Document 8264voVrNr8DV9aZ1gkmVOvYm

I I 96-Hour Acute T DEA saltof PFO ACUTE TOXICITY TO FISH TEST SUBSTANCE Identity:PerfluorooctanesulfonatDeE,A saltm;ay alsobe referredtoas PFOS DEA salt,FC-99, or3M Sample No. 2. (1-Octanesulfonic acid,1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-heptadecafcloumoprdo.-, with,2,2'-iminobis[etha(n1o:l1]),CAS # 70225-14-8) Remarks: Test sample isa mixtureofthetestsubstanceinwater (approximately25% testsubstance and 75% water).Allvaluesreported relatetothismixture.No calculationwsere made toadjustforthe actual concentration,otfhe testsubstance inthetestsample. METHOD Method: EnvironmentalProtectionAgency, EcologicalResearch EPA-660/3-75-009,April,1975. StandardMethods. Type: Staticacute GLP: Yes Year completed: 1979 Species: Lepomis macrochirus Supplier:Osage CatFishedesI,nc.inOsage Beach,Missouri. Analyticalmonitoring:pH and DO /ammonia content Exposure period:96-hours Statisticamlethods: Probitanalysis. Test fishage: Not noted. Length and weight: Average length= 28.6+ 2.17mm. Average weight = 0.60 + 0.15g Loading: 0.2 g fish/L Pretreatment: None Series Test conditions: Dilutionwater: Laboratorywellwater Dilutionwater chemistry: Dissolved Oxygen,: 9.3 mg/L hardness: 255 mg/L as alkalinity: 368 mg/L as pH: 7.8 Conductivity: 50 pmhos/cm CaC03 CaC03 Stock and testsolution preparation:Primarystock prepared in deionizedwater ata concentrationof 150 mg/mL. The test concentrationswere prepared by tr*ansfeninagppropriatealiquots ofthe stockstandarddirectltyo thetestchambers. The test solutionswere noted to foam when stirrinigntoxicantaliquots. Test concentrationswere prepared based on totalsample, not on percentconcentrationofthe testsubstance inthetestsample. Concentrations dosing rate:Once Stabilitoyf the testchemical solutions:Not noted Exposure vessels: 40 litegrlassaquariacontaining30L of test solution. Numbe rof replicateso:ne Number ' of fishper replicatet:en Number of concentrations:sixplusa negativecontrol Water chemistry during the study: Dissolved oxygen range (0- 96 hours): 6.0 -8.4 mg/L (controelxposure) 5.8-8.3mg/L (18mg/L exposure) pH range (0- 96 hours) 8.2-8.3 (controelxposure) 8.3-8.3 (18 mg/L exposure) Test temperature: Temperature heldconstantat22"C throughuse of a water bathfortestvessels. RESULTS Nominal concentrations:Bk control,18,37, 75,160, 320,650 mg/L Element value: 24-hour LC50 = 460 (370-580)mg/L 48-hour LC50 = 370 (290-470)mg/L 96-hour LC50 = 31 (22-43)mg/L 96-hour NOEC = 18 mg/L (C.I.notcalcu- lated) Allelement values-based on nominal concentrations StatisticaElvaluationof Mortality: Probitanalysiswas used to calculatLeC50 valuesand the corresponding confidencelimits. QualityCheck forTest Organism Health: The bluegilslunfishwere challengedwitha referencecompound, AntimycinA. The observed 96-hour LC50 and 95% confidencelimits(C.I.) were withinthe95% confidencelimitsreportedinthe literaturien,dicating thatthefishwere ingood condition. Cumulativepercentmortality: Nominal Test Conc., mg/L 24-hours 48-hours 72-hours 96-hours Neg. Control 0 0 0 0 18 0 0 0 0 37 0 0 10 80 75 0 0 30 90 160 0 0 70 100 320 0 20 100 100 650 10'0 100 100 100- CONCLUSIONS The testsample 96-hour LC50 forbluegilslunfishwas determined to be 31 mg/L witha 95% confidenceintervaolf22-43 mg/L. The 96-hourno observed effectconcentrationwas 18 mg/L. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY Reliability:Klimischranking2. This study,whilewellconducted,lacks analyticadlatafordeterminationofthe testsubstance concentrationinthe testsolutionsand determinationofthe sample purity.There were also gaps inthe measurement ofwater qualitycriterifaora number ofthe concentrationsatgiventime intervals. REFERENCES This studywas conducted by AnalyticalBioChemistryLaboratoriesI,nc.of Columbia, Missourion behalfofthe 3M Company. OTHER Last changed: 5/2/00 Submitted By: Analytical BioChemistry Laboratories, Inc. 7200 East ABC Lane P. 0. Box 1097 Columbia, Missouri 65205 Prepared By: Carl M. Thompson Aquatic Biologist e-*, Alan D. Forb;is Aquatic Supervisor Date 4f Date Approved By: OL ()Jea6alesis A. Ault alit surance :f icer oshunrsaonce YI ylLW aa lIr*at@OoJJr4vohMnasnoalger La rat ry Ma Dat4 Date SUMMARY The acute toxicity of 3M Sample No. 2 to bluegill sunfish (Lepomis macrochirus) was assessed using the methods outlined by the Committee on Methods for Toxicity Tests with Aquatic Organisms (1). Water quality parameters of temperature, dissolved oxygen, pH and ammonia were measured throughout the test and were within acceptable limits. As a quality check, the bluegill sunfish were challenged with a reference compound, Antimycin A. The observed 96 hour LCso a'nd 95% confidence limits (C.I.) were within the 95% confidence limits reported in the literature (2), indicatingthat the fish were in good condition. The results of the four day static fish toxicity study using bluegill sunfishare sti-arizedbelow. Compound 3M Sample No. 2 Antimycin A 96-hour LCSO (95% C.I.) 31 mg/l (22-43 mg/1) 0.000029 mg/l (0.000021-0.000040 mg/1) Also, the results indicated a 96 hour, no observed effect concentration of 18 mg/l. INTRODUCTION This static bioassay was performed at the aquatic bioassay laboratory of Analytical BioChemistry Laboratories, Inc., Columbia, Missouri, for 3M Company, from June 12 to June 16, 1979, as authorized in a letter from 3M Company on June 6, 1979 (Appendix I). The purpose of this test was to determine the 24, 48 and 96 hour LCBO levels for 3M Sample No. 2 to bluegill sunfish (Lepomis macrochirus). The study was performed following the procedures outlined in ABC Protocol Number 7601 (AppendixI). METHODS AND MATERIALS The proeedures for static bioassay, as described in Standard Methods for Examination of Water and Wastewater (3) and Methods of Acute Toxicity Tests with Fish, Macroinvertebrates and Amphibians (1), were used in this experiment. The bluegill sunfish used in the test were obtained from Osage Catfisheries, Inc. in Osage Beach, Missouri. The fish were identified to species using the taxonomic keys developed by Eddy (4). All test fish were held in culture tanks on a 16 hour daylight photoperiod and observed for at least fourteen days prior to testing. Fish culture techniques used were basically those described by Brauhn et. al. (5). A daily record of fish observations during the holding period, along with prophylactic and therapeutic disease treatments, is included in Appendix I. During this period, the fish,received a standard