Document wDgwoeyr1aBRpmxVp52NjNy3E

LRNA33<Z. Study Title Acute Toxicity of L-13492 to the Fathead Minnow, Plaephalva promelas Data Requirements Guideliue Number U.S. EPA-TSCA, Guideline 797.1400 Authors Timothy J. Ward Jeanne P. Magazu Robert L. Boeri Study Coexisted July 13, 1995 Sponsor 3M Company Environmental Laboratory 935 Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55144 Testing Facility T.R. Wilbury Laboratories, Inc. 40 Doaks Lane Marblehead, Massachusetts 01945 T.R. Wilbury Study Number 839-TH Page 1 of 22 BACK TO MAIN T.R. wilbury Study Number 839-TH of 2 2 II. QUALITY ASSURANCE STATEMENT Submitted by: T.R. Wilbury Laboratories, Inc. 40 Doaks Lane Marblehead, Massachusetts Certification: T.R. Wilbury Laboratories, Inc. study number 839-TH, Acute Toxicity of L-13492 to the Fathead Minnow, Pimephalea promelas, was audited by the Quality Assurance Unit for compliance with the protocol, standard operating procedures, and applicable Good Laboratory Practices (U.S. EPA, 1993). The original raw data and a copy of the final report will be archived at T.R. wilbury Laboratories, Inc. for at least 10 years. Quality assurance audits were performed and the findings reported to T.R. Wilbury Laboratories management and the study director on the following dates: Aspect Audited Audit Date Reported to Study Director Reported to Management Protocol: In-life : Draft Report: Final Report: 6 / 8/95 6 / 26/95 7/ 5/95 7 / 13/95 6/ 9/95 6/26/95 7/ 5/95 7/13/95 6/ 9/95 7/13/95 7/13/95 7/13/95 Arthur p. paradice Quality Assurance Officer T.R. Wilbury Study Number 839-TH Page 3 of 22 BACK TO MAIN III. TABLE OP CONTENTS SECTION; I. Good Laboratory Practice Compliance statement II. Quality Assurance statement III. Table of Contents IV. Index of Tables and Figures v. Summary VI. General Information VII. Introduction VIII. Methods and Materials IX. P.esult3 X. References XI. Signature Page XII. protocol Amendments and Deviations Appendix A. Water Quality Data from Toxicity Test PAGE 2 3 4 5 6 7 8 B 13 17 18 19 20 T.R. Wilbury Study Number 839-TH Page 4 of 22 BACK TO MAIN IV. INDEX or TABLES AND riGURES Table 1 Chemical characterization of a representative sample of dilution water used for the toxicity test with fathead minnows, Pimephales promelas, and L-13492 Table 2 Test system summary for the toxicity test with L-13492 and fathead minnows, Plmephale3 promelas Table 3 Survival and sublethal effect data from the toxicity test with fathead minnows, Pimephales promelas, and L-13492 Table 4 Median lethal concentrations (LCSOs) from the toxicity test with fathead minnows, Pimephales promelas, and L-13492 Table A.l Dissolved oxygen concentration, conductivity, pH, and temperature measured during the toxicity test with fathead minnows, Pimephales promelas t and L-13492 Figure 1 Survival of fathead minnows, Pimephales promelas, exposed to L-13492 for 96 hours PAGE 11 16 21 15 T.R. Wilbury Study Number 839-TH Page 5 of 22 BACK TO MAIN - M*.WWifP P The acute toxicity of L-13492 to the fathead minnow. Pimephalaa promolaa, is described in this final report. The test was conducted for 3M Company for 96 hours from June 26 to 30, 1995, at T.R. Wilbury Laboratories, Inc. in Marblehead, Massachusetts. It was conducted by Jeanne Magazu, Jacqueline Nevius, Peter Kowalski, Keven Stevens, Peter Nevius, Robert Boeri, and Timothy Ward according to the protocol developed for T.R. wilbury study Humber 839-TH. L-13492 was supplied by the sponsor. The test was performed under static conditions with five concentrations of test substance and a dilution water