Document Lp5M7YRJ4o0dG092KxLmXyxoz

AR226-2980 Du Pont H L R 592-92 TRADE SECRET Study Title STATIC, ACUTE, 96-HOUR LC50 OF H-19,517 T O FATHEAD MINNOWS. Pimephales prometas Data Requirement Organisation for Economic Co-Operation and Development (O ECD) Guideline for Testing Chemicals: 203 M2C Keith B. Pierson, Ph.D. Stjd y Completed On November 6, 1992 ling Laboratory E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and industrial Medicine Elkton Road, Post Office Box 50 Newark, Delaware 19714 Laboratory ProiecUP Haskell Laboratory Report No. 592-92 Page 1 o f 19 m m r* ' m Du Pont H L R 592-92 G O O D LA B O R A TO R Y PR A C TIC E S T A T E M E N T This;study was cone Sice Standards 2), and MFF Japan Good Laboratory Practice Standards (59 NohSan No. 3850) with the two exceptions noted below. Solubility and stability of the test substance in dilution w ater 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 SO P deviations appear in Appendix l. Areas of noncomoliance (if any) are also documented in the study records. No deviations existed that affected the scientific validity of the study. Submitter: E. I. du Pont de Nemours and Company Sponsor: Du Pont Chemicals E. I. du Pont de Nemours and Company STUDY DIRECTOR: .P * 4 i* o J Keith B. Pierson, Ph.D. Research Toxicologist Environmental Sciences A jfi1/ m *. M o/D ay/Year ,, tS C A C B I not co' GomPan^ Sn&eL0oeS 2I Du Pont H L R 592-92 TABLE O F CONTENTS Good Laboratory Practice Statement.......................... Page ....2 Table of Contents...................................................................... ....3 General Information............................. .......................... ......... ....4 Sum m ary............................... .....6 Quality Assurance Docum entation............................ 7 I. Purpose.................... 8 II. R ecords........... ........................ ......... .................................................. ........................ 8 III. Materials and M eth o d s................................................................................................ 8 A Test Substance......... .................................... 8 B. Dilution W a te r............................................ .....8 C. Animal C u ltu re ....................................... 8 D. Test M ethods............................................................................................................. 9 E. Chem ical A n alysis......................................... 9 F. Statistical A nalysis............................................................ 10 IV. Results and D is c u s s io n ................................ ..................... ...................................10 V. Conclusion 10 VI. Literature C ite d ............................................................................... .......................... 10 VII. Tables.......................................................................................................................... 11 1. Chem ical Characteristics of Haskell Laboratory W ell W a te r.....................11 2. M easured Concentrations of H -19,517 in a 96-Hour, Static, Unaerated Test ............ ......... ............................................. ..............................12 3. W ater Chemistry of the Dilution W ater at Test S ta rt............................... 13 4. Physical and Chemical Characteristics of H -19,517 Test Solutions....... 14 5. Observed Mortality of Fathead Minnows, Pimephales promelas, Exposed to H -19,517 for 96 Hours in a Static, Unaerated T e s t................16 6. Observed Effects of H -19,517 on Fathead Minnows, Pimephales promelas, for 96 Hours in a Static, Unaerated T e s t.................................... 17 7. H -19,517 Concentrations Causing Mortality to 50% (LC5G) of Fathead Minnows, Pimephales promelas, at Specific Tim e in te rv a ls .... 