Document 8Ow9b8R8MK17b2RYNG1RGnKvZ

AR226-2990 Sludyjjfle STATIC. ACUTE, 96-HOUR LC50 OFH-19520 TO FATHEAD MINNOWS, Pjmephales promelas Data Requirement Organisation for Economic Co-Operation and Development (OECD) Guideline for Testing Chemicals: 203 Author Keith B. Pierson, Ph.D. . Study Completed On May 25.1993 Performing Laboratory E. I. du Font de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, Post Office Box 50 Newark. Delaware 19714 Laboratory Project ID Haskell Laboratory Report No. 593-92 Page 1 of 19 fimpanx .Sanitized, Does nol contain TSCA CB6 Du Font HLR 593-92 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT Practice Standards (40 CFR 160 and 40 CFR 792), OECD Prindples of Good Laboratory Practice (C(81)30(Final), Annex 2), and MAFF Japan Good Laboratory Practice Standards (59 NohSan No. 3850) wrth the two exceptions noted below. 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 measured, but these measurements were not made in accordance with Good Laboratory Practice Standards. Protocol amendments and/or SOP deviations appear In Appendix1. Areas of noncompliance (if any) are also documented in thestudy records. No deviations existed that affected the scientific validity of the study. Submitter. Sponsor: E. I. du Pont de Nemours and Company Du Pont Chemicals E. I. du Pont de Nemours and Company STUDY DIRECTOR: /^^ 4. P/'U^ Keith B. Pierson, Ph.D. Research Toxicologist Environmental Sciences SLS /VW/W Mo/Day/Year 2 cmm s^u^' "" ""'""TSCA CBI Du Font HLR 593-92 TABLE OF CONTENTS Page Good Laboratory Practice Compliance Statement.....................................^ Table of Contents........................................................................3 General Information........................>.....<...............<,......>.....................4 Summary.................................................................................6 Quality Assurance Documentation ......................................................7 (. purpose..........................^........^.,................................. 8 II. Records.........................................................................8 III. Materials and Methods .........................................................8 A. Test Substance..............................................................8 B. Dilution Water...............................................................8 C. Animal Test Culture .......................<>......................................89 ED.. ChemMicealthAnoadlyss.is.................................................;.......................................................................9 F. Statistical Analysis ..........................................................9 IV. Results and Discussion.......................................................10 V. Conclusion....................................................................10 VI. Literature Cited ...............................................................10 VII. Tables.........................................................................11 21.. CMheeamsuicreald CChoanrcaecntetrriasttiiocnssoof fHHas-k1e9ll5L2a0byor--ato--ry--W--ell --yV--atae|rin.............11 96-Hour. Static, Unaerated Test.....................V................... 12 43.. WPhaytseircaClhaenmdisCtrhyeomfitchael CCohnatrraoclteartiTsteiscts SoftaHr-1t.9.5..2.0..|.|U._--...--....--...B..^| .... 13 H-1952oN^Bfpn Test Solutions........................................v..............v... ..14 5. Observed Mortality crf^atheadMinnows, Pimephales promelas, Exposed to H-19520|HU|lKfor 96 Hours in a Static, Unaerated Test ...... ^ijjjjjjj^j.^jj...^. .......>.........>.... 16 6. Observed Effects of Fathead Minnows, 7. PH/-m19e5p2/0ajltelllwBoHmMetCeso,fnarc9e6nHtroautrsioinnas SCtaatuics,inUgnMaeorratatelidtyTtoest......... 17 50% (LC50)OTTatneaaTOinnowsP. imephales promelas, at Specific Time Intervals.................................................... 