Document 3exwwd1b1GMpLVyOL2JvLvNBy

AR226-3340 ^0^1^ E.I. DUPONT DE NEMOUKi) & COMPANY INC. REPORT NO: / GL-2001-42 TITLE: Aquatic Toxicity and Solubility of COPY NO. 5 Fliinrotelomer Intermediate AUTHOR: i,Roland C. Hall, Jr. ^ '~~ p" CONTRmUTORS: JThomas V. Robertson, Olver Laboratories DISTRIBUTION: ABSTRACT ONLY Does not contain TSCA CBI ^ GL-2001-42 , Y -/ Report Number Issued ^GL-2001-42J December 5. 2001 Aquatic Toxicity and Solubility of uorotelomer Intermediate Report Written By Period Covered Research Notebooks Work Supervised By Roland C Hall. Jr J December 2000 to May 2001 NA "' J F --\ Kmec j Abstract This report documents a stud^oneJj^Dlver Labatones to determine the the aiqsuoantiec ~~ of^|B^|BB||^luorotelonrcrIntennediate(^'hich mjBB||BUP^> toxicity and solubility The study was conducted in component of the spin agent to be used to conduct a test of the new spin agent in to obtain necessary environmental permits subsequently canceled, however, ^BBBBp^01 order, P^"1 ^ne test ln ^ P1!01 P^"1 was theinfonnaS&n in this report was forwarded to the Line 3 organization to assist in meeting their environmental requirements'"] '"' ^^^^^j| "~ ofUBBBBBII^Flouorotelomer Olver Laboratories determined that the solubility be the order of 6 mg/L at standard temperature and pressure Intermediate appeared to in acute toxicity at concentration near the apparent Their screening tests also indicated no - maximum solubility of 5 mg/L Company Sanitized. Does not contain TSCA CBI .-^\ GL-2001-42 i confidential information and each holder is responsible for its This report contains When no longer needed, please destroy or dispose of in conformance the approval safekeeping. with MP guidelines. No part of this report may be reproduced without of thqilHK^echnical Manager. Ip^^^^ij TABLE OF CONTENTS Introduction Objectives Summary and Conclusions i Patent Situation Program Publication Status Acknowledgement Signature(s) Appendixes Company Sanitized. Does not contain TSCA CBt f~GL-2001-42 I INTRODUCTION made to evaluate the spin agent to be used oj Ilnn ^la^te^2f0W00lyapd1e^ci0si1onplwanats Thetrsapnins-a1g,2e,nDtictohlbperoteesthtevdlevne''aT^n loirmderBtoHo--bta9in ! Fluorotelonier'Tjitermediateand t-he^MMMpilotplant, information was^--- environmental approval to run the tests in J^rherewasno pubiished"o3ta available needed on the aquatic to\icity ofthespm agent ofBB(tttBBB|FluoroteIomerIntermediate, therefore a study was on aquatic toxicity Laboratories to determine the solubility in water and the ~"~ commissioned with CHver toxicityJThetest in the pilot plant was subsequently canceled, however, the meeting rranqfuoartmication in this report was forwarded to the Line 3 organization to assist in ^ | their environmental requirements OBJECTIVE , o i f The purpose of this ot the study was to determine the toxicity of 6 mg/L to Fluorotelomer Intermediate at a maximum concentration in water Cenodaphma and fathead minnows (.Pimephales promelas) SUMMARY AND CONCLUSIONS tests indicated no acute toxicity at concentrations near the theiUH^ The initial screening 6 mg/L In an effort to determine apparent maximum solubility of and chronic toxicity, acute and. chrprug testing was concentrations that results in acute usingg(KlQamendewdith conducted usingofamn eetxhpaannodletdo tiensctresaosluetisoonlusbeirliietys prTehpeareredsult of these (tPeismtsepinhdailceasted that and fathead minnows smalt amounts iBHBQwas not acutely toxic to Cenodaphma solubility limits The 48-hour promelal)at concentrations approaching methanol-aided was greater than 500 mg/L. The no LC5o values for Cenodaphma and fathead minnows (NOEC) for the Three-Brood Cenodaphnia Survival and observed effect concentration 100 ma/L while the NOEC for the 7-Day Fathead Mmnow Reproduction chronic test was Growth chronic test was 500 mg/L These results .suggest that there should ^^^Ulj11- Survival and toxicity due to the presence the maximum be no direct acute or chronic apparent solubility in water of 6 mg/L PATENT SITUATION At the present time we have no plans to tile for a patent Company Sanitized. Does not contain TSCA CB! GL-2001-42 \ PROGRAM No additional testing is planned at this time PUBLICATION STATUS At the present time \ve have no plans to publish the data contained in this report ACKNOWLEDGMENTS Thanks to Tom Robertson and Olver Labortones, Incorporated for their contributions to this study and report. SIGNATURE(S) S.n!zed.Do..n.>c.nt.lnTSCACBI Company GL-2001-42 APPENDIX Appendix A Memo from R Lawrence Hoffman, VicePresidenl., Olver Laboratories to CThomasV. RobertsonjdaleMday 4, 2001 subject |HMBSolubi]ity and Toxicity Deterrmnation Program, Job Number 60175 *as" B "Delermi nation of solubility and aquatic loxicity of DuPont Product ^luorotelomer Intermediate for E E DuPont De Nemours and Company by R ^awrenc'e Hoffman dated May 3. 2001 Company Sanitized. Does iwl cwla'n TSCA CB1 ^ ^ ^s \ v>^^s /"V WpD LABORATORIES Consulting Engineers . Environmental Laboratories 11 "16 South Main Street Blacksburg. Virginia 24060 May 4, 2001 Mr Thomas V. Robertson) 'Development Associate Environmental Affairs E.I du Pontde Nemours and Company, Inc PO Box 27001 Richmond, VA 23261 Re (fffiolubility and Toxicrty ^Determination Program, Job Number 60175 Dear Tom Enclosed for your review and use are three (31copies of the report which describes the performance and results of thefiHJ5olubility and Toxicity Determination program performed by Olver Laboratories Incorporated for E I du Pont de Nemours and Company, Incorporated. This program was performed to support evaluations of the potential for the use of this product by examining its potential effects on wastewater toxicity The program, as described in our December 14, 2000 proposal, consisted of an initial determination of solubility in water and subsequent determinations of acute and chronic toxicity to two common freshwater test organisms As for the standard compliance monitoring performed by Olver Laboratories for DuPont, the toxicrty testing component of this program is reported in a GLP format As indicated on the Material Safety Data Sheet provided with the sample, the material appeared to be only slightly soluble in water As described in the,,report, the solubility of the material in water appeared to be in the order of 6 mg/L at standard temperature and pressure. The solubility in water appeared to increase tojevels of 500 mg/L to 1000 mg/L by adding a small amount of methanol to the|ttB|Tpnoj,to_ addition to water For the reasons referenced in the report, the analysis of the_p1"^ solutions provided inconsistent results and could not be used to confirm the| concentrations used in the toxicity evaluation component of this program. Blacksburg, VA (540) 552-6974 Company Sanded. Does not contain T8CA CBI Mr Thomas V. Robertson "May 4, 2001 Page 2 The initial screening tests indicated no acute toxicrty at concentration near the apparent maximum solubility of 6 mg/L. In an effort to determine the^Oyoncentra- tions that results in acute and chronic toxicity, acute andchronic testing was conducted using an expanded test solution series prepared usingflBBBamendedwith small amounts of methanoi to increase solubility. The results oftnese tests indicated that iBjRvas not acutely toxic to Ceriodaphma and fathead minnows {Pimephales prometas) at concentrations approaching methanol-aided solubility limits. The 48-hour LCgo values for Ceriodaphnia and fathead minnows was greater than 500 mg/L. The no observed effect concentration (NOEC) for the Three-Brood Ceriodaphnia Survival and Reproduction chronic test was 100 mg/L while the NOEC for the 7-Day Fathead Minnow Survival and Growth chronic test was 500 mg/L These results suggest that there should be no direct acute or chronic toxicty due to the presence oflMR81 the maximum apparent solubility in water of 6 mg/L ' I look forward to the opportunity to discuss this with you in more detail and TO answer any questions that you may have As always, please do not hesitate to contact me or David Griffiths Sincerely, RLH/egl < O'(XLUU?MCC R Lawrence Hoffman Vice President Enclosures cc: David W Griffiths, Ph.D., President, Olver Laboratories Incorporated Company Sanitized. Dn^ ^ cnnf^n TSCA CBI STUDY TITLE DETERMINATIONS SOLUBILITY AND AQUATIC TOXICITY OF DuPONT PRODUCT -ffffSHffLUOROTELOMER INTERMEDIATE FOR E. 1. DU FONT DE NEMOURS AND COMPANY DATA REQUIREMENTS DuPONT CONTRACT ORDER NUMBER AUTHOR R. LAWRENCE HOFFMAN STUDY COMPLETED ON May 3, 2001 PERFORMING LABORATORY OLVER LABORATORIES INCORPORATED ENVIRONMENTAL LABORATORIES AND CONSULTANTS 1116 SOUTH MAIN STREET BLACKSBURG, VIRGINIA 24060 for E. I. DU PONT DE NEMOURS AND COMPAN' OLVER LABORATORIES INCORPORATED JOB NUMBER 60-? 75 'Connwv R^W,^ ,,.,,,,, ,,,., ,.,,....,,,, TQ^ pg, OLVER LABORATORIES TABLE OF CONTENTS Page Number REPORT APPROVAL RECORD . . . ' - . . . . . . QUALITY ASSURANCE STATEMENT .. - " - ...... ... ......... ..... .... 1 GENERAL INFORMATION . 2 PROJECT PERSONNEL . ..... ........ .... 3 ......... SUMMARY ... ..... . . . . . . - 4 ............... INTRODUCTION .... ..... METHODS AND MATERIALS ... ........ 1 0 fi----S9OLUBIUTY DETERMINATION ...... ... 4 . ...... . - 4 .... . . . 2 0 JB|l|^QUATlTCOXICITY DETERMINATION .... ..... 4 2.1 TEST ORGANISMS . .4 ... ............ 2.2 TEST SUBSTANCE .,.:..................... 2.3 DILUTION WATER .......... ...... ... 2.4 JOXICTTY TEST PROCEDURES . ....... ........ ..... 2.4.1 GENERAL PROCEDURES . . 2.4.2 TOXICITY RANGE-FINDING TESTS ........ ............. ...... 2.4.3 ACUTE TOXICITY TESTING 24.4 CHRONIC TOXICITY TESTING - - .... . ......... . . . .... .. .. 5 - 6 6 .68 . 8 9 2.5 STATISTICAL ANALYSIS . . . . . . . . . RESULTS 10 ... . . . . .11 ................... ............. 1.0 ||1|SOLUBILITYDETERMINATION .......... ... f --% 2.0 |Bt|TOXlCITY DETERMINATION .... .... 2.1 TOXICITY RANGE-FINDING . . . . .... ........ 2.2 ACUTE TOXICITY TESTING ..... .... ........... 2.3 CHRONIC TOXICITY TESTING . . ... 11 ..... . 12 ......."-1122 ... 14 . Company Sanitized. Does not contain TSCA CB1 LIST OF TABLES Table 1 Table 2 Table 3 Table 4 Toxicity Range-Finding Data Summary Acute Toxicity Data Summary . ... ....... 13 . - 15 Chronic Toxicity Data Summary - Comparison to Standard ... ............ Controls . .. . 16 Chronic Toxicity Data Summary - Comparison to Methanol- ..... ...... Spiked Controls .... APPENDICES 17 ..... . . . . . Appendix 1 Material Safety Data Sheet . . . ... 18 Appendix 2 yl^f, Solubility Determination - Test Protocol . . ... 25 Appendix 3 f-^^M solubility Determination - Analytical Data . iflU ... . . . . . Appendix 4 Toxicity Range-Finding Test Data . . 27 3 2 . . Appendix 5 Appendix 6 Appendix 7 Acute Toxicity Test Data 39 ... ............... Chronic Toxicity Test Data - Pimephales promelas 45 . . . . . Chronic Toxicity Test Data - Cenodaphnia dubia . . . . 67 Appendix 8 Appendix 9 Appendix 10 Appendix 11 Acute Reference Toxicity Test Data . . . . . .93 Chronic Reference Toxicity Test Data . . . . . . . . Dilution Water Quality Analyses Data . . .. . ..101 113 .... .... . Chemical Analyses of Brine Shrimp and YCT Food Mixtures 122 Company Sanitized. Does not contain TSCA CBI Report Prepared by: ^^jo^ ^}J^\^1^L^_______^l^fol Susan Mirlohl Date Bioassay Laboratory Manager Reviewed and Approved for Issue by Vice President and Study Director Company Sanitized. Does not contain TSCA CBI OLVER LABORATORIES QUALITY ASSURANCE STATEMENT Quality Assurance Statement for final report entitled "Determination of Solubility and Aquatic Toxicity of DuPont Producj-t--f^H^|M^ RF"Iu^ orotelomer intermediate for E I du Pont EB^^^^^^SI de Nemours and Company" Audits for the study were conducted on the dates listed below: Items Audited Sample Receipt Range-Finding Test Initiation Range-Finding Test Termination Acute Test Initiation Acute Test Termination Chronic Test Initiation ChTonic Test Renewal Chronic Test Termination Audit Dates 1/11/01 1/25/01:2/01/01 1/27/01; 2/03/01 2/14/01 2/16/01 2/13/01; 2/20/01 2/14/01; 2/21/01 2/20/01; 2/27/01 Date findings reported to management and study director 2/27/01 The information provided within this report was generated in accordance with Good Laboratory Practice Regulations and Protocols. A final review of all data and records indicate the report submitted is an accurate reflection of the study conducted; l Reported by: \ f/Lj^--^ " ^/3/o/ F^ri d. BjoltinghouseDate (Uuahfy Assurance Auditor, Olver Laboratories Company Sanitized. Drifts ^t c'wt^n TSCA CB1 OLVER LABORATORIES Matenal Tested: GENERAL INFORMATION Olver Sample Designation: Sponsor: Study Initiated; Completed: -Experimental Start; End: . Location of Onginal Raw Data and Final Report: 1/12/01:5/03/01 1/25/01.2/27/01 Olver Laboratones Incorporated 1116 South Main Street Blacksburg, Virginia 24060 Page 1 60175 Company Sanitized. Does not contain TSCA CBi OLVER LABORATORIES PROJECT PERSONNEL The study director of this project for E. I. du Pont de Nemours and Company was | \""R. Lawrence Hoffman. ^ 3 The following Olver Laboratories personnel assisted with the various phases of thejJUj ^ ^ aquatic toxicity determination study TITLE INITIALS Susan Mirlohi Stacey Boyles Sheryl Newton Laura Dunham Fran Boltinghouse Bioassay Laboratory Manager Biological Technician Biological Technician Biological Technician Quality Assurance Officer $rq 5L& 5P^ Lrvu? ^<->. Page 2 60175 Company Sanitized. Does nol contain TSCA CBI OLVER LABORATORIES DETERMINATION OF SOLUBILITY AND AQUATIC TOXICITY OF DuPONT PRODUCT'.BBBBUhfcfLUOROTELOMEINRTERMEDIATE SUMMARY Solubility andaguatic toxicity determination studies were performed for the DuPont l l t w o product, facilitate projections and assessments of the potential for toxicity in wastewater that "may contain this material. The solubility determination was performed using a procedure developed by Olver Laboratories based on the substance information contained on the Matenal Safety Data Sheet (MSDS) provided by DuPont.JBBjpolubility in water wasestjmated by establishing a calibration curve for GC/MS analysis. The solubility ofjHfin water was determined to be on the order of 6 mg/L. However, when of|l|Hl| introduced as a methanolic solution, it proved possible to produce what appeared to be homogenous aqueous solutions containing as much as 1,000 mg/L It is believed that these are not true solutions, but rather dispersions or micellar solutions i f The a&ute and chronic toxicity of to Ceriodaphnia dubia and fathead minnows (Pimephales promelas) was assessed utilizing EPA/600/4-90/027F and EPA/60 0/4-91/002 methods^ Tqxjcity testing consisted of initial range-finding tests which included a wide range off^concentrations to establish a general level of toxicity for acute and chronic testing using Ceriodaphnia dubia and fathead minnows. Toxicity range-finding tests were followed by 48-hour static acute Ceriodaphnia and fathead minnow tests, a three-brood chronic Cenodaphma dubia test, and a 7-day chronic static renewal fathead minnow test of||||||| The concentratjonseries used in the toxicity range^nding tests ranged from 0.01 to 1,000 mg/L For acute toxicity testing,^||||||concentratiornansged from 5 to 200 mg/L, white chronic testing included concentrations which ranged from 5 to 500 mg/L Estimated toxicity endpoints (LC5o and NOEC values) were found to be as follows. Toxicity Test Type Toxicity End-points Ceriodaphnia Acute Test Fathead Minnow Acute Test Ceriodaphnia Chronic Test - Survival Ceriodaphnia Chronic Test - Reproduction Fathead Minnow Chronic Test - Survival Fathead Minnow Chronic Test - Growth LCgo >500 mg/L (100% survival at 500 LCgo >500 mg/L (100% survival at 500 NOEC= 100 mg/L NOEC =100 mg/L NOEC = 500 mg/L NOEC = 500 mg/L mg/L) mg/L) Page3 60175 Company Sanitized. Does not contain TSCA CBI INTRODUCTION Solubility determination and aquatic toxicity testing for the DuPont product^B^^^^--H--K--"^'M61"6 performedbYOIverUiboratQnes Incorporated for the E.I. du Pont de fTemours and CornpanylBlBBBIBIyroJamnuary 12^2001 to February 27, 2001. The purpose of o^ll|n this study ^as to detergi'ffitlhe solubility water and to determine the acute and ofjifto chronic toxicity representative freshwater organisms. The toxicity o^Hl^a5 assessed through determination of median lethal concentration (LC5o) values'for acute testing and no observed effect concentration (NOEC) values for chronic testing, using Ceriodaphnia and fathead minnows. METHODS AND MATERIALS 1.0 --yI^^--^B--uSSoSloulubbililiittyy Determination ^^*3 the1abenfllU|UB|BUHBmi All testing was performed using a sample ofJUi provided by DuPont. The sample wa; contained in a 3.63 kg/8 Ib can bearing Solubility tests were performed according to the test protocol that is provided in Appendix 2. mi*1 jaa, 2.0 Jgf^llAAqquuaatic Toxicity Determination t^^^^^^Ol?!?' The toxicity tests were conducted according to "Methods for Measunng the Acute Toxicity of Effluents to Freshwater and Marine Organisms", EPA/600/4-90/027F and "Short-Term Methods for Estimating the Chronic Toxicity of Effluents and Receiving Waters to Freshwater Organisms", EPA/600/4-91/002 The tox-icity testing component of this study was conducted following Good Laboratory Practice general testing and reporting guidelines and the final report was reviewed by the Olver Laboratories Quality Assurance Unit. Copies of the report and original raw data will be maintained by Olver Laboratories. The original signed report and raw data will be archived at Olver Laboratories Incorporated. 