Document MoM3b1MjkDjMMaMY4RYmr2yQk

ACUTE TOXICITY TO FISH (FATHEAD MINNOW) TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonatweh,ich may alsobe referredtoas PFOS, FC-95, or as a component of FC-780B. (1Octanesulfoniaccid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was 2. The testsample isFC780B, identifiebdy the laboratorays "3M Sample 1-3".Currentinformation indicatesitisa mixtureof0.58% PFOS, 14% diethyleneglycolmonobutyl ether,76.7% water,5.5% urea,1.17% Hydroxy foamer, 1.15% sodium octyl sulfate0,.8% polyoxyethylenemonooctylphenylether,0.05% sodium lauryl sulfatea,nd 0.05% benzotriazole. The followingsummafy applies toa mixture with incompletely characterizedconcentrations of impurities.Data may not accurately reflecttoxicityof the fluorochemical component of the testsample. METHOD: Method: Methods forAcute ToxicitTyests withFish,Macroinvertebratesand Amphibians, EPA-660/3-75-009,1975. Type: Acute flow-through GLP: No Year completed: 1980 Species: Pimephalespromelas Supplier: FaftigFishHatchery,Brady,NE. Analyticalmonitoring: Dissolvedoxygen,pH, ammonia, and temperature. Exposure period: 96-hours Statisticamlethods: LC50 valuesand 95% confidenceintervalcsalculated usinga computer program by Stephan,etal.1978. Test fishage: Juveniles Length and weight: Average length 27 mm Average weight 0.34g (wet) Loading: Not given. Pretreatment:None. Test conditions: Dilutionwater: Filterewdellwater Dilutionwater chemistry: Hardness: 255 mg/L as CaC03 Alkalinity: 368 mg/L as CaC03 pH: 7.8 Conductivity: 50 pmhos/cm Lighting: A dailyphotope@od of 16 hours lightand 8 hours dark was maintainedthroughoutthe testingperiod. Stockand testsolutiopnreparationT:he stocksolutiwoans preparedin deionizedwater on a volume/volume basis. Flow4hrough rate:Approximatelyseven volume exchanges ofwater every 24 hours. Stabilityof the testchemical solutions: Not noted Exposure vessels: Glass aquariacontaining30 L oftestsolution. Number of replicates:1 Number of fishper replicate:20 Number of concentrations: fiveplusa blankcontrol Water chemistry during the study: pH range (0-96hours): 7.9- 8.2 (controlexposure) 8.0- 8.1 (1500 mg/L exposure) Temperature range (0-96hours): 21 - 22 "C (controelxposure) 21 - 22 0C (1500 mg/L exposure) Dissolved oxygen range (0-96hours): 7.9- 9.1mg/L (controelxposure) 7.9- 8.6mg/L (1500 mg/L exposure) RESULTS Nominal concentrations: Blank control9,0, 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: Testingwas conducted on the mixtureas describedinthe Test Substance Remarks field.The values reportedapplytothatmixtureand not the fluorochemicalproportionalone. CONCLUSIONS The testsample 96-hourLC50 forfatheadminnow was determined to be > 1500 mg/L. The 96-hour No Observed EffectConcentration(NOEC) was 380 mg/L. The 1500 mg/L concentrationresultedina 15% mortality. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking= 2. The studymeets the criterifaorquality testing.However, the sample puritywas notproperlycharacterizedand the study lacksanalyticaclonfirmatioonfthe amount offiuorochemicalproportion inthe solution. REFERENCES This studywas conducted by AnalyticaBlioChemistryLaboratoriesI,nc., Columbia, MO atthe requestofthe 3M Company, St.Paul,MN, 1980. OTHER Last changed: 6/28/00 ANALYTICAL BIOCHEMISTRY LABORATORIES, INC. P.O. Box 1097 o Columbia,MO 65205 o (314)474-8579 A 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 Fathead Minnows (Pimephales promelas) September 29, 1980 Submitted By: Analytical BioChemistry Laboratories, 7200 East ABC Lane P. 0. Box 1097 Columbia, Missouri 65205 (314) 474-8579 Inc. Prepared By: Carl M. ThompsoY Aquatic Supervisor ii@Frykfrfen J@k'l0 10 JIL&T@ I-r-) A@, Alan D. Forbis Aquatic Biologist Paul Boudreau Biologist Date Dao Date Date Approved By: ,Jamos A. Ault Qu itx-rss?rance Officer L@le,W. Johnson Laboratory ManagetF Date Date TABLE OF CONTENTS Page No. LIST OF TABLES .............................................i.i.......... SUMMARY ................................................................ INTRODUCTION ................................................2........... IIETHODS AND MATERIALS .........................................2 ......... I. Test Fish ...............................................2 ........... II. Test System .............................................2.......... III. Test Compound ...........................................3.......... .IV. Test Procedure-Biological .................................3........ V. Test Procedure-Chemical and Physical .........................3 ..... RESULTS ....................................................4............. LITERATURE CITED .............................................8 .......... QUALITY ASSURANCE STATEMENT ....................................9 ........ APPENDIX I RAW DATA LIST OF TABLES Table No. Page No. 1 The estimated LCSO values with their 95% confidence limits for the 96-hour fathead minnow toxicity study of 3M Sample 1-3 .............................................1 .a.n.d.4...... 2 Chemical characterics of well water at the Aquatic Toxicolou Laboratory of Analytical BioChemistry Laboratories, Inc. (All analyses performed by ABC Labs)..................5.... 3 Water quality measurements during the 96-hour toxicity study of 3M Sample 1-3 to fathead minnows (Pimephales promelas) .............................................6........... 4 Percent mortality of fathead minnows exposed to five concentration of 3M Sample 1-3 during a 96-hour toxicity study ................................................7 ........... 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 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 were: 90, 200, 380, 770 and 1,500 mg/l. 