Document KvYXJ5jZvZ7bO27q1wBaGDBw

463 TRADE SECRET 3x 2! DuPont-12990 Study Title H-25767: Static, Acute, 48-Hour Screening Test to Daphnia magna A u t h o r : Barbra D. Ferrell, B.S . Robert A. Hoke, Ph.D. Study C o m plet ed o n : May 28,2003 P e r fo r m in g L a b o r a t o r y : 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 W ork Request N umber Study Code N umber Company Sanitized. Does not contain Tsc a cm Page 1 of 8 H-25767: Static, Acute, 48-Hour Screening Test to Daphnia magna DuPont-12990 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation o f data obtained from this study. Work Completed by: Associate Scientist Issued by Study Director: A^okt_________ Robert A. Hoke, Ph.D. Senior Research Ecotoxicologist and Manager Date Company Sanitized. Does not contain TSCA O il H-25767: Static, Acute, 48-Hour Screening Test to Daphnia magna STUDY INFORMATION DuPont-12990 Haskell Number: 25767 C om positioJtfl Known Impurities: Physical Characteristics: White liquid Stability: The test substance appeared to be stable under the conditions o f the study; no evidence o f instability was observed. Sponsor: E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study Initiated/Completed: May 13, 2003 / (see report cover page) Company Sanitized. Does not contain TSCA CBi H-25767: Static, Acute, 48-Hour Screening Test to Daphnia magna DuPont-12990 SUMMARY The acute toxicity of H-25767 to the water flea, Daphnia magna (less than 24 hours old) was determined in an unaerated, 48-hour, static test. The study was conducted with 4 concentrations o f H-25767 and a dilution water control at a mean temperature o f 20.7C (range of 20.5-20.8C). One test chamber was used per test concentration with 10 test organisms in each chamber. Based on visual observations, the water control and 1 mg/L test concentration solutions were clear and colorless at test start. The 10 mg/L test concentration solution was clear with a surface film at test start. The 100 mg/L test concentration solution was clear with a surface film and test material on the solution surface and bottom of test chamber at test start. The 1000 mg/L test concentration solution was cloudy with a surface film and test material on the solution surface and bottom o f test chamber at test start. All water quality parameters were within acceptable limits during the exposure. Exposure o f daphnids to the water control and nominal H-25767 concentrations o f 1, 10,100, and 1000 mg/L resulted in 0, 0, 0, 70 and 100% immobility, respectively, at the end o f 48 hours. No immobility or sublethal effects were seen in the water control test organisms. The highest nominal concentration causing no immobility at test end was 10 mg/L. The lowest nominal concentration causing 100% immobility at test end was 1000 mg/L. Nominal H-25767 concentrations were used for calculation of LC50 values. The results are summarized as follows: Nominal concentrations o f H-25767, mg/L 48-hour LC50 (95% confidence interval) for H-25767, based on nominal concentrations, mg/La dilution water control, 1, 10, 100, and 1000 69(28 to 210) H-25767 exhibited medium concern for aquatic hazardb in an unaerated, 48-hour, static acute test using the water flea, Daphnia magna (less than 24 hours old). a Peltier, W.H. and Weber, C.I., Eds. (1985). Methodsfo r Measuring the Acute Toxicity o fEffluents to Freshwater and Marine Organisms. EPA/600/4-85-013. United States Environmental Protection Agency, Environmental Monitoring and Support Laboratory, Cincinnati, Ohio. b 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, ASTMSTP 1179. (W.G. Landis, J.S. Hughes, and M.A. Lewis, Eds.), pp 22-39. American Society for Testing and Materials, Philadelphia. -4- fipafif Sanitized. Does not contain TSCA CBI H-25767: Static, Acute, 48-Hour Screening Test to Daphnia magna DuPont-12990 MATERIALS AND METHODS A. Test Solution Preparation Test substance solutions were prepared by direct addition o f H-25767 to dilution water. Test solutions o f 1, 10, 100, and 1000 mg/L were prepared by adding the appropriate amount o f test material to HLWW in 1-L beakers and stirring for approximately 60 minutes. The water control and 1 mg/L test concentration solutions were clear and colorless at test start. The 10 mg/L test concentration solution was clear with a surface film at test start. The 100 mg/L test concentration solution was clear with a surface film and test material on the solution surface and bottom o f test chamber at test start. The 1000 mg/L test concentration solution was cloudy with a surface film and test material on the solution surface and bottom o f test chamber at test start. B. Dilution Water Dilution water originated from the Haskell Laboratory well which is 378-feet deep and is cased and sealed to bedrock. The hardness o f the HLWW is adjusted to approximately 100-140 mg/L as CaCCL by the flow-proportioned addition o f CaC^. The HLWW is then aerated, passed through a green sand filter to remove iron, and filtered through 50-, 10-, and 5-pm 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 specifications. C. Test Organism Culture Daphnia magna were reared at Haskell Laboratory in 1000-mL Pyrex beakers (10 per beaker at culture initiation) which contained 1000 mL of aerated, filtered HLWW held at 20.8C. Daphnids were fed on a daily basis with a yeast, cereal leaves and trout chow (YCT) mixture (standardized