2.1 Test Organisms Ceriodaphnia dubia and fathead minnows, originally obtained from the U.S. EPA and subsequently cultured at Olver Laboratories, were used for acute and chronic testing. Fathead minnow and Ceriodaphnia cultures were maintained in EPA moderately hard water and the cultunng records indicated that the test organisms were in satisfactory health at test initiation. Cenodaphnia cultures were fed a YCT (yeast-cerophyll-trout chow) and algae (Selanastrum capncomutum) food mixture prepared at Olver Laboratories. Fathead minnow cultures were fed brine shnmp nauplii (Artemia sp. eggs obtainedjrom "Ocean Page 4 B0175 CompanySanitized. Does not contain TSCA CBJ Star International Inc.", Snowville, VT, Lot No. 4400). Test organisms were fed for at least two ho'urs prior to use in testing. Larval fathead minnows used in the chronic testing (Olver Culture No. 1810) were 1-24 hours old at test initiation and the control organisms had an average dry weight of 0 429 mg at test completion. The biological loading factor in the controls was 0.02 g/L. Test organisms were fed 0.1 mL of a concentrated suspension of newly hatched (less than 24 hours old) bnne shrimp nauplii (approximately 700 to 1000 nauplii) 2-3 times daily during testing Larval fathead minnows used in the acute testing (Olver Culture No 1809) were 3 days old at test initiation. The biological loading factor in the controls was 0 05 g/L with an average weight (not dried) of 2.4 mg at test completion. The Cenodaphnia dubia used in the chronic test were obtained from Olver Laboratories cultures produced from EPA-supplied stock. Cenodaphnia used in the chronic testing (Olver Culture Nos. 1-30) were less than 24 hours old, and all within 8 hours of the same age, at test initiation, and the control organisms had an average reproduction of 25.9 (per surviving female) at test completion. Test organisms were fed 0.1 mL of YCT and 0 1 mL of a concentrated suspension of algae (approximately 3.1 x 107 cells) once daily at test solution renewal time. Cenodaphnia used in acute toxicity testing were less than 24 hours old at test initiation and they were not fed dunng testing. The brine shrimp and YCT used in feeding were tested to show that concentrations of metals, pesticides, and PCBs remained within acceptable limits No chemicals were present in the bnne shrimp that made them unsuitable for use as a food source in aquatic toxicity tests. Data for these analyses are included in Appendix 11 Monthly reference tests (Append ices 8 and 9) were performed to ensure that test organism sensitivities and health were within acceptable limits. A sodium chloride reference toxicant (EM Science; CAS #7647-14-5) was used and the results showed fathead minnow and Cenodaphnia sensitivities to be comparable to ranges previously established at Olver Laboratories. 2.2 Test Substance A concentrated sample ofjB^|^;o! ntained in aone-gallon sealecLjpetal container was provided to Olver Laboratonesby the DuPont|||HI----tflH^ac;lity The MSDS provided with the sample identified the material aJj^BPuoruteiomer Intermediate with the following composition: Page 5 60175 'Company Sanitized. Does not contain TSCA CBl Based on the results of the|m|solubility determination, various concentrations of were prepared for use in acute and chronic toxicity testing. The solubility level foij in water was estimated at 6 mg/L, therefore, the theoretical concentrations o^pByjsed in initial range-finding toxicity tests were selected at and below the 5 mo/Lsolubility level Since no toxicity was observed at this level, higher concentrations ofJUpft/ere prepared for use in the acute and chrome toxicity testing. These rangedfrom 5 to 500 mg/L To enhance the solubility of||B|in preparing test solutionslB^Jl/vasinitially dissolved in methanol (Burdick & Jackson, High Purity Methanol, CAS #67-56-1) prior to being mixed theffH in dilution water Test solution preparation for the range-finding, acute and chronic testing is described in Section 2.42 through 2.4.4. In all cases, preparations of standards and test solutions was performed in a manner to minimize loss by volatization to the extent practical. / 2.3 Dilution Water The dilution water used for cultunng, toxicity testing, andjUylest solution preparation was moderately-hard synthetic freshwater prepared using EPA guidelines (EPA/600/490/027F and EPA 600/4-91/002) This dilution water was aerated prior to testing with oilfree compressed air. pH ranged from 7.7 to 8.0, conductivity ranged from 300 336 //mhos/cm, alkalinity ranged from 60 to 64 mg/L, and hardness ranged from 84 to 100 mg/L. Annual dilution water quality analyses data are provided in Appendix 10, 2.4 Toxicity Test Procedures 2.4 1 General Procedures The acute and chronic toxicity tests were performed at 25C 1C by placing the test chambers in a constant temperature water bath. Test organisms were exposed to ambient laboratory light (50-100 foot candles, measured at water surface) for a 16-hour day/8-hour night photoperiod. Transitional lighting (30 minutes, 30-40 foot candles) was included at the beginning and end of the 16-hour light period For the Cenodaphnia dubia acute toxicity tests, plastic beakers filled with 25 mL of test solution were used. For the fathead minnow acute toxicity tests, 600^ mL glass beakers filled with 500 mL of test solution were used. Beaker positions were randomized in the water bath using random numbers. Test organisms were pooled Page 6 60175 Company Sanitized. Does not contain TSCACB1 and placed (five at a time) into each test solution in a random order. Test organisms were less than 24 hours old for the Ceriodaphnia acute tests and within 1 to 14 days old and within 24 hours of age for the fathead minnow acute tests. Routine chemical analyses were conducted. Alkalinity and hardness were measured in the control and highesimptest concentrations at test initiation. Conductivity, was measured in all test concentrations at test initiation and termination. Dissolved oxygen, pH, and temperature were measured daily in one replicate per effluent concentration. All chemical analyses were conducted as per the EPA document "Methods for Chemical Analysis of Water and Wastes" (EPA 600/4-79-020) or APHA, "Standard Methods for the Examination of Water and Wastewater", 18th Edition. Dissolved oxygen was determined by electrode EPA Method 360.1. PASAMETER PH Temperature Dissolved Oxygen Conductivity Alkalinity Hardness TEST METHOD EPA 150 1 EPA170 1 EPA 360 1 EPA 120 1 EPA 3101 EPA 130 2 TEST SOLUTIONS ANALYZED All All All All Control and Highest Concentration Control and Highest Concentration FREQUENCY Daily Daily Daily Beginning/End Beginning Beginning For the Ceriodaphnia chronic test, 30 mL plastic beakers filled with 15 mL of test solution were used. Beaker positions were randomized in a holding tray using random numbers. One Ceriodaphnia neonate (20-24 hours old) was placed in each of ten replicate test chambers per test solution using the assigned positions All test chambers, contained in their holding trays, were placed in the water bath. The number of surviving Ceriodaphnia in each test concentration was determined prior to test solution renewal. Dead Ceriodaphnia were discarded. Test solutions were ^Wv^^ ___ewed daily by transferring each of the original Ceriodaphnia into freshly prepared solutions. The number of Ceriodaphnia neonates, reproduced by the adult female Ceriodaphnia in each replicate test chamber, was counted and recorded at each daily test solution renewal time In the chronic test, Ceriodaphnia were fed 0.1 mL of a concentrated suspension (approximately 3.1 x 107 cells) of algae and 0.1 mLofYCTfood mixture daily, attest solution renewal time. Ceriodaphnia were exposed to ambient laboratory light (50- Page 7 60175 Company Sanitized. Does not contain TSCA CBI 100 foot candles, measured at water surface) for a 16-hour day/8-hour night photopenod with transitional lighting Transitional lighting (30 minutes, 30-40 foot candles) was included at the beginning and end of the 16-hour light period. Temperature was maintained at 25C 1C by placing the test chambers in a constant temperature water bath. For the fathead minnow chronic test, beakers (glass; 600 mL) filled with 250 ml_ of test solution were used. Ten fathead minnow larvae, 1-20 hours old, were placed in each of four replicate test chambers per test solution using random numbers All test chambers were positioned in the water bath using randomization techniques. The number of surviving larvae in each chamber was determined prior to test solution renewal. Dead larvae were discarded. Test solutions were renewed daily by siphoning approximately 80% of the test solution out ofthe^ chamber The original test volume was restored by adding freshly preparedj|BI(s|olutions to the test chambers. 2.4.2 Toxicity Range-Finding Tests o f l j f The initial phase of testing consistedofa series of 48-hour range-finding tests to determine the general toxicity of|B|l|bver a wide range of test concentrations Targeted test solutions, ranged from 0.01 mg/L to 1,000 mg/L Based on the results ofthelB|Bpolubility determination, an initial concentration series was oqU|in selected to perform the toxicity range-finding tests. Since the solubility water was estimated at 6 mg/L, the initial toxicity range-finding tests were performed using a dilution series consisting of control, 0.01 mg/L, 0.1 mg/L, 1 mg/L, and jllH 5 mg/L of Since no evidence of toxicity was observed at these levels, tne^ nig/L||U dilution series was expanded to include targeted 100 mg/LtoU)00 concentrations. To enhance the solubility oi^fffin water, Ulfused to prepare test concentrations greater than 6 mg/L was initially mixed with 500 ,uL of reagent- grade methanol; this mixture was then added to moderately hard water to prepare the test solutions, 2.4.3 Acute Toxicrtv Testing The second phase of testing involved the performance of 48-hour static acute Cenodaphnia dubia and fathead minnow tests. Based on the results of the toxicity range-finding tests, a concentration seges consisting of control, 200 rng/L^ 300 mg/L, 400 mg/L, and 500 mg/L o||fl|||vas used. The highest targetedply concentration used in testing was at 500 mg/L since, above 500 mg/L.^IBBJipniodt appear to reman in solution. At concentrations greater than 500 mg/L small droplets o^HBI^ere observed adhenng to the glassware used to prepare the test solutions. Anadditional control solution consisting of 500 pL methanol/L of dilution water was included in testing to identify any potential toxicity associated with the presence of methanol in solution. Based on the available toxicity data on methanol Page 8 60175 Company Sanitized. Does not contain TSCA CBI (EPA documents: EPA-600/6-91/005 and EPA/600/6-91/003), the concentration of methanol in the test solutions (0 05%) was well below the expected toxic levels (i.e., 2% to 5% for acute toxicity and 0 3% to 0 5% for chronic toxicity) test solutions were prepared by first dissolving the pre-measured volume of concentrate in 500 ^Lofmfithanol prior to mixing in the dilution water The measured concentrations owHR|n test solutions were based on a specific gravity of 1.4 as noted on the matenaLgafetY data sheet provided with the sample The Hlfest acute test dilution series and below. solution preparations are summarized Control Control with Methanol 200 300 400 500 None None 142^L 213 p.L 284 ,uL 355 iiL 2.4.4 Chronic Toxicity Testing The third phase of testing involved the performance of the static renewal Ceriodaphnia dubia and fathead minnow tests. Based on the results of the acute toxicity tests, a dilution senes consisting ofcontrol, 5 mg/L, 10 mg/L, 25 mg/L, 50 mg/L, 100 mg/L, and 500 mg/L of targeted(^(y:oncentrationwsere used for the chronic testing An additional control solution consisting of 500 pL of methanol/L of dilution water was included in testing to serve as a second control for determinations of potential toxicity associated with the presence^" methanol in test solutions Methanol was used as a solvent medium foiHllfprior to mixing of ^pBywrthdilution water and preparation of test solutions^^^ Test solutions were prepared at test initiation and daily chronic test solution f^tock renewals. A 100 mg/L stock soh^o\^1^flfwas usedto prepare the 5 mg/L, 10 mg/L, 25 mg/L, and 50 mg/L|H|^est solutions, solutions were measured volumetrically, dissolved in 500 {A- of puremethanol, and added to premeasured volumes of dilution water. The solutions were gently mixed in a separatory funnel prior to use. Page 9 60175 Company Sanded. Does not contain TSCA CBI Test Concentration (mg/l Control Control with Methanol 5 10 25 50 100 500 >1000 mL dilution water None None 5 mL of 100 mg/L solution 10ml-of100mg/L< 25mLof100mg/L< 50 mL of 100 mg/lJ solution (solution solution 71 ^L 355 {A. 2.5 Statistical Analysis * In light of the analytical difficulties presented^ chronic test data were based on targeted all statistical analyses of acute and test concentrations. Insufficient mortalities occurred in the acute toxicity tests to calculate an LC^. Mortality per test concentration was reported, and the LCgg values were approximated based on the highest test solution used. Statistical analysis procedures described in EPA/600/4-91/002 were used with Toxstat Version 3.5 to perform statistical analyses for the Ceiiodaphnia and fathead minnow chronic tests- For the Cenodaphnia chronic test, survival was analyzed first using Fisher's Exact Test. Normality of distnbution and homogeneity of vanance for the reproduction data were next checked with the Shapiro Wilk's or Chi-Square Tests and Bartlett's Test, respectively. ANOVA and Dunnett's Test or Steel's Many-One Rank Test were used to evaluate reproduction. Any Ceriodaphnia test replicates identified as males were excluded from the reproduction data analyses as per EPA guidelines. For the fathead minnow chronic test, data were transformed using the arc sine [square root (y)] transformation. Survival was analyzed first Normality of distnbution and homogeneity of variance were next checked with Shapiro-Wilks and Bartlett's Test, respectively ANOVA and Dunnett's Test or Steel's Many-One Rank Test were used to evaluate mortality and growth. If any test concentrations showed statistically significant mortalities, they were excluded from the growth data analyses For statistical analyses of growth data, average growth of the organisms was based on the onginal number of larvae, as per EPA method guidelines. Page 10 6Q175 Company Sanitized. Does not contain TSCA CB1 Two sets of data analyses were performedforthe chronic toxicity tests. Test organism survival and reproduction or growth in theflB^est solutions were statistically compared to both the standard control group (with no added methanol) and the methanol control group. RESULTS 1.0 |^BB3oIubility Determination fH9 Attempts to determine the water solubility oljUBfollowedthe approach given in the test protocol that is included in Appendix 2. In brief, excess as received from DuPont - was equilibrated with deionized water in a separatoj^uBnel. After phase separation, an aliquot of the aqueous phase was analyzed for fUcontent by purge and trap gas chromatographic/mass spectrometnc (GC/MS) techniques. The analytical system used for all determinations consisted of a Tekmar Model 3000 purge and trap concentrator interfaced to a Hewlett Packard Model 589DGC, and a VG Trio 1 mass spectrometer Calibration ^andards consisted of samples oigHwn laboratory pure deionized water containingltalQatconcentrations of 80, 40, 20, T.4"4 2, and 2.1 ,ug/L. The standards were prepared by caffifullv adding to 25.0 mi- of water appropriate quantities of a stock^olutipn containing||BJyt a concentration of 40 /^g/mL in methanol. ^fB)w To minimize loss evaporation, the standards were prepared in purge and trap test tubes which were secured to the Tekmar purge and trap unit immediately after preparation. Based on the chromatographic results obtained during calibration.