control at a temperature of 21.9 to 22.3*C. The dilution water was filtered deionized water collected at Marblehead, Massachusetts and adjusted to a hardness of 44 mg/L as CaCO^. Aeration was initiated at 72 hours to maintain dissolved oxygen concentrations above an acceptable level. Nominal concentrations of L-13492 were 0 (control), 130, 220, 360, 600, and 1,000 mg/L. Fathead minnows used in the test were procured from a commercial supplier and acclimated to test conditions at T.R. wilbury Laboratories. After 96 hours of exposure the control fish had an average wet weight (blotted dry) of 0.28 g and an average total length of 32.8 mm. All fish were in good condition at the beginning of the study. Exposure of fathead minnows to the test substance resulted in a 96 hour median lethal concentration (LC50) of 890 mg/L L-13492, with a 95 confidence interval of 600 to >1,000 mg/L. The 96 hour no observed effect concentration (NOEC) is 600 mg/L. f I T.R. Wilbury Study Number 839-TH Page 6 of 22 BACK TO MAIN V I . GENERAL INFORMATION Material Tested; Sponsor: Test Substance Shipped From: Reported Purity: Study initiation Date: Experimental Start Date: Experimental completion Date: Study Completion Date: Archive Location for the Biological Raw Data and a copy of the Final Report: L-13492 3M Company Environmental Laboratory 935 Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55144 3M Company Environmental Laboratory 935 Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55144 100% June 14. 1995 (protocol signed) June 26, 1995 June 30, 1995 July 13, 1995 T.R. Wilbury Laboratories, Inc. Marblehead, Massachusetts * T.R Wilbury Study Number 839-TH Page ? of 22 BACK TO MAIN VII. INTRODUCT XOH This study was sponsored by 3M Company, St Paul, Minnesota. The objective of the study was to determine the 24, 48, 72, and 96 hour median lethal.concentrations (LC50a) of the teat substance to the fathead minnow, Pimephales promelas, a freshwater fish, under static conditions. The test protocol was based on U.S. EPA methods (U.s. EPA, 1993). The report contains sections that describe the methods and materials employed in the study, and results of the investigation. The report also contains an appendix that presents water quality data collected during the test. VIII. METHODS AND MATERIALS TEST SUBSTANCE: The sample of L-13492 (T.R. Wilbury Laboratories Sample Number 502) was delivered to T.R. Wilbury Laboratories on June 14, 19 95 in a cardboard box at ambient temperature. It was contained in a 125 ml plastic bottle. The label attached to the bottle included the following information: **L13492, Hazardous Components/Special precautions: Isopropyl Alcohol, Originator: S.J. Martin -* S.A. Beach, 53-4N-02 2-3E-Q9, 8-5774, 8-7065, 5/4/95, 6/7/95, N2332-1". L-13492 (a thin, clear liquid) was supplied by 3M Company, St Paul, Minnesota. Prior to use the test substance was stored in the dark at room temperature. Test substance stock solutions were formulated in dilution water without the use of a solvent. The test substance was assumed to have a purity of 100% active ingredient and to be stable under storage and testing conditions. Unused te3t substance will be returned to the sponsor. DILUTION WATER: Water used for acclimation of test organisms and for all toxicity testing was deionized water collected at T.R. Wilbury Laboratories in Marblehead, Massachusetts. Water was adjusted to a hardness of 40-48 mg/L as CaC03 and stored in 500-gallon polyethylene tanks where it was aerated and continuously passed through a particle filter, ultraviolet sterilizer, and