Document ZBr3OgG31Zr7z3K0vJGK8Lzr8

AR226-2809 DuPont-12988 TRADE SECRET Study Title H-24750: Static, Acute, 96-Hour Screening Test to Oncorhynchus mykiss Laboratoiy Project ID: DuPont-12988 A u t h o r s : Laurie D. Bouchelle Robert A. Hoke, Ph.D. Study Completed on: May 14,2003 P e r f o 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 C ode N um ber: Company Sanitized. Does not contain TSCA CBF Page 1 o f 8 H-24750: Static, Acute, 96-Hour Screening Test to Oncorhynchus mykiss D u P o n t-12988 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. Work Completed by: ~1 Laurie D. Bouchclle Associate Scientist _________ N ^fD axJ ' D& ' Issued by Study D irector: Robert A. Hoke, Ph.D. Senior Research Ecotoxicologist and Manager Date SanWzett Does norcontain TSCACBI -2- H-24750: Static, Acute, 96-Hour Screening Test to Oncorhynchus mykiss STUDY INFORMATION 9th Collective Nomenclature: Svnonvms/Codes: Submitter's Notebook Nnmberis) Haskell Number- 24750 CAS Registry Number: Purity: Known Impurities. Physical Characteristics: White Solid Sponsor: E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. ' . Study Imtiated/Completed: April 14, 2003 / (see report cover page) In-Life Initiated/Completed: April 14, 2003 / April 18, 2003 DuPont-12988 ComPanY Sanitized. Does not contain TSCA CB! S --------- ---------------- -3- H-24750: Static, Acute, 96-Hour Screening Test to Oncorhynchus mykiss DuPont-12988 SUMMARY The acute toxicity of H-24750 to the rainbow trout, Oncorhynchus mykiss was determined in an unaerated, 96-hour, non-GLP, static test. The study was conducted with 5 concentrations of H-24750 and a dilution water control at a mean temperature of 11,7C. One test chamber was used per test concentration with 10 test organisms in each chamber. Based on visual observations, the dilution water control, 0.1 and 1.0 mg/L test concentrations were clear with no color at test start. The 10 mg/L test ' concentration was clear and colorless with undissolved test material present on the bottom of the test chamber. The 100 mg/L test concentration was clear and colorless with a surface film and undissolved test material present on the bottom of the test chamber. The 1000 mg/L test concentration was cloudy with a surface film and undissolved test material present on the bottom o f the test chamber. All water quality parameters were within acceptable limits during the exposure with the exception of the pH (3.9 --4.2) in the 1000 mg/L test concentration. Exposure of rainbow trout to the dilution water control and nominal H-24750 concentrations of 0.10, 1.0, 10,100, and 1000 mg/L resulted in 0, 0, 0, 0,100, 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-24750 concentrations were used for calculation of LC50 values. The results are summarized as follows: Nominal concentrations of H-24750, me/L 96-hour LC50 (95% confidence interval) for H-24750, based on nominal concentrations, mg/La dilution water control, 0.10, 1.0, 10,100 and 1000 32 (10 to 100) H-24750 exhibited medium concern for aquatic hazard5 in an unaerated, 96-hour, static, acute test using the rainbow trout, Oncorhynchus mykiss. a 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. b 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. -4 - utized. Does not contain TSCA C8t H-24750: Static, Acute, 96-Hour Screening Test to Oncorhynchus mykiss DuPont-12988 MATERIALS AND METHODS A. Test Solution Preparation Test substance solutions were prepared by direct addition of H-24750 to dilution water. The nominal concentrations of test substance solutions were not adjusted for purity. Test solutions were prepared by adding the appropriate amount o f test substance to each test chamber which contained 15 L of Haskell Laboratory well water and stirring for approximately 45 minutes. The water control, 0.1 and 1.0 mg/L test concentrations were clear with no color at test start. The 10 mg/L test concentration was clear and colorless with undissolved test material present on the bottom of the test chamber. The 100 mg/L test concentration was clear and colorless with a surface film and undissolved test material present on the bottom of the test chamber. The 1000 mg/L test concentration was cloudy with a surface film and undissolved test material present on the bottom of the test chamber 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 well water is adjusted to approximately 100-140 mg/L as CaC03 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-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(2) criteria and * specifications. C. Test Organism Culture Rainbow trout, Oncorhynchus mykiss, were received as unfertilized eggs and sperm from Troutlodge, Inc., Sumner, Washington. The eggs were fertilized in the laboratory using the sperm received with the eggs. The fish were held in a 272-L, circular, fiberglass holding tank (84-cm diameter, 49-cm depth) for 28-34 days after hatching in continuously-flowing well water at Haskell Laboratory. During the 7-day period preceding the test, water temperatures ranged from 12.0 to 12.1C (mean 12.1C). 