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AR226-2798 DuPont BLR 61-94 Study Title STATIC, ACUTE, -o-HOUR LC50 OF H-19704 TO BLUEGILL SUKFISH, Lepoils acrochirus Laboratory Project ID "askell Laboratory Report He. 61-94 Data Requirement United States Environmental Protection Agency 40 CFP 797.1400 Fish Acuf Toxicity Test Toxic Substances Control Act Author Keith E. Pierson, Ph.D. Study Completed On 26 August 1994 Perforning Laboratory Haskell .. I. du Font Ie Ne>ours and Company Laboratory for Toxicology and Industrial Elkton Road. Post Office Box 50 Newark. Delaware 19714 Medicine ^Medical Research Project &-- Page 1 of 20 ^^"^"^"...con.a.nTsCACB, GOOD LABORA-m PRACTICE COMPLIANCE STATEMENT This study was conducted in compliance vith EPA PIFRA (40 CFR 160) and EPA TSCA (40 CFR 792) Good Laboratory Practice Standards, OBCD Principles of Good Laboratory Practice (C(81)30(Pinal), Annex 2), and MAFF Japan Good Laboratory Practice Standards (59 NohSan No. 3850) except for the deviations documented and explained belov. 1. Stability and solubility of the test substance under test and storage conditions vere not determined. 2. Tetranin flake food was not tested for pesticide content. The deviation(s) did not affect the integrity of the study. Submit ter: Sponsor: E. I. du Font de Nenour..: and Company DuPont Elastomers t. I. du Pont de Nemours and Company Study Director: Ai/^C 4. P/'CMf^ Keith B. Pierson, Ph.D. Research Yoxicologist CoBpany Representative: Registration Manager Date ^ ffu^osrHf^ ^'S^^.--^^.^ DuPont Bill 61-94 TABU OF LUW1BM1S Page Good Laboratory Practice Compliance Statement 2 Table of Contents General Information .............. Su--ary ............. Quality Assurance Docuaentation .......................... I. ................. II. ................. III. Purpose Records Materials and Methods A. Test Substance .......... B. Dilution Water . . . ............ C. Aniaal Culture . . . D. Test Methods . . . B. Chemical Analysis . 3 4 . 5 6 . 7 7 7 7 7 7 7 8 P. Statistical Methods 8 IV. Results and Discussion 8 V. Conclusion 9 .................... VI. Literature Cited 9 ......................... VII. Tables 10 ....................... Table 1. Chemical Characteristics of Baskell Laboratory Veil ............................ Water 10 ....................... ymSllWI^ Table 2. Water Chemistry of the Dilution Water at Test Start 11 Table 3. Physical^ard Cheaicaj. Characteristics of B-1970& Test Solutions . 12 Table 4. Mortality ot BluegiHSunfish, Lepoais acrochirus, Exposed to B-19704TWU 96 Hours in a Static, Unaerated TeSt 14 Table 5. Behavior of BluefiU, Sunftsh, Lcpoais acrochirus, Exposed to H-19704j4BIMBBfjll!oF*96 Hours in a Staj.ic, Unaeragg Test 16 Table 6. H-19704lfBB--Bdconcentrations Causing . . . . . . . . Mortality-to 50Z (LC50) of Bluegill Sunfish, Lcpoais acrochirus 17 Table 7. Standard Lengths and Wet Weights of Control Fish .............. At Study End 18 VIII. Appendices 19 .................................................. 