Document 65B5JRXYYzRrZVz3Lzd90K4Jm

AR226-2974 TRADE SECRET Study m e STATIC, ACUTE, 48-HOUR EC50 OF H-19.516TO Daphniamagna Data Requirement Organisation for Economic Co-Operation and Development (OECD) .............. Ouideh'ne-fef Testing-CbemiGeds: 202- Author Kevin N. Baer, Ph.D. Study Completed On June 16,1992 Performino Laboratory E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine BktonRoad, Post Office-Box 50 Newark, Delaware 19714 laboratory Prolect.10 Haskell Laboratory Report No. 422-92 Company Sanitized. Does not contain TSOA r ; Page ! o f 18 fr TH IS PASE RESERVED FOR SPECIFIC COUNTRY R EQ tM tCM ENYi f i oes notcontain 2 G&i,. Du Pont HLR 422-92 GOOD LABORATORY PRACTICE STATEMENT This sfei# was conducted accordingto EPA FSFRA Good Laboratory Practice Standards {40 C F B 160), OECD Principles of Good taiorptoiy Practice (C(81)30(FinaJ). Annex 2), andMAFF Japan G ocxHrtory Practice Standards (59 NohSan No. 3850). Solubility and stability of the test substance in dilution water and under storage conditions were not determined. Concentrations of the test substance in dilution water were not measured. Protocol amendments and/or SOP deviations appear in Appendix i Areas of noncompliance (if any) are also documented in the study records. No deviations existed that affected the scientific valkity of the study. Submitter: E. I. du Pont de Nemours and Company Sponsor: STUDY DIRECTOR: Du Pont Chemicals E. i. du Pom de Nemours and Company 'U ^ & g & \ in N. Baer, Ph D. Research Toxicologist Environmental Sciences 6 > f/& f? i Mo/Day/Ysar 'Ifempany .Sanitized. Does not contain TSCACBf Du Pont HLR 422-92 TABLE OF CONTENTS Specific Country Requirements------- Page ......2 Good Laboratory Practice Statement... Table of Contents.-- a f M I M K t S l I Germed information..... .5 Summary .7 Quality Assurance Documentation....................................... ...........................................8 I, Purpose ...... .......9 . Records-- UL Materials and Methods A. Test Substance...... B. Dilution Water........ C. Animal Culture......... D. Test Methods......... E. Chemical Analysis.. F. Statistical Analysis.... ....9 ,...9 ...9 ...9 ...9 10 10 IV. Results and Discussion 10 V. Conclusion ...11 Vi. literature Cited. 11 V. Tables._ .12 1. Chemical Characteristics of Haskell Laboratory Well Water...................... .12 2. Water Chemistry of the Water Control at Test Start............. ................... 13 3. Physic^ and Chemical CfrariiSeristics of H-19,516 Test Solutions........... 14 4. Observed Immobility of Daphnia magna Exposed to H-19,516 for 24 and 48 hours in a Static, Unaerated Test.............................................................. 16 5. Nominal H-19,516 Concentrations Causing immobility to 50% (EC50) of Daphnia magna at Specific Time Intervals................................................ . 17 VIII. Appendix.................................................................. L Protocol Amendments and SOP Deviations....... 