activated carbon. Results of chemical analysis of a representative sample of water are presented in Table 1. TEST ORGANISM: Juvenile fathead minnows, Pimephales promela3, employed as test organisms were from a single source and were procured from Aquatic Biosystems, Fort Collins, Coloradc on April 11, 1995. The species of the fish was verified using an appropriate taxonomic key. T.R. wilbury Study Number 839-TH Page 8 of 22 BACK TO MAIN Table l. Chemical characterization of a representative sample of dilution water used for the toxicity teat with fathead minnows, Pimephalea promelas, and L - 13492. Parameter Unit of Measurement Detection Limit Measured Value pH Hardness Alkalinity Acidity particulate Matter Total organic Carbon Total Phosphorus Nitrate chloride Metals Arsenic Cadmium Copper Fluoride Boron Aluminum Chromium Cobalt Iron Lead Mercury Nickel silver Zinc organochlorine Pesticides plus Toxaphene organophosphorus Pesticides PCB pH units mg/L as CaCOj mg/L as CaC03 mg/L as Caco3 mg/L mg/L mg/L mg/L as N mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L pg/L pg/L pg/L jig/i. n o o -- -- -- 10 1 0.05 1 0.01 0.0002 0.005 0.1 0.5 0.1 0.01 0.03 0.03 0.005 0.0003 0.03 0.02 0.02 0.5 2 0.52 0.5 7.9 44 30 6 ND ND 0.46 0.08 14 HD 0.0002 ND ND ND ND ND ND 0.08 ND ND ND ND ND HD ND ND ND Notes: 1. The pH was measured in a control test vessel at the start of the test. Hardness, alkalinity, acidity, and particulate matter were measured in dilution water at the beginning of the toxicity test. Total organic carbon was measured in dilution water used for this toxicity test. Total phosphorus, nitrate, chloride, fluoride, metals, pesticide, and PCB data were collected during February, 1995, as part of routine biannual water quality testing (these analysis we*-e performed by Pace, Inc., Hampton, New Hampshirej. 2. ND * not detected at or above the detection Limit. - . T.R. Wilbury Study Number 839-TH Page 9 of 22 BACK TO MAIN Prior to testing the fish were maintained under flow through conditions in a 270 liter fiberglass tank or a 40 liter glass aquarium. During acclimation fish were not treated for disease, they Were free of apparent sickness, injuries, and abnormalities at the beginning of the test, and mortality during the final 48 hours of acclimation was less than 3%. During the 14 day period before the start of the test the acclimation temperature range was 22.1 to 22.9*0 and thi dissolved oxygen concentration was always at least 8.0 mg/L. During acclimation fish were fed a dry commercial food (Tetra Min* Staple Food, Lot f TM05) daily except during the 48 hours immediately preceding the test. Fish were not fed during the test. At the conclusion of the test the control fish had an average total length of 32.8 mm and an average wet weight of 0.28 g (loading during the definitive toxicity teat was 0.19 g/L). TOXICITY TESTING: A static screening test was conducted from June 14 to 18, 1995 with five concentrations of test substance and a dilution water control. Nominal concentrations of L-13492 were 0.10, 1.0, 10, 100, and 1,000 mg/L. After 96 hours of exposure there was 100% survival in the control and at 0.10, 1.0, 10, and 100 mg/L, and 20% survival at 1,000 mg/L. The definitive toxicity test was performed from June 26 to 30, 1995, according to T.R. Wilbury Protocol 839-TH (Acute Toxicity of L-13492 To The Fathead Minnow, Pimephales promelas). The protocol, which wa3 signed by the study director on June 14, 1995, is based on procedures of the U.S. Environmental Protection Agency (1993). A summary of the test system