1. Protocol Aaendaents and SQP Deviations 19 Company Sanitized. Does not contain TSCA CBl Material Tested; Haskell Huaber; Purity; Composition; Contaainants: CAS Registry Nmber; Sponsor: Study Initiated/CoBpleted; In-Life Study Initiated/CoBpleted; GHBAL IHFOUtATION Octanolc acid, pentadecafluoro-, saa-aniw salt H-1970A S-^ None reported 3825-26-1 DuPont ElastoBers E. I. du Font de Leiours and Competv Viloington, Delaware 19898 13 August 1993 / 26 August 1994 23 August 1993 / 27 August 1993 Company Sanitized. Does no? contain TSCA CB; DuPont BLR 61-94 suMton H-19704 exhibited low toxicity to bluegill sunfish, Lepoais nacrochirus, in a 96-hour, unaerated, static test vith noinal concentrations of 262, 328, 410, 512, 640, 800 and 1000 Bg/L. The 96-hour LC50 based on nominal concentrations was 634 Mg/L with a 95Z fiducial interval of 567 to 725 ig/L. Work by: tA^y.%^A M. Setfrey T. Turner, Sr. Toxicology Associate t /-' <, A Ho/Day/Year ^^-^UGhM.je- Laurie D. Bouchelle Toxicology Technician ^ / ^ ^ Mo/Day/Year THIS REPORT IS APPROVED FOR ISSUE. /^ ft'^J &. Keith B. Pierson, Ph.D. Research Toxicologist and Study Director EnvironBental Sciences AutrViT ^ W^ Ho/Day/Yeat Company Sanitized. Does not contain TSCA CBl Dates of Inspection: Conduct - 6 23/93 Records, Re:.: rt(s) - 8/9-10/94 Date(s) Findings Reported to; Study Direc::r - 8/10/94 Management - 3/24/94 Reported by:\^_^-_KA_^ J<_ H-^-ll Donna R. Holt Quality Assurance Auditor Ool^ Dat Coinaany.SanitizecL Dcss noi contain TSCA C3: i. nuosx The goal of this study was to assess the acute toxic!ty of H-19704 to Juvenile blueglll sunfish, Lepoais acrochirus. II. RECORDS All data and 'ie final report vill be aalntained either at Haskell Laboratory for Toxicology and Industrial Medicine (Newark, DE) or be archived at the DuPont Records Manageaent Center (Vilaington, OB). A. Test Substance III. HATERIALS AM) METHODS The teat substance is described under General Intonation. Test solutions were prepared by weighing out the appropriate quantity of test substance, adding it to 10 liters of dilution water (see below) and stirring. The test substance appeared to be entirely in solution at study start. Stability and solubility of H-19704 in dilution water were not deterained. B. Dilution water Water (Table 1) originated froa the Haskell Laboratory well. C. Anl--1 Culture Bluegill sunfish, Lepoais acrochirus, were purchased froi either Northeastern Biologists (Rhineback, NT) or Aquatic Research Organises (Haapton, NB). Fish were acclimated in continuously-flowing, well water in 545-L circular, fiberglass tanks for 212 days at a aean teaperature of 21.2C (rangefinder) or for 32 days at a aean teaperature of 22.6'C (definitive study) prior to study start. Pish were fed newly-hatched, live Arteaia sp. (Aquarliu Products, Glen Burnie, ND), frozen adult Artcaia sp. (Golden Gate Brand, Kordon Division, Novaiek, Inc., Hayward, CA) and/or Tetraain* flake food (TetraUerke, Geraany) generally twice daily on weekdays and daily on weekends. Fish were treated with 2-3 parts per thousand salt (NaCI) on days 0-4 of the 212-day acclimation (rangefir.der). Pish were treated with three handfuls of salt (NaCI) on day 0, '200 parts per Billion foraalin for one hour on days 1-3 of the 32-day accliaation (definitive). Sickness, injury and abnoroality were not present at study start. Aniaals were identified by labels on aquaria and test vessels. D. Test Methods |1] Seven test concentrations and a control were used. were 262, 328, 410. 