18 VIII. A p p e n d ix ........................ ................... ...................................................................... 19 1. Protocol Amendments and SO P Deviations................................................. 19 not contain TSC G81 Sanded. 'Does Cowpanf 3 Du Pont H L R 592-92 G EN E R A L IN F O R M A T IO N Material Tested: SMQrmm JHaskalLMimbec Puriiy: Composition: H -19,517 Contaminants: B sglslE i im bsn Not supplied by sponsor None avalad le for this mixture gja?n.S0r; Du Pont Chem icals E. I. du Pont da Nemours and Company Wilmington, Delaware 19898 TSCACBI Sanitized Does not coniato Vi ; Company 4 Company Samtoed. Does not contain TSCA GBl 5 i aai 1 ]* 1 1 _> D u Pont H L R 592-92 STATIC, ACUTE. 96-HOUR LC50 O F H -19,517 TO FATHEAD MINNOWS. Pim&phales promeias SUMMARY H -19,517 exhibited moderate toxicity in a 96-hour, unaerated, static, acute test using fathead minnows, Pimephales promelas. Nominal H -19,517 concentrations w ere 0 ,0 .4 0 9 ,1 .0 2 ,2 .5 9 ,6 .4 1 ,1 6 .0 ,4 0 .0 , and 100 m g/L H -19,517 measured concentrations at 9 6 hours were 0 .0 0 ,0 .1 6 ,0 .5 5 ,1 .8 8 ,4 .0 3 , 7 .4 6 ,1 5 .7 8 , and 48.11 mg/L. The 96-hour LC50 based on measured concentrations at 96 hours vims 4.72 mg/L with a 95% confidence interval of 3.55 to 6 .0 2 mg/L. Work By: __ Robert W. Peterson, B. A. Toxicology Associate A/ # / > i M /D a^/Year Work By: M K ~r Jeffrey T. Turner, Sr. Toxicology Associate ..... tc ij'h A M o/D ay/Year T H IS REPORT IS APPROVED FOR ISSUE. Ci Keith B. Pierson, Ph.D. Research Toxicologist and Study Director Environmental Sciences A/ot/ *72. M o/D ay/Year i i Company Sanitized. Does not contain TSCA CBI 6 SiTM Du Pom HLR 592-92 Q U A L IT Y A SSU R A N C E D O C U M E N TA TIO N Zates of Inspection: Conduct - 7/23/92; 8/5/92 Records, Report(s) - 10/27 - 30/92 r ndinos reported to : Study Director - 10/30/92 Management - 11/6/92 Reported by: Deborah A. Vick Quality Assurance Auditor t'I t , fa t. , IM I , I I INI Date Company Sanitized. Does not contain TSCA CBt 7 Du Pont HLR 592-92 I. PU RPO SE The goal of this study was to assess the acute toxicity of H-1 g *1 7 *n minnows. Pimephafes prom elas. y 9,51 7 10 fathead II. RECORDS All data and the final report will be maintained eithei at Hackoi; 1 , DDuuXpPoonmt RRe^cnodrrdts' n?MafntnaagleMmeendticCineen(tNere(wWairltmi-iDngetloanw,aDree)laowr abree)L. S d at t h " III. M A TER IA LS AND M ETH O D S A. Test S ubstance The test substance is described under General Information Th t :>ct o, was added to dilution water see below) and stirred for 30 minntac v ! ^ Jbstance stock solution. The stock solution ,,a s volumes to make individual lest solutions. Stability and solubility of H fa s r? ,, diluhon water and under storage conditions were n o f d e t e S ' B. D ilu tio n W ater Water (Table 1) originated horn the Haskell Laboratory well. C . A n im al C u ltu re S i 183* PjTMePhL'ss promelas, were reared in continuously flowino well water at Haskell Laboratory. Adult fish resided in 39 L aquaria at 9 rataR maHfti 2 wC nnd-were fed frozen brine Shrimp fArtemia qq Golden Gate Brand, Kordon Division. Novalek, Inc., Hayward C A ^en era l v twirp riaiiu cchhnl,onnrdid0e: ^tileas,ucr<olaarted5nwdi.thnsoaenodn. STuilensdawyi,th sebqgqssVweerree daeer^atseidlSanod ^inSTMtnhiiyto?w ' ' approximately 25C in a water bath. L a r v ^ r ^ T ? 