Document LJRGyYa928047oNkzQ8XxdOBq

ACUTE TOXICITY TO FISH TEST SUBSTANCE Identity: Perfluorooctanesulfonate, DEA salt; may also be referred to as PFOS DEA salt, FC-99, or 3M Sample No. 2. (1-Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-heptadecafluoro-, compd. with 2,2'-iminobis[ethanol] (1:1), CAS # 70225-14-8) Remarks: Test sample is a mixture of the test substance in water (approximately 25% test substance and 75% water). All values reported relate to this mixture. No calculations were made to adjust for the actual concentration of the test substance in the test sample. METHOD Method: Environmental Protection Agency, Ecological Research Series EPA-660/3-75-009, April, 1975. Standard Methods. Type: Static acute GLP: Yes Year completed: 1979 Species: Lepomis macrochirus Supplier: Osage Catfisheries, Inc. in Osage Beach, Missouri. Analytical monitoring: pH and DO / ammonia content Exposure period: 96-hours Statistical methods: Probit analysis. Test fish age: Not noted. Length and weight: Average length = 28.6 + 2.17 mm. Average weight = 0.60 + 0.15 g Loading: 0.2 g fish / L Pretreatment: None Test conditions: Dilution water: Laboratory well water Dilution water chemistry: Dissolved Oxygen: 9.3 mg/L hardness: 255 mg/L as C a C 0 3 alkalinity: 368 mg/L as C aC 03 pH: 7.8 Conductivity: 50 pmhos/cm Stock and test solution preparation: Primary stock prepared in deionized water at a concentration of 150 mg/mL. The test concentrations were prepared by transferring appropriate aliquots 001566 of the stock standard directly to the test chambers. The test solutions were noted to foam when stirring in toxicant aliquots. Test concentrations were prepared based on total sample, not on percent concentration of the test substance in the test sample. Concentrations dosing rate: Once Stability of the test chemical solutions: Not noted Exposure vessels: 40 liter glass aquaria containing 30L of test solution. Number of replicates: one Number of fish per replicate: ten Number of concentrations: six plus a negative control Water chemistry during the study: Dissolved oxygen range (0 - 96 hours): 6.0 - 8.4 mg/L (control exposure) 5.8 - 8.3 mg/L (18 mg/L exposure) pH range (0 - 96 hours) 8.2 - 8.3 (control exposure) 8.3 - 8.3 (18 mg/L exposure) Test temperature: Temperature held constant at 22C through use of a water bath for test vessels. 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.l. not calculated) All element values based on nominal concentrations Statistical Evaluation of Mortality: Probit analysis was used to calculate LC50 values and the corresponding confidence limits. Quality Check for Test Organism Health: The bluegill sunfish were challenged with a reference compound, Antimycin A. The observed 96-hour LC50 and 95% confidence limits (C.l.) were within the 95% confidence limits reported in the literature, indicating that the fish were in good condition. 001567 Cumulative percent mortality: Nominal Test Cone., 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 20 100 100 650 100 100 100 100 CONCLUSIONS The test sample 96-hour LC50 for bluegill sunfish was determined to be 31 mg/L with a 95% confidence interval of 22-43 mg/L. The 96-hour no observed effect concentration was 18 mg/L. