Document Rab89xDrq7Dw6ZenDeGrBpbMz

I DEPARTMENT OF THE NAVY NAVAL RESEARCH LABORATORY W A S H I N G T O N , O .C , 2 0 3 7 8 --1 0 0 0 IN REPLY REFER TO: 3900 Ser 6180-256 26 April 1991 From: Commanding Officer, Naval Research Laboratory To: Commander, Naval Sea System Command (SEA 5122P) Subj : Ref : RESULTS OF QUALIFICATION OF AFFF LIQUID CONCENTRATE FIRE EXTINGUISHING AGENTS MANUFACTURED BY THE 3M COMPANY (a) 3M ltr of 15 Oct 90 Enel : (1) Two copies of subject report. 1. In accordance with reference (a) t the subject products have b e e n , tested in order to determine their compliance with the requirements of MIL-F-24385E(SH). A report covering the work is forwarded herewith as enclosure (1). Copy to: (w/copy of e n d (1)): NAVSEASYSCOM (Code 56Y5, B. Darwin) Naval Weapons Support Centers (Code 304E) Distribution Authorized to US Government Agencies and their Contractors Only; All other requests shall be forwarded to: Commanding Officer Naval Research Laboratory, Wash. DC. THIS INFORMATION HAS NOT BEEN APPROVED FOR PUBLIC RELEASE. US00000299 6180-256A:CRF:tie 26 April 1991 Subj Ref: S S S ? , t 0F q u a l i f i c a t i o n o f AFFF LIQUID CONCENTRATE FIRP EXTINGUISHING AGENTS MANUFACTURED BY THE 3M COMPANY (a) M i l i a r y Specification, "Fire Extinguishing Agent Aqueous Film-Forming Foam (AFFF) Liquid Concentrate 1990 ^ SSa Water'" MIL-F-24385E(SH), November (b) Qualified Products List, "Products Qualified under Aaeiia r Y aSPeClfiCat-n MIL"f "24385, Fire Extinguishing Agent, Aqueous Film-Forming Foam (AFFF) Liquid Concentrate, for Fresh and Sea Water," QPL-24385-So September 1990. 4385 20, ai FF-F2F03Cc ? Cf?%r?vnS ed tbe 3M Company and designated accordanceFwith t r ^ ^ r o c e d u r e s ^ u t l i n e d ^ i n ^ ^ f e r e n c e ^ a ) ^ 03*"6** *n 2- The test data obtained are summarized in the attached tables, reference (';^didate AFFF concentrates met the requirements of (2~ $ (?. Carl R. Fulper Fire Protection Section Naval Technology Center for Safety & Survivability NRL Code 6184 1 Joseph T . Leonard Head, Fire Protection Section Naval Technology Center for Safety & Survivability NRL Code 6184 Attachments 1) Tables l-io 2 ) "Results of Toxicity Tests 96"?OU5' J undulus beteroclitus Flow-Through Acute Laboratory," EA f n a i n I e S t M a t e r i a l s from th Naval Research Engineering, Science and Technology," March 1 9 9 1 Distribution Authorized to US Government Agencies and their Contractors Only; All other requests shall be forwarded to: Commanding Officer Naval Research Laboratory, Wash. DC. THIS INFORMATION HAS NOT BEEN APPROVED FOR PUBLIC RELEASE. US00000300 Tab,e T Concentrate Chemical and Physical Characteristics Test Results (Paragraph 3.3) Characteristic Measured Refractive Index, minimum Viscosity (centistokes) maximum @ S'C minimum @ 25* C Hydrogen Ion Concentration (pH) Spreading Coefficient, minimum Foamabiiity, minimum Fresh Water; Expansion Ratio 25% Drain Time (minutes) Sea Water; Expansion Ratio 25% Drain Time (minutes) Film Formation and Sealabliity Fresh Water; Half Strength Full Strength Sea Water; Half Strength Fufl Strength Quintuple Strength Specifications Requirements i voe 3 Tvoe 6 1.3630 1.3580 20.0 2.0 7.0-8.5 3.0 10.0 2.0 7.0-8.5 3.0 6.0 6.0 2.5 2.5 6.0 6.0 2.5 2.5 No Sustained Ignition (for ail solutions) No Sustained Ignition (for all solutions) Corrosion Generai; maximum -Steel (milii in./yr) -Cu-Ni (milii in./yr) -Ni-Cu (milii in./yr) -Bronze (milligrams) Localized CRES -Steel Total Halides, maximum (ppm) Dry Chemical Compatibility Burnback Resistance, minimum (s) Environmental Impact Toxicity, LQ 0, minimum (mg/l) COD, maximum (m g/l) ^Ot-WCOD, minimum 1.5 1.0 1.0 100 No pits 500 1.5 1.0 1.0 100 No pits 250 360 360 500 1.000,000 0.7 1000 500,000 0.7 Results Type 3 Type 6 FC-203CF FC-206CF 1.3660 1.3589 7.3 3.3 3.9 2.1 8.3 8.1 4.4 6.7 6.7 7.4 6.4 5.6 7.6 7.6 5.0 4.6 Fiashed-NS No ignition No ignition No ignition No ignition No ignition No ignition Fiashed-NS No ignition No ignition <1.5 <1.0 <1.0 <100 No pits 55.6 <1.5 <1.0 <1.0 <100 No pits 31.9 41^ 690 1449 585,000 0.7 3613 295,000 0.7 0) 25 ^ burnback never achieved. Burnback pan pulled at 387 s. and fire was completely extinguished at 416 s. Foam blanket then closed in on hole NS No Sustained Ignition ' Nl No Ignition 1 US00000301 Tbie 2, Stability Test Results (Paragraph 3,3,2) Property Measured Specification Requirement (all solutions! Results Type 3 Type 6 Fresh Sea Fresh Sea Spreading Coefficient, minimum Concentrates (fresh water only) Pre-mixed Solution (fresh water only) 3.0 3.0 4.9 -- 6.9 6.2 -- 7.4 Foamability, minimum Concentrates; Full Strength (fresh and sea) Expansion Ratio 25% Drain Time (minutes) Pre-mixed Solutions; Full Strength (fresh/sea) Expansion Ratio 25% Drain Time (minutes) 6.0 2.5 6.0 2.5 7.4 7.2 6.7 6.3 5.3 4.7 5.6 4.7 6.7 7.2 6.3^ 6.0 5.2 5.3 4.9 4.8 Fiim Formation and Seaiabilrty Concentrates; Half Strength Full Strength Pre-mixed Solutions; Half Strength Full Strength No Sustained Ignition No Sustained Ignition No Sustained Ignition No Sustained Ignition Nl NI Ni NI NI NI NI NI NI NI NI NI NI NI NI NI Fire Performance, 2.6 m2 (28 ft2) Concentrates; Half Strength (fresh/sea) -Extinguishment Time, maximum (s) -Burnback Time, minimum (s) Concentrates; Full Strength (fresh/sea) 45 300 40 42 33 35 421 413 465 390 -Extinguishment Time, maximum (s) _ -Burnback Time, minimum (s) Pre-mixed Solutions; Half Strength (fresh/sea) -Extinguishment Time, maximum (s) _ -Burnback Time, minimum (s) Pre-mixed Solutions; Full Strength (fresh/sea) -Extinguishment Time, maximum (s) -Burnback Time, minimum ($) 30 360 45 300 30 35g 26 28 22 19 502 427 530 480 42 43 44 35 442 511 500 499 26 21 28 22 563 473 545 395 Stratification Concentrates Pre-mixed Solutions; Half Strength Full Strength None None None None None None None None None None None None None Precipitation, maximum {% volume) Concentrates Pre-mixed Solutions; Half Strength Full Strength <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 <0.05 Expansion Ratio corrected for temperature as per NFPA 412 No Ignition 2 US00000302 1ab|e 3. Interagent Compatibility Test Results (Paragraph 3.3.3) for 50/50 Mixture of 3M FC203CE and FC203CF ' Property Measured Foamabillty, minimum Expansion Ratio 25% Drain Time (minutes) Film Formation and Sealabillty Fire Performance, 2.6 rri (28 ff) Extinguishment Time, maximum (s) Burnback Time, minimum (s) Stratification Precipitation, maximum (% volume) Specification Reqirement (all solutions! 