18 VIII. ApPproetnocdoilxA.m..e..n.d..m..e.n.t.s..a..n.d..S..O..P..............................................1199 1. Deviations.............................. I 3 Companx Sanifeed. Does not contain TSCA CB1 Matenai Tested: Du Font HLR 593-92 Synonyms Masks!! Numbec Eurilyi H-19520 TSCACB! 4 eomp^sa"111.^n-0o0e-not contain Du Font HLR 593-92 Sponsor: Study initiated/Completed: tn-Life Study Initiated/Completed: Du Pont Chemicals E. 1. du Font do Nemours and Company Wilmington, Delaware 19898 7-17-92/5-25-93 7-22-92 / 8-7-92 .empany SaniSizeei. Does not coniain TSCA CBf Du Pont HLR 593-92 STATIC, ACUTE, 96-MOUB LC50 OF H-19520 TO FATHEAD MINNOWS, PSmophaiSporsomelas SUMMARY H-19520 exhibited slight toxicity in a 96-hour. unaerated, static, acute test using fathead minnows, Pimephales promelas. Nominai H-19520 concentrations were 0. 75.4.116.179,275,423,650, and 1000 mg/L Measured concentrations at 96 hours were 0.14,72.9,91.7,86.5.243,343,540, and 907 mg/L The 96-hour LC50 based on measured concentrations at 96 hours was 276 mg/L with a 95% confidence interval of 237 -317 mg/L Work By: J^. ^Ujt w. P^Jus^ Robert W. Peterson. B. A. Toxicology Associate ^/W 93 Mo/Day/Year Work By: ,^fe 7^^ r- yeffreyT. Turner, Sr. Toxicology Associate 3/M/fi MoyDay/Year THIS REPORT IS APPROVED FOR ISSUE. ^^JL^W^ Keith B. Pierson, Ph.D. Research lexicologist and Study Director Environmental Sciences )W ^ /wy Mo/Day/Year .GempanX Sanitized. Does not contain TSCA CBB Du Font HLR 593-92 QUALITY ASSURANCE DOCUMENTATION (H-19520) Dates of Inspection: Conduct - 7/22/92 Records, Report(s) - 4/21-22/93 Findings Reported to: Study Director - 4/23/93 Management - 5/24/93 Reported bySZ^K^a^^L Afe-JT Donna R. Hoit Quality Assurance Auditor cSW^ 'Date ,3ompanx Sanitized. Does not contain TSCA CBS Du Pont HLR 593-92 I. PURPOSE The goal of this study was to assess the acute toxidtv of H-19520 to fathead minnows, Pimephaies promelas. II. RECORDS All data and the final report will be maintained either at Masked Laboratory for Toxicology and Industrial Medicine (Newark, Delaware) or be archived at the Ou Pont Records Management Center (Wilmington, Delaware). III. MATERIALS AND METHODS A. Test Substance The test substance is described under General Information. A stock solution was prepared by dissolving the test substance in dilution water (see belo'v) and stirring for 30 minutes. Visual inspection suggested that the test substance was in solution in the stock. Aliquots of the stock solution were added to dilution water to prepare individual test solutions. Each of the above solutions was kept in a container with minimal head space sealed with Saran Wrap9 to avoid potential loss of the test substance through volatility. Stability and solubility of H-195SO in dilution water and under storage conditions were not determined. B. Dilution Water Water (Table 1) originated from the Haskell Laboratory well. C. Animal Culture Fathead minnows, Pimephaies promelas, were reared in continuously flowing well water at Haskell Laboratory. Adult fish resided in 39 L aquaria at approximately 25C and were fed frozen brine shrimp (Artemia sp.. Golden Gate Brand, Kordon Division, Novaiek, Inc., Hayward, CA) generally twice daily, once on Saturday and once on Sunday. Eggs were deposited on polyviny! chloride tiles coated with sand. tiles with eggs were aerated and incubated at approximately 25C in a water bath. Larvae were reared in 39 L aquaria at approximately 21 C and were fed newly hatched Artemia sp. (Aquarium Products, Glen Burnie, MD) generally twice daily on weekdays and daily on weekends. Juveniles were held either in 272-L circular, fiberglass tanks or in 39 L glass aquaria (23-cm depth; 50 (L) x 25 (W) x 31 cm (H)) at approximately 21 C and were fed newly hatched Aflamia so. and/or frozen brine shrimp generally twice daily on weekdays, once on