^Bgappearedto consist of a mixture of three poorly resolved constituents eluting witn^S retention time of between 4.0 and 4.1 minutes. Mass spectra for each of the constituents were virtually Identical in that they exhibited prominent peaks, in decreasing order of intensity, at mass to charge ratios of 77 (100% intensity), 51 (26% intensity), and 69 (14% intensity) Raw data illustrating a typical chromatogram and mass spectrum are included in Appendix 3 Also included in Appendix 3 is the derived calibration curve, which proved to be linear with a coefficient of determination of 0.991. Samples ^----JR of^^H^atu rated water were analyzed for^^B^B^Bksontent on 11 separate ( U occasions, over the course of three separate days. Each run was preceded by the analysis of a mid-point calibration standard containing at a concentration of 0.040 mg/L. Recoveries of the standard ranged from a lowofO.037 mg/L to a high of 0.052 mg/L, and averaged 0.046 mg/L (115% of theory). For reason that are not immediately obvious, the results for the^tyaturated water solutions proved to be poorly reproducible, ranging from a minimum value of 1.15 mg/L to a maximum of 16.7 mg/L. For the entire set of 11 determinations, the average was found to be 6.14 mg/L, with a standard deviation of 5.33 mg/L. Page 11 60175 'Company Sanitized. Does not contain TSCA CB( It is equally puzzling to note that the chromatogram obtained for the^BjIjjsaturatweadter solutions wasdrfferent from that obtained during the analysis oftheTalibration standards Whereas thefllRpeak had onginally appeared as a poorly resolved triplet, the peak for the saturated water solutions took on the appearance of a well-resolved doublet, with one peak eluting at a retention time of approximately 3 6 minutes, and the second with a retention time of 4 1 minutes. Again, mass spectra for the peaks were virtually identical Although it amounts to little more than speculation, possible explanations for the observed behavior o^H^Pin water include the following: ^^^ so,j||j 1. HR'^^y possess sufficient surface activity to form micellar solutions If ^^JR may^end to form monomolecular layers at the air/water interface and/or at the water/glass interface, making it difficult to withdraw representative samples from the bulk "solutions" for analysis. j^^BJRB^Y 2. be sufficiently reactive to undergo degradation (or polymenzation) in the Tieafed inlet of the gas chromatograph. If this were the case, it would not be surprising to see multiple peaks ofvanable intensity. 3 ' Focusing finally on the different peak patterns, and noting that the standards (which appeared as triplets) were analyzed immediately after preparation, whereas the JBMsaturated water samples (which appeared as well-resojyecLdoublets) had aged for up to several days prior to analysis, it may be thatU^is undergoing some unspecified sort of time dependent chemical transformation when allowed to equilibrate with water. Despite the unusual properties exhibited by^^Blinwater, it can be concluded with a high degree of certainty that its solubility is extremelylow, most likely on the order of 10 rug/t or less., 2.0 g^B^^foRxicity Determination 2.1 Toxicity Range-Finding All data generated for the toxicity range-finding tests are included in Appendix 4. The results indicated no acute toxicity to fathead minnows and Cenodaphnia at targeted concentrations ranging from 10 mg/L to 1,000 mg/L (actuaL ^(H owH^f concentration may be less than 1,000 mg/L due to apparent accumulation ^^^ on glassware) Toxicity data for these tests are summarized in Table 1 2.2 Acute Toxicity Testing All data generated for the acute toxicity tests are included in Appendix S^The^ results indicated no acute toxicity to fathead minnows and Cenodapftnia a\ Page 12 60175 Company Sanitized. Does not contain TSCA CBI TABLE 1 TOXiCITY RANGE-FINDING DATA SUMMARY Test ConcentiatiQp11' (mg/L------ --^^^ Control12' 001 0.1 1 0 50 JlOO 1,000^ Toxic;ty End-Point'4' Percent Survival Cenodaphma Fathead Minnow 24-Hour 100 48-Hour 100 24-Hour 100 48-Hour 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 LCso> 1,000 mg/L LCso >1.000 mg/L Notes 1 Targeted test concentrations. 2. Control consisted of EPA moderately hard water and contained '"lol------ 3 Actual(B|PLoncentrationmay be less than 1,000 mg/L based on appearance of feHResidual on glassware used to prepare test solution. 4. LC5o values estimated based on test organism response in highest targeted test solution. Job Number 60175 April 20, 2001 Company Sanitized. Does not contain TSCA CBI 13 OLVER LABORATORIES concentrations ranging from 200 mg/L to 500 mg/L. The survival of Cenodaphnia in the methanol control (7D% survival) was reduced relative to the standard control ^A^ (100% survival) andU|test solutions (90% - 100% survival). No reason for this reduction in survival was identified. The results are summarized in Table depicted, the LCgo values for both tests were estimated based on the highest|PPy test solution used and are reported as greater than (>) 500 mg/L. 2.3 Chronic Toxicity Testing All data generated for the chronic toxicity tests are included in Appendices 6 and 7 and the test results are summarized in Tables 3 and 4. Statisticall^fathead minnow survival and growth were not significantly reduced by exposure tofBtest concentrations ranging from 5 to 500 mg/L This was consistent for comparisons to standard laboratory water (no methanol) controls as well as comparisons to control water spiked with 500 p.L methanol. The no observed effect concentration (NOEC) value for this test was based on the highest test concentration used and is reported as NOEC = 500 mg/L. No unusual behavior was observed by control larvae in the dilution water controls and dilution water with methanol controls. A \nJheCeriodaphnia chronic test, survival was significantly reduced in the 500 mg/L UBLoncentration (40% survival) Cenodaphnia survival and reproduction data were statistically analyzed by comparing the survival and reproduction in each test concentration to the standard control group (with no methanol added) (Table 3) and to the methanol control group (Table 4^Astatistically significant reduction in reproduction was observed at the 25 mg/LHB|test concentration when the data were compared to the standard control group, resulting in a non-continuous dosedependent toxicity response In accordance with standard EPA chronic test data evaluation Bigcedures. the NOEC value for this test may be defined as 10 mg/L, the highesqU concentration under the lowest statistically significant concentration (25 mg/L). Alternatively, test organism reproduction at the 25 mg/L concentration may be considered aberrant and the NOEC may be defined as 100 mg/L, based on improved reproduction (relative to the 25 mg/L concentration) in the 50 mg/L and 100 mg/L test concentrations The reproduction data analyses showed, no statistically significant reduction in reproduction in the 100 mg/L and Icws'lBBf^ concentrations when the data were compared to the methanol-spiked control group (Table 4). The NOEC value for the test when compared to the methanol-spiked control was 100 mg/L. In light of test organism reproduction reproduction in the 5 mg/L, 10 mg/L, 50 mg/L, and 100 mg/L test concentrations, the NOEC for Cenodaphnia is believed to be 100 mg/L. Page 14 6D175 Company Sanitized. Does not contain TSCA CBl TABLE 2 ACUTE TOX1C1TY DATA SUMMARY Test Control12' Control with Methanol01 200 300 400 500 Toxscity EndPoinf4' Percent Survival Cenodaphn/a Fathead Minnow 24-Hour 100 48-Hour 100 24-Hour 95 48-Hour 95 100 70 100 100 100 100 100 100 LCso >500 mg/L 95 100 90 95 LCso >500 mg/L 100 100 100 100 LCso >500 mg/L 100 100 100 100 LCso >500 mg/L Notes. 1. Targeted test concentrations. 2. Control consisted of EPA moderately hard water and contained no|HB| ^^T 3. Control consisted of EPA moderately hard water and contained 500 iA- meth and/l and noB^^B 4. LCgo values estimated based on test organism response in highest targeted test solution. Job Number 60175 May 3,2001 15 CompanySan^d. Does not contain TSCA CBI OLVER LABORATORIES TABLE 3 Test Concenteation01 (mg/L.^9 Control'2' 5 10 25 50 100 ,, 500 Toxicity EndPoints CHRONIC TOXICITY DATA SUMMARY COMPARISON TO STANDARD CONTROLS Toxicity Test Results Cenodaphma Fathead Minnow Percent Survival 100 100 90 Average Reproduction 180 166 171 Percent Survival 98 98 95 Average Growth (mg) 0420 0468 0416 90 13.9*13' 95 0.430 70 152 100 0.442 70 4Q-CT 172 NA'41 100 0.399- 98 0.422 NOEC - Survival = 100 mg/L NOEC - Reproduction = 100 mg/L15' IC^s - Reproduction >500 mg/L161 NOEC - Survival = 500 mg/L NOEC - Growth = 500 mg/L 1C;; - Growth >500 mg/L15' Notes' 1. Targeted test concentrations. 2. Control consisted of EPA moderately hard water and contained nnoof^i^^J^Bf ^^^^^^* 3. Asterisks (*) indicates a statistically significant reduction in survival and/or reproduction as compared to the control group. 4. Not Applicable. Since test organism survival at this concentration was significantly reduced relative to the control group, reproduction data was not included in the reproduction analysis. 5. Reduction test organism reproduction at the 25 mg/L concentration considered abberant NOEC reproduction value based on test organism reproduction in 100 mg/L concentration. 6. IC^;5s values estimated based on test organism response in highest targsted test solution. Job Number 60175 May 3. 2001 ^ 16 Company Sanitized. Does not contain TSCA CBI OLVER LABORATORIES TABLE 4 CHRONIC TOX1C1TY DATA SUMMARY COMPARISON TO METHANOL-SP1KED CONTROLS Cenodaphnia Toxicity Test Results Fathead Minnow Control with MethanoP 10 25 50 100 500 Toxicity EndPoints 100 166 90 17 1 90 139 70 152 70 40't3' 172 NA14' NO EC - Survival = 100 mg/L NOEC - Reproduction =100 mg/L ICas - Reproduction >500 mg/L151 98 0.468 95 0.416 95 0430 100 0442 100 0399 98 0422 NOEC - Survival = 500 mg/L NOEC - Growth = 500 mg/L 10,5 - Growth >500 mg/L Notes: 1. Targeted "test concentrations. 2. Controlconsisted of EPA moderately hard water and contained 500 /^L methanol/L and no-------- 3. Asterisks (*) indicates a statistically significant reduction in survival and/or reproduction as compared to the control group. 4. Not Applicable. Since test organism survival at this concentration was significantly reduced relative to the control group, reproduction data was not included in the reproduction analysis. 5. IC^s values estimated based on test organism response in highest targeted test solution. Job Number 60175 May 3,2001 17 Company SaniEized. Does not contain TSCA CBI OLVER LABORATORIES APPENDIX 1 MATERIAL SAFETY DATA SHEET is Company Sanitized. Does not contain TSCA CB! OLVER LABORATORIES The USDS format adheres to the standards a-nd. regulatory requirements of the United States and. may not meet regulatory requirements in other conn-tries. DuPont Material Safety Data Sheet Page DU004604 J FUJOROTELOKER INTERMEDIATE Be-vised. 12-JAM-1999 CHEMICAL PRODUCT/COHPAMY IDEHTIFICATION ^_a Material Identification f^H^Br 1s a registered trademark of DuPont. Company Identification MAIIOTACTORER/DI STOIBUTOR DuPont 1007 Market Wilmington, Street DE 19898 AONE ITOMBERS . Product Information Transport Emergency Medical Emergency : 1-800-441-7515 : CHEMTREC 1-800-424-9300 : l-BOO-441-3637 CQHPOSITXQIT/INrOBMATIOlSr ON IMGREDIEHTS HAZAHDS IDElTriFICATION Poten-tial Health Effects Eye contact may cause eye irritation with discomfort, tear-ing or blurring of visionBased on related products, inhalation of spray or mist may cause nasal, tharoaj;, or .Inns irritation. Inhalation of large amounts of respiralale particles may be toxic to the lungs. Symptoms may be modest initially, followed in hours by severe shortness of breath requiring prompt medical attention. Carcinog-enicity Information None of the components present in this material at concentrations equal to or greater than 0.1& are listed, by IARC, MTP, OSHA or A.6GIH as a. ca.rcinog'en- 19 Company Sanitized. Does not contain TSCA CBi DUO 04604 DuPont Material Safety Data Sheet page FIRST AID MEASURES First Aid INHALATION If inhaled, artificial remove to fresh, respiration. If air. If not breathing is breathing, difficult, give give oxygen. Call a physician. SKIM CONTACT Flush skin with water after contact. before reuse. Wash contaminated clothing EYE CONTACT In case of contact, immediately flush eyes with plenty of water for at least 15 minutes. Call a. physician. IpGESTIOIT If swallowed, do not induce vomiting. Immediately give 2 glasses of water. Never give anything by mouth, to an unconscious person. Call a physician- Notes to Physicia-as Activated, charcoal mixture may be administered. To prepare activated, charcoal mixture, suspend 50 grama activated charcoal in 400 m.L -water and mix thoroughly. Administer 5 mL/kg, or 350 mL for an average adult. -Tsars" Tiearn'ro TIEXBOTES Flammable Properties Flash Point Method. : Does not ignite : PMCC Hazardous decomposition products including carbon dioxide, carbon "monoxide, hydrogen fluoride, toxic gases or particles may be formed, during combustion. These products may cause severe eye, nose, throat, and lung irritation or toxic effects- Extinguishing Media Water Spray, Foa-m., Dry Chemical, C02. Fire Fighting Instructions Wear self-contained breathing apparatus. equipment. Wear full protective-., ^.nySantee,.^^^^'"TSCACBI DU004604 . DuEont Material Safety Data Sheet (FIRE FIGHTING MEASURES - Continued.) Avoid breathing decomposition products. Page 3 ACCIDENTAL RELEASE MEASURES Safeguards (Personnel) MOTE: Review FIKE FIGHTING MEASURES and HANDLING (PERSONNEL) sections before proceeding with clean-up- Use appropriate PERSONAL PROTECTIVE EQUIPMENT during clean-up. Spill Clean Up Soak up with. sawdust, sand, oil dry or other absorbent ma.tera-al.. Shovel or sweep up. Accidental Release Measures Flush spill area with water. _J------------------------------------------------------------------ HANDLING AND STORAGE Handling (Personnel) Avoid contact with. eyes. Wash thoroughly after handling. Do not store or consume food, drink, or tobacco in areas where they raay become contaminated with this material. Avoid circumsta-ncea that produce respirajble particles unless suitable ventilation, and respirator are used. Storage -~Kavp ccciitaJLnex- tightly closed- Keep away from open flames and heated, surfaces above 200 degrees C (392 deg F) . EXPOSURE CONTROLS/PERSONAL PROTECTION Engineering Controls Keep container tightly closed. Use only with adequate ventilation. Vent heated extruder or dryer fumes outside work area. Do not aerosolize. In spray applications, use airless type pressure spray equipment at less than 60 psi, and exhaust ducts, drip pans, .or .other design .featur&s to minimize worker exposure tr> mists and over spray. Keep away from open flames and heated surfaces above 200 degC (392 degF) . 