601 TABLE 1 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 DDVP Diazinon -? Disyston Methyl Farathion Malathion Ethyl Parathion Concentration 9.3 ppm 8.2 255 ppm 368 ppm 50 umhos/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 <40 ng/1 <20 ng/1 <20 ng/1 <80 ng/1 <110 ng/1 <80 ng/1 4, 002602 mam MU ! . Compound >.3M.Sample No. 6 Antimycin A*** TABLE 2 The Acute Toxicity of 3M Sample No. 6 and Antimycin A to bluegill sunfish (Lepomis macrochirus) ______24 hours_______ >1,000 0.000070 (0.000056-0.000088)** LC50 in milligrams/liter (ppm) ______48 hours >1,000 0.000040 (0.000032-0.000050)** ______96 hours_______ > 1,000 0.000029 (0.000021-0.000040)** *Bioassay as conducted at 22C (1.0), mean weight and length, 0.60 g and 28.6 mm. ;. **95% confidence/interval. ***Antimycin. A standard obtained from Sigma Chemical Company, Type III, crystalline, Lot 125C-0152. C3 m 3 oo -o < C 471 c io <r c 5 00 m TABLE 3 The mortality rates and water quality parameters during the acute toxicity test of 3M Sample No. 6 with bluegill sunfish (Lepomis macrochirus) . mg/l Concentration Percent Mortality Hours 24 48 96 Water Quality __________24 hours___________ Temp. D.O.* NH3*** C mg/l pH** mg/l __________ 48 hours________ Temp. D.O.* NH3*** C mg/l pH** m/l _________96 hours Temp. D.O.* C mg/l pH** n h 3*** mg/l Control 100 0 0 0 22 8.2 '8.3 1.1 0 0 0 22 8.3 8.3 1.2 22 5.8 8.2 0.75 \J ! 22 5.8 8.3 .-75 22 5.0 8.3 1.1 22 3.6 8.3 1.0 180 0 0 0 ; 320 560 000 00 0 - -, 1,000 . 20 20 20 22 8.3 8.3 1.1 22 *'4^7 8.2 0.62 22 3.8 8.2 1.0 ^Dissolved oxygen concentrations - Dissolved Oxygen Probe (Extech Model 8012) used with an Extech Model 671 pH and mV meter. **pH - pH Probe (Fisher Model 13-639-108) used with an Extech Model 671 pH and mV meter. ***Total ammonia concentrations - Ammonia Probe (Extech Model 8002-8) used with an Extech Model 671 pH and mV meter. C O cn" fO aiaviivAv Adoo isaa oOi LITERATURE CITED (1) 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. (2) 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. (3) American Public Health Association. 1975. Standard Methods for the Examination of Water and Wastewater. 14th ed. Washington, DC. 1193 p. (4) Eddy, Samuel. 1969. The Freshwater Fishes. 2nd ed. W. C. Brown Company, Dubuque, IA. 286 p. _* !, (5) 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. (6) Litchfield, J. T . , Jr. and F. Wilcoxon. 1949. A Simplified Method of Evaluating Dose-Effect Experiments. Jour. Pharm. Exp. Ther. 96:99113. (7) National Academy of Sciences. 1971. Water Quality Criteria, 19'72: U. S. Department of Commerce, PB-236 199. 592 p. (8) Food and Drug Administration. Regulations for Good Laboratory Practice. Federal Register, Vol. 43, No. 247, 59986-60025, December 22, 197.8. 7 002605 Quality Assurance Statement for final report #23897 entitled, "Acute Toxicity of 3M Sample No. 6 to bluegill sunfish (Lepomis macrochirus) for Dale L. Bacon, 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 No. 6 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. It was also noted that since the purity of the compound being tested had not been supplied, the LC50 would have to be calculated based on nominal concentrations of the total compound. A final inspection of all data and records on June 22, 1979, indicated that the report sub mitted 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. U . __ JAmets A Ault Quality Assurance Officer Date 8 002606