Document JxNQ175d3VJ6gMGX0BmN7z3v

AR226-2973 TRADE SECRET SludiLDlifi STATIC. ACUTE. 48-HOUR EC50 OF H-19.517 TO Daphnia magna Data Raqmremant Organisation for Economic Co-Operation and Development (OECD) Guideline for Testing Chemicals: 202 Author Kevin N. Baer, Ph.D. Rturiy Completed On June 9, 1992 P^Pr"1"1? Laboratory E. 1. du Font de Nemours and Company Haskeli Laboratory for Toxicology and Industrial Medicine Elkton Road, Post Office Box 50 Newark, Delaware 19714 Laboratory Project ID Haskell Laboratory Report No. 423-92 Company Sanitized. Does not contain TSCA CBS Page 1 of 17 Company Sanitized. Does not contain TSCA CBf 2 Du Font HLR 423-92 GOOD LABORATORY PRACTICE STATEMENT This study was conducted according to EPA F6FRA Good Laboratory Practice Standards (40 CFR 160). OECDMPArFinFciJpalepsanofGGooooddLLaabbooraratotoryryPPraracctitciceeStandards (C(8l)30(Fmal). (59 NohSan No. Annex 3850). 2), and Solubility and stability of the test substance in dilution water 'and under storage conditions v' '9 not determined. Concentrations of the test Protocol amendments and/or SOP substance in dilution water were Appendix not measured. Areas of noncompiiance (if any) are also documented deviations appear in in the study records. No I. deviations existed that affected the scientific validity of the study. Submitter: E. I. du Font de Nemours and Company Sponsor: STUDY DIRECTOR: Du Pont Chemicals E. I. du Pont de Nemours and Company AU^ /^ &W) Kevin N. Baer.Ph.D. Research Toxicologist ^ A / ^ Mo/Day/Year Environmental Sciences Company Sanitized. Does not contain TSCA CBi Du Pont HLR 423-92 TABLE OF CONTENTS Page Specific Country Requirements................................................................................................2 Good Laboratory Practice Stalement................................--....................................................3 TabteofContents-.....................................-..................................................................................4 General lnformation.....................................................................................................................5 Summary......................................................................................................................................6 Quality Assurance Documentation.......................................................... ..............................7 I. Purpose......................-................,..;.......-....... ..............................................................8 II. Records..................................................................................................--....-...............8 III. Materials and Methods...................................................................................................8 A Test Substance.........................................................................................................^ B Dilution Water .............................................................................................................8 C Animal Culture...........................................................................................................8 D. Test Methods.............................................................................................................^ E. Chemical Analysis.................,....-.............................................................................9 F Statistical Analysis.....................................................................................................9 IV. Results and Discussion..................................................................................................9 V. Conclusion..-......-...............................................--...................................................10 VI. Lrterature Cited...................................................................