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 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 estimated LCSO values. TABLE 1: The estimated LCSO 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 i-3 (mg/1) 95% Confidence Lc--In Limits >1500 >1500 >1500 >1500 ------------- In addition to the above LCro values, the results indicated a 96-hour, no observed effect concentration of 380 mg/l. IWMODUCTION 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 3H Sample 1-3 to fathead minnows (Pimephales promelas). The primary objective of the study was to define the acute toxicity of 3M Sample 1-3 by determining the median lethal concentrations (LCSO's) during the course of the test. The study was authorized by Mr. Moe Eluabarawy, 3M Company, in a protocol dated September 15, 1980 (Appendix I) and was conducted from September 25 to September 29, 1980. The term LCSO 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 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). 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 f*or 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 (Rangenls@) 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 0.34 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, Antimycia A. The observed 96 hour LCSO 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 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 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 22*C (10) 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 (220C). 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 assignment 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 concentrations of 3M Sample 1-3: 90 mg/l, 200 mg/l, 380 mg/l, 770 mg/l and 1,500 mg/l. 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. A computerized LC50 program developed by Stephan et. al. (8) was used to calculate the LCso values and their 95% confidenc'e 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 absorbance (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 interpolation 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/l (stock concentration) X 0.25 % dilution for 421 (dilution factor) aquaria #4 aquaria #4 380 mg/l @nominal concentrationl' 3 The nominal concentrations of 3M Sample 1-3 calculated in this manner were: 90, 200, 380, 770 and 1,500 mg/l. The maximum test concentration was set at 1,500 mg/l, as requested by the study sponsor. RESULTS Table I presents the predicted LCSO values and 95% confidence intervals for 3M Sample 1-3. These values were obtained by employing a computerized LCSO program developed by Stephan et. al. (8). TABLE 1: The estimated LCSO 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 LCBO Limits >1500 >1500 >1500 >1500 ------------- In addition to the above LCSO values, the results indicated a 96-hour, no observed effect concentration of 380 mg/l. Mortality data is presented in Table 4. Water quality parameters of temperature, dissolved oxygen, pH and ammonia were measured in the control, low concentration and high concentration throughout the test (Table 3). The dissolved oxygen concentrations 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 TABLE 2: Chemical characteristics of well water at ABC's Aquatic Bioassay Laboratory. Parameter Dissolved Oxygen pH Hardness (CaC03) Alkalinity (CaCO3) 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 a See appendix for individual analyses. 5 TABLE 3: Water quality measurements during the 96-hour toxicity study of 3M Sample fathead minnows (Pimephales promelas). Hours of Exposure 0 24 48 72 96 Temp. ac 22 22 21 22 22 Control b D.O. ML1 pH c 8.5 8.2 9.1 8.1 8.4 7.9 8..6 8.1 7.9 8.1 d NH3 mg/l 0.56 1.6 0.56 1.0 0.64 Low Temp. OC Concentration b D.O. @!@/ i- LIEC d NH3 mg/i 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 High Concentrati( b Temp. D.O. -c mg/i EEC 22 8.4 8.0 22 8.6 8.1 21 8.5 8.0 22 8.3 8.0 22 7.9 8.0 a High concentration water quality was determined in Tank 6 (1500 mg/l 3H Sample 1-3) thro the study. b Dissolved oxygen concentrations - YSI Model 54 Dissolved Oxygen meter and probe. c pH - pH Probe (Corning Model 476022) used with a Corning Model 125 pH and mV meter. d Ammonia concentrations and mV meter. Ammonia Probe (Extech Model 8002-8) used with a Corning Model ON 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/1) 24 Hours of Exposure 48 72 96 Control 0 0 0 0 90 0 0 0 0 200 0 0 0 0 380 0 0 0 0 770 10 10 10 10 1500 15 15 15 15 7 LITERATURE CITED (1) American Public Health Association. 1975. the Examination of Water and Wastewater. D.C. 1193 p. Standard Methods for 14th ed. Washington, (2) Committee on Methods for Toxicity Tests with Aquatic Organisms (C. E. Stephan, Chairman). 1975. Methods for Acute Toxicity Tests with Fish, Macroinverteb rates and Amphibians. Environmental Protection Agency, Ecological Research Series EPA-660/3-75-009, April, 1975. 61 p. (3) Eddy, Samuel. 1969. The Freshwater Fishes. 2ad 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 Antimycia 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 Apparatus 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-FlowBioassays. 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 LCSO. U. S. Environmental Protectlion Agency, Duluth, Minnesota, pre-publication manuscript, 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 Quality Assurance Statement for final report #26241 entitled, "Dynamic 96-Hour Acute Toxicity of 3M Sample 1-3 to Fathead Minnows (Pimephales promelas)," for Mr. Moe 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 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. "@- 0- (Jam@s A. Ault k@@ity Assurance Officer Date 9