commercial fish food (Rangen's) daily until 48 hours prior to testing at which time feeding was discontinued.The bluegill sunfish used for this experiment had a mean weight of 0.60 g and a mean standard length of 28.6 mm. Weight and length measurements were made on a representative group of fish prior to the test and are included in Appendix I. The static -fish bioassay was conducted in 40 liter glass aquaria containing 30 liters of laboratorywell water. Characteristicsof this dilution water are shown in Table 1. These vessels were kept irLa water bath at 220C (1.0). The test fish were acclimated to the dilution water and test temperature and held without food 48 hours prior to testing. Based on the results of preliminary testing with fathead minnows (Pimephales promelas) by 3H Company, six concentrations of the test compound, ranging in a logarithmic series for 18 to 650 mg/l, with ten fish per concentrationwere selected for definitivebioassay. The fisbl were added to the test chambers by stratified random assignment within 30 minutes after addition of toxicant aliquots. The use of 40 liter glass aquaria for this study was authorized by 3M Company (AppendixI). The 3M Sample No. 2 standard was received on June 8, 1979, in good condition. The sample upon receipt was observed to be a yellow liquid and was stored at room temperature (220C). Sample purity was not provided; therefore, the test concentrations were prepared based on the total compound. The test concentrations were prepared by transferring appropriate aliquots of a stock standard directly to the test chambers. 2 The stock standard was prepared in deionized water. The test solutions were noted to foam when stirring in toxicant aliquots. All standard weights and dilution values are listed in Appendix I. RESULTS Table 2 presents the predicted LCSO values and 95% confidence intervals for 3H Sample No. 2 and the reference test against AntimWcin A, a piscicide. These values were obtained by employing the statistical method described by Litchfield and Wilcoxon (6). Mortality rates, test concentrations and water quality data are presented in Table 3. . The dissolved oxygen concentration which stayed between 40 and 100% saturation was considered adequate for testing. The pH values remained consistent with the control throughout the study. The ammonia concentrations were below the toxic limit (7). The study was conducted following the intent of the Good Laboratory Practice Regulations (9) and the final report was reviewedby Analytical BioChemistry Laboratories' Quality Assurance Unit. All original raw data was provided to 3H Company, with a copy retained at Analytical BioChemistry Laboratories. 3 TABLE I Chemical CbLaracteristicsof Well Water at ABC's Aquatic Bioassay Laboratory. Parameter Dissolved Oxygen PH Hardness (CaC03) Alkklinity (C&COS) Conductivity Total Ammonia (NH3) N03-N Ortho-Phosphate Aluminum Arsenic Cadmium Chromium Cobalt Copper Iron Lead Mercury Nickel Zinc DDVP Diazinon Disyston Methyl Parathioa Malathion Ethyl Parathion Concentration 9.3 ppm 8.2 255 ppm 368 ppm 50 umbos/cm <0.05 ppm 0.15 ppm 0.10 ppm <0.01 ppm <0.001 ppm <0.001 ppm 0.001 ppm <0.001 ppm <0.01 ppm 0.012- pp'm 0.009 ppm <0.0001 pps 0.0157 ppm <0.01 ppm <40 