6.0 2.5 No Sustained Ignition 30 360 None <0.05 Results Concentrates Fresh Sea Pre-mixed Solutions Fresh Sea 7,7 7.2 5.4 4.9 Nl Nl 6.8 7.0 4.9 5.3 Nl Nl 30 26 525 505 None <0.05 27 559 None <0.05 24 545 None <0.05 Nl No Ignition 3 US00000303 Tabi 4. Interagent Compatibility Test Results (Paragraph 3 3 3) for 50/50 Mixture of Ansul AFC-5A and 3M FC203CF Property Measnred Foamabiiity, minimum Expansion Ratio 25% Drain Time (minutes) Film Formation and Sealability Fire Performance, 2.6 m2 (28.0 ft2) Extinguishment Time, maximum (s) Burnback Time, minimum (s) Stratification Precipitation, maximum {% volume) Specification Requirement (all solutions} 6.0 2.5 No Sustained ignition 30 360 None <0.05 Results Concentrates Fresh Sea Pre-mixed Saint Fresh Sea 7.1 6.2 5.4 5.3 NI NI 7.2 6.3P 5.5 4.8 Nl Nl 25 26 549 454 None <0.05 23 583 None <0.05 25 505 None <0.05 Expansion Ratio for temperature as per NFPA 412 No Ignition 4 US00000304 Tabje 5. Interagent Compatibility Test Resuits (Paragraph 3.3.3) for 50/50 Mixture of 3M FC206CE and FC206CF ' Property Measured Foamability, minimum Expansion Ratio 25% Drain Time (minutes) Fiim Formation and Sealability Fire Performance, 2.6 mP (28.0 ft2) Extinguishment Time, maximum (s) Bumback Time, minimum (s) Stratification Precipitation, maximum {% volume) Specification Requirement (all solutions* 6.0 2.5 No Sustained Ignition 30 360 None <0.05 Results Concentrates Pre-mixed Solutions fliM ? Sea Fresh Sea 7.3 7.2 5.0 5.5 NI NI 6.9 6.1n) 4.9 4.8 NI Nl 28 29 542 439 None <0.05 26 24 548 538 None None <0.05 <0.05 Expansion Ratio corrected for temperature as per NFPA 4 12 No ignition 5 US00000305 Table 6. Interagent Compatibility Test Results (Paragraph 3.3.3) for 50/50 Mixture of Ansul AFC-5 and 3M FC206CF Property Measured Foamability, minimum Expansion Ratio 25% Drain Time (minutes) Fiim Formation and Sealabiiity Fire Performance, 2.6 m2 (28.0 ft2) Extinguishment Time, maximum (s) Bumback Time, minimum (s) Stratification Precipitation, maximum {% volume) Specification Requirement (all solutions) 6.0 2.5 No Sustained Ignition 30 360 None <0.05 Results Concentrates Pre-mixed Solutions Fresh Sea Fresh Sea 6.7 6.9 5.1 6.1 NI NI 7.4 7.4 5.8 4.4 Nl Nl 29 26 450 485 None <0.05 22 23 552 468 None None <0.05 <0.05 Nl No Ignition i US00000306 , . . . J f 51 7- 'nteragent Compatibility Test Results (Paragraph 333) for one-third/one-third/one-third Mixture of Ansul AFC-5A, 3M FC203CE, and FC203CF Property Measured Foamability, minimum Expansion Ratio 25% Drain Time (minutes) Film Formation and Sealability Fire Performance, 2.6 m2 (28.0 ft2) Extinguishment Time, maximum (s) Burnback Time, minimum (s) Stratification Precipitation, maximum (% volume) Specification Requirement (all solutions) 6.0 2.5 No Sustained Ignition 30 360 None <0.05 Results Concentrates Pre-mixed Soluti Fresh Sea Fresh Sea 7.0 7.1 5.1 4.7 NI NI 7.0 6.8 4.6 5.1 Nl Nl 28 26 467 528 None <0.05 25 28 O) 539 None None <0.05 <0.05 1Ni No ignition () 25% burnback never obtained. Burnback pan pulled at 368 s and fire was completely extinguished at 446 s. ' Foam blanket then closed in on hole 7 US00000307 fnr nnfi thJ a,b!e 8;i, . '? f ragent Compatibility Test Results (Paragraph 3.3.3) e third/one-third/one-third Mixture of Ansul AFC-5, 3M FC206CE, and FC206CF Property Measureri Foamability, minimum Expansion Ratio 25% Drain Time (minutes) Film Formation and Seaiabiiity Fire Performance, 2.6 r r f (28.0 ft2) Extinguishment Time, maximum (s) Burnback Time, minimum (s) Stratification Precipitation, maximum (% volume) Specification Requirement (all solutions! 6.0 2.5 No Sustained Ignition 30 360 None <0.05 Results Concentrates Fresh Sea Pre-mixed Solutions Fresh Sea 6.6 7.0 5.3 6.0 Nl Nl 7.6 7.6 5.5 4.9 Ni Nl 26 23 478 463 None <0.05 26 26 466 506 None None <0.05 <0.05 Nl No ignition 3 US00000308 Tabie 9. Fire Performance Test Results (Paragraph 3.4) .Property Measured 2.6 n t (28.0 ft2) Fire Tests Extinguishment Time, maximum (s) Presh water, Half Strength Full Strength Sea water, Half Strength Full Strength Quintuple Strength Burnback Time, minimum (s) Fresh water, Half Srength Full Strength Sea water, Half Strength Full Strength Quintuple Strength 4.6 m2 (50 ft2) Fire Tests Extinguishment Time, maximum (s) Burnback Time, minimum (s) 40 s Summation, minimum Specification Requirement (all solutions! 45 30 45 30 55 300 360 300 360 200 50 360 320 Results Type 3 Type 6 FC203CF FC206CF 38 40 21 20 34 34 27 22 33 36 450 390 710 567 449 475 498 621 253 370 46 37 402 369 323 328 Table 10. Cap Removed Torque Test Results (Paragraph 5 .1 .1 .1 .1 ) Pass/Fail Criteria Maximum Torque fln-fh) 50 Type 3 FC203CF Results 30 Type 6 FC206CF 31 9 US00000309 FRLEOSWUNOLA-NTTVHSTAREOLSOFTRU9EGM6S-HAHETAOAERUCRCRUIHTAFELLuSAnTdBFOuROXlOaRIsCAMhITTeOTtYeHRrEYoTcEliStuTsS Prepared for: NavaWl RaessheianrgcthonL, aDb.oCr.atory Prepared by: EA EEnAginMeeidSr-pinAa1grt5,lkasSLn,cotMiivecneaRtcroeeyn,glaaiConnniddracl2Tle1Oe1c5ph2enroaltoiognys, Inc. March 1991 EA Project 70003.10 ReportMwwrr 7B US00000310  4 Engineering, Science, and Technology. Inc. EA Project 70003.10 FRLEOSWUNOLA-NTTVHSTAREOLSOFTRU9EGM6S-HAHETAOAERUCRCRUIHTAFELLuSAnTdFBOuROXlOuRIsCMAhITTeTOtYeHRrEYoTcEliStuTsS Prepared for: NavaWl RaessheianrgcthonL, aDb.oCr.atory Prepared by: EA EEnAginMeiedSr-ipnAag1rt5,klasSIn,cotiMivecenaRtcroeeyn,glaaiConnniddraclT2le1Oe1c5ph2enroaltoiognys, Inc. PWroajyencetCtL-MO'.'aMnacgCeurtfo lraegverH, .ABqlu^actkic/PTho.Dxi.cology Laboratory March 1991 S Me*. M i Date R eport N um ber 711 US00000311 1. INTRODUCTION the request of the Naval Research Laboratory, Washington, D.C., BA Engineering, Science, and Technology, Inc. performed flow-through acute toxicity tests on two formulations (type 3 and type 6) of aqueous film-forming foam (AFFF), fire fighting agent. The objective of these tests was to determine the 96-hour median lethal concentration (LC50) of t e test materials to Fundulus heteroclitus (mummichogs) using lethality 1 US00000312 2, METHODS AND MATERIALS 2.1 SAMPLE DESCRIPTION The test samples were delivered to EA on U January, 1991 by courier, ne liter bottles each containing ,ype 3 and typ(s 6 aq,,eous 11m-ormi,,g TChlin J r ' ShiPPed l0gged in' ViSUally *" "<' S**"*' '"e USt y Records, and assigned EA accession numbers (EA sample numbers ATI-017 and ATI-018 respectively,. Based on initial test results an additional one liter of type 6 AFFP was requested in order to 1991su ficient sample volume, and that sample arrived on 5 February amp e #ATl-065). Samples were stored at room temperature (20 C) when n o t in u se . 