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AR226-2979! f -t'rs#*.~j'jfc'i~S?Sk -, -/ 1 te*1"-_B u Pont HLR 584-92 -p***r~t V i; - &fez mmm * J- r I ,s*-ki,). '.`S*^ s e c j s t Study Title ;? STATIC, ACUTE, 96-HOURLG50 pF$1-19,518 TO FATHEAD MINNOWS, Pimcphaies promelas Qsstfiw tflrcfow t Organisation for Economic Co Operation and fyelopment (OECD) Guideline for Testing Chemicals: 23 Author Keith B. Pierson, Ph.D. Study Completed On October 30,1992 Earfarminq laboratory E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Rond, Post Office Box SO Newark, Delaware 19714 Labs atorv Protect ID Haskell Laboratory Report No. 584-92 S5 <0OSx?S> Pge 1 of 18 G& iascmtsKcms-iszmfmimm Ig; P ^ i& OE iS This study was conductedaoc trsCA Good Laboratory Practice Standards (# P R ij|, ID Principles of Good Laboratory Practice {0<81)30{^l !F Japan Good Laboratory Practice Stanfar'*"' rith the two exceptions noted below. S o lu b ility la l^ e in dilution water and under storage o o o d to ^ flM jL , [rations of the test substance in dilution water w ia \ leasurements were not made in accordance ^ - ,, Protocol an_mo_ne_cn_odo_mmi_npeil:niatinscsaernnt(diiwtf/ounatrnn.SoyA)OuaPrredawaelie^sa>oliiOuWdnhoroics/juoilirpmwnpaewennte!aeiHdrispiinr the I. Areas study records. of No . deviations existed that affected the scient! valid# fire study. Submitter: E. I. dii Pont de Nemours a n fCompany Sponsor: Du Porat Chemicals E. I. du Pontd Nemours and Company ) STUDY DIRECTOR: /st : Keith B. Pierson, Ph.D. 3o per iw *. Mo/Day/Year Reses rch Toxicologist Environmental Sciences t*A' 2 Du Pont HLR 584-92 TABLE OF CONTENTS Good Laboratory Practice Statement.................... ............................................. Page .....2 Table of C ontents............ ........\ ...................................................................................... General Information................................. ..................................... ............................ ..4 S u m m a ry .................................. ................................................................................... 5 Quality Assurance Documentation.................................................... . L Purpose ..6B a a o e e o e e B B s e 7 II. Records........... ....................................... .................................. ....................... 7 III. Materials and Methods........................................................................... ..........7 A. Test Substance............................................................................................. 7 B. Dilution W ater.................................................................................. 7 C. Animal Culture .............................................................................................. 7 D. Test Methods............ ............................................................................... E. Chemical Analysis........... ............................................................................ 8 F. Statistical Analysis................................ 8 IV. Results and Discussion.............................. 9 V. C onclusion.................... 9 VI. Literature C ited......................... 9 VII. Tables.............................................................................................................. 10 1. Chemical Characteristics of Haskell Laboratory Well W ater. . . . . . . . . . . . . 10 2. Measured Concentrations of H-19,518 in a 96 Hour, Static, Unaerated T est............................. ....................... .................................. 11 3. Water Chemistry of th$ Dilution Water at Test Start................................ 12 4. Physical and Chemical Characteristics of H-19,518 Test Solutions.......13 5. Observed Mortality of Fathead Minnows, PimephaJes promelas, Exposed to H-19,518 for 96 Hours in a Static, Unaerated Test.............. 15 6. Observed Effects of Hr19,5l8 on Fathead Minnows, Pimephales promelas, for 96 Hours in a Static, Unaerated T e st................................ 16 7. H-19,518 Concentrations Causing Mortality to 50% (LC50) of Fathead Minnows, Pimephales promelas, at Specific Time Intervals.... 