18 18 jPpmpany Sanitized. Does not contain tsca CBf 4  D i Post H L R 422-92 GENERAL llW R M A lim i Do Pont HLR 422-92 w ilt sminante! Not Supplied by sponsor Study Initiated/Completari; IihUfe.Study iflftlated/Cgmpisted; E. I. du Pont de Nemours and Company WHminr*on, Delaware 19898 5-11 -92 / 6-16-92 5-12-92 / 6-7-92 Ppmpany Sanitized. Does not contain TSCA CPI 6 Wmm9 Du Pont MLR 422-92 ! m STATIC, ACUTE, 48 HOUR EC50 OF H-19,516 TO Daphnia magna SUMMARY ss la " andimmoMi!y -- Work By: G &O te- / /* & / Chanes L Hall Aquatic Technician & /2 r Mo/Day/Year THIS REPORT IS APPROVED FOR ISSUE. P . tX J L * Kevin N. Baer,, Pn.D. Research Toxicologist Environmental Sciences b N c/^. Mo/Day/Year Company Sanitized. Does not contain TSCA CB1 7 Du Pont HLR 422-92 QUALITY ASSURANCE DOCUMENTATION (H-19S16) of inspection: Conduct - 8/5/92 Records, Report(s) - 6/15/92 Findings i J q: Study Director - 6/16/92 Management - 6/16/92 Reported by: jn M A b j3 , B re h /? p r Kimberly m Brebner Quality Assurance Auditor >l/t j ' l i3 Date PlMapafif Sanitoad, Does not contain fSCA CBi 8 Du Pont HLR 422-92 I. PURPOSE The goaf of this study was to assess the acute toxicity of H 19,516 to Daphnia magna neonates less than 24 hours old. II. RECORDS Ail data and the final report will be maintained either at Haskell Laborator/ for Toxicology and Industrial Medicine (Newark, Delaware) or be archived at the Du Pont Records Management Center (Wilmington, Delaware). III. MATERIALS AND METHODS A. Test Substance So ree and composition of the test substance ire described under General Information. A solution of each test concentration was prepared by pipetting the appropriate amount of H-19,516 and directly adding it into dilution water for immediate use. The stability and solubility of H-19,516 in dilution water (see below) was not conducted. B. Dilution W 8ter . Water (Table 1) originated from the Haskell Laboratory well and flowed through aquaria containing fathead minnows, Pimephales prometas, prior to use in this daphnia test. C . Animal Culture Parent Daphnia magna were reared at Haskell Laboratory in 250 mL Pyrex beakers (1 per beaker) which contained 200 mL cf filtered dilution water or autoclaved filtered dilution water hek^at approximately 20C. Daphnids were fed two algal species, each species three times weekly during the dilution water renewal process. Neonates0* (less than 24 hours old) were collected from 9-23 day old parent daphnids for use in this te?t. Sickness, injury, and abnormality were not observed. Daphnia maana were identified by labels on the beakers and test vessels. ~ D . Test Methods [1] Five test concentrations and a water control were used. Nominal test concentrations were 65, 108,180, 300, and 500 mg/L. Pyrex beakers (250 mL; 7-cm diameter) containing 200 mL of test solution (6.8-cm depth) were used. Each replicate (four per concentration) contained five daphnids Company Sanitized. Does not contain TSCA CBI 9 Du Pont HLR 422-92 {1 d^hnsd/40 mL) for a total of 20 daphnids per test concentration. Daphnids were a ^ e d to te c o n M andtest beakers using random numbers. Test solutions were held bf iw0eB!L 19' l to l c ^mean 20C), were unaeisted, and vessels were covered with a f S i i i i f 1?' daphnids vwre not fed during this test. A photoperiod of 16 hours light (366-403 Lux) versus 8 fioure darkness was employed wf*h approximately 25 minutes of transitional light (approximately 2 Lux) p r e c e d in g fr .wing the 16 hour iteht interval. Observations were made once daily. The critenon for the effect (immobility) was the inability to swim at least two body lengths in any one direction within 15 seconds after gentle prodding with a glass rod. Dissolved oxygen (YSI Model 54A.. Yellow Springs Instruments, Yellow Springs, OH) pH (Coming Model 610A, Coming Scientific Instruments, Medfield, MA), and temperature (registered American Society of Testing Materials mercury thermometer) ^ f m e i^ h r i^ ^ r o a i^ o ld a t^ ^ b ^ 'n ^ g a j^ th e a n d concentrations alkalinity (Hach kit, Hach Chemical Company, Loveland, CO)?EDT^hardness! (Hach fat, Hach Chemical Company, Loveland. CO), and conductivity (YSI Model 33, Yellow Springs Instruments, Yellow Springs, OH) of ihe dilution water were measured before daphnids were added at the beginning of the test. E . Chemical Analysis Analysis of the active ingredients in H-19,516 were no! conducted. F. Statistical Analysis The 24 and 48-hour EC50 and their associated 95% confidence limits were calculated oy the moving average-angle program [2J. IV. RESULTS AND DISCUSSION An uncovered rangefinding study was conducted using two replicates per concentration five unfed daphnids per replicate, exposed to control dilution water, 1.0, 100,500, and ' 1000 mg/L of H-19,516. Immobilities were 0 ,0 ,0 ,1 0 0 % , and 100%, respectively, were ODSSrveci. An earlier uncovered definitive study was conducted but was repeated using a covered test system. All chemical and physical parameters (Tables 2 & 3) in the definitive te*t were within expected ranges. Nominal test concentrations were 65, 108,180,300, and 500 mg/L All test solutions were clear. Except 500 mg/L was slightly cloudy. Test vessels were coveted to prevent possible volatilization of the components in this mixture. The 48-hour EC50 based on nominal concentrations and immobility (Table 4) was 130 mq/L with a 95% confidence interval of 110 to 150 mg/L (Table 5). J p fip a iij SaMfeed iioes'aol confab TSCA CBi 10 Do Pont HLR 422-92 V. CQNCLUS0H H-19,f1S was allohtSy toxic% t aphni magi neonates (iess than 24 hows old) In an unaerated, sigici, 48-hour test. VI. LSTERATURE CifED 1. OrgansatiorforEconomieCo-OperalionandDevelopment(OECD). Guidatine for Testing Chemicals: 202. 4 Aprii 1984. 2. Peltier, W. H. and C. I. Weber. 1985. Efflw.enfs.tQ.Ffgsiiwster.and.Marina.Qiganisms, EPA/8Q0/4-85/Q1 3. United States Environmentai Protection Agency. Cincinnati, OH., Appencfix E, p. 170. Company SanlHzecI. Doe, Bolconfah TSCA CRI 11 Du Pont HLR 422-92 STATIC, ACUTE. 48-HOUR EC50 OF H-19,516 TO Daphnia magna TABLE 1 Chemical Characteristics of Haskell Laboratory Well Water' Parameter Concentration Parameter Concentration BOD, mg/L COD. mg/L DOC, mg/L TOC2, mg/L Total Kjeldahi N, tr.g/L Ammonia N, mg/L Turbidity, NTU Phenolics, mg/L Color, Co/Pt Solids total suspended, mg/L Aluminum, mg/L Antimony, mg/L Arsenic, mg/L Beryllium, mg/L Cadmium, mg/L Calcium, mg/L Chloride3* mg/L Chromium, mg/L Cobalt, mg/L Copper, mg/L Cyanide, mg/L Iron3, mg/L Fluoride3 <2 <10 2.8 <0.5 <0.2 <0.05 1.6 <0.050 <5 <4 <0.100 <0.060 <0.010 <0.001 <0.002 24.5 6.2 <0.010 <0.020 <0.010 <0.025 0.025 <0.1 Lead, mg/L Magnesium, mg/L MBAS/LAS, mg/L Mercury, mg/L Nickel, mg/L Nitrite3, mg/L Nitrate3, mg/L Phosphate3, mg/L Potassium, mg/L Selenium, mg/L Silver, mg/L Sodium, mg/L Sulfate3, mg/L Sulfide, mg/L Thallium, mg/L Zinc4, mg/L Volatile priority pollutants Acid extractablei priority pollutants Base/neutral priority pollutants Pesticides/PCBs Organophosphate pesticides5, pg/L <0.075 3.8 <0.05 <0.00020 <0.020 <0.1 2.1 <0.1 1.8 <0.005 <0.010 8.0 4.8 <0.05 <0.01 <0.020 ND5 ND5 ND5 ND <0.50 ' Date of sample collection 30 December 1991 unless indicated otherwise, analyses performed at Environmental Testing and Certification Corporation, Edison, New Jersey spate of sample collection 15 January 1992, performed at Du Pont Engineering Test Center Newark, Delaware; 3Date of analyses 13 January 1992, analyses performed at Du Pont Chemicals, Jackson Laboratory, Deepwater, New Jersey; 4Below the Published Method Detection Limit (BMDL); sAnalyses performed at Hazleton Laboratories, Inc., Madison, WlNone detected. jggiBfany Saniiketi iis es fio! contain TSC CB 12 p Du Pont H LR 422-92 STATIC. ACUTE. 