2.2 TEST ORGANISMS Juvenile mummichogs, Fundulus heteroclitus. constituted the test organisms. The test organisms were 92-96 days old at test initiation. The fish were obtained from cultures maintained at Hampton Roads Sanitation District in Virginia Beach, Virginia. separate lots of F. heteroclitus (lot numbers FU-00A and FU-OOAA) were received on 10 and 21 January, respectively. The fish were held at 20 C and 20 ppt salinity and were fed flaked food daily until used in test ng. The mean acclimation temperatures and salinities for lots FU-OOA and FU-OOAA were; FU-00A-19.7 0.7 C and 20.8 0.5 pp, and FU-004A-- 20.2 0.8 C and 20.8 0.6 ppt. There was no mortality observed in either lot of fish during the acciimation period. The mean length and weight of lot FU-OOA organisms vere 22.58 (.1.52) mm and 0.23 (0.05) g, respectively. The mean length an weight of lot FU-00AA organisms were 23.A9 (1.67) mm and 0.27 (0.07) g, respectively. 2 US00000313 2.3 DILUTION WATER The water used for acclimation, holding, and for the flow-through studies Mari T ' ^ l'lal sea"ater- DrP synthetic sea salts (Forty Fathoms, nt*rprls,s' Touson' Maryland) were added to dechlorinated tan waier allowed to age prior to use. A summary of the chemical c aracterrstrcs of Forty Fathoms is presented in Table 1. The source of he makeup water was Baltimore municipal water supply, which upon entry n o t e laboratory was passed through a high capacity activated carbon C r natin U n U Vhich removes chlorine and hexane soluble compounds. 2.4 TOXICITY TEST OPERATIONS AND PERFORMANCE Test procedures used in the two flow-thrnngh studies followed those inSg*"be! in !he P" t0,:1 ATS-FAP-GT-()1 (Attachment I) which is contained * <'"atic Toxlclogy laboratory Duality Control and Standard withCaeuiH ::OC6dUreS Ha"Uai <EA' 1986> EA'S f-otocol in in conformance . guidelines contained in ASTM's E729 procedure, and with EPA (Peltier " e r ' 1985> and Standard Methods (APHA, 1989) documents. The tests employed Masterflex peristaltic pumps which continuously e ivered test solution from the stock solution tanks to splitter assemblies for each test concentration. The fill and dump splitter assemblies divided the solution flow equally to the two replicates ambers for each test concentration. Each test was comprised of five i T T TM 1" " aD`` 3 C0ntr01' The t7Pe 3 AFFF teSt ri" " 115, 240, . ' ' 2,100 pp"' (vnl7vol) and control, while the type 6 APFF test series was 420, 750, 1,350, 2,400, 4,200 ppm and control. * r M h 60 llt,r b,tcl1 o '`n11 test concentration was prepared in a 20 gallon aquarium using 20 pp, Party Fathoms artificial seawater as llution water and control. There were a total of six 20 gallon aquaria, one for each test concentration and control. The masterflex pumps were 3 US00000314 calibrated to deliver teat solution at 40 nl/ain to each concentration's splitter assembly, asterflex silicone tubing was used, and test solution did not come in contact with any pump parts. T h e t e S t charabers were 2 1/2 gallon all glass aquaria, which were calibrated to hold 5 L of test solution at the standpipe overflow. Based on the 40 mi/min flow to the two replicate tanks per concentration, there were more than five 90 percent replacement volumes to each test chamber during a 24-hour period. Each test concentration exposure group consisted of two replicate chambers with 10 organisms each. Mortality observations, pump delivery calibrations and water quality determinations were made daily throughout the 96-hour exposure period, and were recorded on the data sheets. Copies of the original data sheets are included in attachment II. Temperature, dissolved oxygen, pH and salinity were measured in each batch of new test solution daily and for the old test solutions, in at least one replicate of each concentration at 24, 48, 72, and 96 hours. Aeration was provided to the test chambers when the dissolved oxygen-saturation in any of the tanks dropped below 40 percent. The aeration rate was 100 bubbles/minute per EPA guidance. Using the total number of dead organisms per replicate and concentration at the end of the exposure period, the 96-hour median lethal concentration (LC50 value) was calculated by the binomial, moving average, and probit analysis methods as described as Stephan (1977). Copies of the statistical analysis are included in Attachment III. 2.5 REFERENCE TOXICANT TESTS The reference toxicant used was sodium dodecyl sulfate (SDS). The test species were exposed to the appropriate graded concentration series to determine if the lots of organisms were within the acceptable limits of BA's Aquatic Toxicology Laboratory. The test duration was 24 hour and treatments were not replicated. 4 US00000315 26 ARCHIVES riginal data sheets, records, memoranda, notes, and computer printouts are archived at EA's laboratory facilities in Sparks, Maryland. Primary data and associated information vill be retained for a period of 5 years unless a longer period of time is requested by the Naval Research Laboratory, 5 US00000316 3. RESULTS Both flow~through studies required aeration because dissolved oxygen (DO) levels dropped below 40 percent saturation during the test period. Aeration was initiated on test #076 with the Type 3 AFFF on the afternoon of day 1 of the test when DO's of 3.9 and 28.5 percent saturation were measured in the 2,100 ppm replicates. All test organisms were still aiive at that time. Similarly, aeration was initiated on test #094 with Type 6 AFFF on the afternoon of day 1 when DOs reached the trigger value, and all test organisms were alive when aeration was started. All other water quality parameters were within acceptable limits during the tests and a summary of water quality values measured can be found in