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY________________________________________ _________ Reliability: Klimisch ranking 2. This study, while well conducted, lacks analytical data for determination of the test substance concentration in the test solutions and determination of the sample purity. There were also gaps in the measurement of water quality criteria for a number of the concentrations at given time intervals. REFERENCES___________________________________________________ This study was conducted by Analytical Biochemistry Laboratories, Inc. of Columbia, Missouri on behalf of the 3M Company. OTHER__________________________________________________________ Last changed: 5/2/00 001568 Submitted By: Analytical Biochemistry Laboratories, 7200 East ABC Lane P. 0. Box 1097 Columbia, Missouri 65205 Inc. Prepared By: Sty Carl M. Thompson Aquatic Biologist Alan D. Forbis Aquatic Supervisor 7// 3 /net Date Date Approved By: -AviLiilyi (l . l / u J i 1 James A. Ault sOj/alitjuAssurance Officer Date XylMdi. Johnson! Laboratory Manager 1-L-1 Date 001569 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 meas ured throughout the test and were within acceptable limits. As a quality check, the bluegill sunfish were challenged with a reference confound, Antimycin A. The observed 96 hour LCS0 and 95% confidence limits (C.I.) were within the 95% confidence limits reported in the literature (2), indicating that the fish were in good condition. The results of the four day static fish toxicity study using blue gill sunfish are summarized below. Compound 3M Sample No. 2 Antimycin A 96-hour LC5o _______ (95% C.I.)_______ 31 mg/1 (22-43 mg/1) 0.000029 mg/1 (0.000021-0.000040 mg/1) Also, the results indicated a 96 hour, no observed effect concen tration of 18 mg/1. 001570 INTRODUCTION This static bioassay was performed at the aquatic bioassay labora tory 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 LC50 levels for 3M Sample No. 2 to bluegill sunfish (Lepomis macrochirus) . The study was performed following the procedures outlined in ABC Protocol Number 7601 (Appendix I). METHODS AND MATERIALS The procedures for static bioassay, as described in Standard Meth ods 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 day light 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 treat ments, 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 repre sentative 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 laboratory well water. Characteristics of this dilution water are shown in Table 1. These vessels were kept in a water bath at 22C (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 3M Company, six concentrations of the test compound, ranging in a logarithmic series for 18 to 650 mg/1, with ten fish per concentration were selected for definitive bioassay. The fish 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 (Appendix I). 