9T L S 2 ^ 31 approx'mately 21C and were fed newly hatched M s s m so(A ouarium weekend' G,f n Bufniei MD) generally twice daily on weekdays and daily on weekends. Juveniles were held either in 272-L circular, fiberglass tanks or in aquaPaJ 23-cm depth; 50 (L) x 25 (W) x 31 cm (H)) at a p p S a te ly n ln 2 rin d re !?d, new y hatched M e m ia Sk- and/or frozen brine shrimp V generally twice daily on weekdays, once on Saturday and once on Sundav Sickness, injury, and abnormality were not observed. Animals were identified by labels on aquaria and test vessels. laentitied CompanyVnkized. Does ,, a , TSCACB, 8 Du Pont HLR 592-92 D . T es t M eth o d s [1] Seven test concentrations and a water control were used. Nominal concentrations were 0, 0.409, 1.02, 2.59, 6.41,16.0, 40.0, and 100 mg/L. Control fish ranged from 1.7 to 2.4 cm standard length (mean 1.9 cm) and 0.11 to 0.25 g wet weight (mean 0.18 g). Erlenmeyer 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 Wrap to prevent loss of potentially volatile test substance components. The position of test chambers in the water bath was determined by use of random nu ' jrs. Ten fish were added to each replicate using random numbers (one replicate per concentration, total 10 fish per concentration). Control loading was 0.40 g/L at test conclusion. Test solutions ranged from 20.7 to 21.5C (mean 21.0C). Fish were not fed 27 hours prior to nor during the test. A photoperiod of 16 hours light (172 to 215 Lux) versus 8 hours darkness was employed with 25 minutes of transitional light (2 to 3 Lux) preceding and following the 16 hour light interval. Mortality and behavioral effects were noted da: >. Dissolved oxygen (YSI Model 54A, Yellow Springs Instruments, Yellow Springs, OH) and pH (Corning Model 610A, Corning Scientific Instruments, Medfield, MAI were measured in the water control and all test concentrations before fish were added at the beginning of the test and daily thereafter. Temperature was measured (American Society of Testing Materials registered mercury thermometer) in the water control and ail test concentrations before fish were added at the beginning of the test, daily thereafter, arid at the end of the test. Total alkalinity (Hach kit, Hach Chemical Company, Loveland, CO), ED FA 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 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. v E. C h em ical A n alysis Analyses were conducted after the test had ended at the Du Pont Jackson Laboratory at Chambers Works, Deepwater, NJ. Samples and standards were completely combusted in a hvdrogen/oxygen Wickbold Torch. The resulting water was then analyzed for total fluoride concentration using an Orion fluoride ion selective electrode. Concentrations were then calculated from the standards using either a three or four point calibration curve. Company Sanitized. Does not contain TSCA CBI 9 Du Pont HLR 592-92 F . S ta tis tic a l A n alysis The 24-, 48-, 72-, and 96-hour LC50s were calculated by using the moving average angle technique [2]. IV . RESULTS AND D ISC U S SIO N A rangefinding test with five fish per concentration (4.4 L Erlenmeyer flask filled to the top) was conducted with a water conrol and test concentrations of 0.1, 1, 10, 100, and 1000 mg/L. Mortalities were 0. 3. 0, 60,100, and 100%, respectively. Two definitive studies were conducted. Data from the first were discarded because of low dissolved oxygen concer :'ations, and data from the second are reported herein. Nominal concentrations in the definitive test were 0, 0.409, 1.02, 2.59, 6.41, 16.0, 40.0, and 100 mg/L. Measured concentrations at 96 hours were 0.00, 0.16, 0.55, 1.88, 4.03, 7.46, 15.78 and 48.11 mg L. The discrepancy between the measured and nominal concentrations (Table 2; ma< stem from the use of a stock solution in which the test substance was not totally c ssolved. All chemical and physical parameters in the final definitive test (Tastes 3 and 4) were within expected ranges with one exception. The dissolves oxygen concentration in the control and 4.03 mg/L test solutions on day 4 drooped to 58 and 56% of saturation, respectively. This observation was not c' biological significance since fathead minnows can survive at much lower dissoved oxygen concentrations for extended periods. Behavioral