Table 2. The results of the flow-through acute toxicity tests are summarized in Table 3. In the acute test (#076) which was conducted with the type 3 sample there was 100 percent survival in the control and the 115, 240, 49, and 1,000 ppm concentrations. There were no surviving organisms in the 2,100 ppm concentration at test termination. Based on the survival data, the 96-hour LC50 was 1449.1 ppm with a 95 percent confidence interval of 1,000-2,100 ppm. In acute test 91-094, conducted with type 6 sample, there was 100 percent survival in the laboratory control and the 420, 750, 1,350, and 2,400 ppm test concentrations. At test termination there was 25 percent survival f organisms in the 4,200 ppm concentration. The 96-hour LC50 value and 95 percent confidence interval based on the survival data was 3613.0 (2,400--4,200) ppm. The reference toxicant LC50 value conducted on F. heteroclitus lot FU-004 was 7.1 mg/L. EA does not presently have sufficient data points with heteroclitus to calculate a valid acceptability range (minimum 5 values). LC50 values for the previous 3 tests ranged from 11.2 to 15.8 mg/L and are in the same general concentration range as the present test. 6 US00000317 TABLE 1 Parameter Aluminum Antimony Arsenic Barium Bicarbonate Beryllium Boron Bromide Cadmium Calcium Carbonate Cerium Chromium Chloride Cobalt Copper Fluorine Gallium Germanium Iodine Iron Lithium Magnesium Manganese Mercury Molybdenum Nickel Nitrogen Phosphorus Potassium Silicon Sodium Strontium Sulfur (SO.) Thallium Tin Titanium Tungsten Vanadium Zinc Molecular Weight 27.0 121.7 74.9 137.3 61.0 190..80 79.9 112.4 40.0 60.0 140.0 52.0 35.4 58.9 63.5 19.0 69.7 72.6 126.9 55.8 6.9 24.3 54.9 200.6 95.9 58.7 14.0 31.0 39.1 28.1 23.0 87.6 96.1 204.4 118.7 47.9 183.9 50.9 65.4 Forty Fathoms (PPW) 0.06 0.000. 0015 0.12 174.0 0.0020. 12 62.0 0.009 410.0 10.0 0.0007 0.02 18,600 0.0025 0.007 1.9 0.0004 0.00005 0.03 0.03 0.24 1,290 0.008 0.0007 0.005 0.009 0.85 0.04 380 4.5 10,400 12.4 2,600 0.000007 0.006 0.004 0.004 0.0009 0.24 US00000318 TABLE 2 * 0F VATER q u a l i t y r a n g e s FOR 96-HOUR FLOW-THROUGH ----- ----- ^ H L I O X I C I T Y TESTS FOR THE NAVAL RESEARCH LABOR a Test Number EA Accession Number FT-01-28-91-076 ATI-017 FT-02-04-91-094 ATI-018 ATI-065 Temperature (C) PH 18.5-20.2 7.6-8.5 19.2-20.9 7.6-8.4 DO (mg/L) 2.0-8.3(a) Salinity (nrf 1 18.5-20.5 <1.0-8.1(b) 19.2-20.6 1Ca) DayV*1UeAtth ^9 r L ai?d 28,5 percent saturation were measured on uay 1 ' At thls cime, the entire test was aerated. (b) DO values of <1.0 mg/L were measured in test concentrations on Day 1. All test vessels were aerated at this time. US00000319 TABLE 3. RESULTS OF THE USING Fundulus 96-HOUR FLOW-THROUGH heternrlitu* ACUTE TOXICITY TESTS Test number: Sample Description : Accession Number: LC50 values(a) : F T -0 1 -28-91-076(#076) Type 3 ATI-017 FT-02-04-91-094(#076) Type 6 AT1-018/065 (1000-2100)(b) 3613.0 (2400-4200) Exposure Concentration (ppm)-- Percent Survival Control-- 100 115-- 100 240-- 100 21104009000------ 1100000 Control-- 100 420-- 100 750-- 100 1350-- 100 2400-- 100 4200-- 25 (a) EcSTwaues were calculated usins the binomial method. (b) n P*renthesei represent 95 percent confidence intervals. US00000320 .' US00000321 PROTOCOL ATS-FAF-GT-Ol 24. FLOW-THROUGH ACUTE 96-HOUR LC50 ASSAY WITH A FISH 1. TEST OBJECTIVE To assess the toxicity of a test article to a fish species under flow through conditions and determine the 96-hour LC50 using lethality as the test end point. 2. TEST ARTICLE 2-1 Description/Identification Unless otherwise specified, the test article will be supplied by the client. Adequate chemical specifications with special reference to hazardous properties and storage conditions also will be supplied bv the client. 3 22 Methods of Synthesis Information on the methods of synthesis, stability, and data on composi tion or other characteristics which define the test article are on file with the client. 3. EXPERIMENTAL DESIGN 3.1 Test Organisms A fish will constitute the test species as defined by project needs. 3.2 Source Stocks are laboratory reared, obtained from a scientific organism vendor, or field collected using techniques that will be specified in the report. 33 Holding Conditions Upon receipt, fish are transferred to holding tanks which are continu ously replenished with dechlorinated municipal tap water, synthetic sea water, or other appropriate dilution water. The fish are acclimated to laboratory conditions for a minimum of 7 days prior to testing. The laboratory is maintained on a 16-hour light, 8-hour dark photoperiod cycle by a programmable electrical timer. All fish stocks are examined regularly throughout the work week. Dead fish, or those displaying abnormal swimming behavior, discoloration, or pronounced lethargy are removed as observed, and recorded on appropriate log sheets. Fish ETC-T-1Q1 EA Engineering, Science, and Technology, Inc. Effective May 1981 Revision 3, August 1986 Page 1 of 8 US00000322 PROTOCOL ATS-FAP-GT-01 stocks are fed a commercial fish food a minimum of once daily throughout the work week. They are not fed for 8 hours prior to initiating a test. 34 Dilution Water The source of dilution water is the City of Baltimore Municipal Water System. Upon entry to the laboratory, the water is passed through a high-capacity, activated-carbon filtration system to remove chlorine, detergents, and hexane-soluble organic materials. This water source has proven safe for aquatic organism toxicity testing, as evidenced by main tenance of the multigeneration Daphnia cultures, with no evident loss of fecundity. If requested, reconstituted fresh water, synthetic sea water, or other dilution water may be used. ' 3.5 Test Concentration Series The test concentration series to be used for definitive toxicity tests will be selected following preliminary tests to bracket appropriate concentration ranges. A minimum of five concentrations and diluent water controls will be used. If it proves necessary to use a carrier solvent to disperse the test article in aqueous solution, an additional set of solvent controls will be used. All test concentrations and controls will normally be conducted with two replicates. Additional replicates may be used at the client's request. 