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 (22C). Sample purity was not pro vided; 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 001571 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 LC50 values and 95% confidence in tervals for 3M Sample No. 2 and the reference test against Antimycin 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 concen trations were below the toxic limit (7). The study was conducted following the intent of the Good Laboratory Practice Regulations (8) and the final report was reviewed by Analytical Biochemistry Laboratories' Quality Assurance Unit. All original raw data was provided to 3M Company, with a copy retained at Analytical Biochemistry Laboratories. 3 001572 TABLE 1 Chemical Characteristics of Well Water at ABC's Aquatic Bioassay Laboratory. Parameter Dissolved Oxygen PH Hardness (CaC03) Alkalinity (CaC03) Conductivity Total Ammonia (NH3) N03-N Ortho-Phosphate Aluminum Arsenic Cadmium Chromium Cobalt Copper Iron Lead Mercury Nickel Zinc DDVP Diazinon Disyston Methyl Parathion Malathion Ethyl Parathion 9.3 ppm 8.2 255 ppm 368 ppm 50 umhos/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 ppm 0.009 ppm <0.0001 ppm 0.0157 ppm <0.01 ppm <40 ng/1 <20 ng/1 <20 ng/1 <80 ng/1 <110 ng/1 <80 ng/1 4 001573 < / \ ( i i Compound 3M 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)** LC50 in milligrams/liter (ppm) 48 hours 370 (290-470)** 0.000040 (0.000032-0.000050)** 96 hours 31 (22-43)** 0.000029 (0.000021-0.000040)** *Bioassay as conducted at 22C (1.0), mean weight aqd length, 0.60 g and 28.6 mm. **95% confidence interval. ***Antimycin A standard obtained from Sigma Chemical Company, Type III, crystalline, Lot 125C-0152. 001574 Ul (<< << << << < < ) TABLE 3 The mortality rates and water quality parameters during the acute toxicity test of 3M Sample No. 2 with bluegill sunfish (Lepomis macrochirus) . mg/1 Concentration Control 18 37 75 160 320 650 Percent Mortality Hours 24 48 96 00 0 000 0 0 80 0 0 90 0 0 100 0 20 100 100 100 100 Water Quality __________ 24 hours___________ Temp. D.O.* N H 3* * * C mg/1 pH** mg/1 __________ 48 hours___________ Temp. D.O.* n h 3*** C mg/1 pH** mg/1 22 8.4 8.2 0.86 22 7.6 8.3 1.3 22 8.3 8.3 0.78 22 7.8 8.3 1.3 22 8.1 1 8.1 0.90 22 8.0 8.1 0.78 22 7.4 8.3 1.6 _________ 96 hours Temp. D.O.* C mg/1 pH** 22 6.0 8.3 22 5.8 8.3 22 4.8 8.3 n h 3*** mg/1 1.2 1.2 1.5 *Dissolved oxygen concentrations * Dissolved Oxygen Probe (Extech Model 8012) used with an Extech Model 671 pH and mV meter. **pH - pH Probe (Fisher Model 13-639-108) used with an Extech Model 671 pH and mV meter. ***Total ammonia concentrations - Ammonia Probe (Extech Model 8002-8) used with an Extech Model 671 pH and mV meter. O' C/l 0015 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 Pro tection Agency, Ecological Research Series EPA-660/3-75-009, April, 1975. 61 p. (2) Berger, B. L . , R. E. Lennon and J. W. Hogan. 1969. Laboratory Studies on Antimycin A as a fish toxicant. U. S. Department of Interior, Investigations in Fish Control Ho. 26. 21 p. (3) American Public Health Association. 1975. Standard Methods for the Examination of Water and Wastewater. 14th ed. Washington, DC. 1193 p.v (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 Re search Series EPA-660/3-75-011, May, 1975. 45 p. (6) Litchfield, J. T., Jr. and F. Wilcoxon. 