Saturday and once on Sunday. Sickness, injury, and abnormality were not observed. Animals were identified by labels on aquaria and test vessels. iCompanx Sanitized. Does not contain TSCA CB& Du Pent HLR 593-92 D. Test Methods [1] Seven test concentrations and a control were used. Nominal concentrations were 0.75.4,116,179,275,423,650, and 1000 "i^L. Control fish ranged from 1.1 to 1.6 cm standard length (mean 1.3 cm) and G.019 to 0.056 g wet weight (mean 0.030 g). Erienmeyer glass flasks [4.4 liter; 21 (base) x 35 (height) cm] containing 4.4 L of test solution(35-cm depth) were employed as test chambers. Each flask was totally filled to minimize head space and was covered with Saran Wrap9 to prevent toss of potentially volatile test substance components. Ten fish were added to each replicate using random numbers (one replicate per concentration, totallO fish per concentration). Control loading was 0:067 g/L at test condusion. Test solutions ranged from 20.8 to 21.4C (mean 21.0C). Fish ware not fed 27 hours prior to nor during the test A photoperiod of 16 hours light (161 to 215 Lux) versus 8 hours darkness was employed with 25 minutes of transitional light (1 - 2 Lux) preceding and following the 16 hour light interval. Mortality and behavioral effects were noted daily. Dissolved oxygen (YSI Model 54A, Yellow Springs Instruments, Yellow Springs, OH) and pH (Coming Model 61 OA, Coming Scientific Instruments, Medfield, MA) were measured in the water control and all test concentrations before fish were added at the beginning of the test, daily thereafter and at total mortality. Temparatui.. was measured (American Society of Testing Materials registered mercury thermometer) in the water control and all test concentrations before fish were added at the beginning of the test, daily thereafter, at the end of the test and at total mortality. 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 control water were measured at the beginning of the test. A continuously-recording thermometer in a control vessel was used to check for temperature variation during the 96-hour test. E. Chemical Analysis Analyses were conducted after the test ended at the Du Pont Jackson Laboratory at Chambers Works, Oeepwater, NJ. Samples and standards were completely combusted in a hydrogen/oxygen Wickbold Torch. The resulting water was then analyzed for total fluoride concentration using an Orion fluoride ion selective electrode. Concentrations were calculated from the standards using either a three or four point calibration curve. F. ^tetistical Analysis The 24- and 48-hour LC50s were calculated by the SAS Probit method [2]. The 72- and 96-hour LC50s were calculated by using the moving average angle technique (3J. 9 SsroW.SISd^,e^o.coninTSCACB. Dil Font HLR 593-92 IV. RESULTS AND DISCUSSION A rangefinding test with five fish per concentration was conducted with a control and test concentrations of 0.1,1,10,100, and 1000 mg/L Mortalities were 0,0,0, 0,0. and 100%, respectively. Two definitive studies were conducted. Data from the first were considered invalid because of low dissolved oxygen concentrations. Data from the second are reported herein. Nominal concentrations in the definitive test were 0.75.4,116,179.275,423, 650. and 1000 mg/L Measured concentrations at 96 hours were 0.14,72.9. 91.7, 86.5,243,343, 540, and907 mg/L (TaBIe^; The~drscrepancy between the measured and nominal concentrations may stem from use of a stock solution in which the test substance was not truly in solution. All chemical and physical parameters in the final definitive test (Tables 3 and 4) were within expected ranges except on day 4. Dissolved oxygen concentrations dropped as low as 53% (minimum) of saturation at 72.9 mg/L However, these observations were not of biological significance since fathead minnows can survive at much lower dissolved oxygen concentrations for extended periods. Behavioral observations appear in Table 6. The 96-hour LC50 based upon measured concentrations at 96 hours was 276 mg/L with a 95% confidence interval of 237 - 317 mg/L (Tables 5 and 7). V. CONCLUSION H-19520 was slightly toxic to fathead minnows, Pimephales promelas, in a 96-hour, static, unaerated test. VI. LITERATURE CITED 1. Organisation for Economic Co-Operation and De""!opment (OECD). Guideline for Testing Chemicals: 203. 