17 VIII. Appendix.................................................... ................. ................................ is 1. Protocol Amendments and SOP Deviations................................ ........... 1 8 0O9off S8*"1* 3 CtP8** C& Putita ^imposition: I O onim m s^ HAS Registry Number: Sponsor; study initiated/Comoleted: In-Life study initiated/CflfflOifilgd; Du Pont Chemicals E. s.cfu Pont de.Nemours and Company Wilmington, Delaware 19898 8-7-92 / 8-10-92 / 8- 14-92 notcontain XSCACBl S a n tt^ e d . D o e s C om ply 4 ' -v m>$>4,'iwSxlMtei,,- . - [Al:ii<vW. riiSiLISli .<?"t-wp*f'm! Du Pont HLR 584-92 STATIC, TO FAT! 19,518 iromelas jj;*.1 H-19,518 was slightly toxic in a 9--6-hoTu,,r, unseraiid>s ^ i-c--,-a--c-u--t-e--t-o--s-t-u---singf-a--t-b-e--a-d minnows, Plmephakh sl war 0,8.30, 18.5, 41.0,91 1.2 03 ,450i and 1000 mg. _ ^concentrations at 96 hours were 1.51,6.96,15.8, 36.8,87.2,1 8 7 , T--h-e---9-6----h-o--u--r--L-C--5--0-. . based on measured concentrations at 96 hours w ^ j6|>7 mg/L with a 95% fudicia! interval of 44.2 - 84.9 mg/L w Work By: Roheit fl. Peterson.B. A, Toxicology Associate urfiA.i 2 = Ao/uay/Year Work By: _ Wetzel, B. S Associate Mo/Ody/Year Jis8 Aim % THIS REPORT IS APPROVED FOR ISSUE. e. A Keith B. Pierson, Ph.D. Research Toxicologist and Study Director Environmental Sciences 5p Qur?993L Mo/Day/Year Sanitized. Does not contain TSCA CB1 Company Du Pont HLR 584-92 i | QUALITY ASSURANCE DOCUMENTATION (H-19518) nf Inspection: j Conduct - 8/10/92 Records. Report(s) -10/10/92 Findings reported la: Study Director -10/12/92 Management -10/30/92 Reported byj5 j W o j l jf^ W ^ l&f Roberte. Rhea * niiaiitv Assurance Auditor D a te ` Company Sanitized. Does not contain TSCA CBI 6 The goal of this study was.to , minnows, P im ep h ale^ ^^S ^ Alt data and the final report Laboratory for Toxicology and I n d u s t d a i b e archived at the Du Pont Records Management <tenter {Wilmingto&pejaware;. Ilf. MATERIALS AMD ^ A. Test Substance The test substance is described under General Information. The test substance was added to dilution water {see below) and stirre^ fb llb minutes to create a stock solution. Visual inspectionsuggested that trie ^ s u b s ta n c e was in solution in the stock. The stock solution was addidsodilution water in appropriate volumes to make iindividual test solutions. Stability and solubility of H-19,518 in dilution water and urider storage conditions were not determined. j) B. Dilution Water Water (Table 1) originated from the Haskell Laboratory well. C. Animal Culture Fathead Minnows, Pimephales uromelas, were reared in continuously flowing well water at Haskell Laboratory, Adult fish resided tn 39 L aquaria at approximately 25C and were fcd frozen brine shrimp j($rtemia sp., Golden Gate Brand, Kordon Division, Nwalek, Inc., Hayward, GA) generally twice daily, once on Saturday rod once onilunday, Eggs were deposited on polyvinyl chloride tiles coated with sand, lie s with eggs vtfere aerated and incubated at approximately 25C in a w atertith. LaryaiWbre reanfi in 39 L aquana at approximately 21C and were fed newly h a t# ie d -M a s p . (Aquarium Products, Glen Burnie, MD) generally twice m ym prnkdays and daily on weekends. Juveniles were helc efther in 272-L cirCula^ fiberglass tanks or in 39 L glass aquaria (23-cm depth; SO(L) x 25 (W) x 3 i % (H)) at approximately 21C and were fed newly hatch ad Artemia so. and/or frozen brine shrimp generally twice daily on weekdays, oftee on Saturday and once on Sunday. Sickness, injury, and abnormally were not observed. Animals were identified by labels'on aquaria and test v<issels. notcoataia TaCA CBI 7 w m m m m Pont m 5 8 * # f INominal concentrations _ imi fish tangedJam 3.051 g wet weight Ertenmeyer glass------ test solution (35<cra d e M M totally filled to minimize prevent loss of potential yffp j added to each rep licate;^ |p total 10 fish per concentration) Test solutions tanged from 21-1 21(li^e ttt if t tp );x35(heiht);G m ] containing 4.4 L of Each flask was Saran W raf^to Ten fish wens per concentration, _ J j a |3 s i g/L at test conclusion. p ^ | Were not fed 28.5 hours prior to nor during the test. " Iggrs light versus 8 hours darkness was employed sdth 25 miputesLof transifippg light preceding and f.o.l.lowi.ng the 16 hour light in--terv--a*1l Mortality and behavioral effects were noted daily. Dissolved oxygen (YSI Model 54^ Yellow Springs iristajments, Yellow Springs, OH) and pH (Coming Model 616, Coring Scientific Instruments, MedfiekJ, MA) were measured in the water coritfpl and all test concentrations before fish were added at the beginning of the test and d^|y thereafter; temperature was measured (American Society of Ta3sting Materials registered mercury thermometer) in the water control arid aM^estlbonceotm^ns before fish were added at the beginning of the test dally thiiisafter, aridfat the end of the test, Total alkalinity (Hach kit, Hach Cftiemical Company,'Loveland, CO), EDTA hardness (Hach kit, Hach Chem:al Company, Loveland, CO), and conductivity (YSi Model 33, Yellow Springs Inpruimerife, ^ llo w Springs. OH) of the dilution water were measured at the begH'|Ming of thetest. Aabhtinuously-recording thermometer in a control vessel v|is usdd to check for temperature variation during the 96-hour test. E. Chemical Analysis All analyses were conducted after the test was completed at the Du Pont Jackson Laboratory at Chambers! Works, Deepwater, NJ. Samples and standards were completely cam ftltedTn a "hydrogen?o^gen Wickboid Torch. The resulting water was then analyzed for total fluOride cosnncceentration using an Orion fluoride ion selective elect! ride. Concentrations were then calculated from the standards using either aj three dr fbur point calibration curve. F. Statistical Analysis The 24-, 48-, 72-, and 96-hour LCSOs were caicujifedly the SAS Probit method [2]. Company Sanitized. Does not contain TSCA CBI mmmmm I Pont M M m W IS? #lt gwm A rangefinding testwith the top) was conducted 100, and 1000 mg/L Mo)....,, lenmeyer flask filledto M e n s of 0.1,1,10, respectively. Nominal concentrations 450, and 1000 mg/L. Measured co 15.8,36.8,87.2,137,3 9 ^ i measured increase was 3.5, 41.0, 91.1, 203, were 1.51,6.96, Kpected increase in sfic cause for this ibstance was not stance in the water control was noted, but physical parameters in If cher"j ^ 1anci 3 ^ l^ p f e r e within expected ranges except for d i s s o h lg l il^ f t |w f v e d oxygen concentration dropped as low as 49^> Of sM M t;on o h ite ^ 2 ,3, and 4; however, these observations were npf pf fathead minnows can survive at liudh io w e f .^ extended periods. Behaviprai pbseitfatipnsaj^eann^pp 6. The 96-hour LC50 was 61.7 mg/L with a 95% fiducial interval of 44s2 -16p.9 mg/L (Tables 5 and 7) based upon measured conmirations at 96 hours; ) V. O C C L U S IO N H-19,518 was slightly toxic to fatheiid minnows, Pimeppales promelas, in a 96-hour, static, unaerated test, VI. RATURE C iT E i -,*''1 -, 1 Organisation for Economic Co-CRelation and eyilppment (OECD). Guideline for Testing Chemicals: 203. 4 April 1984; 2. Finney, D. J. 1971. probit Analysis, 3rd Ed;, Cambridge University Press ` 333 p. - -r*m S' i 'r/jfj `S'l >~` Com panjrSanitized. Does not contain TSCA CB I Du Pont HLR 584-92 STATIC. ACUTE. 96-HOUR LC50 OF H-19.516 TO FATHEAD MINNOWS, Phmphales prometas l ! TABLE 1 Chemical Characteristics of Haskell Laboratory Well Water1* 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, apparent Co/Pt7 Solids 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 0.8 <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 ne, mg/L Volatile priority pollutants Acid extractable priority pollutants Base/neutral priority pollutants Pestiddes/PCBs Orgnphosphate pesticides5, pg/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 NP6 ND6 ND6 ND6 <0.50 iDate of sample collection 26 May 1992 unless indicated otherwise, analyses performed at Environmental Testing and Certification Corporation, Edison, New Jersey; J a te of sample collection 1 July 1992, performediat Du Pont Engineering Test Center, Newark, Delaware, 3Date of analyses 10 June 1992, analyses performed at Du Pont Chemicals, Jackson Laboratory, Deepwater, New Jersey; 4Below the Published Method Detecbon mit (BMDL). Analyses'performed at Hazleton Laboratories, Inc., Madison, Wl; 6None detected; 7Umfe> based on cobalt/platinum referents. notconla/arsCACat 10 Pont HLR 584-92 " TM * a>eaPmmelaS TABLE 2 nt M-19 51 ,n a l -- Measured C o n c e n tr^ o n ^ U Sam ples taken at time of totial mortality a notcontain TSCACBl l1 Du Pont HLR 584-92 STATIC, LC5JF 1 ^ 1 9 ^ 8 TO FATHEAD M NNOWS, Pmephales promelas TABLE 3 Water Chemistry Of the Dilution Water at Test Start Total Alkalinity (mg/L as CaCQa) 80 EDTA Hardness :(mg/L sis CaC03) 78 Conductivity (lunhos/cm) 170 J 12 m Pont HLR 584-92 STATIC, ACUTE, 98-HOUR LC50 OF H-19,518 TO FATHEAD MINNOWS, Pimephalespmmelas |TABLE 4 Physical and Chenlfcal Characteristics of H-19,518 Test Solutions Measured Concentration (mg/L) Time OHr 24 Hr 48 Hr 72 Hr 96 Hr TTlfiffftlY**1 Oxvoen._C:ciL H2OControl (1.51) 6.96 15.8 36.8 87.2 187 395 997 8.4 8.4 8.3 8.$ 8.3 8.4 8.4 8.4 6.9 7.3 7.2 7.4 7.4 7.5 7.7 7.9 5.2 5.7 6.1 6.6 6.4 6.7 a a 4.8 4.9 5.5 5.8 5.9 6.5 a a 4.4 4.5 5.0 5.1 5.1 a a a Qtt Hz0 Control (1.51) 6.96 15.8 36.8 87.2 187 395 997 7.5 7 .$ 7.5 7.5 7.$ 7.5 7.$ 7.6 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.3 7.4 7.4 7.4 7.4 7.4 7.4 a a 7.4 7.4 7.4 7.4 7.4 7.4 a a 7.4 7.4 7.4 7.4 7.4 a a a Reading was not made because of total mortality. Coi?}, 13 0oesnote.oiitait, f SCA OB/ Da Pont HLR 5P4-92 STATIC AOUTE, 91-HOUR LC50 OF H-19,518 TOFAT&EAD MINn OWS, pm phales pmmelas TABLE 4, Continued Physical and Chemickl Characteristics of H-19,518 y Tent Solutions Measured Concentration (mg/L) Time 0 Hr 24 Hr 48 Hr 72 Hr 96 Hr frmperature. C i20 Control (1.51) 6.96 15.8 36.8 87.2 187 395 997 21.2 21.2 21.2 21.2 21.2 21.3 21.3 21.4 21.2 21.2 21.2 21.2 21.2 21.2 21.2 21.2 21.1 21.1 21.1 21.1 21.1 21 ;1 a Reading was not made because of total mortality. 21.3 21.3 21.3 21.3 21.3 21.4 21.3 21.3 21.3 21.3 21.3 21.3 a Com'," y s ,, >M oes 14 SCACfltf Da Pont HLR 584-92 STATIC, ACUTE, 96-HOUR LC50 OF H-19,518 TO FATHEAD MINNOWS, Plmephales pmmelas TABLE 5 Observed Mortality of Fathead Minnows, Plmephales promelas, Exposed to H-19,518 for ^6 Hours In a Static. Unaerated Test M aam im d Concentration (mg/L) Cumulative Mortality in Percent8 _l---------------- 24 Hours ! 48 Hours 72 Hours 96 Hours Water Control (1.51) 0 0 0 0 6.96 0 0 0 0 15.8 00 00 36.8 10 10 10 10 87.2 50 50 60 80 187 80 80 100 100 395 100 100 100 100 997 100 100 100 100 There were ten fisn per test concentration at test start. 15 STATIC, ACOTEJ . TO FATHEAD MINNOWS TABLE 8 Du Pont HLR 584-92 9,518 Measured Concentration (mg/L) 24 Hours Water Control (1.51) 6.96 15.8 36.8 87.2 187 395 997 0/10 0/10 0/1 c 0/9 0/5 2a/2 b b Number of Fish With Effect 48 Hours 72 Hours 96 Hours 0 0 0 0/9 2a/5 2a/2 b b 0 0 0 3c/9 4a.d/4 b b b 0 0 0 2c/9 2a.d/2 b b b apartial ioss of & jrlibrium. Total mortality. . cLySng on bottom of the test vessels. Erratic swimming. ComPnyS a n ity Does nei contain ?SCAC8J 16 Du Pont HLR 584-92 STATIC, ACUITE, 96-HOUR LC50 QFH-19,518 TO FATHEAD MINNOWS, mephales prometas I TABLE 7 Time 24 Hours 48 Hours 72 Hours 96 Hours LC50 (mg/La) 91.8 91.8 71.9 61.7 5% Fiducial Limits : (mg/La) 61.9 134 61.9 134 50.9 101 44.2 84.9 Slope 3.38 3.38 4.93 5.82 asased upon measured concentrations at 96 hours. Y-intercept -6.63 -6.63 -9.15 -10.4 Goto 11 o e s not rSCA CBi Du Pont HLR 584-92 STATIC, ACllrrE. 96-HOUR LC50 OF H-19,518 TO FATHEAD MINNOWS, Pfmephales pmmelas APPENDIX 1 PROTOCOL AMENDMENTS and SOP DEVIATIONS PROTOCOL AMENDMENT NUMBER ONE a. It Is not necessary to weigh fiih and determine test chamber loading prior to test start. b. Stability and solubility ofthetestsubstance 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 aquati- ] . e. Fish will be starved for 24 hours prior to test start. SOP DEVIATIONS a. Readings were not made to measure the amount of light present during the primary and transition lighting periods. Dissolved oxygen concentrations lower than 60% rm rrred In some test b. concentrations and were not reported to the Study Director. c. W ater bath tem perature was not recorded. d. Tem peratures and equilibrium times for individual test chambers were not recorded. Conn'Patty 18 noj