--...----..........-- ............10 . VII. Tables...........................................................-...-...........--.........................................11 1. Chemical Characteristics of Haskell Laboratory Well Water.......................... 11 2. Water Chemistry of the Dilution Water at Test Start .........................................12 3. Physical and Chemical Characteristics of H-19,517 Test Solutions.............13 4. Observed Immobility of Daphnia magna Exposed to H-19,517 for 24 and 43 hours in a Static, Unaerated Test....... ----....-.--.......................................15 5. Nominal H-19,517 Concentrations Causing Immobility to 50% (EC50) of Daphnia magna at Specific Time Intervals..........--.........................................16 VSH. Appendix...................................-............................................--...............................17 I. Protocol Amendments and SOP Deviations.......................................................17 Company SanlS'zed. Does not contsrn TSCA CBI 4 --"IIT-W-" - - --- - ""--^^^'^^"^"'^^'"^i^lin?^^,'^:^^'^'!''-'0-' -: "y'^'tifT'l-f "i^iMtJ'J "^reTM1""'- ---"-"ywFTT^*"- Dn 423-92 Du Pont ChemScaIs E. I. du Pont d Nemoure and Wilmington, 5-18-92/6-8-92 5-19-92/5-21-92 gsr.'3t3"la';:iT5 5 SUMMARY H-19.517 was moderately toxicity to Daphnia magna neonates (less than 24 hours concentrations were 3.0.6.25, old) in an unaerated, static, 48-houftest. Nominal 100 mg/L The 48-hour EC50, based on nominal concentrations 12.5, 25.0, 50.0, and immobility was 12 mg/L with a 95% fiducial limit of 10 to 15 mg/L. and ,,..., ---- _ / Work By. /P^^&U^^ ''Charles L. Hall] AAqnuuaattiirc. Technician CiH THIS REPORT IS APPROVED F9R ISSUE. )^ A/J^asLh Kevin N.^Baer, Ph.D. Research Toxicologist Environmental Sciences 2- Mo/Day/Year {no* c'5ntam TSCA. CBS 6 nates of Inspection: Conduct - 5/19/92 Records, Report(s) - 6/5-C.92 Findings reported to: Study Director - 6/6/92 Management 6/9/92 Reported by: PiblLl^AQ./^ ___ ilDeborah A. Vibk Quality Assurance Auditor ^ ^ Date Company Sanitized. Does not contain TSCA.CBA Du Font HLR 423-92 I. PURPOSE The goal of this study was to assess the acute toxicsty of H-19,517 to Daphnia magna neonates less than 24 hours old. II. RECORDS All data and the final report will be maintained either at Haskell Laboratory for Toxicology and Industrial Medicine (Newark, Delaware) or be archived at the Du Pont Records Management Center (Wiimington, Delaware). III. MATERIALS AND METHODS A. Test Substance Purity and composition of the test substance are described under General Information. A solution of each test concentration was prepared by pipetting the appropriate amount of H-19,517 and directly adding it into dilution water for immediate use. The stability and solubility of H-19,517 in dilution water (see below) and under storage conditions were not conducted. B. Dilution Water Water (Table 1) originated from the Haskell Laboratory well and flowed through aquaria containing fathead minnows. Pimephales promelas. 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 of filtered dilution water or autoclaved filtered dilution water held at approximately 20C. DapL ids were fed two algal species, Ankistrodesmus laifialUS and Selenastrum caprjcpmutum. at a rate of 75.000 ceils/mL of each species three times weekly during the dilution water renewal process. Neonates (less than 24 hours old) were collected from 25-27 day old parent daphnids for use in this test. Sickness, injury, and abnormality were not observed. Daphnia magna were identified by labels on the beakers and test vessels. D. Test Methods [1] Six test concentrations and a water control were used. Nominal test concentrations were 3.0,6.25.12.5, 25.0, 50.0, and 100 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 CB1 Du Font HLR 423-92 (1 daphnid/40 mL) for a total