21 Company Sanitized. Does not contain TSCA CBI DU004604 DuPont Material Safety Data Sheet (EXPOSTOE COKTROLS/PERSONAL PROTECTION - Continued) Page Peraona-1 Protective Equipment SYE/FACE PROTECTIOISf Wear safety galases or coverall chemical splash, goggles. RESPIRA-TORS Wear' HZOSH approved, respiratory protection as appropriate. PROTECTIVE CLOTHrNG Where there is potential for skin contact have available and. wea-r as appropriate, impervious a-lovea, apron, pants, and jaclcet- Ebcposure Guidelines Applicable Exposure Limits Perfluorobutyle thylone PEL (OSHA) TLV (ACGIH) AEL * (DuPont) None Established None Ssta-bliahed 100 ppm, 8 Hr. TWA * AEL is DuPont's Acceptable Exposu-re Limit. Where govemmentally imposed occnpational exposure limits which ara lower fcba-n the AE1. are in effect, such limits shall take precedence. Exposure Guideline Comments No AEL ha.s been established for this product:. Other products with Eluorinated material components have an AEI. of 0.1 mg/io3 to 1 mg/m3 (8 hour TWA) for respirabia size 3ien-qso3- particles. Air monitoring studies conducted at customer sites indicates that the use of the recommended low pressure (less than 60 psi) airless type, garden type or deck specific hand pump sprayer with spray tip orifice nu n ^nni'm of 0-031 inches in diameter does not produce respirable size aerosol particle concentrations near the AEL. PHYSICAL AMD CHEMICAL PROPERTIES Physical Data Boi-Ling- Point % Volatiles Solubility in Water Odor Form Color Specific Gravity 58 C (136 F) & 100 WT^6 Negligible Ethereal Liquid, Clear Colorless 1.4 760 jam Hg Company Sanitized. D^s not contafn T8CA CBt DUO 04604 DuPont Material Safety Data Sheet Page 5 STABILITY ASTD REACTIVITY Chemical Stability Stable at normal temperatures and. storage conditiona. Incompatibility with. Other- Materials Nolle reasonably foreseea-ble. Decomposition Hazardous decomposition prod-nets including carbon dioxide, carbon . monoxide, hydrogen fluoride, toxic gases or- particles may be formed during combustion. These products may cause severe aye. nose, throat, and lung irritation or toxic effects. Polymerization Homopolymerizati'on will not occur. Copolymerization can occur in the presence of free radical sources, heat or XJV light. TOXICOLOGICAI. IMFOBMATI01T Animal Data Inhalation 1 hour ALC: >7,178 ppm in mice Oral LD5Q: >25,000 mg/kg in rats The product is not a sfcin irritant, ia a mild eye irritant, and is untested for skin sensitization in animals. Repeated exposure to high levels (5 vol'6) produced blood changes (decreased pisfcelets and. increased monocyte) . Testa in animals demonstrate no developmental toxicity. The product does not produce genetic damage in bacterial cell cultures but has not been tested in animals. Tests for carcinogenic and reproductive activity have not been performed. DISPOSAL CONSIDERATIONS Waste Disposal Treatment, storage, transportation, accordance with applicable Federal, regulations. and disposal must State/Provincial, be in and Local 23 Company SanWzed. Does not contain TSCA CBi DUO 04 604 DuPcnt Material Safety Data Sheet Page 6 TRAHSPORTATIOH IMFORMATIOtT Shipping Information Not Regulated as a hazardous material by DOT, IMO, or IATA. REGUIATORY IHFORMATlOU U.S. Federal Regulations TSCA Inventory Stat-aa : Reported/Included. TITIS III HAZARD CLASSIFICATIONS SECTIONS 311, 312 Acute : Yes Chronic : Ho Fire ; Ho Reactivity : Ho Pressure ; No I OTHER rNFORMATIOH HTPA, HPCA-HMIS NPCA-HMIS Rating Health. : 1 Flammability : Q Reactivity : 0 Personal Protection rating to be supplied by user depending on use conditions. The data in this Material Safety Data Sheet relates only to the specific material designated herein and. does not relate to iise in combination with any other material or in any process. Responsibility for MSDS : MSDS Coordinator Address : DuPont Chemical Solutions Enterprise Wil-inington, De. 19898 Telephone : 80Q-441-7515 # Indicates updated section. Tbis information is baaed upon technical information believed to be rel4-alile. It is jmbjBct 1:0 revision as additional knowledge nnrl eKperience is gained. End of MSSS 24 Company Sam-feed. Does not contain T8CA CBI APPENDIX 2 'SOLUBILITY DETERMINATION TEST PROTOCOL 25 Company Sanitized. Does not contain TSCA CBI OLVER LABORATORIES @ . Test Protocol for DetermininaJhe Solubility of i in Water 1 Obtain sample of authentic product from DuPont. No more than 100 mL should be required 2 Calibrate the GC/MS system using samples of the authentic product. At the same time, determine a RRi^onfllpn relation to an internal standard having a retention time similar to that of||H^SinceflMlis claimed to exhibit negligible solubility in water, prepare calibration standards bydissolving carefully weighed quantities of the product as received in a suitable solvent Methanol is suggested. The calibration curve should be constructed using a minimum of four, and preferably five, calibration standards J---- 3 " Equilibratel^^HWith laboratory pure water as follows V^^~ a Place approximately 25 mL of the product as received in a clean 2L separator/ funnel. 6 Add 11_ of laboratory pure water c Shake vigorously to intimately mix the two phases, venting to atmosphere as needed to relieve pressure Allow the phases to separate, then repeat both steps a minimum of five (5) times d After mixing, allow the funnel and its contents to stand undisturbed overnight (to ensure complete phase separation) e Remove a portion of the aqueous phase (upper phase) using a clean transfer ofH^o pipet Place the sample in a 40 mL VOAviaLeauipped with a Teflon-hned cap Make-the transfer quickly to avoid loss the atmosphere When full, seal the vial with its cap such that zero harospace exists after the cap is secured Prepare three such vials for subsequent analysis f After removing analytical samples, measure and record, i) temperature of the aqueous phase, and 11)atmospheric pressure oflH| 4 Analyze all three samples for the presence Quantify concentration in units of mg/L relative to the calibration curve and RRF^sdetermined in Step 2 Safety Considerations The toxicrty oi^ffU although not believed to be high, has not been well-defined All prepara tive steps should ta^e place in a hood Standard and customary measures should be taken to guard against skin and eye contact. Should contact occur, follow the procedures outlined in the attached MSDS 7C 'Company Sanffized. Does not contain TSCA CB? APPENDIX 3 SOLUBILITY DETERMINATION ANALYTICAL DATA 27 Company Sanitized. Does not contain T8CA CBI File Operator Acquired Instrument Sample Name Misc Info Vial Number abundance 110000 100000 90000 80000 C \V012S01\V0012202 BP 22 JAN L 9.02 Tno-1 1000 NG STD 25mls 0 D us-ng <^ ^ Cfis-lQfifl.7~/M -J //t^snKD Acq-Method TIC V00122Q2.D ^ofi^WW \ \ 70000 60000 50000 r^^ y . (irtpo^rn 40000 ^ i 30000 , 20000, i fi 1 10000 1 1 \ 1 n, 0 l- rime-? 3 00 3 20 3 40 3 60 3 80 4 00 4 20 4 40 4 60 4 80 5 00 5 20 5 40 5 60 5 80 6 00 A,bunaance Scan 490 (4 083 mm) V0012202 0 1 25000 H^s^cr^ 20000 15000 10000 -.', 51 5000 69 0 hTl]f/a2-> 3 03 44 , 40 50 57 1,1 60 70 |T108 157^ L n9^ 8800__9900 ^ w 145 100 110 120 130 140 150 160 170 180 190 200 221100, | 28 Company SanH?zed. Does not contain TSCA CBi Response Ratio ^/^ /<? %v LflLcaRffTic'^ Cuftve- 12- / -1 10-1, -i i . S-1 j 6- 2-i 1 ,, r, ^ I ' G -^------,------,------------^------------,------,----------------------------------------------F 0 1 2 3 - 4 Amount Ratio Reap Ratio = 3.49e+000 * Amt + 1.33e+000 Coef of Det (r^2) = 0.991 Curve Fit: Linear Method Name: C: \VQ12501----------2 .M "- Calibration Table Last Updated: Wed Jan 24 07:52:06 2001 Company Sanitized. Does not contain TSCA CBI 29 Fils Operator Acquired Instrument Sample Name Misc Info Vial Number abundance 120000 C:\ HPCHEM\1\DATA\VO119 C1\VOG11S 0 3.D BP 1.9 JAN4 1 13:45 using AcqMethod 25 TMnL^LoX--Q---1---I--^--_ ----HH2200EXTRACT 25mls 0 TIC V0011908D ^^/[}JijRfi-TtQ_ ) (7 . Z>oi.ori^ C)^- 110000 100000 90000 ^f e^^.lr - BCOOO ^ ) 2- 70000 60000 i 50000 40000 ; , : J/jTff^^1 ^?n*urf^ / ^ ^ K ^ -- i-^^ i 30000 . 1 ' 1 | 20000 'll \ \ | M\A \ i 10000 \ A ' ifc ^ ~^-<--j\j 1 '..,', 1 . I"ime-> 0 ^ 1 ....;..-----------..'. 1 i 200 250 300 350 '-' . 1 , '-?---------1 - 1'' ' ' ''1 ' ----------, . .-r1! -^r----n---------n---------T 400 450 500 550 600 550 700 750 800 850 "ibundance 450000 ; 400000 350000 Sean 424 13 533 mm) V0011908 D 7 wf^/tS^scr^on oF B^^^^^ 300000 250000 200000 5 1 150000 '^1- 100000 50000 69 57 n/z-> i'--.r'li|" 0 - 30 38 35 40 44 , S4 82 38 , 'i . '--^TTTTTr^"^^^TTT^-r^T^TT^^- A5 50 55 60 65 70 75 80 85 90 127 95 100 106 113 US KG 95 100 105 110 115 120 125 130 135 140 145 150 30 Company Sanitized. Does not contain TSCA CBI File Operator Acquired Instrument Sample Name Misc Info Vial Number Iftbundance 120000 C.\H?CHEM\1\DATA\V011S01\V0011508.C ' EP 19 JAN 1 13:45 using AcqMethod " 25 MTrLi--^--^^--E20 EXTRACT 25mls ^^^^ 0 TIC V0011908 D /</ (^/ , ^ftTiW. ^^"^ ^z-P^^r/VyV h^O 7 -^ 110000 100000 f f c 90000 80000 / ? 70000 60000 50000 1 ^ ^ ^ ?: R ^ -~'i ^ 1 p.? k}l i i^ M 1 40000 1 1 30000 1 1 I 1 l 20000 fl i 10000 ' '"' ' ' ' ~. i : m ^ ',,, ,!, ,/\ time-> ':1 2 60 1 ~'i1" i ' 3 00 3 20 3 40 ' 3 60 ''^i .1|'--^--p 11-' 1 3 80 4 00 4 20 4 40 4 50 4 '8i 0' "5 0i"0 52i0'.'5. 1- ''.' 40 5 60 ^5 .8.0..6.i0.0 ..,, 6 20 i. ...i.. 6 40 6 60 .,i 6 80 7.0r0. 7..2-0.- abundance -2200000 2000000 T Scan 498 (4 150 mm) V0011908 D t^MS ^71?^H oF 1800000 1600000 51 1 1^----MH^ l 1400000 . 1200000 69 1000000 800000 :.^ 600000 400000 200000 39 0 m/z-> 30 40 100 j 1 J 53 I 93 -)-T>1' 36|, "III 1'r-rA .^ 11 50 60 70 80 90 100 1lu08" ; 110 119 120 157 131 |I Ipp 14|5 169 130 140 150 160 170 2C37 'IB1 , 1p8195 180 190 200 227 219 210 220 230 31 Company Sanitized. Does not contain TSCA CBI APPENDIX 4 TOX1CITY RANGE-FINDING TEST DATA 32 Company Sanitized. Does not contain TSCA CBl OLVER LABORATORIES OLVER LABORATORIES INCORPORATED ENVIRONMENTAL BIOLOGY LABORATORY CLIENT _JhiEaa^ TOXICANT/EFFLUENT TEST SUMMA ACUTE TOXICITY TEST RESULTS Cenodaphnia TEST DATE(S1 l/25-|^-l),| TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION ^.^ hniM-S EPA-MHW OLVER LABS conlinuous Fathead Minnow i/^-i/z.iloi -Z ciay5 EPA-MHW OLVER LABS continuous CHKON1C I()Xlf:IIY7-EST RESULTS Cenocfap/ima TEST ORGANISM AGE DILUTION WATER SOURCE AERATION EPA-MHW OLVER LAB continuous TEST ACCEPTABILITY CONTROL SURVIVAL (s90%) loo'/, lo^/ CONTROL SURVIVAL (^80%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (&60%) AVE NO YOUNG/SURVIVING FEMALES (& 15) STATISTICAL RESULTS SURVIVAL IN 100% (.HI';)VICS+ Cont.-^ Lc50 95% C I METHOD loo'/ > 5 "^ /L rJD-1- N0 loc/ '> 6 ^3/1- M& N[> AVE WT/SURV CONTROL (z0 25 mg) SURVIVAL IN HIGHEST ( ) CONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION"? HOMOGENEOUS VARIANCE? NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH COMMENTS t^Da. Not pt.-v^^m'rvA.Vt. f_Auu- Vo lAj.'j.-f-Kc^-V mwWru\ 11 nEMPLArElDIODENCIIlOIOADMIWTESTSUM WPO OLVER LABORATORIES INCORPORATED ENVIRONMENTAL BIOLOGY LABORATORY TEST SUMMA CLIENT TOXICANT/EFFLUENT PoUo^-^p ACUTE TOXICITY TEST RESULTS JOB NO (:,on5 PERMIT NO ^/A ft^nae-F' n AirtJ "Tt &f-& PAGE .OF_{_ CHRONIC TOXICITY TFEST RESULTS Cenodap/ima Falhead Minnow Cenodaphni TEST DATEtS^ -Z/l-T-Mol -z./i-'z-Jz o| TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION ^^ V^^S EPA-MI-IW OLVER LABS continuous -i I^^C, EPA-MHW OLVER LABS conlfnuous TEST ORGANISM AGE DILUTION WATER SOURCE AERATION EPA-MHW OLVER LAB conlinuous TEST ACCEPTABILITY CONTROL SURVIVAL (^90%) Inn/ Inn'/ CONTROL SURVIVAL (^80%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (^60%) AVE NO YOUNG/SURVIVING FEMALES (^15) STATISTICAL RESULTS SURVIVAL IN 100% (.H^'-i.+C^t--) LCao 95% C I METHOD loo'/. > 1000 I^.^/L N)CS* N& AVE WT/SURV CONTROL (&0 25 mg) loo'/ > lOon -^-q/L. ND ND SURVIVAL IN HIGHEST ( ) CONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION? HOMOGENEOUS VARIANCE? NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH COMMENTS -fr rJ&= Niu4- Cagk-trr.i'^oAjIg. (A\>^. -\-a^$^.-fr;(.l-^ fnor-k\\-t-u'\ 1: JEMI'LAt61UIOBEIICII'OIQADMIM\TESISUMVtfPO Sample Information Clieni r'i^-,n+ Permit No ''""MI a Taxicanl/Efdue nl ^ JBBBi- Sample Type ^~ Grab Collected WA AM/PM Composite Col ecled From AM/PM / To AM/PM / ^a'1-'^ / / (Date) / (Da e) / (Da e) Job No ^o;-75 Sample3 NO TRC hJ/ A . mq/L, Initial p H N/A Initial G) 0 M/A Adjustnnents mq/L V-L P3 o+ 1 . ACUTE TOXICITY T eslInfo malion A nalyst(s ) 5rn,J^^ T eslSpe CIBS P. i^,.n(V^ C ulture N o ^S?'i StartofTest t'10 End of lest y3^ Test Container Size AM^i / AM?F5l 1 ^co W A as Z- d-^S Test Solution Volume 'loo Lenglh (s<; SD) & H:to.o5CM Dissolved Oxygen (Time VVeight (SD) a.L(( A 0.3'- '"5 / / / T est Mod e s-h^Kc- Aeration fJ/fti D ilulion ViValer & Pr\ (r)Hb>j Beq nning Date/Time/Ra e Pho opened 16-hourda y/0-hour Cone or % Rep 5.0 "IS/L A Number of Live Organisms 0 24 48 ID 10 ]0 % Survival /O^/ Dissolved Oxygen (mg/L) 0 24 48 7.S z^ IP . pH 0 24 7.7 ^ 40 '7,7 Temperature (C) 0 24 48 S5 ^5- ^ Initial Alkalinity (mg/L as CaCOa) Initial (mg/L a ^0 /o 1-0 PI^L A 10 16 10 /oo/. 7.?S ^ 1 0 77 'T-."?- 7,7 25 a^ 35- 0 1 n-^ A 10 1^ io 10^ -7.^ M- -70 7-8 1-.^ 7.7 25 ^ 3^ 0 01 <TVL- A 10 10 \0 (GO/, 1^ :?-,^ 7,^ 7-8 ^1' 77 Z5 ^5~ '25' ^"^ f\ 10 f0 )o (OO/ 2^. ^.(. 7/^ 7.5 ^.-7- ^7 Z5 ^ S?^ (pd 8 ^6- Analyst's Inilials/Time 5/n 4'Wn ^ Sfiol ^30 Commenis -^ (& ? i^>-/So -".L Wi?rh<Ani) ^Rr^ '\/// 1 L Condition o( Organisms al TesI lnilialion/12nd H<D,^\TM J 5/y\ ^D 5E^ MH^) , rior-ro^\ , ^ ^ 6/r> UMO ^./v) 5/n c?) -}-^\ ^, '7 , ^Qr^'7 ! \-wo SM Notes C-^^t.Li.^ : T L& 5 o > N\Tcmp]aleslUioOendi\aculerOXTEST WPD Sample Informalion Clieni ^ r^ipnrt Permil No Toxicanl/EHIuo Sample Type Grab Collected ^ M w ^ ^ iQ| L a - c ^ J - AM/PM / / Composile Col ecled (Dale) From AM/PM / / (Da e) To AM/PM / / (Da e) Job No , </-75 Samp! eNo TRC ^/ A Initial p H N/A Imlial I:3 0 AJ/A Ad)uslr"nenis mq/L mq/L -^2. ^ '. ACUTE TOX1CIT T eslInfo rmalion A nalysl(s ) 5rn ;SR(J T est Spe cies C-J^oic. C^ullure hJo \r^iM,<W\ Ln\1^^ A 9e <- i-\ ^^ Lenglh ( stSO) M//>i VVeight( xSD) N/A 1 eslMod e &+^-hc- D ilulion V/Valer & Pf\ (nHiAS Start of Test U'-lO AM/1 EndotTesI ^1-5 Tes Conlamer Size AM/r 3^ Tes Solulion Volume 'Z^ Dissolved Oxygen (T / / ' Aeralion Beq nning Dale/Time/Ra e Pho openod 16-liourda y/0-h Cone or % Rep s.o f'a/L ^ & Number of Live Organisms 0 24 40 ^ 5 5" 5 ? ^5 % Survival /00>< Dissolved Oxygen (mg/L) 0 2.1 48 0 -7-& :M 7,^ ^-T PH TempBralure (C) Imlial Alkalinity Init 24 48 0 24 48 (mg/L as CaCO,) (mg ^ 7.^ 75 a^ 3S ^0 1.0 m^ . A 5 b 5 ^ 5 /^c>7 -78 ^1, 7,^ ^7 ^ ? 5-3 7? Z'5 ^^ ^ 0 ' "V^L A 'S % 5 - ^ (T /DO/ :1SL -?(. 7.^ 7.8 5 ^) q ^ ^-^ 2S IS' <5S- 0 i-'l TV^ A s % 5 I ' 5 5? w b sj ^8 7^ 7^ 7-8 ^-L, 7 ^ 26 ' a^ 95" ^"-VTT,^ A 6 Analyst's Inilials/Time 5 f y 5 ) ^B 5m H .IPR . 5? ^fj fQO/ 7.0 ^([^ s/n 7-L, ^4 7-0 <-6 SR^ SIT) Commanis \/tL /s -X / <s. <\ ^ i- s / d " L /D('-tt,c,nu ^ / /MiW ^ ^ SU) Condition of Organisms at Test Inilialion/l Hnd H<A^VIM , narn~ic,\ . A! I o 175cl/1'i^ J j ^ 7,-] 2.? ?-? a5" (ffQ s'e^ S/n &L6 SlJ Notes Ca,r-h^-tt ^-'-So fl p pp cfn c/ <hs-o l~f-f)^, /\o 'T7> q / a^ ^hs. ^ N \Templ3lcs\G]oOenchtarule\TOXTEST WPO 1- co UJ h- >- i- '4 0 0 X 0 \- <r Ill h3 u < .2 01 ro ro u Q. Q c 0 U ' 0 Q: a ^ 10 (^ J J In 0 ft Us a) ^ 0 \P j;. rr < < t-t c ID D) ;? ^ 1 Q 0 1 m (eD 0 N -0 V) "0 0 > 0 CD c in 7n '55 c 0 m 0 0 ro "E ]3 D 1- '5 '5 0 <) 0 10 -e -0 ^ in co c UJ <D 0) M J 01 ^ c 3 3 0 J: 03 '5^ ra ro K XI (U S ^ E 0 j: U3 .2 'ro D -0 0 0 c c m 0 c Q. r Q ro 01 0 cu m < 03 0. c3 -I? J i 4: 1 ^ ^ '2 .1 ^ -3' 2 e J EJ- c- 1 '/S >u | 5' 5' ^ c c y 0 rt ^ ~m t/) Q: 5 0 Q 'M' <B Z ^ ~W wQ. <B tO M ffl ? +1 H "(O IX IX (D -0 ^ 0 c 0 ^, ^n n3 "s 3 0 t- 1-- < cu C ID "01 c 01 5 ^tn. '3 0) m 0) < J 0 1 -J "0' E tn < r- < < ? $ 0 < Z s9 Z in z0 0 '0c3. -U 0 0) 0. Q E Q E 'ra "ra in 3 .00 ro EC. -0 ~] M c < '5' m n3 - 'a-9 "TO Q S D- < + 2 1 c 0 C. "ra -0 c- a T3 < p E 15 ^0 ^ c 0 (3 T3 <D 0) ^ a Q Z 0 W (U c in 0 a | i ^ 5 0 E 'c E '" c n 0 0) x CD (B OS Q ro 1- t- LO U 0- CO u > n - 3 ca t: u y I U- 3 c i| "c E.c I5 U -: in M '0 " m o c m ^ U X? n" To ei -= 0) E. \^v^ \ ^ \~3\TO -v \ \ fo tO \ CC\ f^\ 0 0 ^d E " 3. 5 < S ~r 1!ra i s5 ^ 03 u ^- (n in 3 m ^r m CM re 0E - ^ 0 ^0 0, CO 0^, a- r^- <r~ S Q- r- ^- 0 ^ C 0) 1 0? XI 0) SE- 0 in tn Q 1^0 CO T T^ r- T CO CM 0 a; c- ? ^< 0 s? E 0 en c ^ 0 tn & -a o) 0 Z 0 ^c^ >^"\ ^ ^- ^ -y CM U^ 0 in LO <; 0. cfi Q) Q: in OS lr\ rt in CA r" r~ C3~ ^- <d- (^3 r^ ^ r'' oC f- >; 0 p c^- Vf\ 1? \s\ V^ \r\ ^> <c c^ s? en 0 U 0 0 8 0 cT e 0 0 \ \ \^ \ ^\ f^ ff- ^-r ^ f& Ln ^ u^ t^ ^-3 r^ OQ r~ ^ t!^ r" &X1 r-' '33 0 x 0 Q ^ ^n \n ir\ Tt in <s: j0 J c3 \ \ v- Company Sanitized. Does not contain TSCA CBI v i Q 3 0 si ft) A Q m VI =tt~ u -J -^ fl JS ^ (( 0 in s '5 z c^ ^ ^ ri^ ^ ^ -Z C ej v-^ i -z- tf fl^. c 1^7 $ -c -^ ic <3' -z <y ta X C-) -z % ? ^ C 'z. &J -3 zl u' -2? ^ 'on c E b L E LO tn 'E 'TO a. C <3^ 0 I f < 1.5 ^- o c LU "E 0 7 c 'm m 1- "5 U] f- l c a c ^ 0 '5 c: . -5 c = ' (-.) Sample Informalion Clieni 'C'yjporA. - nnr Permil No Toxicanl/EKlue Ssmpio Type Grab Collected AM/PM Composite Col ecled From AM/PM / To AM/PM / r * 1 1 (Dale) / (Da e) / (Da e) Job No (soils Sample3 NO TRC. Imlial p H r^^/^ N/A Inilial D 0 N/A Adjusinnerils N/A mq/L mq/L T esl Into malion' A nalysl(5 ) T eslSpeass C ullure f' o" A ae T- Stn.Sfj^ f. fr-m<\q.s Vio^ ti<^\"i' , Length ( KSD) VVeight (xSD) T esl Mod e ?^V<K-V\(- D ilulion V/Valer E^R mW<i PS i ^ I . ACUTE TOXICIT Start of Test -? ;S-0 End of Tesi 35"^ Tesi Container Size Tesi Solulion Volume AM/($Q AMitPR 3 S?^ Dissolved Oxygen (Ti / / / Aeration Beq nning Dale/Time/Ra e Pho openod 16-hourda y/8-ho Cone or % Rep 1000 Tng/^ A Number of Live Organisms 0 24 40 )0 10 ^ Dissolved Oxygen (mg/L) % Survival 0 24 40 0 /o0/. 1&. 7^ ^ ^ pH 24 7.^ Tomperature (C) 48 0 24 40 '15 55 35 0?^ Inilial Alkalinity (mg/L as CaCO,) Inili (mg/ (.^ \0 iOO rnn/L A -LQ to 10 1^7. 1&. 7 0 ^T ^T 7,^ 7.^ ^5 35 ^ C.n^.1 A ;0 (0 /O \^/. 1SL. 7,^/ -7,0 7^ 7,y 7 5 -^5 3^ 35- (AL| .. Analysl's Initials/Time Comments ^rop^ Jm 5^ ^ Condilion o( Organisms al Test Imlialion/i 5nd 5-f^ 5/n S^l SEI^ 5^ S^J $&^ i5'/n V^CflV^M , nor/na\ / 1 H CQ \-^^V/ ^n^ ^i SG.^ ^^ Notes C^-fb^ =^3 L'-'So > l N\Tpmplate<;<BioBencli\3CulelTOX1ESTWPD APPENDIX 5 ACUTE TOXICITY TEST DATA 39 Company Sanitized. Does not contain TSCACBS OLVER LABORATORIES INCORPORATED ENVIRONMENTAL BIOLOGY LABORATORY TEST SUMMA CLIENT DtAftin-tTOX1CANT/EFFLUENT ^^^^^-- Acu,-^ ACUTE TOXIC1TY TEST RESULTS ToXvf-i+a JOB NO PERMIT NO 'n&4i-)3 N/A PAGE I OF ' CHRONICTOXIC1TY T-EST RESULT Cenodgphnia Fathead Minnow Cenodaphn TESTDATE(S) Z/M-'Z./lte/of 2/H-Z/lb/o/ TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION ^'zl^ hours EPA-MHW OLVER LABS continuous 3 d^S EPA-MHW OLVER LABS conlinuous TEST ORGANISM AGE DILUTION WATER SOURCE AERATION EPA-MHW OLVER LAB continuou TEST ACCEPTABILITY CONTROL SURVIVAL (&90%) loo'/ ^/. CONTROL SURVIVAL (i00%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (^60%) AVE NO YOUNG/SURVIVING FEMALES (z 15) STATISTICAL RESULTS SURVIVAL IN 100% (Hgh-.S.t CaAC^ LCso 95% C I METHOD ^57 >5oo ^/L NO'*' \>b> AVE WT/SURV CONTROL {z0 25 mg) 1^7. > 5 00 r"5/L NtS rvb SURVIVAL IN HIGHEST ( ) CONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION? HOMOGENEOUS VARIANCE? NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH iniCMrLA7E>DIOOEMCIHDIOAUMI)llIESlSUMWrD Sample Inform a 11qnj|^^^^^^ Client' Qnflpn j^ Permil No Toxicant/Erflue Afl^ Sample Type Grab Collected Composite Col ected AM/PM From AM/PM / To AM/PM / / / (Dale) / (Da B) / (Da e) Job No /.Ol^ Sample3 NO M/^ | T RC 'i Initial p H Initial C) 0 | Adjustr-nenis ma/L mq/L Cone or % Rep Number of Live Organisms 0 2A 40 % Survival Dissolved Oxygen (mg/L) 0 24 48 fDO ^)OM ^ fl C D 5" 5 5' 5 5" ? 