1949. A Simplified Method 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-238 199. 592 p. (8) Food and Drug Administration. Regulations for. Good Laboratory Practice. Federal Register, Vol. 43, No. 247, 59986-60025, December 22, 1978. 7 001576 Quality Assurance Statement for final report #23893 entitled, "Acute Toxicity of 3M Sample No. 2 to Bluegill Sunfish (Lepomis macrochirus) for Dale Bacon, 3M Company, St. Paul, Minnesota. In accordance with ABC Laboratories intent that all studies con ducted at our facilities are designed and function in conformance with good laboratory practice regulations and the protocols for individual laboratory studies, an inspection of the final report for 3M Sample No. 2 was conducted and found to be in an acceptable form by a member of our Quality Assurance Unit. An inspection of the daily mortality rate of the test organisms prior to the initiation of the study indicated they 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 apt been supplied, the LC5o would have to be calculated based on nominal concentrations of the total compound. A final inspection of all data and records on June 20, 1979, indicated that the report sub mitted to you is an accurate reflection of the study as it was conducted by ABC Laboratories. Should you have any questions relating to the information provided in this statement or the function of our Quality Assurance Unit, please contact me at your convenience. 8 001577 APPENDIX I RAW DATA 001578 14 ANALYTICAL BIOCHEMISTRY LABS Aquatic Bioassay Lab BEST COPY AVAILABLE Acute Toxicity Bioassay D ate Time _ T o x ic a n t : 3 j y i - ~ l - _________ Lab N o. : Z J g q ' l Form ulator: ^sT AD ate Madel it?A ( Tank:-- 3 P Punty: S olvent: 1 Stock Standards F in a l Gross Wt. 3 o .o *o g Tare w t. 0 .o*o g Net w t. 3o 0*0 g A d j. n e t w t . Jo-OCO g D ilu tio n v o l. Too ml C o n c e n tra tio n ( q mg/ml W orkxinhig S ta n d a rd s A liq u o t o f concentrated stock D ilu tio n V o l. _________ ` F in a l C oncentration mg ** a + 0 -*o a je l,opo 5 Or"'J c+") T est Species : Lo t No. Temp 2 Z C . S o u r c e i ^ r y ^ a f / ^ 7i>r- Cfandition Weight: Length.: N o ./V e ssssee lV u7 Vessel S ize : W a te r Chemii stry : u4agJ?4/g o pH: T . g Void Alkalinity :-gfoy3 E ACL Hardness ; 2 ^ ^ ---------------------------------- -------------------------------------------------------------------------------------------------- C oncentration 24 h r - fvup/Ctf m l Dead Obs. Dead Obs. 48 h r 72 h r 96 h r Obs. ttead O bsT De ad Obs. Dead Obs. ^M TCttt- -- o Q CO C5 >37 75 Ik o - 320 5. L? 7^ 15 U p4- 0 0 &D c> n ** 1 -ft 0 O 57 0 o 7 o- t o toCO Z . lL o \ /( 7 (3 0 w \ o Iv |r; / n w O b server Date Ct T y *- - J^r,s ~ - T -- T _____ ____ v __________,_____1_r_t_i.U_c_a_r_ft_r_i_L_ iledi-- S'^<VA<lUw[ t o ____3 g > J p __-----------------\^.y4~ ared by : -C o r r e c te d f o r p u rifty o f' prr3im- a r y s ta n d a rd **Checked w ith class S w eights Checked b y : ~fk<. -jO _____ _ <! 77~a^- 001579 3Wi,i Water Quality O *\ocof5 24 hours J+8 hours 96 hours oncentratiori Tgmp. D.O.* NHS** T gmp, D.O. * NHfi*"* Tgmp. D . O . 1 NH*** Tgmp. D.O.