of 20 daphnids per test concentration. Daphnids were added to the control and test beakers using random numbers. Test solutions were held between 19.8 to 19.9C (mean 19.9C), were unaerated. and vessels were covered with a glass plate. Daphnids were not fed during this test. A photoperiod of 18 hours light (151-172 Lux) versus 8 hours darkness was employed with approximately 25 minutes of transitional light (approximately 2 Lux) preceding and following the 16 hour light interval. Observations were made once daily. The criterion 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 Mode! 61 OA, Corning Scientific Instruments, Medfield. MA). and temperature (registered American Society of Testing Materials mercury thermometer) were measured daily in all replicates of the water control and all test concentrations before daphnids were added at tha beginning and at the end of the test. Total alkalinity (Hach kit, Hach Chemical Company, Loveland, CO). EDTA hardness (Hach kit, Hach Chemical Company, Loveland, CO), and conductivity (YSI Model 33, Yellow Springs Instruments, Yellow Springs, OH) of the dilution water were measured before daphnids were added at the beginning of the test. E. Chemical Analysis Concentrations of H-19,517 in the test solutions were not conducted. c. Statistical Analysis The 24 and 48-hour EC50 and their associated 95% fiducial limits were calculated by the probit method [2]. IV. RESULTS AND DISCUSSION Two rangefinding studies were conducted. The first rangefinder was not covered to prevent possible volatilization of the components and was repeated. The second rangefinding study was covered and 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-19.517. Immobilities were 50,10,100%. 100%. and 100%, respectively, were observed. One replicate of the control was inadvertently contaminated during the test. All chemical and physical parameters (Tables 2 and 3) in the definitive test were within expected ranges. Nominal test concentrations were 3.0, 6.25,12.5, 25.0, 50.0, and 100 mg/L. Test concentrations were slightly cloudy. Test vessels were covered to prevent possible Company Sanj&ed. Does mi, 9 volatilization of the components in this mixture. The 48-hour EC50, based on nominal mg/L with a 95% confidence interval of concentrations and immobility (Table 4), was 12 10to 15mg7L(Tab!e5). V. CONCLUSION H-19.517 was moderately toxic to Daphnia magna neonates (less than 24 hours olc in an unaerated, static. 48-hour test. VI. LITERATURE CITED Co-Operation and Development (OECD). Guideline -or 1. Organisation for Economic Testing Chemicals: 202. 4 April 1984. The Probit Procedure. 1985. SAS User's Guide: Statistics, Version 5 Edition 2. SAS Institute Inc., Cary, NC. Company Sanltfeec,. Does not contain TSCACBl 10 " Du Font HLR 423-92 STATIC, ACUTE, 48-HOUR EC50 OF H-19.517 TO Daphnia magna TABLE 1 Chemical Characteristics of Haskeli Laboratory Well Water1 Parameter Concentration Parameter Concentration BOD. mg/L COD, mg/L DOC. mg/L TOC2 mg/L Total Kjeldahl N, mg/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, ng/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 ND6 N08 NO8 ND6 <0.50 ^DaHQ of sample collection 30 December 1991 unless indicated otherwise, analyses performed at Environmental Testing and Certification Corporation, Edison, New Jersey; ^Da&e of sample collection 15 January 1992, performed at Du Font Engineering Test Center. Newark. Delaware; 3Date of analyses 13 January 1992, analyses performed at Du Font Chemicals, Jackson Laboratory, Deepwater, New Jersey; ^elow the Published Method Detection Limit (BMDL); SAnalyses performed at Hazleton Laboratories, Inc., Madison, Wl; SNone detected. Company Sanitized, Does not contain TSCA CB! 11 TABLE 2 Water Chemistry of the Water Control at Test Start Total Alkalinity (mg/L as CaCOa) 76 EDTA Hardness (mgi-asCaCOs) 75 Conductivity {^mhos/cm) 172 ^PanySanffeecf.Does ^ contain TSCACBi 12 Du Pont HLR 423-92 STATIC, ACUTE, 48-HOUR ECSO OF H-19,517 TO Daphnia magna TABLE 3 Physical and Chemical Characteristics of H-19,517 Test Solutions Start Nominal Concentrations (mg/L) Aa B3 Ca 08 48 Hours Aa B81 C8 D8 Dissolved Oxygen. mgA. H20 Control 3,0 6^5 12.5 25.0 50.0 100 8.9 8.9 8.9 8.9 8.8 8.8 8.8 8.8 8.8 8.8 8.8 8.8 8.9 8.9 8.8 8.9 8.8 8.8 8.8 8.8 8.8 8.8 8.8 8.8 8.9 8.9 8.8 8.9 DH HaO Control 3.0 6.25 12.5 25.0 50.0 100 7.5 7.5 7.5 7.5 7.6 7.5 7.6 7.5 7.7 7.7 7.7 7.7 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.6 7.5 7.6 7.5 7.5 7.6 8.9 8.9 8.9 8.9 8.8 8.8 8.8 8.8 8.9 8.9 8.8 8.8 8.9 8.9 8.9 8.9 8.8 8.8 8.8 8.8 8.8 8.8 8.9 8.9 8.8 8.8 8.8 8.8 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 7.9 *A. B, C, and D represent replicate test vessels containing five daphnids each at test start. .CompanySanitized. Does no! contain' 13 Du Font HLR 423-92 STATIC, ACUTE. 48-HOUR EC50 OF H-19,517 TO Daphnia magna TABLE 3 Continued Physical and Chemical Characteristics of H-19,517 Test Solutions Nominal Concentrations (mg/L) Start ______________ M B8* Cs Da 48 Hours _____________ A3 0s C9 D8 Temperature. C H20 Control 3.0 6.25 12.5 250 50.0 100 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 i9.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.9 19.8 19.8 19.9 19.9 19.9 19.9 19.9 19.8 19.8 19.9 19.9 19.9 19.9 19.9 19.8 19.8 19.9 19.9 19.9 19.9 19.9 19.8 19.8 19.9 19.9 19.9 19.9 ^ start. B, C, and D represent replicate test vessels containing five daphnids each at test CompanySanitized. Does not contain TSCA CBI 14 Du Foot HLR 423-92 STATIC, ACUTE, 48-HOUR EC50 OF H-19,517 TO Daphnia magna TABLE 4 Observed immobility of Daphnia maqna Exposed to H-19,517 for 24 and 48 Hours in a Static. Unaerateti Test Observed Immobility in Percent Start Nominal Concentrations (mg/L) A3 B81 C" D3 48 Hours A8 B3 Ca D8 Water Control 0 3.0 0 6.25 0 12.5 0 25.0 40 50.0 60 100 80 0 0 0 0 0 0 0 0 0 0 0 0 20 0 0 80 100 100 60 80 60 0 0 0 0 0 0 0 0 0 0 0 0 80 20 1 00 60 100 60 100 100 100 100 1 00 100 100 100 100 100 a^, B, C and D represent replicate test vessels containing five daphnids each at test start. '"ot contain TSCACBj 15 Du Poos HLR 423-92 Time STATIC. ACUTE, 48-HOUR EC50 OF H-19,517 TO Daphnia magna TABLE 5 Nominal H-19,517 Concentrations Causing Immobility to 50% (EC50) of Daphnia magna at Specific Time Intervals EC50 (rng/L-*) 95% Fiducial Limits (mg/U1) Slope Y-intercept 24 Hours 46 48 Hours 12 16 200 3.2 -5.3 10 15 5.5 -6.0 'Based upon nominal concentrations. ^s no* contain YSCAC 16 Du Posit HLR 423-92 STATIC. ACUTE. 48-HOUR EC50 OF H-i 9,517 TO Daphnia magna APPENDIX I PROTOCOL AMENDMENTS AND SOP DEVIATIONS There were a total of 6 Protocoi deviations and 4 SOP deviations. Protocol amendments Daphnid cultures will be "eo green algae; species and rate will be at the 1. discretion of the Study D rector. Reason for change: A-gae is food of choice; SOP in progress. 2. Water samples for analys s will not be taken. Reason for change: Sscnsor request. All test concentrations we1-3 covered with glass plates to prevent potential 3. volatilization of the compc-.nds in the formulation. Reason for change: Sacnsor request. 4. Synonyms: change ^flIBIIIIHIIIIIF 1 Reason for chanoe: E -or in protocol. 5. AQ019-E and AQ027-P Aere supporting SOP'S. Reason for change: Tasting SOP in error: revision in progress. AQ023-P and AQ026-P were not filed in QA at the time of this study. 6. Reason for change: De ay in issuing SOP'S. SOP deviations 1. AQ009-E has AQ011 -E "stead of AQ-037-P reference. 2. Culture water bath temperature and hatched date were not recorded. The batch number for bearers were not documented in the study notebook; thus 3. preconditioningof the vessels could not be verified. The date, time, and temperature of when the waterbath was filled was not 4. recorded in the study records.