36 Toxicant Delivery System A solenoid-activated proportional diluter system similar to that des cribed by Peltier and Weber (1985) will constitute the toxicant delivery system. The proportional diluter will be calibrated prior to test initi ation and at test termination. Each test concentration should be at least 50 percent of the next highest concentration in the series. Test vessel calibrations, cycle times, flow rates, and other pertinent data will be reported. 37 Test Vessels and Test Volume Test vessels will be of sufficient volume as to not exceed the organism loading ratio and minimum number of daily tank volume replacements. 3.8 Test Organism Number A minimum of 10 test organisms will be used in each test replicate, con stituting a minimum number of 20 fish per treatment group. Fish will be randomly assigned to each test replicate. ETC-T-101 EA Engineering, Science, and Technology, Inc. Effective May 1981 Revision 3, August 1986 Page 2 of 8 US00000323 PROTOCOL ATS-FAF-GT-Ol 39 Test Environment The test vessels will be maintained at a temperature similar to that at which the fish have been acclimated, but within the recommended test range for that species. Test temperature will not vary more than +2 C from specified acclimation temperature. The tests will be conducted on a 16-hour light, 8-hour dark photoperiod cycle. 3-10 Analysis of Test Concentrations for Test Article If required, test solution replicates may be analyzed for chemical analytical verification of achieved test concentrations. The analytical method and number of determinations will be formulated after consultation with the client. Where chemical analyses are necessary, both nominal and actual measured test solution concentrations will be reported. 3-11 Test Observations , Test organisms will be observed at a minimum of 24, 48, 72, and 96 hours after test initiation to determine number of animals killed by test article concentrations. The study will terminate after completion of the 4-day observation period. The study may be extended, however, at the request of the client. Routine water quality measurements are performed on the test solutions and diluent water controls according to the following schedule: tem perature is measured daily from all test chambers; dissolved oxygen, salinity (when applicable), and pH are measured initially from all test chambers as veil as at test termination, and from the control, low, medium, and high concentration at 24-, 48-, and 72-hour intervals. Specific conductance, total alkalinity, and total hardness are measured from the diluent water and sample (when applicable). Specific conduc tance may also be used for concentration monitoring at the request of the client. Test solutions will not be aerated unless dissolved oxygen concentrations fall below 40 percent of saturation. Analytical methods will be conducted according to APHA et al. (1985) and EPA (1979). Dead fish are removed when observed. At the end of the 96-hour test period, a minimum of 30 surviving fish are removed from the test con tainers , placed in labeled specimen bags, and frozen. The fish are subsequently thawed, measured to the nearest mm (standard length), patted dry, and weighed to the nearest 0.01 g. The average length and weight of the test fish are reported, along with their respective ranges and stan dard deviations. ETC-T-101 EA Engineering, Science, and Technology, Inc. Effective May 1981 Revision 3, August 1986 Page 3 of 8 US00000324 PROTOCOL A TS -FA F-G T-O l 3.12 Data Analysis The LC50 values and associated statistics will be derived by the probit analysis, moving average, and binomial test methods as described by Stephan (1977). Depending on the nature of the data, other methods may be used including the probit approximation method of Litchfield and Wilcoxon (1949), SAS probit analysis (SAS Institute 1979), or graph ical interpolation using the log concentration vs. percent lethality as described by APHA et al. (1985). The methods used will be specified in the report. 4 FINAL REPORT The final report will be prepared to contain the following information: * Objectives and procedures stated in the approved protocol, including any changes made to the original protocol * Identity of the test articles by name or code number and their strength (i.e., quality/purity) and a description of any pretreatment . * Source of the dilution water, its chemical characteristics, and a description of any pretreatment Test concentration series used and duration of the assay * Hean standard lengths and weights of test fish, the respec tive ranges, and approximate loadings of fish to test volume (g/L) * Routine water quality characteristics of dilution water and selected test concentrations * unforeseen circumstances that may have affected the quality or integrity of the study * Signature of the project coordinator, program manager, and. quali ty control officer as authorization of the report. * Location of all archived data and the original copy of the final report at EA Items of data to be included in the report consist of: experimental design and test performance; effects on general appearance of test organisms; morbidity and mortality; tabular presentation and appropriate ETC-T-101 EA Engineering, Science, and Technology, Inc. Effective May 1981 Revision 3, August 1986 Page 4 of 8 ** US00000325 PROTOCOL ATS-FAF-GT-Ol statistical evaluation of water quality characteristics; and survival data where necessary. 5. QUALITY ASSURANCE 5.1 Amendments to Protocol Amendments to the authorized protocol instigated by EA or by the client will be made only after proper authorization. Such authorization is achieved by completion of the Amendment to Protocol form by EA and the client. -2 Standard Operating Procedures Unless otherwise specified, all procedures mentioned in the protocol are subject to detailed Standard Operating Procedures (SOPs) which are con tained in the SOP manuals of the participating departments. These SOPs and protocols follow the U.S. EPA's Good Laboratory Practice Standards (GLPs) (EPA 1983). 