Document VGMRERz6N0V2o1Orya2dQyMg

AR226-2960 DU PONT USE ONLY Study Title STATIC, ACUTE, 48- AND 64-HOUR EC50 0 Du Pont HLR 457-91 [TO Daphnia magna Organisation for Economic Co-Operation and Development (OECD) Guideline for'fes ting Chemicals: 202 Author Kevin N. Baer, Ph.D. Study Completed On June 26, 1991 Performing Laboratory E. I. du Pont de Nemours and Company i~ :nfi iskeil Laboratory for Toxicology and Industrial Me Elk ton Road, Post Office Box 50 Nevark, Delavare 19714 Laboratory Project ID Haskell Laboratory Report No. 457-91 Page 1 of 15 Com paq SanlUized.Dces 9 Du Pont UK 457-91 TOS PAGE IB SB U D HR SPCdPIC COBUKI w w w m m w I I Samlae<l.DoeS n o . o o . n T S C * U 6 Company 0 ! Du Pont HLR 457-91 GOOD LABORATORY PRACTICE STATEMENT This study was conducted according to United States Environmental Protection Agency Good Laboratory Practice Regulations for FIFRA (40 CFR 160). Areas of non-compliance are documented in the study records. No deviations existed that significantly affected the validity of the study. Submitter: E. I. du Pont de Nemours and Company Sponsor: Du Pont Chemicals Study Director: ' I* . / Kevin N. Baer, Ph.D. Research Toxicologist Environmental Sciences 11 MoVDay/Yr nalGOiam T 3 C A C5 Company Saniiized. Does Du Pont HLR 457-91 QUALITY ASSURANCE DOCUMENTATION ['STATIC, ACUTE, 48 AND 64-HOUR EC50 OF STUDY: |TO Daphnia Magna AUDITS: Items Audited Protocol Conduct Records, Final Report Audit Dates 6/19/91 6/19/91 6/25/91 DATE FINDINGS REPORTED TO MANAGEMENT AND STUDY DIRECTOR: 6/25/91 Reported by: Ul & r c.bn Cs Kimberly B. Brebnei Ati'vl 4Pir Appurando Alin n Date Company Sanitized. Does not contain TSCA CM 4 Du Pont BLR 457-91 TABLE OF CONTENTS Page Specific Country Requirements . . . ................. ............... 2 Good Laboratory Practice Statement .................................. 3 Quality Assurance Documentation ............................ 4 Table of C o n t e n t s ........................................................ 5 I . S u m m a r y .............................. . . ; ..................... 6 n. General I n f o r m a t i o n .............. 7 til. Purpose .............................................................. 9 IV. R e c o r d s ............................................................... 9 V. Materials and M e t h o d s ............................................... 10 A. Test M a t e r i a l ........................................... 10 B. Dilution W a t e r ................................................. 10 C. Animal C u l t u r e .......... .. . ............................... 10 D. Test M e t h o d s .......... 10 E. Chemical Analysis ............... 11 F. Statistical Analysis ......................................... 11 VI. Results and D i s c u s s i o n ............................................. 11 VII. C o n c l u s i o n .......................................................... 11 VIII. Literature Cited .................................................. 11 IX. T a b l e s .............................................................. 12 Table 1. Chemical Characteristics of Haskell Laboratory Weil W a t e r ..................................................... 12 Table 2. Water Chemistry of the Water Control at ~%gst Start . . J 3 Table 3. Physical and Chemical Characteristics of Test S o l u t i o n s ...........................................14 Table 4 . ^ Observed Immobility of Daphnia magna Exposed to U f l H B H w or 24, 48, and 64 Hours in a Static, Unaerated T e s t ...........................................15 ) f Du Pont HLR 457-91 { SUMIfKT ______ _ w J vas moderately toxic to Daphnia magna neonates (less than 24 hours old) in arPunaerated, static, 48-hour test. Nominal test concentrations were 0.31, 0.63, 1.3, 2.5, 5.0, and 10 mg/L. The 48-hour EC50 was 8.3 with a 95X fidicial interval of 5.7 to 17 mg/L. Due to absence of 100Z immobility in the high test concentration at 48 hours, the test was extended to 64 hours. As a result, immobility increased in all test concentrations and the 64-hour EC50 was 0.27 mg/L. -y Work By ; n V . Troy Baxter/ Aquatic Technician 6 / z<//f, ' Ho/Day/Yr Study Director: ././Vi //. Kevin N. Baer, Ph.D. Research Toxicologist Environmental Sciences Ho/Dav/Yr THIS REPORT IS APPROVED BOR ISSUE. Kerfin N. Baer, Ph.D. Study