5" ^ S-) 1 S R 3' 70 ^ ^1 ^/- 57 P3 1 o f -Z, T est Infor mation A nalyst(s ) ,?^ / ^ E. ^ T eslSpe cies C'.fnwfoan/iii dubio. C ulture N o ^ridiui'c/iio. 1 A ne c-;?i./ /irf Lenglh (i;<SD) */ //i \^Veighl ( xSD) ^/^ T est Moc e 3.-fc.hc- D ilulion V^aler 'rA /Mt/i-i ACUTE TOXICITY SlarlofTest End of Test /30 AM/ ia.^0 AM/ Test Container Size 30 MI Test Solution Volume 3i'A<l Dissolved Oxygen (Tim / / / Aeration ^/.A Beq nning Dale/Time/Ra e Pho openod 16-hourda y/8-hou pH 0 24 4Q ^ P 1-S 7J Temperalure (C) 0 24 43 as a'' ^5 Initial Alkalinity (mg/L as CaCO,) Inilia (mg/L i-loO pp/^ A 6 c. 0 S 5 ^ 8 ^^ -7.0 ^.f ^ i? ^ ^4 ? ^o/. S 5' v 5" 5 ^^ 25 2? ?5 300 ^M A 6 C- J3 Analyst's Initials/Time Comments 5 ^ ^^ S ? 'I S ^ ^ 5' .f ^' ^ 5; Spi 11.1,0 100 / J^rJ Condilion of Organisms at Test Initiation/I=nd /iM/^ii/ ' 1 S ^ 3 7.0 1-S ^ ^ 7f7 a? i^ 7^ iJ> <,<-6, ^ ^ ^^ norMd-1 ^or' ^ T^ Con+Ti) U - ^ t,Lfi ^ Notes ft)ri)oi/^ Lc-c, ^(/-^QL/+ ^e~; ^ Q n o f aw^OTrPc/ l^ N UemplalBsWioDencWaculBnOXTEST WPD Sample Inlormalioi^^M CPeliremntil NDo uoo^r7^~^^ Toxicanl/Efnue Sample Typa if ^ff^^^^ Grab Collecled Composite Col Bded From AM/PM To AM/PM AM/PM / / / / (Date) / (Da e) / (Da B) A/^f Job No . - SampI eNo hjft TRC Initial p H Initial t3: 0 Adjust ments \/ mq/L mq/L T oslInfo malion A nalyst(s ) ^if>, -R rJ T eslSpe cies ('fryirAa/i/i/c dubiQ- Cculture h o .nrfii/i'cAlO < A^ae < ji./ /in . L englh (xSO) u!A V'Velghl (xSD) vlA T ost Mod e A tc.hc Cdilution \ /Valer CP^ |T-IH^ P3 2.^ o-^ ~&1 ACUTE TOX1CIT Start of Tesi End of Test /30 AM/' Ie? So AM/^ Test Container Size 30^1 Test Solution Volume 9S'^i Dissolved Oxygen (rim / / / Aeration ^//A Beq nning Dalen'ime/Ra e Pho opened 16-hourda y/8-ho Number of Live Dissolved Oxygeri Organisms (mg/L) pH Temperature (C) Cone or % Rep 0 2-1 ^^ % Survival 0 24 4d 0 24 40 0 24 Initial Alkalinity Inilia 48 (mg/L as CaCO,) (my/L 100 SA^\ ^ ' 6 c 0 5" ^' 5^ 5' 5 4 ^/. 5" ? ^5r/ 5" 5 \ s q.T- 7P -{ ? 1-^ 11 ^ ^S- 3.5 (U-o/ A vJ '/foo^L 6 ftfori c. 0 ^ 5" 0>-' ^8 1-3 1.0 ^ ? ^ ~\ 11 2.5 a? ^5 4( ^ 5 7o/ j" $ $ JB )------ :^S S 54 ConN ^ 5 H ? C. ?. D b ^ Analyst's Inilials/Time Commenis > ftCCicliTi+'iUi.f ? ^ 5 S ;^> V viynic ^ < /&0/ 45/ se.^ isA0 SerJ in 4h'1- n ri.1') Conclilion o( Organisms al Tesi Inilialion/E-nd hwl^f)L/ ' =1 S r)^ 1.0 s^ 6.-6 JW Ct,|ymii /i,/-Aia//0 n1^ I ^ ^ .^^a 1 t 11 a? a? -^S SLO ^ 5Liy 5L.& S^ ^"tT"0 1^ >-"Hi-i+ tvvi rl,'Lnl GO Notes ftirfroi/^^ up^ci/rec/ (^pnl4 N \Teiii|'ildlos'DioOoncli\ociilc\TOXTEST WPO Sample Inform alion ----1 Client L^E "^B^ Permit No Toxicanl/Efflue " l -- t Sample Type Grab Collected ^^^^^ AM/PM / / Composite Col ecled (Date) From AM/PM / / (Da e) To AM/PM / / (Da e) Job No &U/75 Sampit6 No TRC ^A Initial p H Initial C3 0 Ad)usl nenis \> mQ/L mo/L i^ ; D-^Z. . ACUTE TOX1CITY Tebt Inrormalion A nalysl(s ) a/n /Lm^ T eslSpe cies C ulture N o A QC p. (i^n-i-el^ fio^ 5 'Jf'~>.s Length (;;SD) c).^6! A o,oC VVeight( sSD) 3 4 A a 2-1 T esl Mod e 5-)*,h'C D ilulion V/ValBr epfl ftlH^i Start o( Test End ol Test f.'.OQ AM/pivd 1^ f^WS Test Container Size Test Solution Volume Cpoo '^So c.yn Dissolved Oxygen (Tim w3 / / / Aeration Beq nning Dale/Time/Ra e Plio openod 16-hourda y/8-hou Cone or % Rep Number of Live Organisms Dissolved Oxygen (mg/L) % 0 21 40 Survival 0 24 48 0 pH Temperature (C) Initial Alkalinity Initia 24 48 0 24 48 (mg/L as CaCOj) (mq/L 5CC Pfm A B In 1 /O 1" 10 fool- 7,B 7.0 ^.?) 7.6 7-5 -^ ^5 Ib 106 1- ^T ^- (f^L ^co pp^ A B 10 10 (0 i60 1 -7-& 7-o 1^ 7. ft 7.^" -^7 ^5 ^^ ^5 10 IQ fO 1 00 1 . 300 Pprr\ A B '7.00 fpm A 6 so Iff /o 10 l6 10 IQ /o IQ ID IQ /o loo't, -7.8 -?-^ (^(P -7.& 7^ 77 25 ^ ?.^ 160 1. 16Q1- ~1'?) 7-0 r^ 7.8 ^ tool- /3^l^'O' 77 % 3>5- 5,5 C-o.v^ 1 A 10 , q c! 6 10 lo 16 Analyst's Inidals/Tinie /rtt.H ^ ^ Comments Condition of Organisms at Test Inilialion/EEnd ^o'' i0&/- 'i/wo ncAl/'huJ "76 7^ ^(P ~7.& 7.6- 77 5/71 i-w Vf^ Sn^ LmT? \^0 norrn^ / H (^l/kJti, norrrvi y 25 ^5t5(T-) LwD ^ Ll^D (eC> Notes Co-rboM^'T ^-=IE >"5oo N \Templales\BioBench\,icule\1 OXTEST WPD Sample Inform alion ^^^ Cheni r-) t^-pjr^T-B^^^B Permit No Toxicanl/Ernue Sample Type Grab Collected m iAM/PM Composite Col ecled From AM/PM / To AM/PM / / / (Date) / (Da e) / (Da e) Job No &OH& SampI sNo Wft | TRC i Initial p H Initial C30 1 . Adjuslinents ma/L mq/L P-> -2 o . ACUTE TOX1CIT /^p Test Information A nalysl(s >m Start of Test ^ on AM^P T est Spe cies p p^in^ (<-,5 EndofTest i 50 AM?Pfy C ullure N o (fio'-l Test Container Size (j^po A 98 5 d-AA)5 Test Solulion Volume '-/^ L englh.,(>xSD) o '-fTi oo'- c-"i Dissolved Oxygen (Tim VVeight ( siSD) 7 i-l -t 0.21 w<\ / / / T es! Moc e S +a.-ft'('. Aeration D ilulion V/Valer &/'fl /v>Hi,\ Beq nning Dale/Tirne/Ra e Pho openod 16-hourda y/B-ho Cone or % Rep Number of Live Dissolved Oxygen Organisms (mg/L) PH % Temperature (C) Initial Alkalinity Initia 0 24 -10 Survival 0 24 40 0 24 48 0 24 40 (mg/L as CaCO,) (mg/L C^n-^ A [mi D^\) % TUD ><!- 1 0 1 |0 loo / 7-8 7?. ^.^ "^fi ^5 7-.T 75 ^6' ^ 10 s0 10 ;0u /. /' Analysl's Initials/Time Comments M6.H 41 l.rtID i r' Condition of Organisms at Test lnilialion/1;nd Lwp s^ WD L^O 5-n UnO wo S's-i 1^0 \^0 Hec.lt-^, , vn/irmc, //HtcJ/f^^ j , no.-rp^i Noleb CiAfbo^ ^ N nBmplalBs\BioOBndi\aciileMOXTEST WPD APPENDIX 6 CHRONIC TOXICITY TEST DATA Pimephales promelas 45 Company Sanitized. Does not contain TSCA CBI OLVER LABORATORIES INCORPORATED ENVIRONMENTAL BIOLOGY LABORATORY CLIENT Q^pon-l- TOXICANT/EFFLUENT. JOB NO PERMIT NO GonS f-VA TEST SUMM PAGE OF ' TEST DATE(S) ACUTE TOXICITY TEST RESULTS Cenodaphnia Fathead Minnow CHRONIC TOXICITY T EST RESUL Cerioclaph TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION EPA-MHW OLVER LABS continuous EPA-MHW OLVER LABS continuous . TEST ORGANISM AGE DILUTION WATER SOURCE AERATION EPA-MH OLVER LA continuo TEST ACCEPTABILITY CONTROL SURVIVAL (a90%) CONTROL SURVIVAL (;>80%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (&60%) AVE NO YOUNG/SURVIVING FEMALES (;> 15) STATISTICAL RESULTS SURVIVAL IN 100% LCso 95% C I ME1HOD COMMENTS AVE WT/SURV CONTROL (zO 25 mg) SURVIVAL IN HIGHEST ( 5oo "ZS/L ) CONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION? HOMOGENEOUS VARIANCE-7 NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH "t^S." Rcf>nitju.cA\oo/(j-oit>-tii im EMPLATF'niOBEIICHlB10ADMIH\TESTSUM WO OLVER LABORATORIES INCORPORATED ENVIRONMENTAL BIOLOGY LABORATORY TEST SUMM CLIENT D">Por-rlJ TOXICANT/EFFLUENT JOB NO PERMIT NO ACUTETOXICin'r TEST RESULTS Cenodaphnia . Fathead Minnow (j p|-T5 ^ - f^, PAGE ' OF__ CHRONIC TOX1CITY TrEST RESUL Cenodaph TEST DATEfSI TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION TEST ACCEPTABILITY CONTROL SURVIVAL (2 90%) EPA-MHW OLVER LABS continuous EPA-MHW OLVER LABS continuous TEST ORGANISM AGE DILUTION WATER SOURCE AERATION - CONTROL SURVIVAL (2 60%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (;>60%) EPA-MH OLVER L continuo AVE NO YOUNG/SURVIVING FEMALES (215) STATISTICAL RESULTS SURVIVAL IN 100% LC^o 95% C I 1? METHOD COMMENTS -$(- Corral l>j&,4r/- i^ir+^-i Ac^e^ nn<;-^^.no\ AVE WT/SURV CONTROL {tQ 25 mg) SURVIVAL IN HIGHEST ( 5oo ^3/1.) CONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION'? HOMOGENEOUS VARIANCE'? NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH ( SOQ^-L 'nc.-zc,'. Rt.pr,iju,Aonif (j-^^H-i o-f- -<4v>c^,o\ PI<- \ Li4<^ o-(- d;lu.+iB^ uj<^.+<-^\ >1 >TEMPLATE'BIOB6HCmBIOAOMimTFSTSUM WPD OLVER LABORATORIES AQUATIC TOX1C1TY TEST SAMPLE FOR STANDARD SYNTHETIC DILUT Client- P^on-^ Effluent/Toxicant _ Synthetic Dilution Wa!??" Testing Requirements 48-Hour 48-Hour 96-Hour ^deratelv Hard Water Acute Cenodaphnia Test Acute Fathead Minnow Test Acute Fathead Minnow Test (MHW) Job Number Permit No Cpc.l-76 P f^>/A >< 7-Day Chronic Fathead Minnow Tes _ 3-Brood Chronic Cenodaphnia Teal ~.Other _________________ TEST DAY Carboy No /Identification Pieparalion Date Time & Day of Use Age of Dilulion Water''' Aihalinily (mg/L) (Acceptable Range 60-70 mg/L) 1 ^ y <2-/3/0/ ^'^m^^/-?/> I '7 ^ Q^l 5 (o0 2 ^--y ^l3^] ^^m^//'/^ 6?(;|d[^ S ^/ 3 =^3 c?/(5 /ol '^am^ ;5/C!l a^ci^J 4 =^3 a/i?/o( ^''"am^a /A./OJ 3 c(dy5 5 ^ ^ a / / t F / D / ;)^oam^a//7/0( 1 rf^ 6 ^^ ^/(p/c 1 "fm/^^ /1^ oi d^ ^0 IpO 60 (p0 Gpo Hardness (mg/L) (Acceptable Range 80-lOOn-K.i/Ll Conductivity {t'rtihos/cm] PH Total Residual Cl, (mg/L) D 0 (mg/L) ^ 30 o 7-? ND' ^ ? ^7 3 o d -7.^ ND 7-^ % 30 "7 -7.^ ND 1 ? ^y^ s^ 7,? NO 7^ ^ 3^/ -7.^ NO -7,9 Continuous 7^ 313 ^.0 NO 7 ? Conlinuon C(O2)MEMaEchNTbSalch mof Sdiylunljt&nnewl.if.a- edrilu-is.tion..no. tw.u.a.s.tee-d.r.I.fso..ra.r-me,o-nraer-Bl-hriaw.ni. lh14la^dhanyrsalfnorlulowriRinignntih7pe-dprweaplaRrfatainodn draaaleaent orade chemicals fEPA/600M-91/002l------------------ NO = Not Detectable OLVER LABORATORIES LARVAL FATHEAD MINNOW SURVIVAL and GROWTH TEST SURVIVAL DATA SHEET CLIENT. PERMIT NO EFFLUENT/T_O_XICANT DILUTION WATER PP^ f^l-W _ PHOTOPER10D 16 hr Day/8 hr Night CONC REP START 1 2 A 10 i<s !o Can^f B C D 10 f6 fft 10 G 10 10 !6 is ^\ JOB NO (lOUS PAGE / of -Z ,, TEST ORGANISM AGE Z- Whf^ TT^ START of TEST _J^ A WP M}JU./I^1 END of TEST 1'^ A M^P ^-^.IJQJOi NUMBER SURVIVING PER DAY 3 4 5 6 7 6 1 0 to ;0 /0 & iQ 0 10 i0 n 10 0 ^ ^ o /0 0 In 10 COMMENTS TEST SOLUTION VOLUME 250 ml TEST CONTAINER SIZE 600 ml A ^ 8 ^L C 0 10 Ift 0 (0 10 \ lo i0 FEEDING 0 1 ml BRINE SHRIMP NAUPLll 2-3 TIMES PER DAY 10 I 0 10 10 ^ ^\ ^ ^ 10 lo !0 0 10 t. 10 10 10 16 t0 10 10 10 10 10 A B 16 C ^L 0 A B ^l C y^lL 0 10 [0 10 / 10 0 10 } 10 16 /0 16 /0 0 /^ 10 10 10 13 10 So }0 10 16 10 10 1.0 ^ 1 t 9 ^ 10 fo i0 [6 i0 \0 9 ^ 10 10 10 10 fo fn /0 10 10 1^ \6 Q 10 \o 10 ^ 10 10 f0 10 1^ /o /0 /6 A ^ 8 ^SL. C 0 10 is 10 0 /Q \o /0 Iff 10 10 i0 10 /0 ]o 10 10 10 10 \6 Q /Q lo 1 10 10 1 i0 10 10 )6 /Q 10 A B lo0^ C D ANALYST COMMENTS 10 10 10 10 PL^ Fij /-k< n /0 [0 0 /0 A' i0 0 /0 10 |0 10 10 10 0 1,6 \D ^ Lrup 'jlO ^ii ^P ^f-^ H^P V-! ^j/j____Qro.anisrn^ 0 /0 /0 h 10 10 lo /^ to /0 ^ L^D ^70 Off/Q/a r i0 (0 ^ hsa.l-i^if ^'f~ '/c's /- 1-11'/-''~ 'n'^n ^r-A ^ornp\r.ris ^ N 'TcniplaicsiBioBenchYchronicSFH^-ISURV WpO 49 Company Sanitized. Does not contain TSCA CBI fleviicd November I MOO LARVAL FATHEAD MINNOW SURVIVAL and GROWTH TEST SURVIVAL DATA SHEET CLIENT ^P'-fR^i-* PERMIT NO EFFLUENT/T_O_XI_C_A_N_T. DILUTION WATER ffl m^^ PHOTOPERIOD 16 hr Day/8 hr Night CONC REP START 1 2 3 A B 5oo ^L. C D 10 /0 /0 10 10 10 lo 1 10 /o {0 10 10 ;o 10 SO /0 A 10 /0 /> Carrkol B 10 10 /0 /0 -y ^i- C ; 10 /Mc-f^'-ino D 10 10 7 ^ 10 In /0 A 10 B 10 C 10 D 10 /y^ ^AMjg^2- JOBNO Ijini 0 TEST ORGANISM AGE. START of TEST 3 W) PPAAGGEE,-2- , of "-2 ^-Zj^-^J < 1-ZH I /5 I ol END of TEST ^ A M I^^^TSffI 0 I 4 5 6 7 COMMENTS f^ lo /0 9 /0 lo /0 /0 )0 /67 9 ^ /o /(? ^ ^ TEST SOLUTION VOLUME. 250 ml TEST CONTAINER SIZE. 600 ml )D 1. /0 !0 lo /^ ^ ^ ? C; ^ f ?^ /0 ^ ? FEEDING 0 1 ml BRINE SHRIMP NAUPLIl 2-3 TIMES PER DAY A 10 B 10 C 10 D 10 A 10 B 10 C 10 D 10 A 3 C D ANALYST COMME:NTS 10 10 10 10 MCH 5^ ^ uriCi 3-oT? 2^D ^KJ ^^ C^l+or-i-^ lfc;0 -C.''/'Yflft',5 ftnif Aff/jpur ^ ^ p i? /T^,,/ y Cf-(- -^e r-f- -i^r/-,(1-/-)0/^ r Comolc^y N 'TcmplilcslBioBcnch'ichromriFHMSURV W?D 50 Company Sanitized. Does not contain TSCA CB1 imsed Noc.-nber I 2000 LARVAL FATHEAD MINNOW SURVIVAL and GROWTH TEST GROWTH DATA SHEET LABORATORIES CLIENT JOB NO _J_01-?2. PAGE f PERMIT NO ANALYST Lm^ EFFLUENT/TOXICANT JHBL___.__- INITIAL BALANCE CALIBRATION CHECK Expected DATE Weight WEIGHED -)6 QO _3^1^1.i^j_ rnq / Actual Weight of ^ ,, Hl_JU_ mg CONC REP FOIL WEIGHT (mg) FOIL WEIGHT + FISH WEIGHT of FISH NO of SURVIVING FISH AVERAGE ^ WEIGHT PER FISH PERCENT SURVIVAL ^-m A . /O ^ H (p /^ ^ / ("'? J5nrrJ B IOkW /// 79 ^r/f 10 C 113 1-0 1/^95 ^ ^z y 0 P/o^ ?$"W '-1 o^ /o .H^2 - 57) 31Z H^ /^>^) // /no / ^ /< /o^) ^- A 9^-W /<?? /^ TI^ / o (Urol B 9^3? ^3^ 3.50 /Q (.^oH) C ^(p.O^ ^S^ ? ^ f D I !o^ (s7 /^[/ 7^ ^ /O 9 ^iH 3^o ^5-3 . 'I/O /^ / /^^ / ^9^//. A /os 9^ /n^,^^ ^ 00 /^ B ^9 /,? ~?3.^^ ^.30 1 ^^/L C f/ ^(0 ^ ^z 5"^.l 10 D ?0 5^ P^^^ H^! IQ .5^6 ^^0 -S^^ ^^ /^/9 / ^ / /ro'/ loo '/ ^5^ A ?5'/93 4 S^ /o /O B '7-f^? ?^ ^5 H ^n 16 ^/L- C ^^. ^ 7^ '/5 H^l 1 6 D "P7-66, ^1 0^ 3 H3 ? "/5-^y ^.5/. ^^/ . 3^3 1^0 / loo /- 1DO'/^0 / A B ^5- ^3^ C D ^656 ^P.^^^9 9?./^ ^3 ?? /o.?77 ^5 ^ 10^. ^H 3^ 5 03 ^ ^ L}.^ 7 /o 7 - 10 .-5^ . S-63 ."y;? .^y^ ^ /- /^^y- ^"/ /c'o / A 75: 5-/ &>.^ ^ ^ /o 5'0 B ^ ^^ ^ 5^ 1 /o IC> ^P^ .^// 7 100 '/ ~ /r/o /- C ^/L/ ^/^ ^ ^i, 3.^ f0 D ~7V "^ 7^5^ ^ ^ /O 377 vyy /^o '/ 1^0 "/ FINAL BALANCE CALIBRATION CHECK. Expected Weight COMMENTS Clllhir/ -f^/ y/0 5'6 00 mg / Actual Weight V'9. "7^ mg y- ^e^pa^e. ^j Q ^ _ 1 u-'c.\'s\-d- ^oci-s.^ o,-. Hv.- nrt'Qi'i-.^i r-i^.ri'o^- o-p la.f^ae_. N '.Tcmplalcs'.B]oBcnch\chronic\FKMGRWTH WPD 51 Rcmscd November I 2000 Company Sanitized. Does not contain TSCA CBI LARVAL FATHEAD MINNOW SURVIVAL and GROWTH TEST GROWTH DATA SHEET LAB 0 RATO R(E S CLIENT ^PU-PO^ ^ JOB NO (1,0,7s' PAGEJL of-2-- _______-____ PERMIT NO ANALYST imD_ EINFITFILAULEBNATL/TAONXCIECACNATLI_BRBAlTBIO|plTCCH--EC._K_~.Expected DATE Weight WEIGHED 5^ 00 SJI^JU- mg / Actual Weight J^UJL-HIS- CONC REP FOIL WEIGHT FOIL WEIGHT WEIGHT of NO of AVERAGE ^ (mg) +FISH FISH SURVIVING FISH WEIGHT PER FISH PERCENT SURVIVAL ^^T ^3^ ^77 //.^ A 9/5^ /O B 76 'a^ ?06'f? 3.^3 I'O /oo C '77 /^ ^ /7 8 ^^ to ^ 0 J3/ /o / ^ A /,^ /O^ 0-5 .^ ^3 /o 7/^ 5(50 B 1^ ^7- ^ /.^ 7 .W -3P3 59?? -33/ ^^ . ^/^ 100 f. 166 /. /06'/- loo /. /oo'/ % /, C ^/L. ^/6. ^37y ^/ ^? /o D ; ^S? S^.?0 ^ ^/ 10 H^ W /^^ /-. /oo 7 A B C 0 A B C 0 A B C 0 A B C D FINAL BALANCE CALIBRATION CHECK Expected Weight __5"g. 0 0 mg / Actual Weight -9'7 1^' m9 . COMMENTS r^j.jHt^-1^ - ^ i _ t^c-^-c^j!. ^m'^r b^sS-^A a^ i'tKL jgy-ie vVi Q.I ng^'P^^-c^- l-Q-yv&Lj-. N 'Tenipl!iics'.BioBcncMchronicifKt[GRWTH WPD 52 Company SanHteed, Does not conlain T8CACBI Rcns.cd November I :000 CHRONIC TOX1C1TY TEST CHEMICAL ANALYSIS DATA SHEET Page_/_ ofY O L V E R LABORATORIES CLIENT _'Di)pQnT-JOB NO IVORS' _____ PERMIT NO EFFLUENTn-OXICANT _ DILUTION WATER KPR IN1T1AL DAY 1 TEST INITIATION DATE ^l/^lol TEST COMPLETION DATE ^ ''-/ft i TEST ORGANISM P 0 ro A (. la. ^______ PHOTOPER10D ^6-HoiirDav/8-Hour Night LIGHT INTENSITY 50-100 ft-c CONCENTRATION CCoon-nfTOjrp(jfW^/f 5^ SQO .^J.a1t- AM^v-:Kaw^^ooO\} DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 COMMENTS PH D.O (mg/L) Temperature (C) Conductivity (^mhos/cm) Hardness (mg/L as CaCOj 7B 1.^ 7~7 7? '^ -7.6 "79 ^^ -^? ^5 ^6" ^5 5o0 3oo ,^?^ w 7-^ ^0 -^ -7-^ 7^ "^ '^^ Sb ^ '^1^ ^13 3/'4 Alkalinity (mg/L as CaCO,) Analyst's Initials 5m SiZ^J LmD U^D -r^ se^ LmP FINAL pH D.O. <mg/L) Temperature (C) Conductivity (^mhos/cm) Analyst's Initials INITIAL 75 (^.0 X5 ^ Sf.^ DAY 1 ^f- 7^ 7.-?) ,^^ ,^ ^ ^^ 1^ 7-J? 5^ 3-c? 3^ 95 ^^ .^.3 33^ ^96 UT]D //nn 5R^ ^/' CONCENTRATION Coh-h'Cll DAY 2 DAY 3 DAY 4 DAY 5 73 ^^ ?>^ ^^ LM7 DAY 6 ft) ^..L ^ W \^\ ^"TH^U ^iiTt-t-r\ l^ ; > ,'f^aa Co^renhccf-iw. j DAY 7 COMMENTS pH D.O. (ma/L) Temperature (C) Conductivity [ymhos/cm) tiardness (mg/L as CaCO,) f-.^ -T? ^ 3^0 ^ 7-^ 7-^ 7^ ^ 2^ ^^' 3oo ^^ 2^ ^ Alkalinity (mg/L as CaCO,) Analyst's Initials fcO L^D LoO 60 S^^J imo FINAL pH -1.5 2?"/ n D 0 (mg/L) Temperature (C) Conductivity (/^mhos/cm) Analyst's Initials G,0 55 3!6 JSW 5n 5 ^ ^^ ^ 3?)0 ^0 ^r?o LniT) N ffEMPLATE'.BIOBENCHlChRONIC'.CMRONCHE V;PD -'?--?? -7.7 7-,^ 7-? ^ 55 w 3^ 1^ <?^ LeC) fco LtYW) ^R^ 7o 7? 7-S? ^-^ ^s ^53)3 ^H ^ q^iot (o0 ^"69 ^M i-rx? 1.^ 7-^ 75 7.6, ^^ ^ /,0 // 0 35 ^-^ 33 ^ ^ ^ 2^/3 w 3^5 ^fJ A/^ i^O t^? (^^ ^^/o^ 1ACCMT Revise November 1 2000 53 Company Sanitized. Does not contain TSCA CBI O L V E R LABORATORIES ^% fk N^iBh^ CUENT T)nppn+ "-" -.in /,n'i'?5'V^ CHRONIC TOX1C1TY TEST CHEMICAL ANALYSIS DATA SHEET page -^ of H N \TEMPIATE'BIOBENCH\CHRONIC.CHRONCHE WPO 54 Company Sanitized. Does not contain TSCA CB1 CHRONIC TOX1C1TY TEST CHEMICAL ANALYSIS DATA SHEET Page of '-i O L V E R LABORATORIES ^ CLIENT T)u p0n+ ~f9HHJL JOB NO hO\^S *- PEEFRFMLITUNEONT_/T_O_X_iC_A_Nf ijBBB-^i DILUTION WATER. gP4 /YTffu7 TEST INITIATION DATE '3-/ (3 /pi I TEST COMPLETION DATE. W-^1 6 j TEST ORGANISM ^ _ <g rg^ <_ jg ^___N_ic_h_F PHOTOPERICO t6-Houf Dav/8-Hour LIGHT INTENSITY 50-IOOft-c INITIAL DAY 1 DAY 2 DAY 3 50 ?P^ DAY 4 DAY 5 DAY 6 DAY 7 COMMENTS pH D 0. (mg/L) Temperature (C) Conductivity (^mhos/cm) Hardness (mg/L as CaCO,) T-r 7-.? Z5 360 zs 7.7 7.^ -^ -7-g 9-? ^5 ^ ^^ ^0 ^>T- ^7 7^ ?.o ^ 7-^ 7J -T^ J? ^ ^'7 '314 Z}3 ^1^ Alkalinity (mg/L as CaCO,) Analyst's Initials UrO 5??^ /(n0 ?^T) W TM Lyw FINAL pH D.O. (mg/L) Temperature (C) Conductivity O-imhos/cm) Analyst's Initials INITIAL 7.5 (^.0 ^5 3n ^i^l DAY 1 7^ -7^ 7.1/ 7-3 ^ .