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 of fish occurred in the holding tank during the 48 hours prior to use for testing. Fish were identified by labels on the holding tanks and test chambers. Company Sanitized. Does not contain TSCA CBI -5- H-24750: Static, Acute, 96-Hour Screening Test to Oncorhynchus mykiss________________________________ DuPont 1298S D. Test Methods Five nominal test concentrations and a dilution water control were used in this study. The nominal test concentrations were 0.10, 1.0, 10, 100, and 1000 mg/L H-24750. 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 test chamber was used per test solution (total of 6 test chambers; 10 fish in each test chamber). Each chamber was covered with a glass plate to prevent the fish from escaping. Addition offish 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 11,7C. A photoperiod of 16 hours light (approximately 264 to 424 Lux) and 8 hours darkness was employed which included 30 minutes of transitional light (11 to 37 Lux) preceding and following the 16-hour light interval. Dissolved oxygen concentration, pH, and temperature were measured in all test chambers (Table 1). 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. ' RESULTS AND DISCUSSION The study was conducted with 5 concentrations of H-24750 and a dilution water control at a mean temperature of 11,7C. One test chamber was used per test concentration with 10 test organisms in each chamber. Based on visual observations, the dilution water control, 0.1, and 1.0 mg/L test concentrations were clear with no color at test start. The 10 mg/L test * ' concentration was clear and colorless with undissolved test material present on the bottom of the test chamber. The 100 mg/L test concentration was clear and colorless with a surface film and undissolved test material present on the bottom of the test chamber. The 1000 mg/L test concentration was cloudy with a surface film and undissolved test material present on the bottom of the test chamber. All water quality parameters were within acceptable limits during the exposure with the exception of the pH (3.9 - 4.2) in the 1000 mg/L test concentration. Exposure of rambow trout to the dilution water control and nominal H-24750 concentrations of 010, 1.0,10, 100, and 1000 mg/L resulted in 0, 0, 0, 0,100, and 100% mortality, respectively, at the end of 96 hours (Table 2). 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-24750 concentrations were used for calculation of LC50 values. -6- Company Sanitized. Does not contain TSCA C B l H-24750: Static, Acute, 96-Hour Screening Test to Oncorhynchus mykiss DuPont-12988 Toxietty data analysis was conducted based on nominal test concentrations using the binomial " 0/ ' / j 16 96' hour LC50 for H`24750' based n nominal concentrations, was 32 mg/L, with a 95 /o confidence mterval of 10 to 100 mg/L. H-24750 exhibited medium concern for aquatic azard in an unaerated, 96-hour, static, acute test using the rainbow trout, Oncorhynchus mykiss. REFERENCES ? E c ^ 2 o T f 7 T i % 0; OPera,i0" and DevelP" " <ECD>' G-Me.me for Testmg American Society for Testing and Materials (ASTM). (1988). Standard Guide for " T 8 ^ CUte J X1City TeStS with Fishes' Macroinvertebrates, and Amphibians. E 729-88a. Annual Book of ASTM Standards, Vol. 11.04. " '/ a C1T ef ' R^ MOrC Ck' R" 311(1 Rabert>W- ( I993)` Assessing ecological hazard under TSCAmethods and evaluation o f data. Environmental Toxicology and Risk Assessment. A ST M ST P 1179 k ilad e^ h i' 8hCS' ^ M'A' LCWiS' EdS )' PP 22' 39' American Society for Testing and Materials, -7 Company Sanitized. Does not contain TSCA CB H-24750: Static, Acute, 96-Hour Screening Test to Oncorhynchus mykiss DuPont-12988 TABLE 1 DISSOLVED OXYGEN AND pH OF TEST AND CONTROL SOLUTION SAMPLES Nominal H-24750 Concentration (mg/L) Water Control 0.10 1.0 10 100 1000 Test Initiation (0 hours) Dissolved Oxygen (mg/L) 10.8 10.9 10.5 10.3 10.3 10.1 _ PH 7.6 7.6 7.5 7.5 7.2 4.2 Test Completion (96 hours! Dissolved Oxygen (mg/L) pH 8.9 9.1 9.0 8.9 9.6a 10.3b 7.3 7.3 7.3 7.3 7.2a 3.9b a Measurement performed at 48 hours due to total mortality, b 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-24750 ' '' Nominal H-24750 Concentration ________(mg/L)________ Water Control 0.10 1.0 10 100 1000 ----------- ------- -------------- Number of Dead Test Organisms / Number of Test Organisms at Studv Start 24 Hours 48 Hours 72 Hours 96 Hours 0/10 0/10 0/10 0/10 8/10 10/10 0/10 0/10 0/10 0/10 10/10 10/10 0/10 0/10 0/10 0/10 10/10 10/10 0/10 0/10 0/10 0/10 10/10 10/10 Company Sanitized. Does not contain TSC CBS