512, 640, 800 and 1000 Bg/L. Noiinal test concentrations Glass aquaria [20 liter; 39.9 (length) x 19.8 (width) x 25.5 ci (height)j Company Sanitized. Does not contain TSCA CBJ ^^jfe&^a.feg^iVw;.;--. ^SJ&S&~, -fcS;.j^ OuPont BLR 61-94 f t containing 10 L of solution (approximately 13.5-ca liquid depth) were employed. Positions of test chambers in the water bath used to aatntain constant teaperatu.-A vere assigned uaing random numbers. Ten fish vere added to each replicate using randoa numbers (two replicates per concentration; total 20 fish per concentration). Control fish ranged froa 1.2 to 2.8 ca standard length (Man 2.1 ca) and 0.0451 to 0.524 g vet weight (Bean 0.228 g). Control loading vas 0.23 g/L at test conclusion. Test solutions ranged from 21.3 to 22.I'C (mean 21.7C). Pish vere not fed approxiaately 24 hours prior to nor during th'j test. A photoperiod of 16 hours light (312 to 344 Lux) versus 8 hours darkness was eaployed vlth 25 minutes of transitional light (<2.15 Lux) preceding and following the 16 hour light interval. Mortality and behavioral effects were noted daily. Dissolved oxygen (TSI Model 54A, Yellow Springs Instruaents, Yellow Springs, OB), pH (Corning Model 610A, Corning Scientific Instruaents, Medfield, MA) and temperature (Arerican Society of Testing Materials registered aercury theraoaeter) were aeasured in each replicate before addition of fish at test start, dally, at total fish aortallty and/or at test end. Total alkalinity (Bach Cheaical Coapany, Loveland, CO), EDTA hardness (Bach Chealcal Corpany, Loveland, CO), and conductivity (YSI Model 33, Yellow Springs Instruments, Yellow Springs, OB) of the control water were measured before fish were added at test start. A continuously-recording thermoaeter in a water-control replicate was used to check temperature variations during the 96-hour test. B. Chemical Analysis Cheaical analyses of the test substance concentrations in water were not perforaed because a suitable method was not available. V. Statistical Methods The 24-, 48-, 72- and 96-hour LC50s and their 95% fiducial intervals were calculated using probit analysis (2]. IV. RESULTS AfO DISCUSSION Protocol aaendments and/or SOP deviations appear in Appendix 1. A rangefinder was conducted. Nominal test concentrations were 0, 0.1, 1, 10, 100 and 1000 g/L. Mortality after 96 hours was 0, 0, 0, 0, 0 and 80Z, respectively. The definitive study was conducted using noainal B-19704 concentrations of 262, 328, 410, 512, 640, 800 and 1000 g/L. We planned to aeasure B-19704 concentrations in test solutions, but the suggested analytical method failed. Preliminary research suggested it was not cost effective to develop a new method. Bence, nominal B-19704 concentrations are reported herein. Con^nv Sanifc". ^ Rsi conia!?l TSCA a" DuPont BLB 41-94 All eh--leal and physical parameters were within expected rang-- (Tables 2-3). f/L The 96-hour LC50 (Tables 4 and 6) was 634 vith a 95X fiducial interral of 567 to 725 ag/L based upon noalnal concentrations. Behavioral observations are reported in Table 5. Fish Retries are reported in Table 7. v. concufsioN H-19704 exhibited low toxicity to Juvenile bluegill sunfish, Lepoais aacrochirus, in an unaerated, static test. The 96-hour LC50 was 634 ag/L (95X fiducial interval of 567 to 725 g/L) based upon noaiinal test concentranons. 