to 1700 to 2100 mg/L total solids) and the green alga, Selenastrum capricornutum, at a rate of 62,500 cells/mL o f culture medium. The-combination of YCT and alga is equivalent to 0.1-0.2 mg total organic carbon per daphnid per day. Neonates used in this test were less than 24 hours old and were collected from the 7th, 10th, and 12th broods o f 21,28, and 28-day old parent daphnids, respectively. Sickness, injury, and abnormalities were not seen and ephippia were not being produced by the parent daphnids. No adult immobility was seen in the cultures used for testing during the 48-hour pretest period. Daphnia magna were identified by labels on the culture beakers and test chambers. Company Sanitized. Does not contain TSCA CBi -5 - H-25767: Static, Acute, 48-Hour Screening Test to Daphnia magna DuPont-12990 D. Test Methods Four nominal concentrations and a dilution water control were used in this study. The n o m inal concentrations were 1, 10, 100, and 1000 mg/L H-25767. Pyrex beakers (250-mL) containing 200 mL of test solution (6.8-cm test solution depth) were used as test chambers. One replicate test chamber was used per test concentration with 10 daphnids in each chamber. The test chambers were covered with a glass plate during the test. Daphnia magna neonates, less than 24 hours old, were used in this study. Daphnids were not fed during the test. Addition of daphnids to test solutions was initiated after mixing of the test solutions was completed. Immobility and behavioral observations were made daily. The criterion for the effect (immobility) was a lack o f reaction to application o f a gentle stimulus. A recirculating water bath was used to maintain mean temperature in the test chambers during the 48-hour test at approximately 20.7C with a range of 20.5 to 20.8C. A photoperiod of 16 hours light (approximately 564 to 740 Lux) and 8 hours darkness was employed which included 30 minutes o f transitional light (34 to 45 Lux) preceding and following the 16-hour light interval. Dissolved oxygen concentration, pH, and temperature were measured in all replicates o f the control and test substance concentrations (Table 1). These measurements were taken at test start, and at test end or at total immobility in a replicate chamber. Test solutions were not aerated during the test and were disposed o f in an appropriate manner at test end. RESULTS AND DISCUSSION The study was conducted with 4 concentrations of H-25767 and a dilution water control at a mean temperature o f 20.7C (range o f 20.5-20.8C). One test chamber was used per test concentration with 10 test organisms in each chamber. Based on visual observations, the water control and 1 mg/L test concentration solutions were clear and colorless at test start. The 10 mg/L test concentration solution was clear with a surface film at test start. The 100 mg/L test concentration solution was clear with a surface film and test material on the solution surface and bottom o f test chamber at test start. The 1000 mg/L test concentration solution was cloudy with a surface film and test material on the solution surface and bottom o f test chamber at test start. All water quality parameters were within acceptable limits during the exposure. Exposure o f daphnids to the water control and nominal H-25767 concentrations o f 1, 10, 100, and 1000 mg/L resulted in 0, 0, 0, 70 and 100% immobility, respectively, at the end o f 48 hours (Table 2). No immobility or sublethal effects were seen in the water control test organisms. The highest nominal concentration causing no immobility at test end was 10 mg/L. The lowest nominal concentration causing 100% immobility at test end was 1000 mg/L. Nominal H-25767 concentrations were used for calculation of LC50 values. Sanitized. Does notcontain TSCA CB| -6 - H-25767: Static, Acute, 48-Hour Screening Test to Daphnia magna DuPont-12990 H-25767 exhibited medium concern for aquatic hazardc in an unaerated, 48-hour, static acute test using the water flea, Daphnia magna (less than 24 hours old). REFERENCES 1. Organisation for Economic Co-Operation and Development (OECD). Guideline for the Testing o f 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 o f ASTM Standards, Vol. 11.04. c Smrchek, J., Clements, R., Morcock, R., and Rabert, W. (1993). Assessing ecological hazard under TSCA: methods and evaluation o f data. Environmental Toxicology and Risk Assessment, ASTM STP 1179. (W.G. Landis, J.S. Hughes, and M.A. Lewis, Eds.), pp 22-39. American Society for Testing and Materials, Philadelphia. "*pany Sanlfeerf. Does not contain TSCA CBg H-25767: Static, Acute, 48-Hour Screening Test to Daphnia magna DuPont-12990 TABLE 1 DISSOLVED OXYGEN AND pH OF TEST AND CONTROL SOLUTION SAMPLES Nominal H-25767 Concentration (mg/L) Test Initiation (0 hours) Dissolved Oxygen (mg/L) PH Test Completion (48 hours) Dissolved Oxygen (mg/L) pH Water Control 8.4 7.8 8.5 7.8 1.0 8.4 7.9 8.6 7.8 10 8.4 7.9 8.7 7.9 100 8.4 7.9 8.7 7.9 1000 8.4a 7.7a 8.6a 7.4a a Measurements taken at 24 hours due to total immobility in a test concentration. TABLE 2 IMMOBILITY OF Daphnia magna IN AN UNAERATED, STATIC, ACUTE, 48-HOUR TEST WITH H-25767 Nominal H-25767 Concentration (mg/L) Water Control 1.0 10 100 1000 Number o f Dead Test Organisms / Number o f Test Organisms at Study Start 24 Hours 48 Hours 0/10 0/10 0/10 0/10 10/10 0/10 0/10 0/10 7/10 10/10 agoes gvotcontain TSCAOKI -8 -