5.3 Reference Toxicant A reference toxicant bioassay, utilizing sodium dodecylsulfate (SDS) or other appropriate reference toxicant, will be used as an internal quality check of the sensitivity of each batch of test organisms. Testing will be prior to or concurrent with the test or following the acclimation period for newly acquired organisms. The results of each test will be compared with historical toxicological information of reference toxicant tests performed at EA on the species, to determine if the results are within acceptable limits using the control charts outlined in Peltier and Weber (1983). 5.4 Quality Assurance Evaluation The study described in this protocol may be subject to internal audit by the Quality Assurance Unit. A quality control officer will be respon sible for monitoring each study to assure the client that the facilities, equipment, personnel, methods, practices, records, and controls are in conformance with EA's QC program and EPA's GLPs. 5 .5 Inspection by Regulatory Authorities In the event of an inspection of EA by an outside authority during the course of the study, the client will be consulted before inspectors are permitted access to any of the project records or the experimental areas. STC-T-101 EA Engineering, Science, and Technology, Inc. Effective May 1981 Revision 3, August 1986 page 5 0f g US00000326 PROTOCOL ATS-FAF-GT-Ol 5.6 Archives Copies of project-specific records shall be transferred to the client promptly after the project is completed or as negotiated and budgeted with each client. Original primary data will be retained by EA for 5 years. Primary data include laboratory data sheets, records, memo randa, notes, photographs, microfilm, and computer printouts that are a result of the original observations and activities of the study and which are necessary for the reconstruction and evaluation of the study report. 5.7 Location This study will be conducted by the Aquatic Toxicology Services Department of the Scientific Operations Division of EA Engineering, Science, and Technology, Inc. at the Mid-Atlantic Regional Offices, Sparks, Maryland. 6. SPECIFICATIONS FOR FLOW-THROUGH ACUTE TOXICITY TEST WITH FISH 6.1 Basic References American Public Health Association et al. 1985. Standard Methods for Examination of Water and Wastewater, 16th edition. APHA, Washington, D.C. 1,268 pp. American Society for Testing and Materials (ASTM). 1980. Standard Practice for Conducting Acute Tests with Fishes, Macroinvertebrates, and Amphibians. ASTM Designation: E729-80, Philadelphia, Pa. pp. 285-309. EA Manual T-101. 1981. Aquatic Toxicology Studies, Quality Control and Standard Operating Procedures Manual. Revised, August 1986, (Internal document prepared by Aquatic Toxicology Services, EA Engineering, Science, and Technology, Inc.) Peltier, W.H. and C.I. Weber. 1985. Methods for Measuring the Acute Toxicity of Effluents to Freshwater and Marine Organisms. EPA-600/ 4-85-013. U.S. Environmental Protection Agency, Cincinnati, Ohio. U.S. EPA. 1983. Toxic Substances Control; Good Laboratory Practice Standards, Title 40 CFR Part 792. Fed. Regist. 28(230):53922-53944. ETC-T-101 EA Engineering, Science, and Technology, Inc. Effective May 1981 Revision 3, August 1986 Page 6 o f 8 US00000327 62 Test Specifications Test organism; Fish age: Fish length: Temperature: Light quality: Light intensity: Photoperiod: Diluter and test chamber flow rate: Aeration: Loading ratio: Dilution water: Test container: No. of concentrations: Test animals per container: PROTOCOL ATS-FAF-GT-Ol A fish species to be specified in final report All fish for any one test will be selected from the same age-class The largest fish will not be more than 50 percent longer than the smallest fish To be specified Wide-spectrum fluorescent lights 50-100 f.c. 16-hour light, 8-hour dark Continuous flow-- a minimum of five volume replacements per test chamber during a 24-hour period None, unless dissolved oxygen falls below 60 percent saturation in first 48 hours of the test or below 40 per cent saturation thereafter In general, test chambers may not contain greater than 0.8 g/L biomass Dechlorinated tap water, reconstituted fresh water, synthetic seawater, or appropriate receiving water 19- or 38-L all glass aquaria Minimum of five concentrations and a control in duplicate series 10 (if loading is exceeded, more replicates or larger aquaria may be used) ETC-T-1Q1 EA Engineering, Science, and Technology, Inc. Effective May 1981 Revision 3, August 1986 Page 7 of 8 US00000328 Feeding regime: Test type and duration: End point: PROTOCOL ATS-FAF-GT-Ol Animals are not fed during the test unless fish are too small to survive 96 hours without feeding. Feeding is terminated 48 hours before test initiation in the lot from which organisms are obtained Rangefinding-- 24-96 hours Definitive-- 96 hours Reference toxicant-- 24 hours Lethality-- defined as cessation of opercular movement and inability to respond to external stimuli (gentle prodding with glass rod) ETC-T-101 EA Engineering, Science, and Technology, Inc. Effective May 1981 Revision 3, August 1986 Page 8 of 8 US00000329 ATTACHMENTS II orafe s a n s r ->, US00000330 Atpjalie Organisi Acme Toxicily Fast Data Street Project No.: _ / O C>'ly / 0 IfcS F : S la li^ F l w li>fOHj|lt..ftui)e\Ami TestArtide: F><t Piar*.. h i Test C ontainer, ri Client: Alq f>fZttt.L_ TesiVolume: S* P QC TesiNo.: f T~ OJCf/^l- TestOnidllon:_ "~1(& llS SainpleNo.: _ p|~Tt*"Ol^ _______ DloiionNo.:_SLlu ^ H TEST O H iA N tS M tS C o m m u ti Nam e; . A i n t i i i i l i j L i i f j i i __ _ S a e n titrc Natile: j - , t u r P i o / f i 'k t ; l ol No : . p M O O cf / \ _ __ Soutee: liyl <**1S _______ A cdim alion : f o ^ . o r Arie *}(& s|3ys Biiytiinuiy Date - ^ ! `l Em litiy Date. A lk d iin ily i llafiiness: S a m jilu C o n d u c liv liy lime: J J V_ Iime. D ilueitl US00000331 ACU E TOXIC! fV TEST OBSEUVATION JATA SHEET ......... i Ho .. . _ 7 o l . c ;,3 Client Q/U< iict /o OC lesi No. . Tesi A ilk le P i i <2 Sample No. 1 r i 7 - O S i OA D iju u jtl __ _____ o X - - * l t l< ~K . O litom i No A3 _ loci t ki((.miMii - i ` l ( ! - ! E l ^ l r t - ' l.o l Ufi. H i - o tv * A Acclim ation Pelim i / '( A cclim atim i Temo Rai ne Mean 1 CM I le t ! ....... --...... fie l 1qm 1. Ending D i n _____________________ Ini..J_i.71 T im e ________ T eil: Statiti. fcfowy tlwougfi. Renewal l esi C oni anici H1lj[ll-lv)iS Test Volum e _ 'lest D o ta tim i __________ / (, ' ) ' t Im i liin ji: lianjju __________________________ Cunceiilialtrui Ol % -IioaLs... "/a o 1 su Imi Conlalnai No. .. Hata . 