48-HOUR EG50 OF H-19,516 TO Daptmia magna TABLE 2 Water Chemistry of the Water Control at Test Start Total Alkalinity (mg/L as CaCQ3> 78 EBTA Hardness (mg/L as CaCOs) 76 Conductivity (mhos/cm) 180 J Company Sanitized. Does not contain TSCA CBI i 13 Du Pont HLR 422-92 STATIC, ACUTE, 48-HOUR EC50 OF H-19,516 TO Daphnia magna TABLE 3 Physical and Chemical Characteristics of H-19,516 Test Solutions Nominal Concentrations (mg/L) Aa Start Ba Ca Da 48 Hours Aa Ba Ca Da Dissolved Oxygen, mg/L H2 O Control 65 108 180 300 500 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.2 m H2 O Control 65 108 180 300 500 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7 9.1 9.1 9.2 9.1 9.1 9.1 9.1 9.1 9.1 9.2 9.1 9.2 9.2* 9.2* 9.2* 9.2* 9.2* 9.2* 9.2* 9.2 9.2* 9.2* 9.2 9.2* 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 8.0 Bx 7.9* 7.9* 7.9* 7.9* 7.9* 7.9* 7.9* 8.0 7.9* 7.9* 8.0 7.9* A, B, C, and D represent replicate test vessels containing five daphnlds each at test start. Measurements taken at 24 hours due to total immobility. d ife s a . nor contei,, TSCa 14 f 1 j Da Pont MLR 422-92 STATIC, ACUTE. 48-HOUR EC50 OF H-19,516 TO Daphnia magna TABLE 3 Continued Physical and Chemical Characteristics of H-19,516 Test Solutions Nominal Concentrations (mg/L) A* Start B* C Da m 0 Control 65 108 180 300 500 20.1 20.1 20.1 20.1 20.1 20.1 20.1 20.1 20.1 20.1 20.1 20.1 20.1 20.1 20.1 20.1 2C.1 20.1 20.1 20.1 20.1 20.1 20.1 20.1 48 Hours Aa Ba Ca D 19.8 19.8 19.8 19.6* 19.8* 19.8* 19.8 19.8 19.8 19.8* 19.8* 19.8* 19.8 19.8 19.8 19.8* 19.8* 19.8 19.8 19.8 19.8 19.8* 19.8 19.8* A. B. C. and D represent replicate test vessel? containing five daphnids each at test 11 ` Measurements taken at 24 hours due to total immobility. jponipi " 'SM Rfeed.o^ obi I5 Da Peat HLR 422-92 STATIC, ACUTE, EC50OF magna TASLl 4 Observed f Observed Immobility in Percent Nominal Concentrations (mg/L) Aa 24 Hours Ba Ca Da Water Control 0 0 0 0 65 0 0 0 0 108 0 0 0 0 180 100 100 100 100 300 100 100 100 80 500 100 100 80 100 48 Hours Aa Ba Ca Da 0000 0000 20 0 20 0 100 100 100 100 100 100 100 100 100 100 100 100 sta|^At B* ^ and D represent replicate test vessels containing five daphnids each at test .company Saniffeed. Doss net contain TSCA c a r 16 Du Pont HLR 422-92 STAfi, H-19,518 OF mm s Nominal H-19316 Comienif$&ShsCaustefImmobility to 50% <EC50) of Daphnia magna at Specific tim e intervals Time EC50 (mg/L) 95% Confidence Interval (mg/La) Slope Y-intercept 24 Hours 48 Hours 140 130 120 170 110 150 NAb NA NA NA "Based upon nominal concentrations. bNA means not available; parameters could not be generated by the moving average angle method. Company Sanitized. Does not contain TSCA CBI 17 2>u Pont HLR 422-92 STATIC. ACUTE, H-1&S18 TO APPENDIX 1 PROTOCOL AMENDMENTS AND SOP DEVIATIONS There were a total of 2 Protocol deviations and 1 SOP deviations. 1. discretion of the Study Director. al8a* S P * and rate will be at the BeasotLfor Change: Algae is food of choice; SOP in progress. 2. Water samples for analysis will not be t :ken. Season for change: Sponsor request. SQP deviations 1. Beakers were acclimated for 48 hours. Season for Change: Test was delayed for 24 hours. iB gipang Sanitized, Does not contain tsca C lt 18