4 April 1984. 2. Finney. D. J. 1971. Probit Analysis, 3rd Ed., Cambridge University Press. p.333. 3. Peltier.W. ' and C.!. Weber. Eds. 1985. Methods for measuring the acute toxidty of effluents to freshwater and marine organisms. EPA/600/4-85/013. United States Environmental Protection Agency, Environmental Monitoring and Support Laboratory, Cincinnati, Ohio. Appendix E. 10 companySanitized. Does not contain TSCA CBI Du Pont HLR 593-92 STATIC, ACUTE, 96-HOUR LC50 OF H-19520 TO FATHEAD MINNOWS, Pimephales promefas TABLE 1 Chemical Characteristics of Haskell Laboratory Well Water1 Parameter Concentration Parameter Concentration BOD, mg/L <2 Lead. mg/L <0.075 COD, mg/L DOC, mg/L TOC^mg/L <10 1.4 1.8 Magnesium, mg/L MBAS/LAS, mg/L Mercury, mg/L 3.7 <0.05 <0.00020 Total Kjeldahl N, mg/L Ammonia N, mg/L 0.8 <0.2 Nickel, mg/L Nitrite3, mg/L <0.020 <0.1 Turbidity, NTU <1.0 Nitrates, mg/L 1.9 Phenolics, mg/L <0.050 Phosphate3, mg/L <0.1 Color, apparent Co/Pt7 <5 Potassium. mg/L 1.7 Solids Selenium, mg/L <0.005 | Total suspended, mg/L <4 / Aluminum, mg/L <0.100 Silver, mg/L Sodium, mg/L <0.010 8.2 Antimony. mg/L <0.060 Sulfate3. mg/L 5.0 Arsenic, mg/L <0.010 Sulfide, mg/L <0.10 Beryllium, mg/L <0.001 Zinc, mg/L <0.058 Boron4, mg/L 0.020 Volatile priority Cadmium4, mg/L <0.002 pollutants ND6 Calcium, mg/L 24.3 Add extractable Chloride3- mg/L 6.2 priority pollutants ND6 Chromium, mg/L <0.010 Base/neutral Cobalt, mg/L <0.020 priority pollutants ND6 Copper, mg/L <0.010 Pesticides/PCBs ND6 Cyanide, mg/L <0.025 Organophosphate Iron3, mg/L 0.001 pesticides5, ng/L <0.50 Fluoride3, mg/L <0.1 : tDate of sample collection 26 May 1992 unless indicated otherwise, analyses performed : | at Environmental Testing and Certification Corporation, Edison, New Jersey; ^ate of sample collection 1 July 1992, performed at Du Font Engineering Test Center, Newark, Delaware; 3Date of collection 10 June 1992, analyses performed at Du Pont Chemicals, Jackson Laboratory, Deepwater, New Jersey; ^elow the Published Method Detection Limit (BMDL); SAnalyses performed at Hazleton Laboratories, Inc.. Madison, Wl; SNone detected; 7Units based on cobalt/platinum reference. ) . . l 1 ^mpany Sanitized. Does not contain TSCA CB& Bu Pont HLR 593-92 STATIC. ACUTE, 96-HOUR LC50 OP H-19520 TO FATHEAD MINNOWS. Pimephales promelas TABLE 2 Measured Concentrations of H-19520^_ in a 96-Hour, Static, Unaerated T( Nominai Concentration (mg/L) Measured Concentration (mg/L) Start End Control 75.4 116 179 275 423 650 1000 0.00 69.7 93.3 132 209 278 455 815 - 0.14 72.9 91.7 86.5 243 343s 5403 907a ^Samples taken at time of total mortality. 12 .ompanySanitized. Does not contain TSCA CB( Du Pont HLR 593-92 STATIC, ACUTE, 96-HOUR LC50 OP H-19520 TO FATHEAD MINNOWS, Pimephales promelas TABLE 3 Water Chemistry of the Control at Test Start Total Alkalinity (mg/L as CaCOa) 80 EOTA Hardness (mg/L as CaCOa) 85 Conductivity (nmhos/cm) 175 13 .CompanySanitized. Does not contain TSCA CBI Du Pom HBLR 593-92 STATIC. ACUTE, 96-^01^1-050 OF H-19520 TO FATHEAD ^INMOWSrSwnflp?ates promelas TABLE 4 Physical H-195; icsof est Solutions Measured Concentration (mg/L) Time OHr 24 Mr 48 Hr 72 Hr 96 Hr Dissolved Oxyoan. mp/L Control (0.14) 72.9 91.7 86.5 243 343 540 907 8.5 7.3 6.7 6.3 5.3 8.5 7.4 6.6 5.9 4.8 8.6 7.4 6.7 6.4 5.4 8.6 7.4 6,7 6.4 5.3 8.5 7.5 6.6 6.8 5.3 8.6 7.9 7.2 6.8 8.5 8.1 -a 8.5 8.2 - - oa Control (0.14) 72.9 91.7 66.5 243 343 540 907 7.5 7.4 7.4 7.4 7.4 7.5 7.4 7.4 7.4 7.4 7.5 7.4 7.4 7.4 7.4 7.5 7.4 7.4 7.4 7.4 7.5 7.4 7.4 7.4 7.4 7.5 7.4 7.4 7.4 - 7.5 7.5 . . ( 7.5 7.5 " - - "Dash (-) indicates that reading was not made due to total mortality. 