^-^ ,^^ ^ ^^ 5,0 3-0- ^0 ^ ?-^ ^ 3/^ ^ 33) 336 ^^ /^D 4nn ^^ ftft IX) CONCENTRATION_____^-> t?PJ^, DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 ^ ^^ ^53^fff ImD DAY 7 COMMENTS pH D.O. (mg/L) Temperature (C) Conductivity Cumhos/cm) Hardness (mg/L as CaCO,) "r^ 7? Z5 ^00 7^ 7^ ^^ 7J 7^ ^ ^5 ^^ ^^ 3^o ,^-7- ,^7 7^ ^,0 7 ^ 7.? 7-? z? ^h '^S a^ 3)V W 3/^ Alkalinity (mg/L as CaCO,) Analyst's Initials ^Two ^!?^ VnO L^T) ^^ ^ LmO FINAL 1 pH 7-^ ^3 "7^ l'?.^l 7.^ 73 -M D 0. (mg/L) Temperature (C) 3A (oQ ^0 ^ ^^ ^5 ^.0 3-9 ^.0 5.f ^5 ^'7 15- fl5' Conductivity (^mhos/cm) 3o0] 710 Analyst's Initials SP^ ^0 N \TEMPl_ATeBIOBNCH\C'riRONlQCrlRONCrlE WFO 2 j 5A^ A^ ^^( 55 331 A^ m \^0 ^0 1^9 ------------------= Revise Novemtier 1. 2000 Company Sanitized. Does not contain TSCA CB5 CHRONIC TOX1C1TY TEST CHEMICAL ANALYSIS DATA SHEET Pane 01 OLVER LABORATORIES ^^ ^ CLIENT J^lJLl2fl.aL(BBBtf- JOB NO t^'1^-5- * EPFEFRLMUITENNTO/TO_X_I_CA_N_T__I---g_--_--_ DILUTION WATER: tiffi-Wlf^ INITIAL DAY 1 TEST INITIATION DATE ^//3/Q/ TEST COMPLETION DATE ^^OfO} TEST ORGANISM PHOTOPERIOD f prc.-r\ ^/fl.?. 16-Hour Dav/8-Hour Might LIGHT'lNTENSITY 5Q-100ft-c CONCENTRATION OCX) f (? fY\ DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 COMMENTS pH D.O (mg/L) Temperature (C) Conductivity (^mhos/cm) Hardness (mg/L as CaCOj Alkalinity (mg/L as CaCO:,) Analyst's Initials FINAL pH D.O (mg/L) Temperature (C) Conductivity (^mhos/cm) A-natyst's Initials INITIAL 7^ 7? ^ Soo 'U 6^ LhiO 15 6.0 ;?5 317 ^J DAY 1 1.Q '7-^ 7? '7-^ 7^ -?-^ ^5 ^^ ^t^ '^ 7,? y/o 7? -'^ T-^ 3-5 ^ 16 3 oo 30"^- ,^0-?' 3H 31^ ^ HI )OL/ /oty (?-^ ^s IOQ 6>5 6^ ^ ((70 (,0 ^ ^i-J ftnO w? 5W S^^i Lmo 7^ 3..? 9.^ ^1^ frr'n '?5 ^7 ^ ,3/,Z //7?^ 7.3 -73 7^> 7.3 3.? 3 A ^.k 0.^ ^^ ^ ^ % 331 3^0 <2)^ ^ 5^ M LrtiD i^n CONCENTRATION f 06 y j7 rr\ DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 "' COMMENTS pH D.O (mg/L) Temperature (C) Conductivity (^mhos/cm) Hardness (mgn- as CaCD,) ^r z? ^.5- 300 7 ^ 7-^ 7? 7'^ 7.? ^7^ 7.? 7-^ ^ ^^ ^5 ^ 3 oo ,^"?- 30-?- 3N ZO -^ 7,? 7^ ?5 ?.5' 3B ^ Alkalinity (mg/L as CaCO,) Analyst's Initials LMO ^PN FINAL pH ~1^ 17 ^ D 0 (mg/L) Temperature (C) Conductivity (^mhos/cm) CoO ,3^ 55 ^ m -^0^ 5W ^0 Analyst's Initials N \TEMPLATBBIOBENCH\CHrlOHIC',CHRONCHE WPO LmD l7 3 ^ ^^r ^^ 4nQ ?/nn ?^ 5-^ 73 -7-3 vo 3-^ 36 ^ 3^ 3?f ^f?fv' A^ 73 ^ ^')" ,^^ L^O irnp 7^ 4-7 ^ 3^ t^D Revise November 1. 2000 Sfi Company Sanitized. Does not contain TSCA CBS -"'"'"-" _ _ _ _ _ _ _ _ _- --___~Q"c ~^4-o--. ^-n o^ ^HSi^ _J- - L " Co^npc<r\san ---^ -1 'TO S'V^A0.'^ A. TEST FOR NORMALITY OF SURVIVAL DATA Q-'^va^s_____ Title File ^''-'^''^I^TtCIB--l --To--xJiciiy-? promelas Survr/al-2/13/01 Transform ARC SINE (SQUARE ROOT(Y) Shapiro - Wilk's TesC for Normality D = 0 1560 W = 0 8283 Critical W W = 0 8960 (alpha 0 01 , M = 23) = 0 92-10 (alpha = 0 d5 , M = 28) Data FAIL normality test (alpha = 0.01) Try another transformation. Warning ; - The first three homogeneity tests are sensitive to non-normality and should not be performed with this data as is B. TEST FOR HOMOGENEITY OF VARIANCE Title -F-Lle. :^f |(HS ^ Dupontl----Bj^TTooxxicicity-P GIB^^I promelas Survival-2/13/01 Transform AfiC 2 INS (SQUARE ROOT(Y)) Hartley's Test for Homogeneity of Variance Bartlett's Test for Homogeneity of Variance These two tescs can not be performed because at least one group has zero variance Data FAIL to meet homogeneity of variance assumption Additional transformations are useless ^^-^^ '^^-'yy Olver Labnrjtones Inco rpor.iti.-d 11.16 South Main Street ^SAiS Blackshurg, Virginia 2-1060 ^-^W> Phniie (5401 552-5548 FAX (5.40) 552-1715 q-7 'Company Sanitized. Does not contain TSCA CBI t>c^r^ fr,n^\nSiS- Co rrx ?&.<-, s^n --p S-Va-^a/r^ CirA-gA S .j C. STEEL'S MANY-ONE RANK TEST Title File DuponC-lBB|Toxi=ity-P promelas Survival-2/13/01 ^--------A Transform A5C SINE (SQUARE ROOT(Y) Steel's Many-One Sank Test - Ho Control-sTreatment GROUP IDENTIFICATION TRANSFORMED MEAN RAN-K SUM CRIT. V?J_,UE SIG DF 0.05 Ccntrcl 5 m9/L 10 mg/L 25 mg/L 50 mg/L 100 mg/L 500 mg/L 1 3713 1 3713 1335B 1 3305 1 4120 1 4120 13713 18 00 1750 16 00 20 00 20 00 18.00 10 00 1000 10 00 10 00 10 00 10.00 4 00 400 4 00 4 00 4.00 400 Critical values are 1 tailed ( k = 6 l ,,1% Olver Laboralones [ncorporjied 1116 South M-iin Street ^ Blacksburg, Virginia ;-(06U Plionc lS-401 552-5548 FA.X 15401 552-1715 58 Company Sanitized. Does not contain TSCA CBI t^O-Vc. (\no.\^c.i'S - _) Cjr^fiaiv-)^,^--n ^^a.^c^cij C-.,-.4yolS A. TEST FOR NORMALITY OF GROWTH DATA Title File Dupor^B^B^^H^^--^^--^--^ FToxicity-P ^WUMP^ ^ promelas Growth-2/13/01 Transform- Shapiro - Wi-k's Test for Normality NO TRANSFORMATION D W = 0 0809 0 9'U-l Critical W = 0 8960 (alpha = 0 01 , H = 2S) W = 0 9240 (alpha = 0 05 , N = 28) Data PASS normality test (alpha = 0 01) Continue analysis B. TEST Title File FOR HOMOGENEITY OP VARIANCE *- > DupontjpmyToxicity-P ^P^^lA promelas Growth-2/13/01 Transform NO TRAMS FORMAT ION Barclett's Test for Homogeneity of Variance Calculated Bl statistic = 1 2700 (p-value = 0 3733) Data PASS Bl homogeneity test at 0 01 level. Continue analysis Critical 3 = la 8119 (alpha = o 01, dr = 6) = 12 .5916 (alpha = 0 05, df = 6) ^^^^^^ Olvcr L-iboratones Incorporaled ^Z^^^ ll16 South Mam Street Ss;iv^ BPhlaocnfces(b5u4r0g), 5V5i2rg-Sin5i4aS 2F.1A06X0 (540) 552-1715 ,-q Company Sanitized. Does not contain TSCA CBI ^)Ci.\^x. ^.QciAu'^iS- Caf-^o<<<'.';,p^ -p3 ^A-Q.^^cifd C-o^-Vfa]^ 1 C. ANALYSIS OF VARIANCE OF DRY WEIGHT DATA Title File DupontV-M^BBBl^BT9o^xicity-P prornelas Growth-2/13/01 Transform NO TRANSFORMATION ANOVA Table SOURCE DF SS MS F Between 6 0 0117 0 0020 0 50o5 Within (Error) 21 0 0809 0 0039 Total 27 0 0926 (p-value = 0.7965) Critical F = 3 8117 ,, = 2 5727 (alpha = 0 01, df o.21) (alpha = 0 05, df 6,21) 'Since F < Critical P FAIL TO REJECT Ho All equal (alpha = 0.05) D. DIMNETT'S TEST Title File Dupon't'iW^Bfl--H --To--xiHc ity-P promTrealanssfoGrmrowth-2/13/01 NO TRANSFORMATION Dunnett ' s Test - TABLE 1 OF 2 Ho Control<Treatment GROUP IDENTIFICATION TRANSFORMED MEAN MEAN CALCULATED IN ORIGINAL UNITS T STAT SIG 0 05 1 Control 0.4198 0 4198 2 5 mg/L 0.46B3 0.4683 -1 1053 3 10 mg/L 0.41SO 0.4160 0 0855 4 25 mg/L 0 4295 0'4295 -0.2222 5 50 mg/L 0 4415 0 4415 -0 4957 6 100 mg/L 0 3985 0 3985 0 4B43 7 500 mg/L 0 4217 0.4217 -0 0456 DunnetC critical value = 2 4SOO (1 Tailed, alpha = 0 05, df [used] = 6,20) '^ (Actual df = 6,21) f^-^r^f Olvcr Ljboratnri&s Incorporated 1116 South Main Street yy-?i^WS Sl'";!"1'"1'!;. VirE'"'3 24060 pllnne y401 3s2-5548 ^X (s-40' 532-1715 60 9WI ----........ r. J -.-..,, ,,. ^SL '5'tc^j.cVd C<i^-\-,^i\ S Title File Dupont-lrt^BrToxicity-P promelas Growth-2/13/01 . 'Ctef|ft Transform NO TRANSFORMATION DunneCt's Test - TABLE 2 OF 2 Ho Control<TreatmeniC GROUP IDENTIFICATION SUM OP REPS MIN SIG DIFF (IN ORIG UNITS) % OF CONTROL DIFFERENCE FROM CONTROL 1 Control 4 2 5 mg/L 4 0 1079 25 7 -0.0485 3 10 mg/L 4 0 1079 25 7 0 0038 4 25 mg/L 4 0 1079 25.7 -0 0097 5 50 iTig/L 4 0.1079 25 7 -0 0217 6 100 mg/L 4 0 1079 25 7 0 0213 7 500 mg/L 4 0 1079 25 7 -0 0020 ; INHIBITION CONCENTRATION (IC^s) ESTIMATE Title File Cupont|k----B Toxic ty-P " tl------N promelas Growth-2/13/01 Transform NO TRANSFORMATION GPJ? IDENTIFICATION MEAN SMOOTHED MEAN CONCENTRATION 1 Control 2 5 mg/L 3 10 mg/L 4 25 mg/L 5 50 mg/L 6 100 mg/L 7 500 mg/L ICp estimate witn p = 25 is > 0 41SB 0 4583 0 4160 0 4295 0 4415 O 39B5 0.4217 500 0000 0.4440 0.4440 0 4290 0 4290 0.4290 0 4101 0 4101 0 0000 5 0000 10 0000 25 0000 50.0000 100.0000 500 0000 ^/^f ^^^'5^^ OIvcr L^borjiorna Incorpornted 1116 South Main Sireet y^'SAiSt Bijcksburg, Virginia 24060 <'y"?W pllme Is'"! S^-Sa-lS F.IX (540) 552-1715 61 Company Sanitized. Does not contain TSCACBl Qci.i-c l^nc^ijC,',;, - Camo^f\<,,^n "VT! rO'g.'W-io.noV C-on-pc^s A. TEST Title File FOR NORMALITY OF SURVIVAL DATA f ^*fc C u p o nM '" pBH. Tox 'flBMft i c i i y - P promelas Survival-2/13/01 Transform ARC SINE (SQUARE ROOT (/) Shapiro - Milk's Test for Normality D = W = 0 1627 0 B625 Critical W W = 0 3960 (alpha = 0 01 , M 28) = 0 9240 (alpha = 0 OS , N = 28) Data FAIL normality test (alpha = 0 01) Try another transformation. Warning 1 - The first three homogeneity tests are sensitive to non-normality and should not be performed with this data as is B. TEST FOR HOMOGENEITY OF VARIANCE Title File Dupon"tB'lfjUiIfTfotxicity-P promelas Survival-2/13/0:1. Transform ARC SINE (SQUARE ROOT(Y)) Hartley's Test for Homogeneity of Variance Bartlett's Test for Homogeneity of Variance These two tests can not ce performed because at least one group has zero variance Data FAIL to meet homogeneity of variance assumption Additional transformations are useless- ^O^-'I^^ ^"V^ ^^^ i^-Xv' Olver Laboratories Incorporated 1116 South Main Street Blackjiburg, Vu-guua 24060 _ i Phone {S4Q) 5S2-5S48 FAX (540) 552-1715 CompanySanitized. Does not contain TSCA CBi OoAo. r^ncA-i.Ci.i'a- Cg.r^oo.v'.^^n -Vn rrig-'^o-nQ^ ''-o'--'V''-ri\S C. STEEL'S MAIIY-OME RANK TEST Title Pile Dupont Tcxic.Lty-P promelas Survival-2/13/01 Transform ARC SINE(SQUARE ROOTiY)) GROUP Steel's Many-One Rank Test IDENTIFICATION TRANSFORMED MEAN Ho Control<Treatment RANK SUM CRIT VALUE SIG DF 0 05 Control w/ MeOH 5 mg/L 10 mg/L 25 mg/L 50 mg/L 100 mg/L 500 mg/L 1 3305 1 3713 1 3358 I 3305 1 4120 1.4120 1 3713 20 00 19 00 13 00 22 00 22 00 20 00 10 00 10 00 10 00 10 00 10 00 10 00 4 00 4 00 4 00 4 00 4 . 00 4 00 Critical values are 1 tailed ( k = 6 ) 1 /f%f^ Olver Laboratories Incorporated 1116 South M-iin Streel J^SS Blacksburg, Virginia 24060 6 3 /y-^W ^'o"11 (S-Ml) 552-554S FAX (540) 552-1715 Company Sanitized. Does not contain TSCA CB1 _______0<xVc> ^r^o^Cu3 ^s- Cci-f\c^.r\\o(^ -Vn rog-"Vnar>^\ C^fiv\s___________ A. TEST FOR NORMALITY OF GROWTH DATA TideFile Dupon^.tp|BH|lTJoIxicity-P promelas Growth-2/13/01 Transform NO TRANSFORMATION Shapiro - Milk's Test for Normality D = 0 0799 W = 0 9786 Critical W W = 0.8960 (alpha = 0 01 , N = 23) = 0.924C (alpha = 0 05 N = , 28) Daca PASS normalicy test (alpha = 0 01) Continue analysis B. TEST FOR HOMOGENEITY OF VARIANCE Title File Dupontfldl^pToxicity-P promelas Growt-i-2/13/01 ^f--------P Transform NO TRAMS FORMAT I OH Bartlett's Test for Homogeneity of Variance Calculated Bl statistic = 1 1843 Data PASS Bl homogeneity test at 0 01 level (p-value = 0 9776) Continue analysis Critical B = 16 8119 (alpha = 0 01, df = 6) = 12 5916 (alpha = 0 05, df = 6) .^, % Olver Laboratories la(.orporated 1116 South Main Street .^SS BIacksburg, Virginia 24060 f'y^'W phone ls<()> SS^SS'18 PAX (5-40) 552-1715 Company 6 4 Sanitized. Does not contain TSCA CB1 '^cx-'rc.. ^n&A'j^i.s-- Cc.nN-c><-.<-\<,sn -Vo ^^ f'0<.-TViQ.rio\ C<L^~vt)\'- C. ANALYSIS OF VARIANCE OF DRY WEIGHT DATA Title Flla Dupon^fC^BMHhIH.'oIxicity-P promelas Growch-2/13/01 Transform . NO TRANSFORMATION ANOVA Table SOURCE Dp SS MS P Between 6 0 0131 0.0022 0 5754 Within (Error) 21 0 0799 0 0038 Total 27 0 0930 (p-value = 0 7456) Critical F ^ = 3 8117 ==2 5727 lalpha = 0.01, df = 6,21) (alpha = 0 05, df = 6,21) 'Since F < Critical F FAIL TO REJECT Ho. All eqnal (alpha = 0 05) D. DUMNETT'S TEST Title File * Dupont<|^t^|TQxicity-P ^tt------N promelas Growth-2/13/01 Transform NO TRANSFORMATION Durmett's Test - TABLE 1 OF 2 Ho - Control<;Treatment GROUP IDENTIFICATION TRANSFORMED MEAN MEAN CALCULATED IN ORIGINAL UNITS T STAT SIG 0 OS 1 Control w/MeOH 0 4068 2 5 mc/L O 46B3 3 10 mg/'L 0 4160 4 25 mg/L 0 4295 5 50 mg/L 0 4415 6 100 mg/L 0 39B5 7 500 mg/L 0 4217 0 4068 0.4683 0 4160 0-4295 0.4415 0-3985 0.4217 -1 4103 -0 2121 -0 5217 -0.7963 0.1892 -0.3440 Dunnett critical value = 2 4600 (1 Tailed, alpha = 0.05, df [used] = 6,20) (Actual df = 6,21) ^^Yt-^S^^ l,r'7^''^^f-^<f^r y^^ Olver Laboratories Incorporated UUU16) S.loutlhi Mam Street Blacltoburg, Virginia 24060 Phone (540) 532-S548 FAX (540) 552-1715 65 Company Sanitized. Does not contain TSCACBI DcL-V-a, (\ ,-M-.\,;C,,< -l ^1'cr^ -,,,'>.> r,,^ ~G rTi'^'^id.nol U-.riT'rolS Title File Dupont i^B--B--Q--Tofaxitcf ity-P ^BBB^I^^^^^^' prome-as Growth-2/13/01 Transform MO TRANSFORMATION DunnetC's Test - TABLE 2 CF 2 Ho Control<Treatmern: GROUP IDENTIFICATION NUM OF REPS MIN SIG DIFF [ IN ORIG UNITS) % OF CONTROL DIFFERSNCS FROM CONTROL 1 Control w/MeOH 4 2 5 mg/L 4 3 10 mg/L 4 4 25 mg/L 4 5 50 mg/L 4 6 100 mg/L 4 7 500 mg/L 4 0 1073 0 1073 0 1073 0 1073 0 1073 0 1073 26 4 26 4 26 4 26 4 26 4 26.4 -0 0615 -0 0093 -0 0228 -0 0348 0 0082 -0 0150 ; INHIBITION CONCENTRATION (lC;s) ESTIMATE Title File Dupc^n^ ^f^1---^^T--^|iBc^ity-P promelas GrowT:h-2/13/01 Transform NO TRANSFORMATION GRP IDEOTIFICATICH MSAN SMOOTHED MEAtI CONCENTRATION 1 Control w/MeOK 0 4068 0.4375 0 0000 2 5 ni9/L O -1683 0 4375 5.0000 3 10 mg/L 0 4160 0.4290 10.0000 4 25 mg/L 0 4295 0.4290 25.0000 5 50 mg/L 0 44:15 0.4290 50.0000 6 100 mg/L 0 3985 0.4101 100.0000 7 500 mg/L 0 4217 0 4101 500.0000 ICp estimate with p = 25 is > 500 0000 ^^X-^%|/ Olver Laboratories tDCOrpor-lted '^f^f 1116 South Main Strea Blacksburi;, Viryinu 24060 Phone (S40) 552-5548 FAX (540) 552-1713 6 6 Company Sanitized. Does not contain TSCA CBI APPENDIX? CHRONIC TOXiClTY TEST DATA Ceriodaphnia dubia 67 Company Sanitized. Does not contain TSCA CBI OLVER LABORATORIES OLVER LABORATORIES INCORPORATED ENVIRONMENTAL BIOLOGY LABORATORY CLIENT O^Pon^ TOXICANT/EFFLUEN JOB NO PERMIT NO ^ un5 ^J/A TEST SUMMAR PAGE OF ACUTE TOXICITY TEST RESULTS TEST DATEfS) Cenodaphma TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION EPA-MHW OLVER LABS continuous TEST ACCEPTABILITY CONTROL SURVIVAL (iQO%) Fathead Minnow EPA-MHW OLVER LABS continuous -,. .. DILUTION WATER SOURCE AERATION CIIKONIC IOX1C11Y r EST RESULTS Cenodaphnia ^2o-2/n;o (-6 l^rs. EPA-MHW OLVER LABS continuous CONTROL SURVIVAL (iQO%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (&60%) loo'/ 10 W AVE NO YOUNG/SURVIVING FEMALES (;> 15) 1 Q-O STATISTICAL RESULTS AVE WT/SURV CONTROL (i0 25 mg) SURVIVAL IN 100% SURVIVAL IN HIGHEST ( Soo ^^/^ CONC Ho/ LCso NUMBER OF REPLICATES i0 NUMBER OF ORGANISM/REPLICATE l 95% C I NORMAL DISTRIBUTION? Vti HOMOGENEOUS VARIANCE? ^ METHOD NOEC METHOD Ejnne,1-1 '5 NOEC SURVIVAL 100 "a/ NOEC REPRODUCTION/GROWTH lo '"a/i-'"' '"'" COMMENTS Ifiy Rfp^^Jhin j^ nj\<-^. np'-< iOor^fi- QJ-A ^^A^. 5i, R^procivM-h'tn/CrvOt-i^, ex, noo-c-nn-,1-tinMAjs. Jo^.-A.-^^^A-tn't +o^i'LitM Tc^on^t . A S-bv.hi.-hLtJ.I^ 3jgrtifi'<p^ , ^ . _ ^ ^ Cooi-cn-Vf^hbrii ^onl-nn-'^n^'Qni Bo.'i-i.A. nn t^- -h^- Mo&C. I i o^Lfsfc-h'tiri biZ-lit*j<A \o iw, o-l- -b^- 0 rr^te.a..n.n-.-.iw l\/r--((bbrr.^Tr>><<^^i<ic-^.. c<r. A bg. ^1 3^^ <=< ^g'J i's,-^< lu T<f'^v\~^-c5 llllEMPLATe'niODEHCH'BIOAOMIIHTCSrSUMWPO OLVER LABORATORIES INCORPORATED ENVIRONMENTAL BIOLOGY LABORATORY CLIENT Plkporrl-1 TOXICANT/EFFLUENT JOB NO AoHJL PERMIT NO _N/^ TEST SUMMARY PAGE ; OF I TEST DATE(S) ACUTE TOXiCITY TEST RESULTS Ceriodaphnia Fathead Minnow CHRONIC TOXICITY T EST RESULTS Ceriodaphiiia 2/ZO-^/o/ TEST CONDI DONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION TEST ACCEPTABILITY CONTROL SURVIVAL (a90%) EPA-MHW OLVER LABS continuous EPA-MHW OLVER LABS continuous TEST ORGANISM AGE DILUTION WATER SOURCE AERATION -^- CONTROL SURVIVAL (a 80%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (^60%) 1-& Ins. EPA-MHW OLVER LABS conlinuous lo/ (0 8o/' AVE NO YOUNG/SURVIVING FEMALEB(s15) 15.^ STATISTICAL RESULTSSURVIVAL IN 100% LC^ 95% C I f METHOD COMMENTS ->! Con-VfQ^ V>lG.\i;j- u^^ ^A^ f^tV^nO \ AVE WT/SURV CONTROL (s.O 25 mg) SURVIVAL IN HIGHEST ( 5on ""</i) CONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION? HOMOGENEOUS VARIANCE? NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH TC-ZS; P.t^>^^>cLu.LA^Cl'>/&vo'l+t-> 1-1 O/. 1 0 ; Y.s MO 5-^1:5 100 "^1/L. lOo '"3/L '>Sot> "'3/1 ( Soa^kL Q-^ fA>jiJyio./ia\ P-r \ \-'\'^-r '^'s- d\\^'non <^'.V^-') inlFMI'LftlEMJIODEHCl^OlOADMIH^TESTSUMVtTD n OLVER LABORATORIES AQUATIC TOX1CITY TEST SAMPLE USE FOR STANDARD SYNTHETIC DILUTION Client D^n-b - Effluent/Toxicant Synlhelic Dilution Wale' Testing Requirements 'IB-Hour 40-1-lour 9G-Hour Moderately Hard Water Acute Cenodaphnia Test Acule Fathead Minnow Test Acute Fathead Minnow Test fMHW) Job Number Permit No &o)~]a, Page 7-Day Chronic Fathead Minnow Test -2^- ' 3-Biood Chronic Cenodapliina TesI Olher _________________ TEST DAY Carboy No /Identification Preparalion Dale Tune & Day of Use Age of Dilution Water"' 1 ^3 2)^0 ( '^SrnltCTZ /20/(?l ' ^'J 2 3 rf3 i n o\ ll^'ga/pmZ /2l /Ol ^3^5 z|^Toi ll^'am/Q^- /12./0] 1. dtt-^i 3 ctc^l& -1 ffH 2. 1') o> (|^n/fl /23/c 1 ^ dai.|4- 5^ ^v a^i'? /oi '^/pn^/^O) ^ Sd^r G ^7 s^/^/ ,, / am/g^^/^/O; y Lt* ^ ^ ^J Alkalinity (mg/L) (Acceptable Ranqe 60-70 mg/L) 6^ ^ ^ LT- ^c^ G ? Haidness (mg/L) (Acceptable Range 80ToOmp/Ll ConduclivilY (ffmhos/cm) pH Total Residual Cl, (mg/L) 0 0 (mg/L) Aeration /DO 33>(e ^8 ND- 7 B Continuous 100 ^ ^e ND ^6 Continuous loo ^ 1-S ND -i.8 Continuous ^13l? --i <? NO ^-8 Continuous ^ol ^ 15- -/^ ND ? ? Continuous n 31^ ^.0 NO -^ Continuous COMMENTS (1) Synthetic dilution water is prepared with laboratory deiomzed water and reaaent araue chemicals fEPA/600/i-9l/002) (2) Each batch of dilution v/aerjs not used for more than 14 davs (ollowina the preparation dale NO = Not Detectable Cenodaphnia dubia SURVIVAL AND REPRODUCTION RECORD CLIENT __L>'A{ _ PERMIT NO W. EFFLUENT/TOXICANT .4 DILUTION WATER PP> -nm^" t-'MUIUl-'cr^iuu iiCONCENTRAT10N REP .