i3 presented in Table 2. The test was conducted at a target temperature of 22 2*c with five concentrations of test substance and a dilution water control. Nominal concentrations of the test substance were: 0 mg/L (control), 130, 220, 360, 600, and 1,000 mg/L.Appropriate amounts of test substance were added directly to dilution water in test vessels to formulate test media without the use of a solvent. Twenty fathead minnows were indiscriminately and equally distributed among two replicates of each treatment. The test was performed in 20 liter glass aquaria that contained 15 liters of teat solution (water depth was approximately 18 cm). Test vessels were randomly arranged in a water bath during the 96 hour test (a random numbers table was used to select the location of each vessel) and they were loosely covered. A 16 hour light and 8 hour dark photoperiod with a 15 minute transition period was automatically maintained with cool-white fluorescent lights that provided a light intensity of 28 footcandles. Aeration was initiated after the 72 hour dissolved oxygen concentration measurements to maintain dissolved oxygen concentrations above acceptable levels. T.R. Wilbury Study Number 839-TH Page 10 of 22 Table 2 Test system summary for the toxicity test with l -13492 and fathead minnows, pimephalea promelaa. Species: Pimephalea promelaa Age/size of Test organisms: juveniles/-).28 grams, wet weight Test Duration: 96 hours Test Vessels: 20 liter glass aquaria Depth of Test Media: 18 cm Number of Replicate Test Vessels: 2 Organisms per Replicate: 10 Temperature Range: 21.9-22. 3C Minimum Dissolved Oxygen: 4.2 mg/L Photoperiodi 16 hours light and 8 hours dark with a IS minute transition period. Light Intensity: Water Quality Measurements: End points: Acceptability Criteria: 28 footcandles Dissolved oxygen, pH, temperature, and conductivity daily in each test vessel. Temperature mcfasured at least every hour in 1 test vessel. Hardness, alkalinity, acidity, and particulate matter in the dilution water. Dilution water analysis is summarized in Table 1. 24, 48, 72, and 96 hour LC50; 96 hour HOEC. *10 control mortality and acceptable temperature range. T.R. Wilbury Study Number 839-TH Page 11 of 22 BACK TO MAIN The number of surviving organisms and the occurrence of sublethal effects (immobilization, loss of equilibrium. Loss of reflex, excitability, discoloration, or change in behavior ]> were determined visually and recorded initially and after 24, 48, 72, and 96 hours (mortality and sublethal effects were monitored twice each day). Dead test organisms were removed when first observed. Dissolved oxygen (YSI Model 57 meter; instrument numbers 1), pH (Beckman model pHI 12 meter; instrument number 124), conductivity (Cole Parmer meter, instrument number 94), and temperature (Beckman model pHI 12 meter; instrument number 124, probe pHT-1) were measured and recorded daily in each test chamber that contained live animals. The temperature in one test vessel was recorded at least hourly during the test. STATISTICAL. METHODS: Results of the toxicity test were interpreted by standard statistical techniques. Computer methods (Stephan, 1983) were used to calculate the median lethal concentrations (LC50s). All calculations were performed by the author using nominal concentrations of test substance and the number of live fathead minnows. The no observed effect concentration is the highest concentration of test substance that did not cause any toxicant-related mortality or sublethal effects. ' T.R. Wilbury Study Number 839-TH Page 12 of 22 IX. RESULTS