4 ggmpanx Sanitized. Roes not contain TSCA CBI Du Pont HLR 593-92 STATIC, ACUTE, 96-HOUR LC50 PN-19520 TO FATHEAD MINNOWS, Pimephales promotes TABLE 4, Continued Phhyyssicka^laannddChh9emmjircaKl Cjlhiaariacteristics of H-19520Q------------lTeSsotlutions Measured Concentration (mg/L) Time OHr 24 Hr 48 Hr 72 Hr 96 Hr Temperature. C Control (C 14) 72.9 91.7 86.5 243 343 540 907 21.4 21.4 21.3 21.3 21.3 21.2 21.3 21.2 21.1 21.1 21.1 21.1 21.1 21.0 21.1 21.1 20.9 20.9 20.9 20.9 20.9 20.9 .a 20.8 20.8 20.8 20.8 20.8 20.8 - - 20.8 20.8 20.8 20.8 20.8 - - - "Dash (-) indicates that reading was not made due to total mortality. 15 Doesno^containTSCA CB1 .J ampanx Sanitized. Du Pont HLR 593-92 STATIC. ACUTE. 96-HOUR LC50 OP H-19520 TO FATHEAD MINNOWS. Pimephales promelas TABLE 5 Observreed Moirtality of Fathead-Mti nnows, Pimephales promelas. Exposed to Hl-1-915;9520(UBHR))rr 96 Hours in a Static, Unaerated Test Measured Concentration (mg/L) (Cumulative Moirtality in Percerit3 24 Hours 48 Hours 72 Hours 96 Hours Control (0.14) 0 0 0 0 72.9 0 0 0 0 81.7 0 0 0 0 86.5 0 0 0 0 243 10 10 10 10 343 80 80 100 100 540 100 100 100 100 907 100 100 100 100 'There were ten fish per test concentration at test start. 16 Companx Sanitized. Does not contain TSCA'CBl Du Pont HLR 593-92 STATIC. ACUTE, 96-HOUR LC50 OF H-19520 TO FATHEAD MINNOWS. PimepfSUes promelas TABLE 6 Observed Effects of H-1 Pimephalffs promelas. for Fathead Minnows, Unaerated Test Measured Concentration (mg/L) INumber of Rsh With Effect 24 Hours 48 Hours 72 Hours 96 Hours Control (0.14) 72.9 91.7 86.5 243 343 540 907 0/10 0/10 0/10 0/10 0/9 ia.b/2 .c 0/10 0/10 0/10 0/10 0/9 1b/2 - - 0/10 0/10 0/10 0/10 0/9 - - - 0/10 0/10 0/10 0/10 0/9 - - - ''At the surface. ''Partial loss of equilibrium. ^ash (-) indicates that no reading was made because of total mortality. l 7 .Company Sanitized. Does not co.^'i i S-^H ^ Du Font HLR 593-92 STATIC, ACUTE. 96-HOUR LC50 OF H-19520 TO FATHEAD MINNOWS, Pimephailes pmmelas TABLE 7 ^ P W H-195; IjConcentrationCsausing IMortality to 50l% (LC50) Of Piitheaa Minnows, iftimephales promelas, at Sipedfic Time 1ntervals Time LC50 (mg/U) 95% Limits (mg/L*1) Slope Y-intercept 24 Hours 299 2606 346 14.2 -35.2 48 Hours 299 260C 346 14.2 -35.2 72 Hours 276 237d 317d N/Ab N/A 96 Hours 276 237'1 317<1 N/A N/A "Based upon measured concentrations ^ot applicable. Slope and Y-intercept angle technique. ^S^o fiducial interval. W/o confidence limit. at 96 hours. not calculated by the moving average R 18 Sosr'pany Sanitized. Does not contain TSCA CB( Du Pont HLR 593-92 STATIC. ACUTE, 96-HOUR LC50 OF H-19520 TO FATHEAD MINNOWS. Pimephales promelas APPENDIX 1 PROTOCOL AMENDMENTS and SOP DEVIATIONS PROTOCOL AMENDMENT NUMBER ONE a. ft is not necessary to weigh fish and determine test chamber loading prior to test start. b. Stability and solubility of the test substance in dilution water and under storage conditions will not be determined. c. Samples will be taken from each replicate at test start and end for chemical analysis of the test substance concentration. d. At test end surviving fish will be sacrificed according to aquatic SOl e. Fish will be starved for 24 hours prior to test start. SOP DEVIATIONS a. Dissolved oxygen concentrations on day 4 were less than 60% of saturation, and this observation was not reported to the Study Director. b. The temperature of the water bath was not recorded. c. Temperatures and equilibrium times for individual test chambers were not recorded. PROTOCOL AMENDMENT NUMBER TWO a. The test start date on the short-term study protocol sheet (7-23-92) was incorrect. The start date should have been 7-22-92 as noted in the correction made by Robert Peterson on 7-21-92. 19 Sanitized. Does not contain TSCA CBI .^panx