ag/l <20 nsll <20 ng/l <80 ng/l <110 ng/l <80 ng/i 4 Compound 3H Sample No. 2 Antimycin A*** TABLE 2 The Acute Toxicity of 3M Sample No. 2 and Antimycin A to Bluegill Sunfish (Lepomis macrochirus) 24 hours 460 (370-580)** 0.000070 (0.000056-0.000088)** LCSO in milligrams/liter (ppm) 48 hours 370 (290-470)** 0,000040 (0.000032-0.000050)** (0.0 *Bioassay as conducted at 22*C (1.0), mean weight aAd length, 0.60 g and 28.6 mm. **95% confidence interval. ***Antimycin A standard obtained from Sigma Chemical Company, Type III, crystalline, Lot 125C-01 TABLE 3 The mortality rates and water quality parameters during the acute toxicity test of 3M Sample No. 2 with bluegill sunfish (Lepomis macrochiru mg/l Concentration Control 18 37 75 160 320 650 Percent Mortality Hours 24 48 96 0 0 0 0 0 0 0 0 80 0 0 90 0 0 100 0 20 100 100 100 100 Temp. oc 22 22 22 22 24 hours D.O.* ESLI pH** 8.4 8.2 8.3 8.3 Water Quality NHS*** mg/l Temp. oc 48 hours D.O.* mg/l pH** 0.86 22 7.6 8.3 0.78 22 7.8 8.3 NHI-F-W mgll 1.3 1.3 8.1 8.1 0.90 8.0 8.1 0.78 22 7.4 8.3 1.6 Temp OC 22 22 22 *Dissolved oxygen concentrations Dissolved Oxygen Probe (Extech Model 8012) used with an Exteth Mode **pH - pH Probe (Fisher Model 13-639-108) used with an Extech Model 671 pH and mV meter. 'k*Total ammonia concentrations Ammonia Probe (Extech Model 8002-8) used with an Extech Model 671 pH LITERATURE CITED' (1) Committee on Methods for Toxicity Tests with Aquatic Organisms (C. E. Stephan, Chairman). 1975. Methods for Acute Toxicity Tests with Fish, Macroinvertebrates and Amphibians. Environmental Protection Agency, Ecological Research Series EPA-660/3-75-009, April, 1975. 61 p. (2) Berger, B. L. , R. E. Lennon and J. W. Rogan. 1969. Laboratory Studies on Aatimycin A as a fish toxicant. U. S. Department of Interior,Investigationsin Fish Control No. 26. 21 p. (3) American Public Health Association. 1975. Standard Methods for the Examination of Water and Wastewater. 14th ed. Washington, DC. 1193 (4) Eddy, Samuel. 1969. The Freshwater Fishes. 2nd ed. W. C. Brown Company, Dubuque, IA. 286 p. (5) Brauhn, J. L. and R. A. Schoettger. 1975. Acquisition and Culture of Research Fish: Rainbow Trout, Fathead Minnows, Channel Catfish and Bluegills. Environmental Protection Agency, Ecological Research Series EPA-660/3-75-011, May, 1975. 45 p. (6) Litchfield,J. T., Jr. and F. Wilcoxon. 1949. A SimplifiedMethod of Evaluating Dose-Effect Experiments. Jour. Pharm. Exp.. Ther. 96:99113. (7) National Academy of Sciences. 1971. Water Quality Criteria, 1972., U. S. Department of Commerce, PB-236 199. 592 p. (8) Food and Drug Administration. Regulations for.-.GoodLaboratory Practice. Federal Register, Vol. 43, No. 247, 59986-60025, December 22. 1978. 7 Quality Assurance Statement for final report #23893 entitled, "Acute Toxicity of 3M Sample No. 2 to BluegillSunfish (Lepomismacrochirus)," for Dale Bacon, 3M Company, St. Paul, Minnesota. In accordancewith ABC Laboratoriesintent that all studies conducted at our facilitiesare designed and function in conformancewith good laboratorypractice regulationsand the protocols for individual laboratorystudies, an inspectionof the final report for 3M Sample No. 2 was conducted and found to be in an acceptable form by a