Insoluble material was not noted during the test. Biological and water quality data generated by the acute toxicity test are presented in Table 3 and Appendix A, respectively. The measured concentrations of dissolved oxygen ranged from 4.2 to 8.9 mg/L (mean - 7.3 mg/L), the temperature ranged from 21.9 to 22.3C (mean 22-l*C>, conductivity ranged from 170 to 210 jjmhos/cm (mean = 190 pmhos/cm), and pH ranged from 7.2 to 7.9. One hundred percent survival occurred in the control exposure and these fathead minnows did not exhibit any sublethal effects. The 24, 48, 72, and 96 hour LC50s for fathead minnows exposed to L- 13492 are presented in Table 4. The 96 hour LC50 for fathead minnows exposed to L-13492 is 890 mg/L, with a 95% confidence interval of 600 to >1,000. The slope of the 96 hour dose response curve could not be determined with these data. The 96 hour no observed effect concentration (NOEC) is 600 mg/L. T.R. Wilbury Study Number 839-TH Page 13 of 22 BACK TO MAIN Table 3. Survival and aublethal effect data from the toxicity teat with fathead minnows, Plmoph&lea promelaa, and L-13492. Nominal Concentration of L-13492 (mg/L) Rep. Number alive 0 24 48 72 96 hr hr hr hr hr Number affected 0 24 48 72 96 hr hr hr hr hr 0 (control) 130 220 360 600 1,000 i 10 10 10 10 10 2 10 10 10 10 10 1 10 10 10 10 10 2 10 10 10 10 10 1 10 10 10 10 10 2 10 10 10 10 10 1 10 10 10 10 10 2 10 10 10 10 10 1 10 10 10 10 10 2 10 10 10 10 10 i 10 3 3 3 3 2 10 7 5 3 3 00000 00000 00000 00000 0 000 0 00000 0 0 00 0 0 0 0o 0 00000 00000 0 123 3 02 133 Note: Affected fathead minnows were lethargic at 48 and 72 hours and they were lethargic and exhibiting a loss of equilibrium at 24 and 96 hours. T.R. Wilbury Study Number 039-TH Page 14 of 22 BACK TO MAIN Figure 1. Survival of fathead minnows, Pimephales promelas, exposed to L-13492 for 96 hours. T.R. Wilbury Study Number 839-TH Page I S of 22 BACK TO MAIN Table 4. Median lethal concentrations (LC50s) from the toxicity test with fathead minnows, Pimephalea prometlaa, and L-13492. 95 percent confidence limit Calculation method 24 hours 48 hours 72 hours 96 hours 1,000 mg/L 940 mg/L 890 mg/L 890 mg/L 600 to mg/L 600 to >1,000 mg/L 600 to >1,000 mg/L 600 to >1,000 mg/L Binomial Binomial Binomial Binomial ................ - ..........................- ..................... oo o A T.R. Wilbury Study Number 839-TH Page 16 of 22 BACK TO MAIN mm,mm X . REFERENCES Stephan, C.E. 1983. Computer Program for the calculation of LC50 Values. U.S. EPA. Duluth, MN. Personal Communication. U.S. EPA. 1993. Toxic Substances Control Act Test Guidelines; Final Rules. Section 797.1400. U.S. EPA. 1993. 40 CFR Part 792. Toxic Substances Control Act (TSCA); Good Laboratory Practice Standards. Code of Federal Regulations, Title 40, July 1, 1993. | 1- T.R. Viilbury Study Number 839-TH Page 17 of 22 xi. sianatuRs pack ; 't/Vim/ 1- s>'i\C Jeanne P. Magaiu f Study Director" Aquatic Toxicologist idlj / Robert L. B e e n Coauthor -? a /p.'i'"" /(j peraer L. Kowalski Aquatic Toxicologist 7i;<nT .I A /V.,-.,` ( y ' i j I k _ 1 For Jacqueline 1M. JNevius Aquatic Toxicologist 4\/ Vr Keven M. Stevens Biologist "?1' y J7i:< L peter c. Uevius Biologist Mi, Timothy J. Ward Author ? ;//;M T.R. Wilbury Study Number 839-TH Page 18 of 22 BACK TO MAIN BACK TO MAIN BACK Table A .1. Dissolved oxygen concentration, conductivity, pH, and temperature measured during the toxicity test with fathead minnows, Pimephales promolaa, and L-13492. Nominal Concentration of L-13492 (mg/L) Rep. Dissolved Oxygen (mg/L) Conductivity (pmhos/cm) ---------------------------- ------------------- ------- -- 0 24 48 72 hr 96 0 24 48 72 96 hr hr hr b a hr hr hr hr hr hr 0 (control) 130 220 360 600 1,000 i 2 1 2 1 2 1 2 i 2 1 2 8.8 7.5 6.9 6.1 8.2 8.5 8.8 7.9 7.4 7.1 8.2 8.4 17 0 170 170 180 180 170 170 170 170 180 8.8 7.9 6.9 4.9 6.4 7.4 8.8 7.7 6.8 6.1 7.0 7.7 180 100 170 180 180 180 180 170 180 180 8.8 7.4 6.8 6.0 7.7 8.3 8.8 7.4 6.4 5.7 7.4 8.3 180 180 170 180 180 180 180 180 180 180 8.8 7.3 6.2 5.1 6.3 7.1 8.8 7.6 6.4 5.9 6.9 7.7 180 180 180 190 190 180 ISO 180 190 190 8.8 6.9 5.9 4.2 6.2 7.5 8.8 7.5 6.1 5.3 r.l 7.6 190 190 190 190 190 190 190 190 190 190 9.9 8.1 7.8 7.1 7.7 7.3 8.9 7.8 6.6 5.1 6.4 6.5 200 210 200 210 210 200 210 200 210 210 Note: Aeration was initiated in each test vessel following the 72 hour measurements. The "b** measurements were made before aeration and the "a* measurements were made 5 hours after the initiation of aeration. T.R. Wilbury Study Number 839-TH Page 21 of 22 BACK TO MAIN Table A.l. Continued. Nominal Concentration of L-13492 (mg/L) Rep. pH Temperature (*C) --- -------------------- ----------------------- 0 24 48 72 96 0 24 48 72 hr hr hr hr hr hr hr hr hr 96 hr 0 (control) 130 220 360 600 1,000 1 2 1 2 1 2 1 2 1 2 1 2 7.9 7.5 7.4 7.2 7.8 7.9 7.5 7.4 7.2 7.8 7.9 7.6 7.4 7.2 7.6 7.9 7.6 7.4 7.2 7.6 7.9 7.5 7.4 7.3 7.7 7.8 7.5 7.4 7.2 7.6 7.9 7.5 7.4 7.3 7.6 7.9 7.5 7.4 7.3 7.6 7.8 7.5 7.4 7.3 7.6 7.8 7.5 7.4 7.3 7.6 7.8 7.6 7.5 7.3 7.7 7.8 7.6 7.5 7.3 7.6 22.1 22.1 22.2 22.1 22.2 22.1 22.1 22.1 22.1 22.2 22.0 22.2 22.1 22.2 22.2 21.9 22.1 22.1 22.1 22.2 22.0 22.1 22.1 22.1 22.2 22.0 22.1 22.2 22.1 22.2 22.0 22.1 22.2 22.1 22.3 22.0 22.1 22.2 22.1 22.2 22.0 22.1 22.2 22.1 22.2 22.0 22.1 22.2 22.1 22.2 22.0 22.1 22.1 22.1 22.1 21.9 22.1 22.1 22.2 22.2 T.R. Wilbury Study Number 839-TH Page 22 of 22 04/01/2001 ENVIRONMENTAL LABORATORY ARCHIVED FINAL REPORT BACK TO MAIN LAB REQUEST NO. N2332 REQUESTOR NAME: SAB/S.J.MARTIN DEPARTMENT: 3048 PROJECT NO: BI703004UQ DATE RECEIVED: 06/07/1995 DESC: L -13492 ECOTOXICITY EVALUATION CONTRACT LAB(S): TRWIL EXP COMP DATE: 08/30/1995 DATE COMPLETED: 09/02/1995 PROJECT LEAD: PHONE NO: 3M FAX NO: 651-778-6176 Tests conducted following 40CFR 797; methods 1050, 1300, 1400. Sample desc.: L-13492, 45% solids, from S. Martin. SAB 6/7/95 Sample is a clear liquid. Lot confirmed as Lot #2 as per S.J. Martin, of 3M SCD Lab on 7/18/95. SAB Results from 48-hr Daphnia magna acute toxicity test: Test Cone., mg/L 0 (control) 13 22 36 60 100 Percent Immobilization and/or Mortality 24-hrs Exposure 48-hrs Exposure 00 00 55 0 55 0 100 - W 7o 100 sab 7/31/95 Results from 96-hr Fathead minnow acute static toxicity test : Test Cone. mg/L 0 (control) 130 220 360 600 1000 24-hrs exp 0 0 0 0 0 50 Percent Mortality 48-hrs exp 72-hrs exp 00 00 00 0o 00 60 70 96-hrs exp 0 0 0 0 0 70 sab 7/31/95 Results from 96-hr algae growth inhibition test: Test cone. mg/L 1.0 2.0 4.0 8.0 16 EC50 95% C.I. sab 9/1/95 Percent change from Control 24-hrs exp. 48-hrs exp. 72-hrs exp. 02 -2 44 17 27 16 17 38 22 42 55 63 51 61 15 14 7.1 (9.8 - >16) (8.2 - >16) (4. 7 - 11) 96-hrs exp. 0 30 50 56 80 4.9 (3.5 - 6.7) SAMPLE DATE CODE DESCRIPTION RESULT * = CONTRACT LAB DATA 04/01/2001 LAB REQUEST NO. N2332 SAMPLE DATE CODE ENVIRONMENTAL LABORATORY ARCHIVED FINAL REPORT CONTINUED DESCRIPTION RESULT 1 05/04/95 L-13492 Lot 2 TRWIL 48-HR-EC50 *DAPHNIA ACUTE TEST TRWIL 96-HR-LC50 *FISH ACUTE TEST - 96HOUR TRHIL IC50-ALGAE *ALGAE ASSAY - IC50 34 MG/L 890 MG/L 4.9 MG/L * = CONTRACT LAB DATA BACK TO MAIN MDL or (95% C.I.) 2 MDL or (95% C.I.) (30 - 39) (600 - >1000) (3.5 - 6.7)