0 Date * -ff? - ....... ___ M ____ 38 39 Dal* * 3? 53 22 O d * i ;>o p ______ A ( ~1; io 3f 6 0 -- j Ih 3 B l i ___ Ho * Nuniliai ol l Ivo Oiyaiiiitnipai Day yus 3 A Data Data 31.- '/o No & 3 :i H o j a L| 4 i^ 5 40 4 l ~* ut 7 ci 4 6 t </ 6 Data Data ^ ! ' . ---- -- fllii, Imiti -- ------ -t-vtj_ _ IrT^. ili'*?. . Hz' i r r i s o __ US00000332 AQUATIC T O X IC O L O G Y BENCH SHEET Project No.: *7 L 1O Client: v o i Ktb S fc Y U ^ c tv QC Test No.: - ^ T - 0 2 , 0 ^ 1 - Date/Time ; ,hh) i I'-CC U P/r/<?: 'b A c tiv ity .Citi Investigator _ r* (i*'Ctr2i,c<.cv-l5 jZ k C ^ i~ -I'JLC -- n, '-c cl o o. A-\ cxvcirrc-p. L j .va. f"*'bk ^ZOi) .pp* *\ .v t '+^ iUi.^."felC^ ^L. t- r?r ^ rei'>'<)-r V ^ 4ici'Aini -.^y ,ncT I' _ .1 ,J Vn A b O D 5pm. \>-i. i t ' 1- "A A-lS~ r ir t \a - . \ , a j r -++&, i;a('ac >k. Zi. ct.:sK.iZr , i1l'cu 1 1' zhhi i. 5C f / J rfWl k 11 1 '<%tj . rS rl -I-ii y US00000333 AQUATIC TOXICOLOGY LABORATORY BENCH SHEET 7PROJECT MO: ______ C C ' i )C CLIENT: a Tct, TEST NO: f r- /' C ^ H TEST SET-UP TEST INITIATION TEST CONCENTRATION SERIES Date/Time Initials Activity /*//*''f\ I@3o0 KSH Dilutions made HflO Li>,iiSH t e s t Vessels filled `$ i O 1351? 4 S Transferred organisms /ZSH Head Counts COMMENTS: Test Cone. Volume Test (Z or mg/1) Materiali ml) Final Volume Ccrdrrc^. Q 60 X -75V '350 ^CrO 7-31 S^ S'- c (VV-c ! / I2TESMEDIATE DILUTION PREPARATION AND FEEDING Dilution Preara cion Food: Feeding US00000334 A,,"iai,: Mianism Acut Toxicity h.s( n,lta US00000335 Project Nu,: 7 0Oj ? m __ Tust A r t ic le ; ......] " : f C lie n t: M i jf., f /%.!% QC Tesi No.: i it'\ ip 1 ^ Samle No.: A f | " < D ilu tio n N o.: __ ly-' > 1t Aquatic Oryuiiism Acute Toxicity Test Data Sheet lE S f: S tu tii^jF lo w Um niyh, tteoewal _ 5 jt Just C ontainer i ^ 'J i Test Volum e: J T * <-.* Tesi D uration: 'it k,. .... _ TEST D lU iA N IS M . \ C om m on Naine: /T ** ^^ _j S c ie n tific Name: f i ; >_ ! t"* Lot N o. : ___f u ` ou ^ Source: | li s . ,i* C ;,\ 1C .T ;>-, , i> Ueylnitmt) Dal e ` .M . 1mu:. ! 1 1| ' Ending Date: .. * * / . 1( i line: 1 ' '1> Sample A lka lin ity: _ D ilnenl Hairiness: A cclim ation: .. u i A u e ^ l ... days C o n d u c tiv ity : US00000336 Test DataAf|u<ilc Organism Acute Toxicity Sheet Project No.. ---- ---- ------------ 1 E S I: S la iic v fjo w tiirouyh) Renewal TEST ORGANISM T- A" ' C' * : 1 4 Tesl Com ainei: r f- X ..- , , a i e , u : ----------T t Volum e: _ 5 , 4 . . . _____________________________________ W,, , ,, : F <U _, QC Test No.: F T cJltf <t i o Sample No.: /tT I ' f Q _____ D ilutio n N o .: ____ Test D uration: Tt, ^ ' * Lo t No.; I a I)!>/ Source: IV x ^ y ^S *t> Acclim ation. _ L i_ J .iy .i_ or Aae ? L. days Beginning Date: 1 ^ 3 ! 1 ` l ime: / >Y E nding Date: __ T im e ; _______ Sample A l k a l i n i t y ................... Diluent Hardness: ________ C o n d u c tiv ity : Cone, or % C&.siurd 1 Con No. fi Nun ber o f Live Organi sms k 0 24 48 72 96 H T jf " fi - nfL* - fi 6 *n y _ 4 _ _ ft m o l `p fi 6 i O/U),* A ____ A ll Tests Tam peate re (C Dissolved Oxygen (m g/liier 24 48 72 9fi X . 24 48 72 96 t l '1 f / f m #3 I f. 4! lU 1 Z L - il 2 J u 7 . 1 ?.S m m ill. 7 -7 7 V 7. n . 6 -7 7 .2 f r y M 1*1,0 i n IU r / . t f ) i (Sf.l m ii.T 1 -i 6 ,5 6-1 5 7 735 . 0 7 1 7 , 3 7. s' in i f i M- 7 -*/ 65 J U l- o Sail or Brackish Water Tests I VpH . 24 48 7? 96 Salinity (ppt| 74 48 72 96 / 24 AU V/ nU m ,1 % ?. K /l it 'i in II2 / ? / , (K \ f i 4, U s | 7 70 ... ` .. * i ' .1 fa o vL 7,4 *0 f*?.? 1 1 '/ n i 10 0 h . ifa H -1 li L ?.iif . o a.o .7 7 I e i 1-2 hs Iffa 111 " III n s m JJ -- (r-0 y * Vi S) v * 7-K U M -S ftr - L I 76 -- f.?.S M S -- -- 1 "1 77 -- -- 1 6 . 6 r--M*. II n Instrument Number Correction Factor Time investigators n% ,* a AW m > 7 . a a. n 6 ? ^ (,1 9 6 9n 40 . //. 7fi'/ 7//0 7 'i 1 ? U.5 'd.S - , i $ ?___ e,! (3.7lOl ma iMl. I, IS j J i i m HI .r/i/ti llVo fs l*|S5r ts lJt' /-' |JLV ill 0 IfVI ito |D . - 6S tLjZlt pH : f(il`>l is r J it US00000337 I'imi- iH,. . I ; o 1 _ ( >>h:Cileni _____ 1_-Og :ii.i ___ OC lei No. r t f.'l'i ) Im i A n kle , l__.......... .;__: Siiiitl* No. . D ilu itili____ IHIinmt Ho. Jll 1 `.I_________ 1 !~'5___ l ^ iA z m _______ C on ten ti Ilm OI % r . c . d .( Im Coniai! No. D il* 71 A C U E rottici TY TEST OUSEHVATION DATA SHEET l .- l llni-MHMII i ' . ' t i l j '____ _ t<>iN r y* - 1 _________ Ill|iimiiu| I l.m- Eniliiifj Udii __ l lll lu'l li, I Ime lime Accliiiialioti l'ttlm l _____I . j >1y ______ A cdin ull ii ( tu p .______________________ le d : Siali:, 1 la w lliaii|j| llenewal Toil Container < .'.O- I flange _ _ _ _ _ _ _ _ M im ___ ____________ IO : lei ________________ (lei lu x . . Teil Volume _ Imi Om alimi le u lenii: lliiiy g :i(p 1t : D ali i F I .* U si* 2 V . . 110 Comjilsluil r ' j 11 >' I K <______ _ N u minti u l U v a O tia iilu iil l l Day a4 J 7lH D ili D ali 6 0 . S U 1 i l ` y n;>o ----------------H ------ .A ---------------------- . I , f, . | V ^ - - r r----1--J----f-t--1----- - " V-- t ' . :) i ..., --n " - 7 'i V- st. 1 'V VI 'j 1 Hu 7J G 2,1 37 l o 70 S -- > -17 35 HO m ---- ..V f -- 1 1 I in il. Tim `rs US00000338 AQUATIC TOXICOLOGY LABORATORY BENCH SHEET PROJECT MO: 7OC;'- V CLIENT: TEST NO: Aa.-,, I o , n l : t, TEST INITIATION Date/Tlme Initials Activity 1 '-IC C ^ Dilutions made Test Vessels filled `3 ! W5 K 3 r Transferred organisms Head Counts COMMENTS: TEST SET-UP TEST CONCENTRATION SERIES Test Cone. (Z o r m g /1 ) > f f * Volume Test Material(ml) 0 .'1 -lv Final Volume CO u ItO ff>.t 'f 'tO f ' iVtlO f '* - 1J t ; [`'{." r+ -i.-f ,J> !s/ V . b V rSTE S M E D M E DILUTION PREPARATION AND FEEDING Dilution Preparation Food: Feeding Day Time I n i t i a l s 1 _0 I H S yjLin Si1 1030 1 Sample/Diluent Day A"'* 01 7^ 5*IV/P ^ I'O i%%. 0 1 /j IQS'/w*' 2 Initials Initials 3 4 IOS*? 5 IS y S H r4'TI-On //5io^ S, 3 ...34............II""'.....*................ X b y Initials Amt Amt Amt 1. ' ...... -- US00000339 AQUATIC TOXICOLOGY 3 ENCH SHEET 'i*ject -,o. ,,,,, f s e o . i 0 - .rinr- in f / ** rfts/ f t * - / . . b C Test No.; _ , F T . : i c " f f, I / r<n* " CtlYttV /LS pf "O G F^ F si ,'`j O O -'-S ' <7 -i i ' - k . ,/ - H* ** "+*~J, 1 ,T% 1*0 .