member of our QualityAssuranceUnit. An inspectionof the daily mortality rate of the test organismsprior to the initiationof the study indicatedthey were in good health and should not bias the observed mortality in the study. It was also noted that since the purity of the compound being tested had not been supplied, the LCSO would have to* be calculated based on nominal concentrationsof the total compound. A final inspectionof all data and records on June 20,-1979,indicatedthat the report submitted to you is an accuratereflectionof the study as it was conducted by ABC Laboratories. Should you have any questions relatingto the informationprovided in this statementor the functionof our QualityAssuranceUnit, please contact me at your convenience. a A.-Ault (fq:tlliiAtsysuranceOfficer :71131 Date 8 APPENDIX I RAW DATA 14 ANALYTICAL BIOCHEMISTRY LABS Aquatic Bioassay Lab Acute Toxicity Bioassay Date: Time: -Toxicant: 3m--Z- Lab No.:?, 36ti Tank: g62 Purity-: Formulator: E2gE5E'@-.@Date Made: -Solvent: Stock Standards Workihg Standards Yinal Gross Wt7-.- Tare wt. 0.060 9 Net wt. -Adj. net- Wt. ipo-ose g yo.'age i* Aliquot of concentrated stock Dilution Vol. Final Concentration Mg/ Dilution vol. 7@oo ml Concentration-LAa mg/ml (Balance Calibration L"-**) + 0-496 w ls.0007 c-lw4fj ------------------------------------------------------ ----------------- n Test Species: Lot No.- 1674 Temp.: a!oc -Source f@@22 kam..t 1 4@lg@ t: - Len gth.: no./vesseiv u 70w"" Vessel Size: !1!2--e Void: 4 !b5 Water Chem3.stry:LL4".gr.@ pH: 7. 1? Alkalinity: &d@LBardness: 297.5 ------------------------------------------------------ ------------- r_ACO -s -e-@6 .. Concentration tinolt/-O. ml Dead Obs. 24 hr 48 hr 72 Fir 96 hr I ead Obs. -!De eaal d Obs. Dea ZYs-. -Tead'Obs. Dead'Obs. C2 3-1 75 (go -3zo C2 0 0 1 -7 II-or.In ir) Observer Date Remarks: -L C7- i Cl-T T iE n" 14 P 4ared by: --,@Correctedfor **Checked with ,A-h4 @ur:Vty oY pzy:rmarystandard class S weights Checked by: "a@@ c LA.CXA Water Quality 24 hours 48 h:)urs oncentratiori T8mp. D.O.* -NH mg/l C m& @l PH** mg /i T8mp. D.O.* -C M/1 pH** NH*3** TSmp.-D.O.* mg /1 C ma/I PH** NmHg*3*/*i T8 t 7- is-7 I.3 K-4 9.zIn 7, 93 1/.3 at 7S 3zo z z. o -?k7.li v-.3 *Dissolved oxygen concentrations Dissol@ved Oxygep Probe (Ey'.tec-hModel 8012) used w 671 p4 and mV meter. **PH - PH Probe (Extech Model 200C) used Oith an Extech Model 671 pH ahd mV meter. ***Ammonia concentrations - Ammonia Probe (Extech Model 0002-8.)used with an Extech meter. ANALYTICAL BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Biodssay -.roxicantTest Species -21 11. Lab Number -@Exposure period 2A Probit Analysis Work Sheet LOoo@T.@@ Date tested Date reported C@,v5 Temperature Water quality /'7!q Lk..;o.P-I4U:2. Concentration No. dead Obseri;ed % Expected % total no. mortality mortality 10-E Contribution to ChT tc) U/ lc> to /to (!214,q (g!@.11 6cj 0 cq 0 -0-0170 animals K -30 LC04 LC so LC16 S LC /Lc 84 so + LCso/LC 16 (o;o 2 S @or3?