Document 0Jy540zxVDbmznBKwJJ79z9mJ

AR226-2814 DuPont-14096 TRADE SECRET Study Title H-24750: Static, Acute, 48-Hour Toxicity Screening Test with Daphnia magna Authors: Barbra D. Ferrell, B.S. Robert A. Hoke, Ph.D. Study Completed on: January 14,2004 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-14096 Work Request Number:jW K j Study Code Number: Sponsor: E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. Company Sanitized. Dost imi. < . es not contain TSCA CBI t Page 1 of 8 H-24750: Static, Acute, 48-Hour Toxicity Screening Test with Daphnia magna DuPont-14096 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. Work Completed by; (fykJyL0^h huuM nB-aJrb_raTDV.T?F_e_rreIlfl, tB3.S. Associate Scientist (4 Zotifi Date Issued by Study D irector: Robert A Hoke, PhD . Senior Research Ecotoxicologist and Manager Date 0 m Company Sanitized. D oes "t contain TSCA CBI -2- H-24750: Static, Acute, 48-Hour Toxicity Screening Test with Daphnia magiia STUDY INFORMATION 9th Collective Nomenclature: Svnonvms/Codes: DuPont-14096 Submitter's Notebook Numberis): Haskell Number: 24750 CAS Registry Number: Puritv: 99.6% Known Impurities: Not supplied by the sponsor Physical Characteristics: White Solid ' Stability: The test substance appeared to be stable under the conditions of the study; no evidence of instability was observed. Study Initiated/Completed: December 29,2003 / (see report cover page) In-Life Initiated/Comnleted: December 29,2003 / December 31,2003 Company s an/i/; ,2ed- Coes not cntain Tsnurmm. -3 - H-24750: Static, Acute, 48-Hour Toxicity Screening Test with Daphnia magna DuPont-14096 SUMMARY The acute toxicity of H-24750 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 of H-24750 and a dilution water control at a mean temperature of 20.4C (range o f 20.0 - 20.8C). One test chamber was used per test concentration with 10 test organisms in each chamber. Based on visual observations, the dilution water control and 0.10, 1.0, and 10 mg/L test concentrations were clear and colorless with no undissolved test material at test starts The 100 mg/L test concentration was slightly cloudy with undissolved test material settled on the bottom o f the test chamber at test start. All water quality parameters were within acceptable limits during the exposure. Exposure of daphnids to the dilution water control and nominal H-24750 concentrations of 0.10, 1.0, 10, and 100 mg/L resulted in 0, 0, 0, 10, and 20% immobility, respectively, at the end of 48 hours. No immobility or sublethal effects were seen in the dilution water control test organisms. The highest nominal concentration causing no immobility at test end was 1.0 mg/L. The lowest nominal concentration causing 100% immobility at test end was greater than 100 mg/L. The results are summarized as follows: Nominal concentrations of H-24750, mg/L 48-hour EC50 for H-24750, based on nominal concentrations and immobility, mg/L dilution water control, 0.10,1.0, 10, and 100 greater than 100 H-24750 exhibited low concern for aquatic hazard(I) in an unaerated, 48-hour, static acute test using the water flea, Daphnia magna (less than 24 hours old). Company Sanitized. D oes not contain TSCA CBl -4 - H-24750: Static, Acute, 48-Hour Toxicity Screening Test with Daphnia magna DuPont-14096 MATERIALS AND METHODS A, Test Solution Preparation Test solutions of 1.0,10, and 100 mg/L were prepared by adding the appropriate amount of test material to Haskell Laboratory Well Water (HLWW) in 1-L beakers, stirring for approximately 5 minutes, sonicating for approximately 45 minutes, and stirring for approximately 5 minutes. The 0.10 mg/L test solution was then prepared by adding 100 mL of the 1.0 mg/L test solution to 900 mL HLWW in a 1-L beaker. After the preparation of the 0.01 mg/L test solution, all test solutions and the dilution water control were stirred for an additional 15 minutes. The dilution water control and 0.10,1.0, and 10 mg/L test concentrations were clear and colorless with no undissolved test material at test start. The 100 mg/L test solution was slightly cloudy with undissolved test