observations appear in Table 6. The 96-hour LC50 based on measured concentrations at 96 hours was 4.72 mg/L with a 95% confidence interval of 3.55-6.02 mg. L (Taoies 5 and 7). V. CONCLUSION H-19,517 was moderately toxic to faiheao minnows, Pimephales promelas, in a 96-hour, static, unaerated test. V I. LITERATURE CITED 1. Organisation for Economic Co-Opers: on and Development (OECD). Guideline for Testing Chemicals: 203 4 April 1984.2 2. Peltier, W. H. and C. I. Weber, Eds. 1985. Methods for measuring the acute toxicity of effluents to freshwater and -narine organisms. EPA/600/4-85/013. United States Environmental Protect on Agency, Environmental Monitoring and Support Laboratory, Cincinnati, C-nio. Appendix E. Company sanitized. Done no, contain TSCA CBI 10 Du Pont HLR 592-92 STATIC, ACUTE, 96-HOUR LC50 OF H-19,517 TO FATHEAD MINNOWS, Pimephales promelas TABLE 1 Chemical Characteristics of Haskell Laboratory Well Water1* Parameter Concentration BOD, mg/L COD, mg/L DOC, mg/L TOC2, mg/L Total Kjeidahl N, mg/L Ammonia N, mg/L Turbidity, NTU Phenolics, mg/L Color, apparent Co/Pt7 S o lid s Total suspended, mg/L Aluminum, mg/L Antimony, mg/L Arsenic, mg/L Beryllium, mg/L Boron4, mg/L Cadmium4, mg/L Calcium, mg/L Chloride3- mg/L Chromium, mg/L Cobalt, mg/L Copper, mg/L Cyanide, mg/L Iron3, mg/L Fluoride3, mg/L <2 <10 1.4 2.9 0.3 <0.2 <1.0 <0.050 <5 <4 <0.100 <0.060 <0.010 <0.001 0.020 <0.002 24.3 6.2 <0.010 <0.020 <0.010 <0.025 0.001 <0.1 Parameter 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 Zinc, mg/L Volatile priority pollutants Acid extractable priority pollutants Base/neutral priority pollutants Pesticides/PCBs Organophosphate pesticides5, ng/L Concentration <0.075 3.7 <0.05 <0.00020 <0.020 <0.1 1.9 <0.1 1.7 <0.005 <0.010 8.2 5.0 <0.10 <0.058 ND6 ND5 ND5 ND6 <0.50 1Date of sample collection 26 May 7992 unless indicated otherwise, analyses performed at Environmental Testing and Certification Corporation, Edison, New Jersey; 2Date of sample collection 1 July 1992, performed at Du Pont Engineering Test Center, Newark, Delaware; 3Date of collection 10 June 1992, analyses performed at Du Pont Chemicals, Jackson ' Laboratory, Deepwater, New Jersey; '`Below the Published Method Detection Limit (BMDL); 5Anaiyses performed at Hazleton Laboratories, Inc., Madison, Wl; 6None detected; 7Units ' based on cobalt/piatinum reference. Company Sanitized. Does not contain TSCA CB! Il Du Pont HLR 592-92 STATIC, ACUTE, 96-HOUR LC50 OF H-19,517 TO FATHEAD MINNOWS, Pimephales promelas TABLE 2 Measured Concentrations of H-19,517 in a 96-Hour, Static, Unaerated Test Nominal Concentration (mg/L) . Measured Concentration (mg/L) Start End Water Control 0.409 1.02 2.59 6.41 16.0 40.0 100 0.00 0.40 0.76 2.38 3.87 9.86 18.85 53.76 0.00 0.16 0.55 1.88 4.03 7.46a 15.78 48.11 aSamples taken at time of total mortality. Company sanitked. Does of contain TSCACBi Du Pont HLR 592-92 STATIC, ACUTE, 96-HOUR LC50 OF H-19 517 TO FATHEAD MINNOWS, Pimephales promelas TABLE 3 W ater Chemistry of the Dilution Water at Test Start Total Alkalinity (mg/L as C aC 03) 79 EDTA Hardness (mg/L as CaC03) 77 Conductivity (iim hos/cm ) 170 Con>pany Sanitized. c oes not contain TSCACS1 Du Pom HLR 592-92 STATIC, ACUTE, 96-HOUR LC50 OF H-19,517 TO FATHEAD MINNOWS, Pimephales promelas TABLE 4 Physical and Chemical Characteristics of H-19,517 Test Solutions M easured C o n c e n tra tio n (mg/L) D^solved Oxvoen. mg/l H20 Control 0.16 0.55 1.88 4.03 7 46 15.78 48.11 m H20 Control 0.16 0 .5 5 1.88 4.03 7.46 15.78 48 { t Time Q Hr 24 Hr 48 Hr 72 Hr 96 Hr 8.4 7.0 6.4 6.0 5.2 8.4 7.0 6.4 6.1 6.0 8.4 7.2 6.7 6.3 5.7 8.4 7.2 6.6 6.3 5.4 8.5 8.5 7.4 7.7 6a.9 6.4 a 5a.0 8.4 8.0 a a a 8.4 8.0 a a a 7.6 7.4 7.4 7.4 7.0 7.6 7.4 7.4 7.4 7.0 7.6 7.4 7.4 7.4 7.1 7.6 7.4 7.4 7.4 7.1 7.7 7.4 7.4 7.4 7.1 7.6 7.4 a a a 7.6 7.5 a a a 7.6 7.5 a a a aReading was