1. test. 2. 333 p. 40 CFR 797.1400. VI. LITBATuRB CITBD Toxic Substances Control Act. Fish acute toxicity Finney, D. J. 1971. Probit Analysis. 3rd Ed., Cambridge Uni-.. Press. ' C.mp.S,-ze<.D.esn..c.n."TSCACB BOD, g/L COD, g/L DOC, g/L TOC2, g/L Total Kjeldahl N, mg/L A--onia N, g/L Turbidity, NTU Phenolics, g/L Color, apparent Co/Pt3 Solids, total suspended, mg/L AluninuB, g/L Antimony, ag/L Arsenic, g/L Berylliic, g/L Boron, mg/L Brfide, g/L CadBiiw, g/L Calclua, g/L Chloride, g/L ChroBiuB, g/L Cobalt, g/L Co"per, g/L Cy nide. g/L Iron, g/L Fluorlde, g/L <7 <0.2 <0.2 <0.01 <O.G1 <0.04 <0.5 <0.01 24.5 6.9 <0.05 <0.05 <0.02 <0.005 <0.1 0.1 Lead, g/L HagnesluB, ag/L MBAS/LAS, Bg/L Mercury, ag/L Nickel, g/L Nitrite, og/L Nitrate, ag/L Ortho-Phosphate, Potassium, Eg/L Selenium, og/L Silver, mg/L Sodlua, Bg/L Sulfate, ng/L Sulfide, ng/L Zinc, ng/L Bg/L <0.1 3.8 <0.02 <0.00020 <0.05 <0.1 0.5 <1 1.8 <0.005 <0.02 8.7 5.2 <0.10 <0.04 Volatile priority pollutants ND4 Acid extractable priority pollutants ND5 Base/neutral priority pollutants NO5 Pesticides/PCBs W Organophosphate pesticides, vg/L ND* lD&te of s--ple collection 12 May 1993 unless Indicated otherwise, analyses performed at Lancaster Laboratories, Lancaster, Pennsylvania; 'Date of sample collection 23 March 1993, analyses perforoed at Lancaster Laboratories, Lancaster, Pennsylvania; 'Units based on cobalt/platinun reference; None detected; level of detection ranged froa < 5 < 100 vg/L; 'None detected; levels of detection ranged fron < 10 to < 25 ug/L; "None detected; levels of detection ranged froa < 10 to < 100 ug/L; iflom detected; levels of detection ranged froa < 0.01 to 4 ug/L; None detected; levels of detection tanged froo < 0.01 to < 0.1 ug/L. 10 p,,,pan^ SanSt^d. Do^.ol con^ ^CA CBS DuPont HLR 61-9* Static, Acute, 96-Hour LC50 of H-19704 To Blueglll Sunfish, Lcpoaia --crochirua TABLE 2 Vater Chtnistry of the- Dilution Hater at Test Start Total Alkalinity (ng/L as CaCO)) 79 EDTA Hardness (ag/L as CaCO;) 76 Conductivity (unhos/cm) 165 11 Company Sanifasd. Does not contain TSCA CBl -.^Sifca-f; TABIf 3 Physical and Cheaical Characteristics of 3-19704 Test Solutions Nominal Concentration (ng/L) 0 A B 24 A B Hours 48 A I > 72 A I 1 96 1 L Dissolved Oxygen1' (Bg/L) - Control 8.5 8.5 7.8 7.9 7.5 7.,6 7.3 7. 5 7.,1 2d2 8.5 H.5 7.U /.(! 7.(, 7. B 7.A 7. 5 7,,0 328 8.5 8.5 8.0 8.1 7.6 7.,6 7.3 7,.3 7,.0 410 8.5 8.5 7.9 /.9 7.5 7,,7 7.1 7..0 7,,1 512 8.5 8.5 8.0 8.0 7.7 7 ,7 7.3 7..2 7,.0 640 8.5 8.5 7.9 7.8 7.5 7,.6 7.0 7,.2 6..9 M 800 8.5 8.5 8.1 8.1 7.6 7,.5 6.9 7,,0 6 ,7 1000 8.5 8.5 8.2 8.1 7.7 7,.6 --- 7,,0 -- pB Control 262 328 410 512 640 800 1000 7.4 7.4 7.4 7.4 7., 7.3 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.5 7.4 7.4 7.2 7.1 7.3 7.4 7.3 6.9 6.9 7.3 7.2 7.2 6.9 6.9 7.3 7.3 7.2 7.0 6.9 7.2 7.2 7-2 7 .4 7.3 7 .4 7 .4 7 .3 7.4 7 .3 7 .4 7 .4 7.3 7 .3 7 .3 7 .4 7.3 7 .4 7 .3 7 .4 7.3 7 .4 7 .3 7 .2 7.2 7 .2 7 .2 7 .3 7.3 7 .3 7 .3 7 .2 --- 7 .1 '' All fish had previously died so no reading was aade. theoretical saturation at 22C was 8.8 mg/L. Company Sanitized. Does not co t TABLB 3 CONTINUED Physical and Chemical Characteristics