material settled on the bottom of the test vessel 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 of the HLWW is adjusted to approximately 100-140 mg/L as CaCC>3 by the flow-proportioned addition of CaCl2. The HLWW is then aerated, passed through a green sand filter to remove iron, and filtered through 50-, 10-, and 3-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(2) and ASTM(3) 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 approximately 20.5C. Daphnids were fed on a daily basis with 2 mL/L of a yeast, cereal leaves and trout chow (YCT) mixture (standardized to 1700 to 2100 mg/L total solids) and the green alga, Selenastrum capricomutum, at a rate of 62,500 cells/mL of culture medium. The combination of YCT and alga is equivalent to 0.1 -0.2 mg total organic carbon per daphnid. Neonates used in this test were less than 24 hours old and were collected from the 4thbrood of 12-day old parent daphnids. 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. D oes not contain TSCA CBI -5 - H-24750: Static, Acute, 48-Hour Toxicity Screening Test with Daphnia magna DuPont-14096 D. Test Methods Four nominal concentrations and a dilution water control were used in this study. The nominal concentrations were 0.10, 1.0, 10, and 100 mg/L H-24750. Pyrex beakers (250-mL) containing 200 mL o f 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 of reaction to application of a gentle stimulus. A recirculating water bath was used to maintain mean temperature in the test chambers during the 48-hour test at approximately 20.4C with a range of 20.0 to 20.8C. A photoperiod of 16 hours light (approximately 591 to 810 Lux) and 8 hours darkness was employed which included 30 minutes of transitional light (23 to 52 Lux) preceding and following the 16-hour light interval. Dissolved oxygen concentration, pH, and temperature were measured in all replicates of the dilution water control and test substance concentrations. These measurements were taken before daphnids were added 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 CONCLUSION Exposure of daphnids to the dilution water control and nominal H-24750 concentrations of 0.10, 1.0, 10, and 100 mg/L resulted in 0, 0, 0,10, and 20% immobility, respectively, at the end of 48 horns. No immobility or sublethal effects were seen in the dilution water control test organisms. The highest nominal concentration causing no immobility at test end was 1.0 mg/L. The lowest nominal concentration causing 100% immobility at test end was greater than 100 mg/L. The 48-hour EC50 for H-24750, based on nominal concentrations and immobility, was greater than 100 mg/L. H-24750 exhibited low concern for aquatic hazard(1) in an unaerated, 48-hour, static acute test using the water flea, Daphnia magna (less than 24 hours old). not contain TSCA CBl Sanitized. D oes Compaq -6 - H-24750: Static, Acute, 48-Hour Toxicity Screening Test with Daphnia magna DuPont-14096 REFERENCES 1. 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. 2. Organisation for Economic Co-Operation and Development (OECD). Guideline for the Testing of Chemicals: 203, 17 July 1992. 3. 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. TSCACBi Sanitized. D oes not contain C om ply -7 - H-24750: Static, Acute, 48-Hour Toxicity Screening Test with Daphnia magna DuPont-14096 TABLE 1 DISSOLVED OXYGEN AND pH OF TEST AND DILUTION WATER CONTROL SOLUTION SAMPLES Nominal H-24750 Concentration (mg/L) Test Initiation (0 hours) Dissolved Oxygen (mg/L) pH Test Completion (48 hours) Dissolved Oxygen (mg/L) pH Dilution Water Control 7.6 7.3 8.7 8.0 0.10 7.5 7.6 8.6 8.0 1.0 7.5 7.5 8.7 8.0 10 7.6 7.5 8.7 7.9 100 7.8 7.2 8.8 7.8 TABLE 2 IMMOBILITY AND SUBLETHAL EFFECTS OF Daphnia magna IN AN UNAERATED, STATIC, ACUTE, 48-HOUR TEST WITH H-24750 Nominal H-24750 Concentration (mg/L) Dilution Water Control 0.10 1.0 10 100 * effects given in superscript a daphnid lethargic Number Immobile (Number Affected*) / Number at Study Start 24 Hours 48 Hours 0/10 0/10 0/10 037 l0 04a/10 0/10 0'7l0 03710 14710 227 l0 Company Sanitized. D oes not contain TSCA CBI -8 -