* NH*** TM g /]- ml_ pH** M -Z l mg/1 pH** m g J/l mg/1 pH** m g J/l mg/1 pH* m g V l Z.'L % -7 9 - 2 . (.3 &A. QJZki. ?,k S 3 /.3 it- 6 ' 0 . 3 /. 2. ! n. 1 1 <*.G /. o z ~t~ jl 0-T *7.s f .3 /.3 jS~. <y 5 - 3 A Z . J i . 2Zl. z *- s .0 ~-^t Q - --- <.2 /, / Z **- M - 0 . 9c? 2 t. EZ- V i 'zi' 7. V -it? -X ^ 4 S.3 3 " f.3 A fe jqo "3 /. rror 3 G Cr//i* ^ D i s s o l v e d oxygen concentrations - Dissolved Oxygep Probe (Extech Model 8012) used with an Extech Model 671 pH and mV meter. Or? C * * p H - pH Probe (Extech Model 200C) used With an Extech Model 671 pH arid mV meter. ***Ammonia concentrations - Ammonia Probe (Extech Model 8002-8) used with an Extech Model 671 pH and mV meter. 11 ANALYTICAL^ BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay -Toxi cant 3 frVy Probit Analysis Work Sheet -------- ---- Date tested - tyf-z. - (o/,(0 h ^ Test Species G u d I^ Date reported !p /(n Lab Number '?S~3y^c\^ ^____________ Temperature___ 'ZZP C Exposure period bi/<^ Water quality lo-gXff Concentration i ( No. d e a d / ! total no. Observed % mortality %E x p e c t e d | Contribution mortality J"E ; to Chi rokrTO-C- v Of(C> ioO 1 Otto 12,0 ! <?/ /O i -O - ! ------ O ,1 O f> i 0 .0 0 0 L hA 7 / 4 _______ ______ " . | i o - o g a____ (0/(0 ! / ...________ / / ______ 2 4 ____ * > ! -o t ? o ____ 11 41 .* ; !i Y^l animals 30 Total contribution to Chi_____ O) l K LC8i 2 l Chi*- contribution X total animals = /-ffp to Chi ,K Chi2 (p-.05) for (K-2)_J_deg. of freedom LC SO dL^o LCi6 " '$~LO S - LC /LC _ + Le /LC,, 81 SO_____ SO *6 s - Abn+,c 4TfXc o?/zo - /fr Confidence limits (.05) for LCsq N1 2^0 , f LCeSO,, = S 2-77 / N f F - = /,2.S LC50 / f LCS0 =* lower limit " 3*70 2 r \r - W x LC50 001581 l ANALYTICAL_BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay -Toxicant 3 ^ Probit Analysis Work Sheet Date tested_^ Test Species Date reported Lab N u m b e r __ -exposure period Concentration ... - CO^STjrZBL^ ____ l( o - ?20 _________ 2 ,6 0 Temperature__ - . ' P J Z P C __________ kvt4 8 ________ Water quality [s j j U l Q. t | No. ; : 'V I I 1 dead , ,otBl no- O/ C>/ t o 2 -/iO J / t O ______ ' / Observed % mortality Expected % mortality 1 Contribution "E ! to Chi o -- i -- 1--------l f.n ... ( CD j O -oc''D Z.O / ______l o ____ in| i l o o f a h _____ ________ : I o ooq ~! 1! t^ r a l animals K 22. 4- - l.C8* 44 o LCS0 LCu ?1Q IS? O S -* L CS/kL C 50__+__L_CSO/IC., / L : i . .' / !\ T otal c o n trib u tio n to Chi________ O 0 ___________ Chi2- c o n trib u tio n X to ta l animals to Chi K 0 Chi2 (p -.0 5 ) fo r (K -2) l deg, of freedom * ______ S- s * y & * S o_ = /.*} Confidence lim its (.0 5 ) N1 - II O ~ f lc50 s2- " / \ f F - . LCS0 / f i-CSjQO * lower lim it = 2 ^ 0 LC50 X f LCg j j * upper l im i t Analysis by: 'fUir-Uso - ? 7 Q , T ^l fyi f_ ? 7 o *y ^ j/} \ Cfiecked By: J d ^ j l J j D k P a t e-.j J & f . , * 0015^ il ANALYTICAL^ BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay ^Toxicant .3-00: Test Species Lab Number Probit Analysis Work Sheet Date tested, .; fo/fn- - (>/{(r>/-7Cf _Qry f' II ^ *"*} Date reported _________________ Temperature... . Water quality [joJUL# 4f-^ O : Concentration j No. dead Observed % Expected % J j C ontribution ! ----- p ------------------- 1 ... C nKj-rRi)<-- to ta l no. O / ( m ortality m o rta lity j0-E ! . j ___ | to Chi _-- !Z 1 I------------- ln--" /--n\<*n------------ 1 --1-1------------------- 5^/10 ............ 