of B-19704J Test Solution Noiinal Concentration (g/L) Temperature (C) Control 262 328 410 512 640 800 1000 0 A B 24 A Hours 48 BA B 72 96 A BA 22.1 22.1 22.0 22.1 22.1 22.1 22.1 22.0 22.1 22.0 22.1 22.1 22.0 22.1 22.0 22.0 21.6 21.6 21.6 21.7 21.7 21.6 21.6 21.6 21.7 21.6 21.6 21.6 21.7 21.6 21.6 21.6 21.8 21.7 21.7 21.7 21.7 21.7 21.7 21.8 21.9 21.7 21.7 21.7 21.8 21.7 21.8 21.8 21.6 21.5 21.4 21.5 21.6 21.5 21.5 .I--* 21.5 21.5 21.4 21.4 21.6 21.6 21.5 21.6 21.5 21.4 21.3 21.4 21.5 21.4 21.4 -------- All fish had previously died so no reading was made. Company Sanitized. Does not c .'(' '' . '-, ^S!''W ^ ^ :' .;'i' "' ^s^J, ;? . DuPont HLB 1-94 f Static, Acute, 96-Hour LCSO of H-1^704 To 1 Bluegill Sunfish, Lepoals nacrochims TABL2 4 W------f Mortalityr of Bluegill Sunfish. IfWatis acrochims, Expos ed to H-19704 Jror 96 Hours in a Static, unaeratedTest Mortality In Percent Noainal Concentrations (t/l.) 24 Water Control A* 0 B* 0 Total 0 |w> 262 A 0 B 10 Total 5 328 A 10 B 10 Total 10 410 A 10 B 20 Total 15 Tioe (Hours) 48 72 96 0 0 0 0 0 0 0 0 0 0 0 0 10 10 10 5 5 5 10 10 10 10 10 10 10 10 10 10 10 10 20 20 20 15 15 15 A and B represent replicates; each replicate contained 10 fish (total 2 : fish per test concentration) at study start. W w ^^ u DoesnotcontainTSCACBl Company Sanit.aed. . Does n -.;i-^. .- .-,,,...;,.- ^^^^^ P . y- n,..^^;^ ?' ^ , | 1 ^-- % .' DuPont HUt Sl-94 Static, Acute, 96-Hour LC50 of H-197C34 To Blueglll Sunfisb, LepoBis acroehlrus TABLE 4 aMtUIHW Mortality of Bl uegillSunfish, Lenglis iacrochirus, 1 Exposed to B-1970401 B9)^or 96 Hours in a Static; UnaeratecTTest Mortality in Percent Noainal Concentrations (Bg/L) 24 Time (Hours) A4H8 -7"2' - -96 512 A" 30 30 30 30 B" 30 30 30 30 & Total 30 30 30 30 640 A 40 40 40 40 B 40 40 40 40 Total 40 40 40 40 800 A 70 70 70 70 B 60 60 60 60 Total 65 65 65 65 1000 A 90 B 90 Total 90 10090 95 100 100 90 90 95 95 A and B represent replicates; each replicate contained 10 fish (total 20 fish per test concentration) at study start. ft . .^.^.^uiaiu^ Company Sanitized. Does no'; contain TSCA CSI ^.^^.-...^J ^TB^^^Sl^ws^'a^^r B" '"';"'S'SSli-S,. l-ff'.II'i'ti' r-ll'"r.---v^. f1' " -"> -- i^w-sa?/^^!? DuPont BLR 661^^44 Static, Acute., 99<66--Hour LC50 of B-19704 T<o9 BBlulugf(lill Soranfish, Lepoalis aaccrroocchilruas UUM5 B4ehavlor of Bluegm junfish, Leggais acrocb irus, Exposed t o H-1970A' "xor 96 Hoturs in a Scat rc'ttiaeratenrest | NoBinal Concentrations (g/L) Water Control A B 262 A B 328 A B 410 A B Nuiber Shoving Behavior/Total Number of Fish Alive I ii 24 0/10 0/10 0/10 0/9 0/9 0/9 i' Tiae (Hours) ii, 48 0/10 0/10 72 96 f f, ------------------------ 1: ^ 0/10 0/10 * 0/10 0/10 t 0/10 0/10 0/10 i 0/9 0/9 0/9 i t. '1 } 1 0/9 0/9 0/9 0/9 0/9 0/9 0/9 0/9 0/9 0/9 0/8 0/8 0/8 0/8 i AB 00//77 00//77 00//77 00//77 :I, ii 640 ;? I "" A 0/6 0nf/t6- 0/6 0/6 ''| B . 0/6 0/6 0/6 0/6 'y 800 A ,a/3 3la-//3I 3->-.-/3 3- -/.3- 1 B 4/4 4"/4 4-/4 4"/4 1 i 1000 A B rr//ii ----b r/i ----b r/i -- r/i Lethargic. ^ "All fib had previously died. 16 L . . .,,ii.a,firi&.*.,,,.^-.i Company Sanitized. Does not contain TSCA CBI \- --^i J Tine LC50 <Bg/L") 24 Hours 647 48 Hours 634 72 Hours 634 96 Hours 634 952 Fidui:ial Litits (Bg/L) (fflg/L-) 575 746 567 725 567 725 567 725 Based upon nooinal concentrations. Slope 4.83 5.09 5.09 5.09 Y-Intercept -13.6 -14.3 -14.3 -14.3 17 .