(J 4 T ( '-. i a i t w c O -' y,(Jc >-*' -/ C> 'u,, *.), e ," Sv.""*. * <-*. " o i ` " p Oui, 'rf C 'u - 1 - T t'j f b O. 1' 7 ; i .-a Ig / r"..ii ,, l b ' l`S V C y--1 f ^ r- r-. ^ 1 J . .L , 3 (5 i i -V ? .-w .r Ji 'nvwmoiior r- ^ U 11 l/r/5 l/l /^ l 2 /. /F / Ao"b 5,- T" " /I.* ^ ^I .IT :'t-i -Tla \ p !! -fUk j-n 2 ,V'~ -'-rb L - ,r x ^ 1reciti*. *ji~ ^.<- itA '^ C,, ' - -fV^:- Jh,vil*" x>'M. i b 3cT t> 0 oT- i c-o pf'V.)1^ ne G )C,c5c' '.: O L>^D A f es f Ai j2.-^4-jo( 5"% ) 5 2 1 -o% 52 Is- i*0 - ^ s f %t- >7*0 F V H. 55) / L-S fx/v IbSb n 'if "D ci f <ji n , i ,i%$ &(Nr* m, ;,, H f0>? 6,.o < Y v s * \ fj*** c i>*H| c .*t fc'v / . :& a +*+ /<- cS S*. ye US00000340 Length-Weight Data Sheet A/Ciuii I c ..'-gI' ^ Project No<s).: ~70003>Jo ocTestMnkt F t ~ot ; -o^ Investigator: _ T im e, D ate: JL Test Lot No.; y^<4 - C OH_____________ Ct/VI r1 t 11 o ` Individual No. 1 2 3 4 5 6 7 3 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25: 26i 27 28 29 30 Weight (g> ..... Renne Mimo s.n. o07 (9. aS* c -J?$ O I 2 G- j(* o a ^ e-2H C. iS o-a<?..... O -a: 0- Q2 G * 57 o -a7 CM? o -a ^ O ! t c -a3 * !7 o-ao 0-s.H o a I G-(S 0 17 9 *3 a. O- J35- 0)5 . -'f'O .U . .3-S" 0,^3m Standard Length imml X i *o fr Z X -li 23-21 22 - 6 2 ^5) . 7 - J2.t7.OS X c -C9 53-95 3 io 3.1 3 Z 3 3-U? . a .7 6o a a a t34 i"0 JU 72. as-#> 20 3fc 0 1fc S a 1 1o 21-H9 ........ A 3 - x i a a* 9.1 209a JJ o n _.aL3L-o5 M -~ !h 2 o.o232033-.-.311k l4I $" . i fStG^hi US00000341 Length-Weight Data Sheet l Vi ( Project No(s).: 7 /V C 1 : QC Test No (si.: Test Soecies: L o t N o . : .... " r "T" ~ " ^ v / -- 1 --,, ' - C c". ^ . --* . .m e. Date: L- j> 7 '/ ` . / 7 I . Q.CH In d iv id u a l No. 2 3 4 5 6 7 8 .9 10 11 52 13 14 15 16 17 18 , 19 ... 20 21 22 23 24 25 26 27 28 29 30 W eight (g) . 0. \S n^ - 7 \ r , *>. '- T ~ _ ^ <" Standard Length (mm) 77, r a -- --------------- --Q 7- * ' - ' 1 V u * a-'-". Range Mean S.D. L V C . :7 7. : i r< v -2 1 7 '. 3 / C !. / *> i " " 7 -'t " ?~ 77 V . ~ .xo ^ ."-S V/ , j 3 = -- _____________ Q . 2 0 ____________ /- r o , r& -7 1 3 u 2 1. 7 - 7 O . 3f - V 'l _ 0 .2 ' <V -J '* -? x , 0 - ,'*7 L /C " G ."2.(2? .3 7 7 7 7 0 .3 C 0-7 ? 1 3 "7 1---------------- ^ * 1/ X V XX 7 '7 . 7 7 2Z- ^ 1 .7 V. ci f 7 V . '-LG . v/ v 7 7 - 7 -V 7 V x .'7 1? 3 1 7 -7 V i C- 7 / 7 .^ < 23. /7 I . (5 -^ 7 C -cll ^ 7 . V e! 1. ( s i *> US00000342 Chain-of-Gustody Record EA ENGINEERING. SCIENCE. ANO TECHNOLOGY INC. Client:. Aquatic Toxicology U boratofy 15 Loveton Circle Sparks (Baltimore), Maryland 21152 AidijcP Qe-S/a^xk Project N o . : __ T y d S - / t; FOR; GNFFtCB' SE.O NLY Spodesto bo tests: -- - a maca . .. CadfldMlHila&%x . --------- POlt-w-r - an I p nmift CL MmpflaftH Otter AAcutc CChwnic BBJosee*Hsialiart ------------------- -Accession Number (office us only J ll' i Grab Collection Start End Composite Date/Time Qate/Tlme (Including Site. Station Number and Outfall Number) H /2, Number/Volume ____ _______ ' Relinquished By: Date/Time Received By: Date/Time Relinquished By: Date/Time Client Purchase Order Number: Comments: neeefeNrt'Bp ' x L a X LLaboratory: ataffftm aL/s^fa) / O W Sampie^SMppetf By: (circle) Rnf.Be, Pdro. CP? AWsorrt O ther. Air Bill Number:. WM -Report Ptwualon SdnpkrTsmpersturst a t Recato: Pleas Read Sampling instructions on Back of Form c A-Ctefit/Samptaf US00000343 US00000344 ******************************************************** [190,192]STEFZZ.BAC 14-Feb-91 09:31 AW TEST NAME: NAVAL RESEARCH F. HETEROCLITUS FT-02-04-91-Q94 96-HR LC50 *********************************************************************^^ CONC. 4200 2400 NUMBER EXPOSED 20 20 NUMBER DEAD 15 0 PERCENT DEAD 75 0 BINOMIAL PROB.(PERCENT) 2.06947 LESS THAN 0,001 THE BINOMIAL TEST SHOWS THAT 2400 AND 4200 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT. AN APPROXIMATE LC50 OF 3613.01 IS OBTAINED BY NONLINEAR INTERPOLATION BETWEEN 2400 AND 4200 WHEN THERE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE METHOD NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. COMPARE RESULTS WITH ORIGINAL DATA TO SEE IF THEY ARE REASONABLE. [190,192]STEFZZ.BAC 14-Feb-91 09t30 AM TEST NAHES NAVAL RESEARCH F. HETEROCLITUS FT-01-28-91-076 96-HR LC50 ir*********************************************************************** CONC. 2100 1000 NUMBER EXPOSED 20 20 NUMBER DEAD 20 O PERCENT DEAD 100 0 BINOMIAL PROB.(PERCENT) LESS THAN 0.001 LESS THAN 0.001 THE BINOMIAL TEST SHOWS THAT 1000 AND 2100 CAN BE USED AS STATISTICALLY SOUND CONSERVATIVE 95 PERCENT CONFIDENCE LIMITS BECAUSE THE ACTUAL CONFIDENCE LEVEL ASSOCIATED WITH THESE LIMITS IS GREATER THAN 95 PERCENT. AN APPROXIMATE LC50 OF 1449.14 IS OBTAINED BY NONLINEAR INTERPOLATION BETWEEN 1000 AND 2100 WHEN THESE ARE LESS THAN TWO CONCENTRATIONS AT WHICH THE PERCENT DEAD IS BETWEEN 0 AND 100, NEITHER THE MOVING AVERAGE METHOD NOR THE PROBIT METHOD CAN GIVE ANY STATISTICALLY SOUND RESULTS. COMPARE RESULTS WITH ORIGINAL DATA TO SEE IF THEY ARE REASONABLE. US00000346 17A m e r n m * W R U T / lf l.S Ii RETTEST Bitmap :LI-ENT 'I,|`:V REPORT/ANALYSIS REVEE RECORD ,, LEVEL .'JO. \i ' ,-j .. . _______ DATE SUBMITTED PROJECT NUMBER. 7 _ -L :l 3 ' . DATE DUE TO CLIENT r- ?50GRAM fv ,c 7 ' ~ f c x 'C QC TEST NO. AUTHOR TYPE ANALYSIS_________ REPORT/ANALYSIS CHECKLIST OA/QC ITEM REVISEES DATE Samples/data collected, transported,^ -- and received according to study plart^ _ , requirements. ' Ao[ c ,t I. Samples prepared and processeti ing to stuay plan requirements. accord C \- v ' ,U . - 0 1 '- V . . Y Samples and data collected and analyzed using calibrated equipment. 4 Quality control activities completed. ivi vV<~ r _' A* A,, A 5- Data checked for correctness, source identification, and data confirmation. . '' . / / 6 - Calculations checked: - Hand calculations checked by review - Documented and verified computer programs used. - 1 Results reviewed for compliance vich \study plan requirements. -- ,.vVv.x` T a .- - 'C ,..;-- - > lcu.G ! 3. Reported results and facts checked ,,--- -,. rJr 'i : !' . \ - .1 o. against original sources. v t- u - - .V . . 9. Figures and tables checked for correct presentation of data. ? -?/ 10. Commentary reviewed and resolved. AUTHOR DATE /TbA//. H a Jtj/ -, . 11. All study plan and quality assurance/conerel requirements have been FILED BY PROJECT MANAGES EA-QA-Q7