9 o 1(0 I> 4 ! fM ! 1 /(f. | 0 .0 ? ___ /k rO * ! ^ /ro 7 t> f [r i / 9 ! I a.oZcy l o ( n < f ) j q q 'i o | c>-ae>o } i! / ii ' ^ a l animals K l-C8lf LC 50 LC** = 3J. JLL Ar T o tal c o n trib u tio n to Chi Chi2- c o n trib u tio n X t o ta l animals = 2.C?Q _________ to Chi K Ch! (p -.0 5 ) fo r (K -2) - deg, o f freedom ... ...... ... ^ 0 -eifr*-i C T fc/n/* s - lC**/LCs * LCs / LCu ' s- ___ = (.no Confidence lim its (.0 5 ) fo r LC$o N' ~Z 0 _______ f l c 50 s2-77 / n Tn 7" , A3^ LC50 / f LCS0 lower lim it = ~t TL. % Kr LC50 3 ( C~ZZ ~ 4 5 9521- C. I / 0015S3 BEST COPY AVAILABLE 0015S4 iO. 32.376. \ RITHMIC NORMAL.^ ' M STOCKDIRECTr*{ CODEXBOOKCO.r/ WOOD.MAS.0206/ 14 ' ANALT l CATTUITI CTID'libx'KY LAbb Aquatic Bioassay Lab Acute Toxicity Bioassay Tew*cant: /\.A. *v* % vs. A 'Formulator y'JtwJT CbCOCK Stana 4rqs Final G ross W t . ___2 0 g Tare wt. n.OO g Net wt. _ Adj. net wt. 3 0 WS.g JO -- g* D i l u t i o n voi. (a o ml Concentration^?. 3q m Date . - ^ 2 Time: { . ' 0 0 f Tank: Lab No. :^ 4re^jt. l o t i c i Purity : Pate M a d e : . .... Solvent: Working Standards Aliquot of concentrated stock Dilution Voi. - Zoo' Final Concentration O. 0 0 / 5 **) mg [ 1er cn Test Spee<cies : J / i U Lot No. l < 2 SourceO_ sku*. (^TyNtoariei Condit ion : odi Temp.: 2 i V ______ Weight: fl, x <a Length. : Z4<? '"No. /VeSsSseji :6 ____ Water Chemi stry : ^ VesselJSize: 2 0_____ Void pH: 7 . ! Alkalinity : ___________ _ f y Hardness :<f.f -------------------- Y a - r - - - ---------------< Concentration 24 hr - -S-/V ml Dead Obs. 'Dead Obs. A SEVckJ utei.-,L b /.$ o 0 3.00001% A /& 0 60 s.osiMz. a .3* 0 e> 3X o 3.noooi'b A Sfc 1 & ,S(. 1 Of 0 0 0 /0 A __________ s _ 0- Q O /X A /.o /. 3 3 T 6 l- % if 48 hr Dead Obs. o o O o 0s 5 $ 72 hr 96 hr Dead O bs. Dead Ob s . <9 O O _JL i 2-- |5 s 5 ?5 i : 5$ Dead - )bserver 5ate Remarks : il) Prepared by: ^Corrected **Checked U-- tfrL. ct s rr (e /h t/M - CT 4h . or ary standard Checked by: 0015S5  11 ANALYTICAL^ BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay v AIwxicant Test Species Probit Analysis Work Sheet _____________ Date tested jfy/ff - Date reported ^______ Lab N u m b e r ____ Temperature ________ Swposure period * C o n c e n tra tio n ft/y J J p r- C o A j n r i o < ^ ' C?.Oo<So?tL O '0&oCClo 3 4 - hr<___________ W a t e r quality <D *i j No. dead , | / to ta l no. v & / to i lO Observed % m ortali ty O 0 r .f) 1 Expected % I I Contribution m o r ta lity j "* ; to Chi jj { | \0 1 1 *0 0 1 CO z> ' 'Z'eD ! O I o >00 a .00& t o ToU^-. animals ' K LCgi| * Q-Otecfo I 1 3J23. (& / tO f C Q / f r W V ____ 2 - ___ i V - C 4 - C ,_____ / i! / <! / i T o ta l c o n trib u tio n to Chi______ Chi2- c o n trib u tio n X to ta l animals = to Chi K o . S . Chi2 ( p -.0 5 ) f o r (K -2) I deg; o f freedom ** 3 *T" tC o .o**rTO 50 -------- -- LCie O 00 & t S 9-* LCft /LC 8** SO + LC_ / 50 LC\.6, s = - ^ T a g e o ^ g 7^ -- Tauio^^t /r Confidence limits (.0 5 ) for LCso N1 - \ Q _________ r LC50 - S2- 77 / NTn 7 = t $ Z.