not made due to total mortality. Coa* ^ Sana 14 CB|contain TSCa Du Pont HLR 592-92 STATIC, ACUTE 36-HOUR LC50 OF H-19,517 TO FATHEAD MINNOWS, Pimephales promelas TABLE 4, C ontinued Physical and Che~ :ai Characteristics of H-19,517 Test Solutions Measured Concentration (mg/L) Temperature. C H20 Control 0.16 0.55 1.88 4.03 7.46 15.73 48.11 Time 0 24 Hr 48 Hr 72 Hr 96 Hr 20 ~ 20 ~ 20.8 203 20 ~ 20-5 20 8 20 E 20.9 20.9 20.9 20.9 20.9 20.9 20.9 20.9 20.8 20.8 20.8 20.8 20a.8 a a 21.5 21.5 21.5 21.5 21.5 a a a 21.4 21.4 21.4 21.4 21a.4 a a aReading was not made due :: :::ai mortality. Company Sanitized. Coes not contain TSCA C8t Du Pont HLR 592-92 STATIC, ACUTE, 96-HOUR LC50 OF H-19 517 TO FATHEAD MINNOWS, Pimephales promelas TABLE 5 Observed Mortality of Fathead Minnows, Pimephales promelas Exposed to H-19,517 for 96 Hours in a Static, Unaerated T e st' Measured Concentration (mg/L) Water Control 0.16 0.55 1.88 4.03 7.46 15.78 48.11 Cumulative Mortality in Percent3 24 Hours 48 Hours 72 Hours 96 Hours 0 0 0 0 0 . 100 100 100 0 0 0 0 20 100 100 100 0 0 0 0 20 100 100 100 0 0 0 0 20 100 100 100 a There were ten fish per test concentration at test start. ttiflp w y S iftStteiA S W not contain T8C A CBI I6 Du Pom HLR 592-92 STATIC, ACUTE, 96-HOUR LC50 OF H-19 517 TO FATHEAD MINNOWS, Pimephales promelas TABLE 6 Observed Effects of H-19.517 for 96 Hours on Fathead Minnows, in a Static, Unaerated Pimephales Test p-omelas, Measured Concentrations (mg/L) Water Control 0.16 0.55 1.88 4.03 7.46 15.78 48.11 Number of Fish With Effect 24 Hours 48 Hours 72 Hours 0/10 0/10 0/10 0/10 1a/10 1b/10 c c c 0/10 0/10 0/10 0/10 3b/8 C C c 0/10 0/10 0/10 0/10 3b-8 C c c aLying on bottom of the test vessels. bPartiai loss o f equilibrium. cTotal mortality. 96 Hours 0/10 0/10 0/10 0/10 3b/8 C C c Company Sanitized. Does not contain TSCA GBI Du Pont HLR 592-92 STATIC, ACUTE, 96-HOUR LC5Q OF H-19 517 TO FATHEAD MINNOWS. P m e p ^ p r a m /Is TABLE 7 Fathalw kl*' Concentrations Causing Mortality to 50% (LC50) of Fathead Minnows, Pimephales promeias, at Specific Time ir Time 24 Hours 48 Hours 72 Hours 96 Hours LC50 (m g/La) 5.48 4.72 4.72 4.72 95% Confidence Limits (mg/La) Slope 4.24 7.09 N/Ab 3-55 6.02 N/A 3.55 6.02 N/A 3.55 6.02 N/A Y-intercept N/A N/A N/A N/A ^Based upc~ measured concentrations at 96 hours angle te c h iiq u f S' pe ^ Y`in,ercepi "* calculated by the ng average Company Sanitized. Does not contain TSCA CBI Du Pom HLR 592-92 STATIC, ACUTE, 96-HOUR LC50 OF H-19 51 ~ TO FATHEAD MINNOWS. Pimephales promelas A P P E N D IX 1 P R O T O C O L A M EN D M EN TS and SO P DEVIA TIO N S PROTOCOL AMENDMENT NUMBER ONE a' f not necessary to weigh fish and determine test chamber lose ng prior to icsi stan. b. Stability and solubility of the test substance in dilution water anc jnder storage conditions will not be determined. c. Samples will be taken from each replicate at test start and end chemical analysis of the test substance concentration. w d. At test end surviving fish will be sacrificed according to aquatic c DP AQC29- P-002. ~ e. Fish will be starved for 24 hours prior to test start PROTOCOL AMENDMENT NUMBER TWO a. The test substance concentrations were changed to facilitate th- -veighing process and w ill be: 0.409, 1.02, 2.59, 6.41, 16.0, 40.0, and 10C ~ig/L SOP DEVIATIONS a. Temperature of water bath was not recorded. b. Temperatures and times to equilibria of individual test container* .vere not recorded. c Maintenance was not performed on balance L1642 when calibra: on was out of Standard Operating Procedure range. d. Which pH calibration buffers were used was not recorded durinc oH meter calibration on 7-25-92, 7-26-92, 7-27-92, and 8-5-92 through 8-9-92. e. Study Director not notified in some test solution. on 8-9-92 of low dissolved oxyqen co centration c fnpany Sanitized. Does not contain TSCA CBI I9