-0 2 Confidence linits(.05)for LCSO Ni f LC so @ s 2-77 LCso / f LCso lowerlimit- 3-?0 LC so X f LC so upper limit= -690 Analysis by: Cli@cked-By: Total contributionto Chi Chi2- contribut;onX total animals /-go to Chi K 2 Chi (p-.05) for (K-2)---L@eg.o'f freedom LC So -T-,90 -?o I;ko Y4,2 \L Date: 11:@ 73 d@@4 rf (Title) I 4z421?q (Dat@) ANALYTICAL BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay -foxicant '3pv-l Test Species Lab Number -icxposure period Concentrat*ion e_ (060 Probit Analysis Work Sheet Date tested - (0,/tz Date reported Temperature @Zt--717r4 - --g .2-c-r ;IVs Water qualit 0 No. dead Observed % total no. mortality Expected % O-Z: Contribution wortality to Chi 4Q 6)/.I)1 @n 1 1 !TE .1 - ---------- t,-zi animals 30 K LCB4 LC 320 LCI6 s LC 84 /LC50 + LC so ifLCI6 2 s @320 -@ 2 Confidence limits (.05)for LCso Ns 10 - f LCso S 2 - 7 7 N-i t -Z g. LC SI) LC so LCSD X f LC so lower limit= upper Iimi t - +-70 - Analys is by: L;&2E@@ -@ cli-ecked By: Total contributionto Chi Chi2- contr!bl+tionX total animals to Chi K 2 Chi (P-.05) for (K-2)_Lde-9. of freedom I& @- LC50 - -&, ct4 Date: (rit --170 95!.VCt. 1. I (Date) ANALYTICAL BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay -ToxicantTest Species Probit 3 nn-,-2, 72:>I CA p Analysis Work Sheet Date tested LOT 1g-7r4 Date reported Lab Number. Temperature -Exposure period (o k;,S Water qualit -7 -7- -7-OC' Concentration nAJ No. dead Observed % Expe@cted % Contribution total no. mortalityll mortality 10-E i to Chi &9 90 64 If# 17-a 10 C) rwe? C.2 animals K t-C04 LC so LCIR, s LC /LC + LC /LC 84 -50 so 16 Total contribution to Chi- Chi2- contribution X total animals (ioc) to Chi K di 2 Chi (p-.05) for (K-2)_I=deg. of freedom s -70 Confidence limits. (.05) for LCso Ni C) - IFLC so S2-77 N-. q62I%v- LC 50 LC So / LC 50 , lower limit = 7ZZ- LC 50 X f LC so = upper Iimi t - "3 - Aralysis by: i @ia me) ==IW'& . - Checked- By: Date: 42 ;e- 3;G2co (Title)- 95% c.I. f (Date) C2, p ul r4 CR to 4@6 a) -4 a" a 0 Vol 0 loo 0 I-H it T::p LT i,T-r. 1 7tt rift! i7[ @7L W: ITIQ .1 rF- T-1 IITF -LL IT 40 co 0 7 Ld -1 L -i:il@ioo -;7 I I' tii 17T.rFl.r' 1:@ it T.4 1 IT: t LLLL[I 14' 0 -TT- T 00 fill Ti, :j 00 7-T 0 14' Al@ALX'I'ICAZ7-BTUCTMn-l@).LL*AjB\Z>Y Aquatic Bioassay Lab Acute Toxicity Bioassay Date: -@eIIII79 Tc%..,pcant: -Formulator* -A Lab No.: ate Made: Time: Kt 00 ank: Tpurity: solvent oc Sta s Working Standards Final Gross Wt.- 30 - 9 Aliquot of concentrated stock Tare wt.- Dilution Vol. /ov Net wt. Final Concentrationo.oo/,s mg Adj. neT wt. yo ---@g* Dilution vol. t-o0 ml ConcentrationQ.3o mg7mi (Balance Calibratio ----------- --------------------------------------------------------- Test Species: Source _DD$.,,, -No. /Vesse.1:5- Lot No. 45'79 Cond:Lt3.on: @lood( vessel-'Size: I-T Te mp 2 I-Ioc Weight: fl. 'S@alk@-Length. Vo3.d:q Water Chemistry: A,o pH: 7.1 Alkalinity: -Hardness:qs ---------------Z------------------------------------6-A-(--US -------------- Concentration rpl 4&&l Dead Obs. 2-4 hr 48 hr -*Dead-(@-b-s. Dead Obs. 72 hrDead Obs. 96 hrDead Obs. DeadfO s, r,@. L 0 /Is OMO I'd A- /Is 0 0 3.06@;b -3Z. A & I)- 0005!6 .3t .3-L 0 10 01000/0 A I-t) C5 o. OoOlk A )bserver )ate Remax,ks: T &14 Pre_pared b Checked by: 10 e@orrected f,tik @@i @@FS&O rfriknary standar -d **Checked u<l;eh tAas w e it-_th)s rest Specie,ses4 Lab Number E,..posureperiod Concentration 0 A.7Tr?