^,,p,,^= /o3 1 (A) Day 1 0 2(B) 0 3 (C1 0 L-on-VrO ^ 4(D) 0 5(E) 0 e(F) 0 7(G) 0 B (H) 0 I 3(1) 0 10 (J) 0 0 TOTAL Day 2 0 0 0 0 0 0 0 0 0 0 0 CONCENTRATION /. Survival ^0 P J i -^ / \ ( ]y\t.-i^"o^ - 'S~CKI -<J>L TOTAL REP 1 (Al 2 (B) 3(C1 4(D) 5(E) 6(n 7|G> B(H1 9 (1) 10 (J) Day1 0 0 (3 /X n 0 0 0 0 0 0 0 Day 2 0 0 .--. 0 0 0 n 0 0 0 0 CONCENTRATION REP %Survival=/Q5 1 IA> 2(B) 5 "^/L- 3(C) 1 (Dl 5(E) 6 (F) 7 (Gl S (H) 9 (I) 10 IJ) TOTAL. Day 1 0 0 0 0 0 D 0 0 0 0 0 ^i, Day 2 0 0 0 0 0 0 0 0 c 0 0 f^y M \TEMPlATE1.310aENCH\CHRON^C'.COREOP.OD WPD JOB NO 6c''75 PAGE. ofjL TEST ORGANISM AGE _^-~L\^-(^t^-^ 'S ^n _) START of TEST jLl^-_AM^r_2_/_2Q./^J_ END of TEST 3'^ A M lST.L-JJ3Jj^- Day 3 0 0 0 0 0 0 0 0 0 0 0 REPRODUCTION CO UNTOAY Day 4 i-if S 5 ^ Day5 & 0 C> 0 Dayfi 4 Q*~ S ^ Day 7 H I* ^ ^ ^ [n ! 1 /. 1 ^ H S b H 3 ^ Z9 0 S~ ft-*- .-< 0 t l 0 ^1 0 12- 0 /5 41 C^- Day3 0 0 0 0 0 0 C 0 0 REPRODUCTION COUNT/DAY Day 4 ^ 5 Day5 0 0 Day6 IP ^ Day 7 .$' 3- -- -- .-- .-- 4 0 ^ ^ ^ ^ ~^ 6> 3 ^ 0 3 5 0 ^ ^ C. 0 ID L/ -7 0 0 ^ 0 ^ 3 36' ^ 3o ^^ Day 3 0 0 0 0 0 0 0 0 0 0 0 M. REPRODUCTION COUNT/DAY Day 4 5 Day5 > Day 6 g Day 7 H 5 0 if ^ Q f, ^ ) ? H C? ? H ^ G) 1 ~1 0 ^ ^ lO 3 G 0 ^ 3^'i 5 ^5 0 3^ StiJ Jg.-J L, h 3 b 9 ^? ^ ^ 1. ^ ^L^ 71 TOTAL '? ^ J7 /^ ?? /7 S.I n ^^ asi ?o COMMENTS TEST SOLUTION VOLUME 15ml TEST CONTAINER SIZE 30ml FEEDING 0 1 ml T/C + 0 1 ml ALGAE/DAY t S<AI+ ^BO^S TOTAL ^ 1^ 0 '7 ^3- 7 7 3^0 ?/ /^- <43 COMMENTS TOTAL !1 n 1(0 n jQ /^ /'/ /7 ^ /5 ^^ 5'?J COMMENTS X = DEAD - a DIED PREVIOUSLY S/X = ? OF YOUNG PRODUCED BEFORE ADULT DIED Hc-nca ;une ~-0 :"CO ^any Sans&ea, sggg ^ ^^. Cenodaphnia dubia SURVIVAL AND REPRODUCTION RECORD CLIENT___, PERMIT NO ^^A _ EFFLUENT/TOXICANT. D| I1L1 U1 1TIIIIOHNM WVVA/-.T1 E. R, E.>p..f.l, n^Lj PHOTOPE--R--IOD-' -1"6- n--r 'D"la--y/.8'OhrMNii.ghi CONCENTRATION R'"E'"P" ./.S,,n^,=q0 1(A) Day1 0 Day 2 0 21ICd3\ 0 0 3(C) r> 0 \0 4 (01 0 0 r^^/, . 5(E) D 0 6 (H 0 0 7(G) 0 0 8/(un\) 0 0 j. 9 (11 0 0 10 (Jl 0 0 TQTAl---- 0 0 CONCENTRATION /. Survivals ^0 Z'S rn^y, ~ TOTAL REP 1 (A) 2(81 3 (C) 4(01 S!E.l S (PI 7 (Gl B (H) 9 (11 10 (Jl Day1 0 0 0 0 0 0 0 0 0 0 0 Day 2 0 0 0 0 0 n 0 0 0 0 0 '-- JOB NO 6io)"T5 PAGE -2- arji. TEST ORGANISM AGE ^.Z-W hfj- C^i/^-f S hf i-} START of TEST __/'-3C _AM/l^i ^ / 't-SloT END of TEST 3^ A.M-(Q^ - .. ' -' Day 3 0 0 0 0 0 0 0 0 0 0 0 R,,E--BP--RO--D--UC.--DO.-N..c-~o,u>J,N-T^/,D^A.^Y Day 4 3 5 DayS 0 :? Day6 /. 0 Day 7 -i1l ^ 3 0 IP // L/ ^ 1 ^ S 0 b .? 3 ^ 0 11 ^ fc o ^5 8 3 3 0 10 0 8li. 1^ 3 (. 0 11- 3n 3fo y~(c> Day 3 0 c 0 0 0 0 n 0 0 0 0 RiEPRODU CT10N CO UNT/DAY Day 4 ^ ^ .5 $ 5 / ^ 3 3 ^ 3^ DayS 0 Q 0 0 0 ^ 3 A <rt ^ ^ DayS Q 0 b- ^ /, /, ^ ; ^ 0 1 ^ DayT /. ^ 0 0 ^ 0 /,, o/X z /o ^^y TOTAL 13 /^ /^ /?? /? 19 ^0 17 i^ a/ 111 COMMENTS TEST SOLUTION VOLUME 15ml TEST CONTAINER Sk= 30ml FEEDING 0 1 ml T^C -r 0 1 ml ALGAE/DAY TOTAL 1IS0 . 15 c! u 1^ 1^ IH N 11 \\ 13 ^ COMMENTS CONCENTRATION /. Survivals 7-0' ^0 ^5/L, TOTAL ANALYST REP 1 (A) 2 (81 3 (0 4. (01 S (E1 6 (F1 T (Gl S (HI 9 (11 10 (J) . Day 1 0 0 0 0 0 0 0 0 0 0 0 y-& REPRODUCTION COUNT/DAY Day 2 Day 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 n ----[ 0 0 5UI 0 0 0 ii-ft^- Day 4 H 5 fc ^/ 5 ^ <^ ^ 5 ^ ^ '"ttr-J DayS ^ 0 0 S^-o 0 0 6 ^ 5 3 3f -r^ Day 6 ^ 7 ^ ^ 5' 0 z 0 ^ 0 3& .^A Day 7 3. 5' ^ ^ 3 /./;< ^ -o3]/ 10 1% ^ ^Pr NATEMPlATglBIOBEMCHVCHRONIOCOREPROCWPO TOTAL 13 11 15 1 ^ 13 to ?3 ^ /^T- n 15?5^(^ COMMENTS X = OSAO - =01ED PREVIOUSLY ?.'X=# OF ''OUNG PRODUCED BEFORE ADULT DIED . . .-- 'n 'ni-in 77 Company Sanitized. Does not contain TSCAC8J ^^ ^ O L V E R LABORATORIES ^ NO^;-f^i^ii?3^^- NT^| CLIENT PERMIT M^^^^^^^^^^^J ny^2oi >ii\M, EFFLUENT/TOX1CA DILUTION WAT=R Epa "T)-^^ PhOTOPERIOD 16 hr Dav/a hr Night CONCENTRATION REP .^Mry.^l-'?^------ 1 (Al Day 1 0 Day 2 0 Day 3 D 2((0B1) L? .o 0 3(C) 0 0 0 4(Dl 0 0 0 \OQ "5/1- - 5 IE) 0 0 0 6(F) 0 0 0 7(G) 0 8/(Unti 0 0 i sin 10 (J) 0 0 0 0 0 0 0 0 0 TOTAL 0 0 0 Ceriodaphnia dubia SURVIVAL AND REPRODUCTION RECORD j^ !^ JOBNO ^sH5 P AGE ^ Of 5 E ST ORGANISM AC35 <-24 hf^- C^iH., - 3 IT''- ) START of TEST l 3Q AM/1@1 -3. 1 1-0 j Of N Oof TEST AM/> Z-- 10\ REPRODUCTION COUNT/DAY Day 4 DayS Day 6 Day? ^ 5> 5 5 5 ^ ^ 5 5 S ^ ^ ^ ^ ^ S' (9 /Z 0 9/x 0 7^ (7 // 0 ^ <9 ;0 (; ? 0 Q /y. 4 5 0 (o ^ 3 0 q ^ ^y^ 0 ^ TOTAL 3,9. /'? /<,, ^0 J7 / '? / L, ^ ;5 /^ na COMMENTS TE5T SOLUTION VOLUME 15ml TEST CONTAINER SIZE 30ml FEEDING 0 1 ml TVC 0 1 ml ALGAE/DAY CONCENTRATION % Survivals 40 SDL^ -^/L REP 1 fA) 2/I(3B*) 3 1C) 4(D> 5(E) 6 (F) 7(G) 8(H) 9 (11 HL!-" TOTAL . Day 1 0 0 0 0 0 0 0 0 0 0 0 Day 2 0 0 0 0 0 0 0 0 0 0 n Days 0 0 0 0 0 0 0 0 0 0 0 RiEPRODU CTION CO UNT/DAY Day 4 Day5 Dayfi Day 7 3 0 1ij // 7" 5 <9 /2. 5 0 '-1 L 5" 5 0/X ^ 0 7 ,? 3 fc 0 ?/^ 5 ^ 3 P/X ^ $ ^ t5' 0 /o/X 0 '?// ^ s ^3- 33 0 io /K /g ^ TOTAL /)<0/' a^ /1? /o ^ /<, ^ ^ /g f^ (74 COMMENTS I CONCENTRATION /. Survivals REP 1 (Al 2 IB) 3 (C) 4 1D1 Day 1 Day 2 Day 3 REPRODLJCTION C OUNT/DA y Day 4 Day5 DayS Day 7 TOTAL COMMENTS <=OEW =DlED PREVIOUSLY 5 (El 6 (F1 7 |G^ 8 (HI | 9 (1) 10 (Jj | #/X = ?OF YOUNG PRODUCED BEFORE .IQULTDIED TOTAL ANALYSI N \TS-yPH're.E10BENCH\CHRCMIOC3R6PHOQ WPO j[6(A | .<(?^ I-^,\J 6i-r> Sft. td 'Company'SamKzetS. Does not contain TSCA CB! ^cv^fl J"n --0 2000 ^S? O L V E R LABORATORIES ^ CLIENT JOB NO PERMIT NO ^(pcn EFFLUENTH-OXICANT DILUTION WATER. EPS INITIAL CHRONIC TOX1C1TY TEST CHEMICAL ANALYSIS DATA SHEET Paee or ^ ^ TnH^J DAY 1 CONCENTRATION DAY 2 DAY 3 TEST INITIATION DATE. Z-/2-0/P/ TEST COMPLETION DATE '2-h^lo I -TrEe-Se-Tr nOoR^GAAMNIiSCMk^i/^\ n r-.Jd_r.tL,_^7Ic^T15.u. bt. i c. PHOTOPERIOD 16-t-iQUf Dav/8-hour Niqm LIGHT INTENSITY 50-100 rt-c / " i C-pp-t-ro 1 i / ( - ' / !'i;'^-'i- f^<di^rof \ ) DAY DAY DAY DAY 4 5 6 7 COMMENTS pH D 0. (mg/L) Temperature (C) Conductivity (^mhos/cm) Hardness (mg/L as CaCO,) -j Alkalinity (mg/L as CaCOJ Analyst's Initials FINAL PH D.O (mg/L) Temperature (C) Conductivity (^imhos/cm) Anaiysfs Initials 7.6 78 7-8 ^ 8 " ^5 ^ 3^ S^b 5m SLfr \ ^ ~i^ ^ 3?3 ^ ^ ^ M xs 35"3 SL6 ^ S :? 8 -7 ^ ^0 -{ e ^s ^S 1-? ^^ ^ ^-5 TL5 33L. 3l5 ^5 ^5 a? 31$ 3 i5 ^?? SLft ^lA Se-^ S-^ 6L& ^ :?- ^-T- 7.'5 ^-6 ^S as S^lH 3^ 5^b j^rJ ^ 1,D A^ 39^ ^^ :? I. L-^ 15 33'i- 6/-a. -^-u L.X ^^ ^ ^ INITIAL DAY 1 CONCENTRATION LooTro DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 COMMENTS PH D 0. (mg/L) Temperature (C) Conductivity (^mhos/cm) Hardness (mg7L as CaCO,) Alkalinity (mg/L as CaCO;) Analyst's Initials FINAL pH D.O (mg/L) Temperature (C) Conductivity (^mhos/cm) Analyst's Initials Z8 78 2-5 3^ (00 ^ ^ ^ ^ ^f-M ^5- 383 aLfi ^7S ^l ? 15" 3^L 100 u^ &(J> ^8 ^-8 2.5" 53L. |oo 1.^ ^.& ^ ^? A? 3l? ^<Z ^f.Z .!:(-& -7^ 7'? ^6 315 ^ Lp'9. S"^ ?0 1 ? 7? - 1-0' ^5 a-5' 3^ 3l? ^3- /DO ^ LO 5/Zfv SUS + D-te -Yr^-.LnWi i'-'&<ci-f<Li\0^ 3M Tt ^ ^ T.T 7.^ ^.^ ^-l, 1^ ^2 ^ ""?.o i,^ L-^l I? 25' 3^ ^ 35 ^ 3^ 3?^ 3^^ w 3^- 33^ ^Lfl- 5LA -r^N ^eiJ 6L& ^fr 74 ^=======J Revise Novemter 1 2000 ^ 3 ^ O L V E R LABORATORIES CLIENT JOB NO -frnl-l PERMIT NO EFFLUENT/TOXICANT DILUTION WATER: INITIAL CHRONIC TOXiCITY TEST CHEMICAL ANALYSIS DATA SHEET pase "z- o f _H DAY 1 d- DAY 2 DAY 3 TEST INITIATION DATE 'i-JT-oj o) TEST COMPLETION DATE a- 7?? /oj TEST ORGANISM (Vnodcphn'ia rl/,hi(^ PHOTOPER10D 15-Kour Dav/9-Hour Night LIGHT INTENSITY 50-100fl-c to ^'a/t- DAY 4 DAY 5 DAY 6 DAY 7 COMMENTS pH D.O. (mg/L) Temperature (C) Conductivity (/-icnh os/cm) Hardness (mg/L as CaCOj Alkalinity (mg/L as CaCOj Analyst's'Initials FINAL pH D.O. (mg/L) Temperature (C) Conductivity (^mhos/cm) A-nalyst's Initials 7.6 -7-8 ^5 3^ 5m ^ ^ ~\ ^ ^ 3^ SLB ^ s 1 ? I? 3.SL, SL& I "i ^ 4 ^5 35^ 5U!> ^ ? 1 ? -LI ^.0 1 2 ^ ^ :? ? -7 S U ^ ? ^? ^ A5 A5 0') 3^ 3l? b\6 315 3lS 5L&- SLfi^ ^'rlJ ^r^ .u6 =1 ^ -7^ -7-^ ^ (/ T^ '^ :! ' ") ^ '? .o (, S (/-'] ^ ^ 35" a? 31 ? w 391 322 33? ^A 5^ ,S^ &LA ^Lf>- INITIAL DAY 1 CONCENTRATION __5_/r"3/L_ DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 COMMENTS pH D 0. (mg/L) Temperature (C) Conductivity (^mhos/cm) Tiardness (mgTL as CaCOJ 1-8 ~'-S 2-5 3^<o ^? S' ^ s ^ 33- L, ^ ^) ? ^.f ^0 To1 1 ? 7.? 1.^ 2-S 3.S- 3^ ^5 33L 3l ? 3i$ 315 -\ S ^8 X5' 3US' Alkalinity (mg/L as CaCOs) Analyst's Initials ^n^ iLb SL6 -SLA- ^^J Sr^ 516 FINAL pH 1 1 ~\ ^ D 0 (mg/L) 1 ^ ^ ^ Temperature (C) Conductivity (,-mhos/cm) ^r 3S"^ IS" ^53 Analyst's Initials 5i& ^(A H \TEMPl.-i,TBBIOBENCH'.CliRON]C.C.-lRONCHe WD T- ^ -^ ' ^ W SL& -7.S 7,g :] <- 7 l- -7,0 "1,0 L ? ^/-f ^5 ^5 7? 3.S" 346 3^1 33^ 33? W ^fJ ,^A- SL^ 75 Company Sanitized. Does not contain TSCA CBI Revise Novernber 1 2000 CHRONIC TOX1CITY TEST CHEMICAL ANALYSIS DATA SHEET Pass ^> of ^ O L V E R LABORATORIES CLIENT JOB NO PERMIT NO EFFLUENT/TO_X_ICANT DILUTION WATER. INITIAL pH D 0 (mg/L) Temperature (C) Conductivity (^mhos/cm) Hardness (mg/L as CaCOJ DAY 1 7.6 7-8 ^5 3^ a ;-^^-//. TEST INITIATION DATE. ^-/2-0/oi TEST COMPLETION DATE- i /J?/o< TEST DR(-;ANISM r',n0fjnnh^o. cjufan, PHOTOPERIOD 16-Hour Dav/8-Hour Night LIGHT INTENSITY _50-100ft-c CONCENTRATION 50 ^7l_ DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 1 ^ s l-s 1 S 7^ ? ^ ^ a' ^ -//- -7S ^ ^'? ^? ^? 2,5 ^6 ^5 331^ 331, 31? 3i6 3^ DAY 7 ^8 1 ;?-^ 3-'3 32S- COMMENTS Alkalinity (mg/L as CaCOj Analyst's Initials 5/n SL6 5LA 5U^ ^F^ JP^ 5LSY FINAL pH 0.0. (mg/L) Temperature (C) Conductivity (^mhos/cm) Analyst's Initials ^ ^ 3 ^ 3.8^ ^L& ^ ^ 4 IS ^5, ^ T^ T1 ?.? 3-=)t/ SU^ ^.3 (, J ^^ 3V? W 7-? 1-0 ^5 355 ^r^ ^ i- ^ :! 3? 33^ ^Lft ^ IP ^ ? 2? 33"!- $^- INITIAL DAY 1 CONCENTRATION 2.5 rn^ /1 DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 COMMENTS pH 0.0 (mg/L) Temperature (C) Conductivity [urnhos/cm) 'Hardness (mgTL as CaCO,) 1-8 7-S ^5 3^ :? 0' -\ 8 IS 33.1-, ^^ 1-8 3.5 33 L, :?-^ -) c! ;! S 7.S IS- ^? y.o ZB ol^ ^?-? ^i? ^5" 315 Bi5 3i^ S3.S" Alkalinity (mg/L as CaCOj Analyst's Initials ^m SL& FINAL pH D 0. (mg/L) Temperature ("C) ^ q M ^-5- ^ ^ "I "1 15 Conductivity (umhos/cm) 3&^ 358 Analyst's Initials ^ 6L&- N-\TEMPLATcl610BeNCH\CHRCNlC.ChP.CNCHE.WPQ 5L& ^l.^ ^^ J-^ ^ ? ^.^ '7,? :) i. :? 1 -7.a- ~7,0 U 1 ?.^ ?5 ^^ 7.5 ^-^^ 34^ W 330 6L& ^^rJ 5^ 5^> 76 A<-6 l ^ ^ L-S z5 3MI 5LA Revise November 1 2000 i- Doss "of confepn TSCA CBi CHRONIC TOX1C1TY TEST CHEMICAL ANALYSIS DATA SHEET Pae of ^ INITIAL pH 0.0. (mg/L) Temperature (C) Conductivity (^imhos/cm) Hardness (rng/L as CaCO^) Alkalinity fmg/Las CaCOJ Analyst's Initials FINAL pH D.O. (mg/L) Temperature (C) Conductivity (^mhos/cm) Analyst's Initials INrriAL DAY 1 7.8 7-8 ^5 3^ % (/"i 5rr> DAY 2 "?-? IS as 33(/ ')(, L'-l ^ DAY 3 ^^ ^ ? Si 331/ ^1' ^ 6LA DAY 4 ^4-?^ ^/2?/0< TEST INITIATION DATE j - - TEST COMPLETION DATE TEST ORGANISM /VncrhpnmG.rincuc. "HOTOPERlOC 16-Hoiir Dav/8-Hour Night LIGHT INTENSITY 50-100rt-c 5 00 ^^IL. DAY 5 DAY 6 DAY 7 COMMENTS ^ S ^ 2 ^LS 3lb f0^ (o^ AA/ -) '1 ?i.Q '/-? ^^ .1$ ^7 ^^ 3i5 h ^ lo^ ^4 G)^ ^^J, 5^ ^ ^-S ]LS" ^ i^ (DO ^ led SLIV hi S5/.-J + 1i ^t bol-'-- +f'^tLnl=< 'iiCcK-.-ti.M, 4 ^ ^ =1-? -7.3 '/7 :? I/ ~\ Z ^4 (. 1 '7.0 Y-Lo L ^ ^.s 1$ ^? ^ ^^ 7.^ 3^ ^? 3^ 5 3^(. ^n 52.0 ,U& ^(^ ^Lfr 5N ^r^ 5Lfr DAY 1 CONCENTRATION DAY 2 DAY 3 DAY 4 /i^'0 ^a/t- DAY 5 DAY 6 ^ t. to.T55- ^2.2 ,->L^ DAY 7 COMMENTS pH D.O (mg/L) Temperature (C) Conductivity (^mhos/cm) Tiardness (mg7T- as CaCO,) 1-8 ~f-8 2-5 3^ -7? :?? 2-? SSL 1-& ^ 8 ^ 3,31- 7-? 7.<:? ^S -7<? ?,0 1^ l' ^5 ^5 li^ 316 3i^ Alkalinity (mg/L as CaCOj Analyst's Initials ^ 5Lft FINAL pH -1 ^ ^^ D.O (mg/L) ~] 3 ^ H Temperature (C) Conductivity (^mhos/cm) "t5 I? 34? 3LZ Analyst's Initials ALA ^LA- N ffEMPLATBBIOBeNCHXCHRCNIC'.CHRONCHE WPO &LA 5Lfc ^^ 5/ZA/ ^-? -7.4 ^ 0 ^.0 ^ ^ 3^2. 343 SL^- 5tJ 7-) G,^ ^ 3^ l^ ^ ^ L, L. ?? 3^ &L^ 77 Comply S-ar-Ac-; ?-^ ^ ^ a? S'x5' 5L6 ^-L. ^-^ 2? 3^ ^>- Revise November 1. 2000 DoA&IXnc^S^- Cc.-npft^on -'IT? Tl-la- T^.jita.^ Ccn^t A. FISHER'S EXACT TEST FOR THE DETERMINATION OP SIGNIFICANT DIFFERENCES IN TEST ORGANISM MORTALITY Fisher's Exact Test NUMBER OF IDENTIFICATION CONTROL 5 mg/L ; TOTAL ALIVE 10 10 20 DEAD 0 0 0 TOTAL ANIM?-LS 10 10 20 Critical Fisher's value (10,10,10) (alphaaO 05) is 6 0 b value is 10 Since b is greater than 6 0 there is no significant difference between CONTROL and TREATMENT at the 0 05 level Fisher's Exact Test NUMBER OP IDENTIFICATION CONTROL 10 mg/L TOTAL ALIVE DEAD 10 0 9 1 19 1 TOTAL ANIMALS 10 10 20 Critical Fisher's value i'10,10,!C) (alpha=0 05) is 6 0 b value is 9 Since b is greater than 6 0 there is no significant difference between CONTROL and TREATMENT at the 0 05 level N^X'^%/ ^^^-/y$ ^ ^^V Olver Laburatunes Incorporated 1116 Souill M.iin Sireet Bla<.ksburi;, Virginja 2-1060 phaM (540) SS2-33-*5 t^X (540) 552-1713 Company Sanitized. 78 Does not contain TSCA GBI l^oAtA ^^g-\^5iS-CaiT.pft<,'\,^^-VD'I^J.L. 54<AnAo.if^ Con^oi Fisher's Exact Test NUMBER OF IDENTIFICATION CONTROL 25 rng/L ALIVE 10 9 DEAD TOTAL ANIMALS 10 10 TOTAL 19 20 Critical Fisher'3 value (l0,10,"-0; (alpha=0 05) is 6 0 b value is 9 Since b is greater than S 0 C-iere is no significant difference between CONTROL and TREATMENT at the 0 05 level Fisher's Exact Test NUMBER OF IDENTIFICATION ALIVE DEAD TOTAL ANIMALS CONTROL 10 10 50 m9/L 10 TOTAL 17 20 Critical Fisher's value (10,13,10) (alpha=0 05) is 5 0 b value is 7 Since b is 9reater than 6 0 there is no significant difference between CONTROL snd TREATMENT at the 0 05 level r^f$. ^y-5A^S^, Oh'er L^boraloritis Incorporjtcd lllfi Sumh Main Sired Bl-icksburg, Virginia 2^060 Phone (3-10) 552-5S-18 FAX {540) S52.1715 Company 79 Sanitized. Does not contain TSCA CBI t>\-\q ^ncA'-y5^_ Carr^Q.<s5.;i/-i % 't^-a- '^AevM^o/A Con-V^I Fisher's Exact Test NUMBER OP IDENTIFICATION CONTROL 1 100 mg/L TOTAL ALIVE DEAD 10 0 7 3 17 3 TOTAL ANIMALS 10 10 20 Critical Fisher's value (10,10,10) (alpha=0 05) is 6 0 b value is 7 Since b is greater than 6 0 there is no significant difference between CONTROL and TREATMENT at the 0 OS level Fisher's Exact Test NUMBER OF IDENTIFICATION ALIVE DEAD TOTAL ANIMALS CONTROL 10 0 10 500 mg/L 4 o 10 TOTAL 14 6 '* 20 Critical Fisher's value (10,10,10) (alpha=0 05) is 6 0 b value is 4 Since b is less than or equal to 6 0 there is a significant difference between CONTROL and TR3ATMENT at the 0.05 level. ^^^^y^r T^^y^ $^'S^ -^^'L^A^V- Olver Laboratories Incorporated 1116 South Main Street Bl^cksburi;. Virginia 24060 M-,_- ic <m c^ cs <Q civ .elm ;^ i^is Company bU Sanitized. Does not contain TSCA CBl t)c<V^ r^no\sS>;i- Ca/no&tN'Son ^ro '^a- S-V^rn\(yA C^n-'n^l ----------------^-------------,-- GROUP Summary or Fisher's Exact Tests IDENTIFICATION NUMBER EXPOSED NUMBER DEAD CONTROL 10 5 mg/L 10 10 (ng/L 10 25 mg/L 10 50 mg/L 10 100 mg/L 10 500 mg/L 10 SIG 0.05 ^/. Olver Laboratories Incorporated ^y^ 1116 South M.iin Streel ^ Bl-ickiburg, Virginia 24060 81 Phone (S-<0) 552-554S FAX (54U) 552-1715 Company SanHized. Does not contain TSCA CB< Qo>V<\ kna^b^- O^P'-fi'So^-i "^ '\V^t- ty^nAcMiA C^-Vrol B. CHI-SQUARE TEST FOR NORMALITY OF DATA Title File. Dupon _Toxicity-C dubia Chronic-2/20/01 Transform- Chi-Square Test for Normality NO TRANSFORMATION INTERVAL EXPECTED OBSERVED <-1 5 4 0200 4 Actual and Expected Frequencies -1 5 to <;-0 S -0 5 to 0 5 14 5200 14 22.9200 27 s.0.5 Co 1 5 14 5200 12 . >1 5 4. 0200 3 Chi-Square = 1 ,412 ^Critical Chi-Square = 13 277 = 9 488 (p-value = O B370) (alpha = 0.01 , df = 4) (alpha = 0.05 , df 4) Data PASS normality test (alpha = 0 01) Continue analysis C. TEST FOR HOMOGENEITY OF VARIANCE -Title File DupontyJUJ T^xicity-C dubia Chronic-2/20/01 ^^^HHI Transform NO TRANSFORMATION Bartletc's Test for Homogeneity of Variance Calculated 31 statistic = 11 0373 (p-value = 0 0506) Data PASS Bl homogeneity test at 0 01 level Continue analysis Critical B = 15 0863 (alpha = 0 01, df = 5) = 11 0705 (alpha = 0 05, df = 5) ^^^^^$ ^^-^y? __ .i,SS l^^NV OIver Ljboralonei Incorporated 1116 South Main Street Blacksburg, Virginia 2-4060 pllo"e (s4()) 352-554S FAX (5.10) 552-171S Company 82 Sanitized. Does not contain TSCA CBI J^-V^ ^^cils^^- C^rAfi^<\'^on -^-D 'IV^a. ') D. SUMMARY STATISTICS ON REPRODUCTION DATA Title File ^S. Cu' ponti----^ To-Kicity~ -C ^--------A dubia Chronic-2/20/01 Transform 5'^oAa^d Cor^o^ -MC TRANSFORMATION Summary Statistics on Data TABLE 1 of : GRP IDENTIFICATION H 1 Control 10 2 5 mg/L 10 3 10 mg/L 10 4 25 mg/L 10 5 50 mg/L 10 6 100 mg/L 10 MIN 14 0000 15 0000 13.0000 9 0000 10 0000 8.0000 MA;( 22 0000 19.0000 21.0000 19 0000 23 0000 22.0000 MEAN 13 0000 16 6000 17.1000 13 9000 15-2000 17.3000 Title Fll'e Duponn------H. Toxicity-C dubia Chronic-2/20/01 fjpBffl Transform HO TRANSFORMATION Summary Statistics on Data TABLE 2 of 2 GRP IDENTIFICATION 1 Control 2 5 mg/L 3 10 mg/L 4, 25 mg/L 5 50 mg/L 6 100 mg/L VARIANCE 7 7778 1 3778 6 7667 10 3222 11 9556 14 8444 SD 2 7889 1.1738 2.6013 3.2128 3.4577 3.8528 SEM 0 8819 0.3712 0.8226 1.0160 1.0934 1.2184 C V t 15 4937 7 Q710 15.2122 23 1138 22 7479 22 4003 ^ (^^ v\v</y ""^^i^ O"'lve~ r L--jbo--ra-t--on-a- I--nc-o-rrp--ora-t--ed 1116 South MMJjmin Street <%5?