Director i 6 Company Sanitized. Does not contain TSCA CBJ Material Tested: Medical Research Number: Haskell Number: CAS Registry Number: Du Pont BLR 457-91 Other Codes: Synonyms : Composition: Purity: Submitter's Notebook Number: Sponsor: Submitted By: Study Schedule: Notebooks: Du Pont Chemicals E. I. du Pont de Nemours and Company Wilmington, Delaware lq898 Du Pont Chemicals E. I. du Pont de Nemours and Company Ui Imi nrr>rn DplavrP 19803 Study Initiation Date: 6-17-91 Experimental Start Date: 6-18-91 Experimental Termination Date: 6-21-91 Study Completion Date: 6-26-91 7 T S C A CB! ;ed. Does noi conta " Company Sanier % Haskell Laboratory Cost Code: Distribution: Du Pont HLR 457-91 A'* * ZLJ 8 Company Sanitized. Does not contain TSCA CB % H I . PURPOSB Du Pont HLR 457-91 The goal of this study was to assess the acute toxicity of to Daphnia Magna neonates less than 24 hours old. All data and the final report will be maintained either at Haskell Laboratory for Toxicology and Industrial Mdicine (Newark, Delaware) or be archived at the Du Pont Records Management Center (Wilmington, Delaware). IT's 9 Company Sanitized. Does not contai TSCA CB! A. Test Material V. MATERIALS AND METHODS Du Pont HLR 457-91 Source and composition of the test substance are described under General Information. A 10 mg/L stock solution was prepared in dilution water (see below) and used immediately. Stability and solubility of the test substance in dilution water were not determined. B. Dilution Vater Water (Table 1) originated from the Haskell Laboratory well and flowed through aquaria containing fathead minnows, (Pimephales promelas) , prior to use in this daphnid test. C. Animal Culture Parent Daphnia magna were reared at Haskell Laboratory in 2-L pyrex beakers (25 per beaker) which contained 1.8 L of filtered dilution water held at approximately 20C. Daphnids were fed 25 mg/L (dry weight) of a mixture of frozen Purina trout chow and Fleischmann's yeast three times weekly during the water renewal process. Neonates were collected from approximately 29 day old parent daphnids for use in this test. Sickness, injury, and abnormality were not present. Daphnia magna were identified by labels on the beakers and test vessels. . D. Test Methods [1] Six test concentrations arid a dilution water control were used. Nominal test concentrations were 0.31, 0.63, 1.3, 2.5, 5.0, and 10 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 (1 daphnid/40 mL). Daphnids were added to the control and test beakers using random numbers. Test solutions were held between 20.0 and 20.6C (mean 20.3C) and were uriaerated. Daphnids were not fed during this test. A photoperiod of 16 hours light versus 8 hours darkness was employed. Dissolved oxygen (YSI Model 54A, Yellow Springs Instruments, Yellow Springs, OH) and pH (Corning Model 610A, Corning Scientific Instruments, Medfield, MA) were measured in one replicate of the Water control and all test concentrations before c^phnids were added at the beginning, and at the end of the test (64 hours). Total alkalinity (Hach kit, Rach 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. Temperature was measured using a registered American Society of Testing Materials mercury stem thermometer. 10 Company rrofconfaht T S C A Si R. Qiemical Analysis Du Pont HLR 457-91 Chemical analysis of the test substance vas not conducted. P. Statistical Analysis The 48- and 64-hour EC50 was calculated using a probit method 12 J. VI. RESULTS AND DISCUSSION A rangefinding test was conducted using the following concentrations; 0.0 (control), 0.1, 1.0, 10, and 100 mg/L. Immobilities of 20, 0, 20, 100, and 1002!, respectively, were observed. All chemical and physical parameters in the definitive test (Tables 2 and 3) were within expected ranges. The observed immobility at 24, 48, and 64 hours is presented in Table 4. Daphnids were considered immobile if they wete unable to swim within 15 seconds after gentle prodding with a glass rod. All daphnids in the high test concentration (10 mg/L) were lying on the bottom of the test vessels at 48 hours. After testing these daphnids for immobility, 60% were considered to be immobilized. The remaining daphnids were clearly effected but were observed to briefly swim in the water column after gentle prodding and then return to the