- ----O.oooo ?t ANALYTICAL BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay Probit Analysis Work Sheet A Date tested L4 Date reported-- (Olf4l-ZC, Temperature Water qualit No. dead Observed % Expected % total no. mortality mortality ID-Z Contribution to Chi Tot@,-animals K LCi;4 = tc - 0.0040 so LCI6 = 0 -0000 s LC84 /LC so-+ LC so/LC 16 2 oo-oc-0r-r -cZ 40 2 Confidence linits (.05) for LCso Total contributionto Chi Chi2- contribut-lonX total animals to Chi K 2 Chi (P-.05)for (K-2)_J_deg;of freedom LC 50 S2!-77 \4-N LC so ,c LC so , lower I;Mit -C.Cowj;/o LCSD X f LCSO - upper limit -0-ooosislz .%Vnalv,,isby: 0=4Y@ Checked- By: L(:,So Date: a/ (Title) er (Date)t ANALYTICAL BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay rt-jxi6ant Test Specieess Lab Number E,.cposure period Probit Analysis Work Sheet Date tested 9'7r4 Date reported AI I LA v-s Temperature T)- W ater qualit Concentration 92 -0000370. Oooo No. dead Observed % total no. mortaliity Expectted. % D ror ality I Contribution to Chi 0/to 10 3, 1)! !@21 0-000 I to/to tOO t/97.4@ . Tok._ animals K LC84 ma @LC 0 so LC16 s LC 84/LC so + LC so/LC 16 Total contribution to Chi Chi2- contribution X total animals to Chi K 2 Chi (p-.05) for (K-2)_Ldeig. of ;Freedom -30 2 Confidence linits (.05)for LC50 Ng to ol * V LC50 S2-77 LC so / LC50 0 lower Iimit =0.06$*O 52LC so X f LC50 , upper limit u&...wcoj;o -40kr LC So v%nai%,sis by:Chec@ked By: )NJ v- " @'t itle) 1r) Date: r/7 &f 11" .95%c.1. (Date3 .. ......... ANALYTICAL BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay lvxicant Test Specie!S_j@ LLM Lab Number L-Aposure period Concentrat .on Probit Analysis Work Sheet 4 Date tested Date reported Temperature 7,zc2 C q(a hrs Water quality-f?pt-d^ No. dead Observed % Expiected % total no. mortality mortality 0 00 0 -L te> to qg. -7q I Contribution to Chi 3 0.07-0 animals K LC84 '-LC5 0 = e.0000 :-30 Total contributionto Chi I rs Chi2- contribvtionX total animals Chl K 2 Chi (P-.0t5o) for (K-2)_Ldig. of frae6om LC16 = 0- s LC 84 /LC 50 + LC 50/LCIG 2 s -F 2 Confidence limits (.0.5)for LC50 Nit to 2-77 N 'f LC s so (.-5:1i@ 9 LCSC / LCSO , lower limit =O.Oba&Zl LC so X f LCso = upper limit -d.Ooaozto ,Anal%,sisby: (game) Checked- By: 4!@-z9 JAI, LC so Date: C.I. 7@ IS (Dat'e) go, *04 goo 0 WW -HIT T, ir LL if 44 . III r7 Ill.[I*i .,I f I 111 fill fill 111 11 L 2 c r iE 7t:t@: @i4 riIILi Ll 1 j-,.L. I II lit! t .-r: TTr 0 31 I I I I ltil III 7 LL IIII I t 1I1f II +1 LI LI@j. . L 47 i' @.. ..4 L T@T' 1 ::Lt lt7' pE, L7 .0 t rL ANALYTICAL BIOCHEMISTRY LABS AQUATIC BIOASSAY LAB TEST FISH MEASUREMENTS@ Test-. Test Speeccaisies: Source: iL. -6-P-C AC Lot .1, Date lieastired Group Measueurrsee*dd:-. % ----------------------------------- -------------------- ... . . ......... .. ............. . Fish No. Standard I;e'yigt'h(m7n) -7 Weight o. 4 3 4 o-4o 0 7 10 Mean Standard (mm) Mean Weight (g) 0 Remarks.- Prepared By: Checked By:k/--//@- Species:--Source:_O,-X,, -omments: ABC Aquatic Research Laboratuz,y Fish Requisition and Disposition Record Date -s -No. Rec. Designated Use*:* Lot c- Tank #:-4-= Size and Disposition Record Length Weight Length Weight Date (mm) Used Date (mm) (gm) Use )ate Drug Used Treatment Record Conc. Time Results )ate t!Dead Comments Daily Record Date #Dead Comments ell4l e? f9 Date #Dead Commen pep ol ol-lire