Document zoYRj0jM43dDMO0q5MRY4QOEg

AA n u . W ! 473 ,4i6-35^/ DuPont-13347 TRADE SECRET Study Title H-25914: Static, Acute, 96-Hour Toxicity Screening Test with Oncorhynchus mykiss Authors: Laurie D. Bouchelle Robert A. Hoke, Ph.D. Study Completed on: August 1,2003 Performing Laboratory: E.I. du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 Laboratory Project ID: DuPont-13347 Con*>aliy Sanitized. Dom no( contain TSCA CBl Page 1 of 8 H-25914: Static, Acute, 96-Hour Toxicity Screening Test with Oncorhynchus mykiss DuPont-13347 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. W ork C om pleted by: l Laurie D. Bouchelle Associate Scientist / 1003 Issued b y Study D irector: Robert A . Hoke, PhJD. Senior Research E cotoxicologist and Manager D ate .Company Sanitized. Does not contain TSCA Cl -2- H-25914: Static, Acute, 96-Hour Toxicity Screening Test with Oncorhynchus mykiss STUDY INFORMATION Substance Tested:] ft Svnonvms/Codes: H-25914 Submitter's Notebook Number's): Haskell Number: 25914 CAS Registry Number: Composition: DuPont-13347 Physical Characteristics: Amber liquid Stability: The test substance appeared to be stable under the conditions of the study; no evidence of instability was observed. Sponsor: E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study Initiated/Completed: July 21, 2003 / (see report cover page) Company Sanitized. Dees not contain TSCA cm H-25914: Static, Acute, 96-Hour Toxicity Screening Test with Oncorhynchus mykiss D u P o n t-13347 SUMMARY The acute toxicity of H-25914 to the rainbow trout, Oncorhynchus mykiss was determined in an unaerated, 96-hour, static test. The study was conducted with 4 concentrations of H-25914 and a dilution water control at a mean temperature of 11.5C (range of 11.3-11.9C). One test chamber was used per test concentration with 5 test organisms in each chamber. Based on visual observations, the dilution water control and the 0.1, 1.0,10 and 100 mg/L test concentrations were clear and colorless with no precipitate at test start. All water quality parameters were within acceptable limits during the exposure. Exposure of rainbow trout to the dilution water control and nominal H-25914 concentrations of 0.10, 1.0, 10, and 100 mg/L resulted in 0,0, 0,0, and 100% mortality, respectively, at the end of 96 hours. No mortality or sublethal effects were seen in the dilution water control test organisms. The highest nominal concentration causing no mortality at test end was 10 mg/L. The lowest nominal concentration causing 100% mortality at test end was 100 mg/L. Nominal H-25914 concentrations were used for calculation of LC50values. The results are summarized as follows: Nominal concentrations of H-25914, mg/L 96-hour LC50 (93.7% confidence interval) for H-25914, based on nominal concentrations, mg/L(3) dilution water control, 0.10, 1.0, 10, and 100 32 (10 to 100) H-25914 exhibited medium concern for aquatic hazard^ in an unaerated, 96-hour, static, acute test using the rainbow trout, Oncorhynchus mykiss. Company Sanitized. Does not contain TSCA CB1 H-25914: Static, Acute, 96-Hour Toxicity Screening Test with Oncorhynchus mykiss D u P o n t-13 347 MATERIALS AND METHODS A. Test Solution Preparation Test substance solutions were prepared by direct addition of H-25914 to dilution water. The nominal concentration of test substance solutions were not adjusted for purity. Test solutions were prepared by adding the appropriate amount of test substance to each test chamber which contained 15 L of Haskell Laboratory well water and stirring for approximately 35 minutes. Test solutions were clear and colorless with no precipitate at test start. B. Dilution Water Dilution water originated from the Haskell Laboratory well which is 378-feet deep and is cased and sealed into bedrock. The hardness of the well water is adjusted to approximately 100-140 mg/L as CaCCL by the flow-proportioned addition of CaCl2. The well water is then aerated, passed through a green sand filter to remove iron, and filtered through 50-, 10-, and 0.45-jim filters to remove particulates. The water is heated or chilled as appropriate and distributed through aged polyvinyl chloride piping. The dilution water is analyzed twice yearly for major anions and cations, metals, total organochlorine and organophosphate pesticides, and polychlorinated biphenyls. The dilution water meets OECD(1) and ASTM(2) criteria and specifications. C. Test Organism Culture Rainbow trout, Oncorhynchus mykiss, were purchased from Thomas Fish Company, Anderson, California. The fish were held in a 272-L, circular, fiberglass holding tank (84-cm diameter, 49-cm depth) for 32 days in continuously-flowing well water at Haskell Laboratory. During the 7-day period preceding the test, water temperatures ranged from 11.0 to 11.8C (mean 11.3C). The fish were fed freshly-hatched, live brine shrimp (San Francisco Bay Brand, Inc.) and AquaMax Starter Fingerling 300 5D03 (PMI Nutrition