^rW' LC50 LC5o / f LCS0 * lower limit * O'ooeejb 0015S6  11 ANALYTICAL BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay T o x i c a n t ---~ Probit Analysis Work Sheet ^ ___________ Date tested rfL -^(o/t Test Species j d L j t<Sijr Date repo r t e d_ j/f ________ Lab Number M Temperature Ihxposure p e r i o d ^ ___________ W ater quality T i p c*^. Concentration K \ C ffdP r d /O T /7 /3 'C <?.cwo<K O .e e o o ^ l ftt j No. dead , * 7 t o t a ' a* Observed % m ortality :-v O /i O 1 0 no tO ^ 2 -// ? Expected % j Contribution m o r ta lity P " E to Chi - -- r J ____ ' t __ ! < o \ O 'OOO __ ~^sz> 1 O ! >.e*> Q .Q it& U ) 1j* [ 0 / \C> / / lo o fn ^ 1! d1 .5 / ?i .T o t^ . animals K I Jl LCgi^ a .o*ix>~z- 't.c so = ------o-c--*-*-4 o LCie 1 S *- * LC8</LC 50__+__L_CSO/L C1,6, T o ta l c o n trib u tio n to Chi_____________ O ______ ChJ2- contribution X to ta l animals O 4 ^ % to Chi K Chi2 (p -.0 5 ) fo r (K -2 )_ J _ jd g . o f freedom ** T~_ fO<*>*k coati ro^fO^ ^<00*071a /, 3 0 Confidence lim its ( .0 5 ) f o r LC$o * - ____ id_____ Y LCS0 - S2- 77 / V f iF = A "2 - 5 LCS0 / f LCS0 lower l i m i t = 4 0 V r LC50 0 - 0 Q t a o O.Qtfoe^C 1 ' QPfr.l. 001557  11 ANALYTICAL^BIOCHEMISTRY LABORATORIES Aquatic Bioassay Laboratory Acute Toxicity Bioassay Toxicant Test Species Lab Number Probit Analysis Work Sheet COT -is a a Date tested Va - Date reported_ b fa f-v t Temperature___ Exposure period C o n c e n tra tio n Y tf/tP " f A STfZ *- ' _ __ o.>ooo \ V anim als K LCS^ Q%o&ooH-'Z~ X C 5o ** faCOOG'Zi^ L C j e = Q .a*e^> Z-0 W a t e r quality f2pra*\ J i z jD_ j No. dead y j to ta l no* Observed % m ortality :v 0 //e> > Expected % m ortality 6 -- 1~ 1 Contribution ! to Chi i-- /(O 30 ( O i tn\ o - t z - o / / o ________ U n ____ 1 to 1__ o><3 4 -0 .___ j (<* ? (O /fo il! 2 T ! >*<3 < 2 0 / / i1 *1 .4 ; / !J 3 g-- . -- 7*---- 1 T o ta l c o n trib u tio n to Chi_____ C2 / & & ___________ C h i*- c o n trib u tio n X to ta l animals to Chi K ( g o Chi2 (p -.0 5 ) f o r fK -2 ) ( dq. o f freedom 'S S - L C 8a i1/ L C Sc O,, + L C 50/ L C 1.6ft - -- -> t<5<0^5- , #0*00.f 'S = "*'T~g***2-c? ` = Confidence li m i ts ( . 0 5 ) f o r LCs (jo 1 5 $ 8 rs M ++ ri.Lij_'.ilaJj u l l'i'il BESTcopy AVAILABLE /O0/O fOMfg /toi Y 0015S9 BESTCOPY AVAILABLE \ O' analytical biochemistry labs AQUATIC BI0ASSAy WB TEST FISH MEASUREMENTS, Fish No. 1 2 3 M S :s 7 8 9 10 ~Mean Standard Length (mm) Mean Weight (g) Remarks: 1 0 Standard ` `' Length '(mm) Weight ________ : <gr --t--- --- TM 27 Jo 2.4 3o ?o 5c? ' Z(o Q*l& 8 0.4-Z. O.4o o. $g________ .O*(oz* Q.(p? ________ Q'~TZ~. _____ 7^2- ----f-S-.h.-=--2-.-nVt -- 0 `(pc> ^S.h.-o. l*= * P r e p a r e d By: Checked /? r 001590 BEST CO PY AVAILABLE ABC Aquatic Research Labor!tory ish Requisition and Disposition Record Species :g}u4J,/l ________________ Date Source : , / A ie_ik_s_______________ N o - Rec- ./O O P -- ~ ` _____________ ^Designated U s 7f Tank #: / e : _______ _ Comments : Date w Length (mm) Size and Disposition Record Weight (gm) V. f # Used Date Length (mm) Weight (gm) if Us 3ate Drug Used Treatment Record Cone. Time Result: Daily Record