^^cn^I^0^ e^ppaamniS^a-nti'tlyr1..-1-:0"' ---- DuPont ELB (1-94 Static, Acuf, 96-Hour LC50 of H-19704 To Bluegill Sunflfh, Lpoal acrochirua TABLE 7 Standard Lengths and Vet Weights of Control Pish At Study End Replicate A A A A A A A A A A B B B B B B B B B B NiiBber 1 2 3 4 5 6 7 8 9 10 1 2 3 4 5 6 7 8 9 10 Standard Length (ca) 2.3 2.1 2.1 1.9 2.3 2.3 1.2 2.6 1.9 2.2 2.8 2.2 2.1 2.1 1.8 1.7 1.6 2.6 2.3 2.0 Vet Weight ) 0.256 0.194 0.176 0.159 0.262 0.244 0.0451 0.415 0.183 0.224 0.524 0.221 0.226 0.202 0.142 0.100 0.103 0.456 0.269 0.153 18 TSCACBl Sanifeed. Does rr,ot contain Company mssa i PKOTOCOL AMBBMBRTS AMD SOP DBVIATIWS PROTOCOLANBNDNBNTNUMBER ONE a. The ratio between test concentrations is 0.8. b. Test iiirbers. concentratijns should (Note: Randoa be assigned to test vessels using randoc vessels in water bath numbers are also used to assign location and to assign fish to test vessels.) of test c. Take added at saaples for cheaical analysis test start and at test from each replicate before fish are analysis. end. Saaples should be refrigerated prior to d. Because of the snail (not 48) hours. size of the bluegills, they will be starved fcr 24 e. TOC, COD performed. and paniculate analyses of the dilution vater will not be f. Purity and stock checks will not be performed. f. pH and DO readings should be made on study start, dally thereafter, at study all replicates prior to adding fish at replicate. end and/or at total nortallty :a any h. Dead fish in the controls do not i.^ed to be weighed and measured. 1. Test is static (not static reneval). rje. pliMceaatseusrealtensgtuthdsy eanndd. weights of all surviving fish in both of the control 19 ...c^ainT^AC^ , .-mpf; ii^1 - -on-^56"^ DuPont HUt 61-94 Static, Acute, 96-Bour LC50 of H-19704 To Bluegill Sunfish, Lcpomis macrochirus APPBOIX 1 (CONriNUBD) PROTOCOL AHBXMHfTS AW SOP DEVIATIONS CONTINUED PROTOCOL AMENDMENT NUMBER TWO Ve planned to measure H-19704 concentrations in each test solution. Samples for analysis were taken and the analyses vere attempted. The analyses failed and preliminary research suggested that it was rot cost effective to develop a new Bethod. The samples vere therefore discarded and nominal concentrations reported. Since the purpose of this study was tc provide acute mortality data fov designing a bioconcentration test, this procedure was considered acceptable. In 1994 Haskell Laboratory adopted a new systee for handling financial affairs. As a result, it was necessary to develop a new set of test codes for aquatic studies. Hence, the study code (226A) is incorrect. The correct new study code is 226. PROTOCOL AMENDMENT NUMBER THREE This study was performed under SOP AQ018-T-002 (not AQ018-T-001 as shown in the upper righthand corner of the Short-Term Study Protocol Sheet). The current name of the sponsoring business is DuPont Elastomers (the previous naire was Polyers for which Poly was the abbreviation which appears on the Short-Term Study Protocol Sheet). SOP Deviations a. Test concentrations were not assigned to test chambers using random numbers. 20 ^ mpany Sanitized. Does not contain TSCA CBI