International, LLC) twice daily on weekdays and once daily on weekends and holidays. Sickness, injury, and visible abnormalities were not present in fish in the holding tank nor in the fish used for testing. Less than 0.5% mortality was seen during the 48 hours prior to use for testing. Fish were identified by labels on the holding tanks and test chambers. D. Test Methods Four nominal test concentrations and a dilution water control were used in this study. The nominal test concentrations were 0.10, 1.0, 10, and 100 mg/L H-25914. Test chambers were stainless steel aquaria [30 (length) x 30 (width) x 30 (height) cm] which held approximately 15 L of test solution (26-L maximum volume; 17.5-cm test solution depth). One -5 - Company Sanitized. Does not contain t w a c H-25914: Static, Acute, 96-Hour Toxicity Screening Test with Oncorhynchus mykiss DuPont-13347 /""n test chamber was used per test solution (total of 5 test chambers; 5 fish in each test chamber). Each chamber was covered with a glass plate to prevent the fish from escaping. Addition of fish to the test solutions was initiated after mixing of the test solutions was completed. Mortality and behavioral observations were made every 24 hours and at test end. Dead fish were removed from the test chambers during daily observations. Criteria for death were the absence of opercular movement and lack of reaction to gentle prodding. At test conclusion, all fish were humanely sacrificed using MS222 (3-aminobenzoic acid ethyl ester). A recirculating water bath was used to maintain mean temperature in the test chambers during the 96-hour test at approximately 11.5C with a range of 11.3 to 11.9C. A photoperiod of 16 hours light (approximately 267 to 497 Lux) and 8 hours darkness was employed which included 30 minutes of transitional light (72 to 159 Lux) preceding and following the 16-hour light interval. Dissolved oxygen concentration, pH, and temperature were measured in all test chambers. These measurements were taken before fish were added at test start, every 24 hours thereafter, and at test end. Test solutions were not aerated during the test and were disposed of in an appropriate manner at test end. E. Results and Conclusion Exposure of rainbow trout to the dilution water control and nominal H-25914 concentrations of 0.10, 1.0, 10, and 100 mg/L resulted in 0, 0,0, 0, and 100% mortality, respectively, at the end of 96 hours. No mortality or sublethal effects were seen in the dilution water control test organisms. The highest nominal concentration causing no mortality at test end was 10 mg/L. The lowest nominal concentration causing 100% mortality at test end was 100 mg/L. The pretest loading weight for the fish used in this test was 0.2805 g/L. The 96-hour LC50for H-25914, based on nominal concentrations, was 32 mg/L. H-25914 exhibited medium concern for aquatic hazard(4) in an unaerated, 96-hour, static, acute test using the rainbow trout, Oncorhynchus mykiss. Company Sanitized. Does not contain TSCA L01 -6- H-25914: Static, Acute, 96-Hour Toxicity Screening Test with Oncorhynchus mykiss D u P o n t-13 347 REFERENCES 1. Organisation for Economic Co-Operation and Development (OECD). Guideline for the Testing of Chemicals: 203,17 July 1992. 2. American Society for Testing and Materials (ASTM). (1988). Standard Guide for Conducting Acute Toxicity Tests with Fishes, Macroinvertebrates, and Amphibians. E 729-88a. Annual Book of ASTM Standards, Vol. 11.04. 3. Stephan, C.E. (1997). Methods for calculating an LC50. Aquatic Toxicology and Hazard Evaluation, Proceedings o f the First Annual Symposium on Aquatic Toxicology, ASTM Special Publication 634 (F.L. Mayer and J.L. Hamelink, Eds.), pp 65-84. Philadelphia. 4. Smrchek, J., Clements, R., Morcock, R., and Rabert, W. (1993). Assessing ecological hazard under TSCA: methods and evaluation of data. Environmental Toxicology and Risk Assessment, ASTM STP1179. (W.G. Landis, J.S. Hughes, and M.A. Lewis, Eds.), pp 22-39. American Society for Testing and Materials, Philadelphia. Company Sanitized. Does not contain TSCA CBi H-25914: Static, Acute, 96-Hour Toxicity Screening Test with Oncorhynchus mykiss D u P o n t-13347 TABLE 1 DISSOLVED OXYGEN AND pH OF TEST AND DILUTION WATER CONTROL SOLUTION SAMPLES Nominal H-25914 Concentration (mg/L) Test Initiation (0 hours) Dissolved Oxygen (mg/L) pH Test Completion (96 hours) Dissolved Oxygen (mg/L) pH Dilution Water Control 0.10 1.0 10 100 10.8 10.7 10.5 10.6 10.6 7.3 7.3 7.3 7.3 7.3 8.6 7.3 8.5 7.3 8.9 7.3 8.4 7.3 9.4a 7.4a a Measurement performed at 24 hours due to total mortality. TABLE 2 MORTALITY OF Oncorhynchus mykiss IN AN UNAERATED, STATIC, ACUTE, 96-HOUR TEST WITH H-25914 Nominal H-25914 Concentration (mg/L) Dilution Water Control 0.10 1.0 10 100 24 Hours 0/5 0/5 0/5 0/5 5/5 Number Dead / Number at Study Start 48 Hours 72 Hours 0/5 0/5 0/5 0/5 0/5 0/5 0/5 0/5 5/5 5/5 96 Hours 0/5 0/5 0/5 0/5 5/5 Company Sanitized. Does not contain t sc CBi