bottom of the test vessels. All mobile daphnids in the control and lower test concentrations appeared normal, i.e., actively swimming in the water column. Due to the lack of 100% immobilization in the high test concentration at 48 hours, the study was extended to 64 hours (overnight). As a result, daphnids in all test concentrations (excluding' controls")' exhibited immobility ranging from 40%.in the low test concentration (0.31 mg/L) to 95% in the high test concentration (10 mg/L). Again, all control daphnids appeared normal. The 48-hour EC50 was 8.3 mg/L with a 95% fiducial interval of 5.7 to 17 mg/L. The 64-hour EC50 was 0.27 with an undefined lower 95% fiducial limit and 1.1 mg/L upper 95% fiducial limit. VII. CONCLUSION (less tFIn F.C50 was 8 yj/as moderately toxic to Daphnia magna neonates lours old) in an unaerated, static, 48-hour test. The 48-hour mg/L, based on nominal concentrations and immobility. VIII. LITERATURE CITED 1. Organisation for Economic Co-Operation and Development (OECD). Guideline for Testing Chemicals: 202. 4 April 1984.2 2. Finney, D. J. 1971. Probit Analysis. 3rd Ed., Cambridge University Press. 333 p. 11 y Company Saniiaed. Does not contain TS CA CBI Chemical Character! TABLE 1 Du Pont HLR 457-91 oratory Well Water1 Parameter Concentration Parameter Concentration BOD2, mg/L COD, mg/L DOC3, mg/L TOC4 , mg/L KjeldahT N, mg/L Ammonia N, mg/L Turbidity, NT0 Phenolics, mg/L Color, Co/Pt Solids total suspended, Aluminum, mg/L Antimony, mg/L Arsenic, mg/L Beryllium, mg/L Boron5, mg/L Cadmium, mg/L Calcium, mg/L Chloride6 , mg/L Chromium, mg/L Cobalt, mg/L Copper, mg/L Cyanide, mg/L Iron* mg/L Fluoride6 , mg/L mg/L <2 13 1.5 <0.5 0.2 <0.05 <1.0 <0.050 <5 <2 <0.100 <0.060 <0.010 <0.001 <0.020 <0.002 25.0 6.3 <0.010 <0.020 <0.010 <0.025 0.053 <0.1 Lead5 , mg/L Magnesium, mg/L MBAS/LAS, mg/L Mercury, mg/L Nickel, mg/L Nitrite6 , mg/L Nitrate6 , mg/L Phosphate6, mg/L Potassium, mg/L Selenium, mg/L Silver, mg/L Sodium, mg/L Sulfate6, mg/L Sulfide, mg/L Thallium, mg/L Zinc5 , mg/L Volatile priority pollutants Acid extractable priority pollutants Base/neutral priority pollutants Pesticides/PCBs Organophosphate pesticides7, pg/L <0.005 3.5 <0.05 <0.00020 <0.020 <0.1 1.6 <0.1 1.8 <0.005 <0.010 8.1 4.8 <0.05 <0.01 <0.020 NDB ND8 ND8 ND8 <0.50 iDate of analyses 17 October 1990 unless indicated otherwise, analyses performed at Environmental Testing and Certification Corporation, Edison, New Jersey; 2Date of analyses 13 December 1990, analyses analysis performed at Environmental testing and Certification Corporation, Edison, New Jersey; 3Muan value; "Date of analyses 24 July 1990, performed at Du Pont Engineering Test Center, Newark, Delaware; 5Below the Published Method Detection Limit (BMDL); 'Date of analyses 9 November 1990, analyses performed at Du Pont Chemicals, Jackson Laboratory, Deepwater, New Jersey; 7Date of analyses 23 October 1990, analyses performed a*' Hazleton Laboratories, Inc., Madison, WI; None detected. 12 tscacbi Sanitized. Does not contain Company Pu Pont HLR 457-91 TABLB 2 Water Chemistry the Vater Con ;ol at Test Start 6 Total Alkalinity (mg/L as CaC03 ) 82 EDTA Hardness (mg/L as CaC3) 75 Conductivity (umhos/cm) 170 13 TS' tto5so*0. Company Saniti*0-DoeS TABLE 3 Du Pont BLR 457-91 Characteristics of utions Nominal Concentrations (mg/L) Tint (Hours) Water Control 0.31 0.63 1.3 2.5 5.0 10 Dissolved Oxygen (mg/L) 0 8.8 8.7 8.7 8.7 8.7 8.7 8.7 64 8.7 8.6 8.6 8.6 8.5 8.4 8.4 pH 0 7.6 7.6 7.6 7.7 7.7 7.7 7.7 64 7.6 7.7 7.8 7.9 8.0 8.0 8.0 14 Compaq TABLE 4 Observed Immobility of Daphnia magna Exposed to] for 24, 48, and 64 Hours In a Static, Unaer! Percent Cumulative Observed Immobility Nominal Concentrations (mg/L) 24 Hours A B C D 48 Hours A* B C D 64 Hours A* B* C D Vater Control 0.31 0.63 1.3 2.5 5.0 10 0000 0000 0000 0000 0 0 20 0 0 20 20 20 20 20 40 40 0 000 0 00 0 0 0 20 0 0 00 0 0 20 20 20 20 40 20 40 40b 60b 60 80 0000 20 60 60 20 100 60 60 80 80 60 100 60 60 100 100 100 20 60 100 100 80 100 100 100 A, B, C and D represent replicate test vessels containing five daphnids each. bAll "mobile" daphnids were lying on the bottom of the test vessels and vere nobile only after gentle prodding with a glass rod (see Discussion for further comments). Du Font HLR 457-91 Company SanH;zsd. Does not confai" T?>On CB?.