\S\ Blacksburg, Virginia 24060 3 'W^W Pt1"" (S-10) 552-5548 FAX (540) " 552-1715 Company Sanitized. Does not contain TSCA CB1 r t^a.Yt^ (\nc.\>j<<>Cs- CQ n\p^<\'^on ~rc HTSZ. -S-Ta^a^ E. ANALYSIS OF VARIANCE OF REPRODUCTION DATA Title File r(^ Ducontt-------- Toxicitv* -C ^1^4^--r ----f--k )] dubia Chronic-2/20/01 Transform NO TRANSFORMATION AHOVA Table SOURCE DF SS MS F Between 5 113 9333 22 7867 2 5775 Wir.nm (Error) 54 477 4000 B- 8407 Total 59 531 3333 (p-value = 0 0366) Critical F 1 = 3 3769 = 2 3861 (alpha == 0 01, d = 5,54) (alpha == 0 05, df = 5,54) Since F > CriC-cal F RSJECT Ho All equal (alpha = 0 05) F, DUNNETT'S TEST Title. ^sa-3 OupoinilBBI- Toxicicy-C dub-La Chronic-2/20/01 Transform. MO TRAHSFORMATION Dunnett's Test - TABLE 1 OF 2 Ho Control<Treatment GROUP IDENTIFICATION TEAiNSFORMED MEAN MEAN CALCULATED IN ORIGINAL UNITS T STAT SIG 0 05 1 Control 18 .0000 18 0000 2 5 mg/L 15 6000 16 6000 1 0529 3 10 mg/L 1'7 1000 17.1000 0 6768 4 25 mg/L 13 9000 13 9000 3 0834 * 5 0 mg/L 15 2000 15 2000 2 1057 6 100 mg/L 17 2000 17 2000 0 5016 Dunnecc critical value = 2 3100 (1 Tailed, alpha = 0 05 , df [used] = 5,40) (Actual^d = 5,54) ^^<^-^^^ ^/^^\^\<f 'yy'^W Olver Laboratories Incorporated I'll.6 South Main Sireet Black5burg, Virgimj 24060 pli"e (540) 55^-55-IS F-^X (5^0) 552-1715 84 Company Sanitized. Does not contain TSCA CB1 t>ojrQ ^DcA^S^-i _ Cgjrtvpcn-'.'So A -^o '^J- S^^O-CCi ^0 '-T^ \ Title File Dupont[riR Tsxicity-C. ^'B------ft '^^^t DunneCt' s Test - TAELS dubia Chrcnic-2/20/01 Transfo'-m- NO TRANSFORMATION 2 OF 2 Ho-Control<Treatment GROUP IDENTIFICATION Md OF REPS M;N SIG DIFF (IN ORIG UNITS) s OF CONTROL DIFFSRENC2 FROM CONTROL 1 Conrrol 10 "t 5 mg/L 10 3 0716 17 1 1 4000 3 10 mg/L 10 3.0716 17 1 0 9000 I 25 mg/L 10 3.0715 17 1 4 1000 5 50 ing/L 10 3 0716 17.1 2 8000 6 100 mg/'L 10 3 0716 17.1 0 8000 G. INHIBITION CONCENTRATION ESTIMATE Titl^. File GRP Dupont -Sonyl Toxicity-C zoncenc I3ENTIFICA.TION dubia Chronic-2/20/01 Transform. NO TRANSFORMATION MEAM SMOOTHED MEAN CONCENTRATION 1 Control 18 0000 18.0000 0 0000 2 5 mg/L 16 6000 16.8500 5.0000 3 10 nig/I-, 17 1000 16.8500 10.0000 4 25 mg/L 13.9000 15.S250 25 0000 5 50 mg/L, 15.2000 15 9250 50 0000 6 100 mg/L 17 2000 15 9250 100 0000 7 500 mg/L 17 4000 15-9250 500 0000 ICp es cimata with p = 25 is > 500 0000 ^ ^,,'%_ .^^k -^yXv^ OIvcr Ljborjtorics (nLorporaled 1.116 South Mjm Sirecl BIacksburg, Virginia Z4060 Phone (540) 552-35'IS FAX (540) 552-1715 85 Company Sanitized. Does not contain TSCA CBI DCJ-VG. r^neAu.SiS -- CarrvQg.n.snn V? Cc^-Vyofe uriV -ci^.^ p~)g-yoa.'^lJ\(''^0':'-"') A. FISHER'S EXACT TEST FOR THE DETERMINATION OF SIGNIFICANT DIFFERENCES IN TEST ORGANISM MORTALITY Fisher's Exact Test NUMBER OF IDENTIFICATION DEAD CONTROL w/MeOH 5 mg/L ALIVE 9 10 TOTAL. ANIMALS 10 10 TOTAL 19 20 Critical Fisher's value (10,10,1) (alpha=0.05) is negative no significant difference b value is 0 Fisher's Exact Test NUMBER OF IDENTIFICATION ALIVE CONTROL w/MeOH 10 cng/L DEAD TOTAL ANIMALS 10 10 TOTAL 18 20 Critical Fisher's value (10,10,9) (alpha=0.05) is 4 0 . . b value .-s 9 Since b is greater than 4 0 there is no signj-ficaa.t difference between CONTROL and TREATMENT at the 0.05 level f^^^^i ^^^ OIvcr Laboratories Incurporjted 1116 South Main Street BpIhac"k:snb^urog6, )sVisrg^ini^a -24F06T0 x^ .52-1^ ^^^w^. Does not contain TSCA CBI T^oA^- I^Q'-V^SvS- Car!\pc.<^^^, -Vo Cor,-Vx;\<> L.^.o SoJ^^L c-A^o Yng-'H-^ Fisher's Exact Test NUMBER OF IDENTIFICATION CONTROL w/MeOH 25 mg/L TOTAL ALIVE DEAD 99 11 16 2 TOTAL AMIMALS 10 10 ' 20 Critical Fisher's val-e (10,10,9) ialpha=0 05) is 4.0 b value is 9 Since b is greater tnan 4 0 there is no significant difference between CONTROL and TREATMENT at the 0 05 level Fisher's Exact Test NUMBER OF IDENTIFICATION ALIVE DEAD TOTAL ANIMALS CONTROL w/MeOH 9 1 10 50 mg/L .7 3 10 , 1 5 - 1 TOTAL 20 Critical Fisher's value (10,10,9) (alpha=0 05) is 4 0. b value s 7 Since b is greater than 4 0 there is no significant difference between CONTROL and TREATMENT at the 0 05 level ^i^i^-^^^ ^^-y% ^W^iS Olver l-iboratories Incorporated 1116 Soutli Main Street S13'21"''"1^, Virginia 24060 Phone (540) 552-55-4S FAX 1540) 552-1715 ComplSyanded. Does not contain TSCAOBi -- - - - -- ^ -- T)o-Vg- ^noA^'s- Comptx-nso ri V<= Carf^^ L^'-T^ 'Sao^- o-cHjjA mi.!r^Q.^o\ Fisher's Exact Test NUMBER OF IDENTIFICATION ALIVE DEAD CONTROL w/MeOH 9 1 7 3 100 ma/L ; TOTAL 16 4 TOTAL ANIMALS 10 10 20 Cnc-cal Fisher's vslue (10,10,9) (alpha=0 05) is 4 0 b value is 7. Since b is greacer than 4 0 there is no significant difference between CONTROL and TREATMENT at the 0.05 level. Fisher's Exact Test NUMBER OF IDENTIFICATION ALIVE DEAD TOTAL ANIMALS CONTROL w/MeOH 9 1 10 4 6 500 mg/L 10 TOTAL 13 7 20 Critical Fisher's value (10,10,9) (alpha=0 05) is 4.0. b value is 4. Since b is less than or ecrual to 4 0 there is a significant difference between CONTROL and TREATMENT at the 0.05 level. XSS-5i%^ ^^^^^r ^A, ^'y^' Olver Laboratories Incorporaled 1116 South Main Siretl BPhlaocnletib(5u4r0g), 5V5i2r-g5i5n-i1a8 2F^A06X0 (540) 552-1715 88 Company Sanitized. Does not contain TSCA CB1 'Do.V^. ^-n^W^S-Corner (^0^1 Vo rc.^f:Vm\ ^"V^ S'oo-^L ci-cS^-gcil T^.g.-^\-gt..-^o\ GaOUP Suinmary of ?isher's Sx.a.ci Tests IDENT;FICAT;ON NT3MBER EXPOSED NtTMBER DEAD CONTROL w/MeOH 10 5 m9/L 10 10 ms/L 10 25 mg/L 10 50 mg/L 10 100 mg/L 10 500 mg/L 10 SIG 0 05 Olver Laboratories Incorporated 1116 SouUi Main SirEtt Blacksburg, Virginia 24060 Phone (5'40) 5S2-5548 FAX (5-10) 552-1715 g 9 Company Sanitized. Does not contain TSCA CBI ______OoA-0- P-i^o-l^SO _ Camoo.o'^,^ ^ro Cfln\r\ V^V^- 5oo ^<-H- O.^A'Stj me.-^nQ.oo^ B. CHI-SQUARE TEST FOR NORMALITY OF DATA Title File Dupor-t-a----^i Toxicity-C dunia Chronic-2/20/01 =cer Transform SO TPANSFORMATION Chi-Square Test for Normality INTERVAL EXPECTED OBSERVED <-1 5 4 0200 5 Actual and Expected Frequencies -1 5 to <-0.5 -0 5 to 0 S 14 5200 13 22 9200 25 >0 5 Co 1 5 14 5200 11 >1 5 4 0200 3 Chi-Square = 2 6361 , Critical Chi-Square = 13 277 9 488 (p-value = 0 6204) (alpha = 0 01 . df = 4) (alpha = 0 05 , df = 4) Data PASS normality test (alpha = 0 01) Continue analysis C. TEST FOR HOMOGENEITY OF VARIANCE Title ' , DupontMpBB(|Toxicity-C File seer dubia Chronic-2/20/01 Transform NO TRANSFORMATION Bartletc's Test fcr Homo9eneity of Variance Calculated Bl statistic = 20 1697 Data FAIL Bl homogeneity test at 0 01 level (p-value = 0 0012) Try another transformation Critical B = 15 0863 (alpha = 0 01, d = a) = 11 0705 (alpha = 0.05, df = 5) ^^^-'^y^ '^^^ %?\^S ^y"5%V Olver Laboratories Incorporated 1116 South Main Street s10'*"1""^. ''^e'"" ^i^o P110"6 t540! 532-55-48 FAX (540) 552-1715 9 0 Company Sanitized. Does not contain TSCA CBS t>3.-V&. \r].^-<ss- Q.mo&.^a^, -Vo Cs"n^\S ^wV 5ao -u.^- <s.AA(A rrut-rUa.no^ D. SUMMARY STATISTICS ON REPRODUCTION DATA Title File 3upont-flH^ Toxicity-C dubia Chronic-2/20/01 ^^~cer Transform NO TP^ANSFORMATION Summary Statisc-cs on Data TABLE 1 of 2 GRP IDENTIFICATION N 1 Control w/MeOH 10 2 5 mg/L 10 3 10 mg/L 10 4 25 mg/L 10 5 50 mg/L 10 6 100 mg/L 10 MIN 0 0000 15 0000 13 0000 9.0000 10 0000 8.0000 MAX 22 0000 19 0000 21 0000 19 0000 23 0000 22.0000 MEAN 14 3000 16.SOQO 17 1000 13 9000 15 2000 17 2000 Title. File Duponf^^ Toxicity-C. dubia Cbronic-2/20/01 ^ '?cer Transform: NO TP-ANS FORMATION Summary Statistics on Data TABLE 2 of 2 GRP IDENTIFICATION 1 Control w/MeOH 2 5 mg/L 3 10 mg/L 4 . 25 mg/L 5' 50 mg/L 6 100 mg/L VARIANCE 36 S778 1 3778 6 7667 10.3222 11 955S 14 8444 3D 6 0562 1 1738 . 2 6013 3.212B 3.4577 3 8528 SEM 1 9151 0.3712 0.8226 1 0160 1.0934 1 2184 C V % 42.3512 7 0710 15.2122 23-1138 22.7479 22.4003 & -- f^ A< ^^W Olver L^boraioris.s Incorpor.iled 1116 South Main Street Blacksburg, Viri.'mi.i ZW6Q ptlolle 1540) "^-"<s FAX {540) 552-1715 91 Company Sanitized. Does not contain TSCA CBl DoVa- ^noA^s^ Ca^p<w5,&^ -Vo CoV>-As '^r^ Spa ^.L c^Ago myV^a/iQ' E . STEEL / S MANY-ONE RANK TES3T Title. File r \ tXiponig^ml Toxicity-C scsr dubia Chroni C-2/20/01 Transfonn N0 TRANSFORMATION Steel's Many-One Sank Test Ho Control<sTreac-nent GROUP IDENTIFICATION MEAM IN . ORIGINAL UNITS RANK SUM CRIT VALUE SIG DF 0 05 1 Control w/MeOH 14 3000 2 5 mg/L 16.6000 119 00 75 00 10.00 3 10- mg/L 17 1000 121 00 75 00 10 00 4 25 ing/L 13.9000 96 00 75 00 10 00 5 50 mg/L 15 2000 104 50 75. 00 10 .00 5 100 mg/L 17.2000 122 00 75 00 10 -00 Critical values are 1 nailed ( k = 5 ) A F . INHIB 1 TION CONCENTRATION ESTIMATE Title File E U P O n t1* B- B > B^^ Toxicny-C zcenc dubia Chroni C-2/20/01 Transform : N0 TRANS FORMAT I ON GRP IDENTIFICATION MEAM SMOOTHED MEAN CONCENTRATION . 1 Control w/MeOH 14.4000 16 0333 0 0000 2 5 mg/L 16.6000 IS 0333 5 0000 3 10 mg/L 17 1000 16 0333 10 0000 4 25 mg/L 13 9000 15.9250 25.0000 5, 50 rog/L 15.2000 15 9250 50.0000 0 100 mg/L 17.2000 15.9250 100 0000 7 500 mg/L 17 4000 15 9250 500.0000 1C? estim ate with p = 25 is > 500.0000 OIver Laburatonra [Dcorporjicd 1116 Soutli Main Street Blacksburg, Virginia 24060 Phone (540) 552-55-JS FAX (540) 552-1715 92 "Company Sanitized. Does not contain TSCA CBi APPENDIX 8 ACUTE REFERENCE TOXICITY TEST DATA Company Sanitized. Does not contain TSCA CB1 OLVER LABORATORIES OLVER LABORATORIES REFERENCE T DATA SUMM PAGE __ TESTING MONTH: _. J^nuAyU 2-OQl REFERENCE TOXICANT: 5o^i'u^ CLtilori'A.^ ACUTE TOXIC1TY TEST RESULTS ' >.. TEST DATEfS) TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER ' SOURCE AERATION Certodaphnia l/ic.- l/iilol Fathead Minnow l/H-'/l^lo/ < 7.4Viu^ EPA-MHW OLVER LABS Continuous 1 J^.y EPA-MHW OLVER LABS Continuous CHRONIC TOXIC1TY TEST RESUL ^ Cerlodaph TEST ORGANISM AGE DILUTION WATER SOURCE AERATION EPA-MH OIVER LA continuo TEST ACCEPTABILITY CONTROL SURVIVAL (&90%) loo'/, Ion'/. CONTROL SURVIVAL (s80%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (s60%) AVE NO YOUNG/SURVIVING FEMALES (&15) STATISTICAL RESULTS SURVIVAL IN 100% f~ LCso 95% C I METHOD o'/. o'/ 2.6 3/L S.3) S/L 2.Z - Z-B 3/L 4.^ -<..i a/L 5p(rmtL-i-htt<^>, P-fart: AVE WT/SURV CONTROL (&0 25 mg) SURVIVAL IN HIGHEST ( )CONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION? HOMOGENEOUS VARIANCE? NOEC METHOD NOEC SURVIVAL NOEC. REPRODUCTION/GROWTH COMMENTS, p^-fy^c^. -Voy,<a^ -^-\- y^.irt^ u^r-^ "^I'-HH^ t^ cXr<c-^-A '^njt.^ OLVER LABORATORIES REFERENCE DATA SUM PAGE TESTING MONTH: fc'Or^P.yn ZnQl ^J ACU7 E TOX1CITY TEST RESULTS REFERENCE TOXICANT: Spc^um C^Voq'Ae. CHRONIC TOX1CITY TEST RESU TEST DATEfSI TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION Cerladaphnia X/;u-/ifr?bl / ^M hones EPA-MHW OLVER LABS Conlinuous Fathead Minnow ^-z-//(-/ol i ^ -2. EPA-MHW OLVER LABS Continuous . TEST ORGANISM AGE DILUTION WATER SOURCE AERATION Ceriodap EPA-M OLVER continu TEST ACCEPTABILITY CONTROL SURVIVAL (^90%) i oo7. loo'/ CONTROL SURVIVAL (&80%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (s60%) AVE. NO. YOUNG/SURVIVING FEMALES (215) STATISTICAL RESULTS SURVIVAL IN 100% C Hi'jh*>+ llonC..) LCso f 95% C I METHOD AVE WT/SURV CONTROL (z0 25 mg) d/. O'/. 1.\ 9/L 11.1 9/L SURVIVAL IN HIGHEST ( )CONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE /. 8 - 2.? 3/i. ;0.2.- /2.0^/ Pyx^;+ ^ipcQfm-.r~y<.^V<- NORMAL DISTRIBUTION? HOMOGENEOUS VARIANCE? NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH COMMENTS: R<.-ferc-nci -\^^\ u-H 4<i.-t-Y<^U i-K '^<^<.L^\^^ -H^_g.K^<--H d, ^-ck^^ e-s OLVER LABORATORIES REFERENCE DATA SUM PAGE TESTING MONTH: ^CMCJ^ 7.00 I REFERENCE TOXICANT: Soch'u^ CV^t.ri^'L ACUTE TOXIC1TY TEST RESULTS TESTDATEfSI Cerlodaphnia 3)ib-^lp,lol Fathead Minnow 3]zi- Zj^ilol TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION ^ Z.4 VioufS EPA-MHW OLVER LABS ^ Continuous 2 d-^S EPA-MHW OLVER LABS Continuous TEST ACCEPTABILITY CONTROL SURVIVAL (i90%) I OC.7. I oo7. STATISTICAL RESULTS SURVIVAL IN 100% (."'.Sh-t-t- C.WC-) LCso 95% C I METHOD o7, !.(.3/L l-^ -I. 8 3;L Pxb:4- o7. I). 4 3/L lo.fc - IZ.z 3}i CHRONIC TOXIC1TY TEST RESU Cerlodap TEST ORGANISM AGE DILUTION WATER SOURCE AERATION i CONTROL SURVIVAL (s80%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (i60%) AVE NO, YOUNG/SURVIVING FEMALES (i 15) AVE.WT/SURV CONTROL {i0 25 mg) SURVIVAL IN HIGHEST ( )CONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION'? HOMOGENEOUS VARIANCE? NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH EPA-M OLVER L continu COMMENTS: -^L isLQ- I o-APFi-200 1 TITLE: Test Date 11/20/2000 01/19/2001 01/19/2001 01/22/2001 01/24/2001 02/14/2001 03/16/2001 Ceriodaphnia dubia Acute Test Reference Toxicant Sodium Chloride Test Name Mean Mean+2SD Mean-2SD LC50 1.73 2.48 2.48 1.68 1.53 2.14 1.55 2.11 2.23 2.09 1.98 2.01 1 94 3.17 310 2.99 2.90 2.84 2.78 1.04 1.36 1.20 1.06 1.17 1 10 OLVER LABORATORIES. INC 97 'Company "SanWzed. Does not contain TSCA CBt lb-ADr-2001 Ceriodaphnia dubia Acute Control Chart Sodium Chloride Reference Toxicant 8 7 6 ; STs IS Z. ^ 0 m 03 .2. r^" 1 ----------------- ^^------------^^-------------------------------- - ^^-------^-^^ ^----------- .. ^^---, -- i t 0 i i i l ' 11/20/2000 01/19/2001 01/24/2001 03/16/2001 01/19/2001 01/22/2001 02/14/2001 LC50 ------ Mean ------ Mean+2SD ------ Mean-2SD OLVER LABORATORIES, INC 98 Company Sanitized. Does not contain TSCACBa I 6-APR-ZOO I TITLE: Test Date 11/20/2000 12/18/2000 12/19/2000 12/19/2000 12/20/2000 01/17/2001 02/14/2001 03/29/2001 FATHEAD MINNOW ACUTE TEST Reference Toxicant. Sodium Chloride (o/U Test Name Mean Mean+2SD Mean-2SD LC50 11.7 9.9 108 12 11.4 11 4 11.1 11.4 10.80 10.80 11.10 11.16 11.20 11.19 11.21 13.35 .12.60 13.00 12.82 12.70 12.66 12.49 8.25 9.00 9.20 9.50 9.70 9.81 9.93 99 "Company Sanitized. Does not contain TSCA CBI OI-VER LABORATORIES. INC lo-Apr-2001 FATHEAD MINNOW Acute Control Chart Sodium Chloride Reference Toxicant 11/20/2000 12/18/2000 12/19/2000 12/19/2000 12/20/2000 01/17/2001 02/14/2001 03/29/2001 LC50 Mean Mean+2SD Mean-2SD OLVER LABORATOR.IES, INC 100 Company Sanitized. Does not contain TSCA CB1 APPENDIX 9 CHRONIC REFERENCE TOXICITY TEST DATA 1 01 Company Sanitized. Does not contain TSCA CB1 OLVER LABORATORIES OLVER LABORATORIES REFERENCE DATA SUM PAGE TESTING MONTH: Jc\n^^s Z.OO I REFERENCE TOXICANT: &odi'um_ C^T^C TEST DATE(S) ACUTE TOXIC1TY TEST RESULTS Cerlodaphnia Fathead Minnow TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION EPA-MHW OLVER LABS .Continuous- EPA-MHW OLVER LABS Continuous CHRONIC TOXIC1TY TEST RES Cerlod l/?-l/ TEST ORGANISM AGE DILUTION WATER SOURCE AERATION IB-"^ EPAOLVER conti TEST ACCEPTABILITY CONTROL SURVIVAL (2 90%) CONTROL SURVIVAL (^80%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (^60%) iUD 10 / oo AVE NO YOUNG/SURVIVING FEMALES (t15) STATISTICAL RESULTS SURVIVAL IN 100% LCso ,- 95% C I METHOD AVE WT/SURV CONTROL (&0 25 mg) SURVIVAL IN HIGHEST ( 4.o.3A.)CONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION? HOMOGENEO'US VARIANCE? NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH (^^^^^^^^M P>g-<e-j-<.n^t- -\oy.>'c;-V>j -\-<..-4 r^s^VU W.TC i^rtVi'n -Tn<. ^xy<.i^<A r^nj<'. ^. O )0 1 Ye J S+t 2.0 O.Z LABORATORIES REFERENCE DATA SUM PAGE TESTING MONTH; f<.V>y^o,,rM 2pt? ^ REFERENCE TOXICANT: ^oi;u^ &V.\<.nlt. A(;m1: TOXICI [Y TEST RESULTS If!,; '.I-'''1'-:'! Cerlodaphnia Fathead Minnow IbSl CUNUIIIUNy TEST ORGANISM AGE DILUTION WATER SOURCE AERATION EPA-MHW OLVER LABS Continuous EPA-MHW OLVER LABS Continuous CHRONIC TOXICITY TEST RES ' Certoda 2./-,^/ TEST ORGANISM AGE DILUTION WATER SOURCE AERATION 1-8 EPA-M OLVER conlin TEST ACCEPTABILITY CONTROL SURVIVAL (a90%) STATISTICAL RESULTS SURVIVAL IN 100% LCso . 95% C 1 METHOD CONTROL SURVIVAL (iBO%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (^60%) AVE NO YOUNG/SURVIVING FEMALES (t 15) AVE WT/SURV CONTROL {t0 25 mg) SURVIVAL IN HIGHEST (3- /^JCONC NUMBER OF REPLICATES NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION? HOMOGENEOUS VARIANCE? NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH loo (0 loo 2-7 O/ 1 ( Y y Dnnn 1.5 o.iq COMMENTS- Rt-(<yLn<-- -+<>Xi'<-vVu -^r^- Y^l-oo\"lt--*> ^-it-r^. >-Mi''H^i/-> +^A. e-'^.WsA-si-A t-c^E.s. OLVER LABORATORIES REFERENCE DATA SUM PAGE TESTING MONTH: Mo^L'n 2pt> I REFERENCE TOXICANT: ^oj.'ur-A C^i-^-.'^^ ACU I E T UX1CI rY TEST RESULTS il^l il.MI ;;;. Cerlodaphnia Fathead Minnow TEST CONDITIONS TEST ORGANISM AGE DILUTION WATER SOURCE AERATION EPA-MHW OLVER LABS Continuous EPA-MHW OLVER LABS Continuous CHRONIC TOXICITY TEST RES Cerloda 3/15-3/ TEST ORGANISM AGE DILUTION WATER SOURCE AERATION 1-6 EPA-M OLVER contin TEST ACCEPTABILITY CONTROL SURVIVAL (290%) STATISTICAL RESULTS SURVIVAL IN 100% LC 95% C 1 METHOD COMMENTS: Ri4v^"<-*- -y'x^'\^ -\^\- i^^^ i^<.^. CONTROL SURVIVAL (a80%) NO OF CONTROL FEMALES % CONTROL FEMALES WITH 3 BROODS (a60%) AVE NO YOUNG/SURVIVING FEMALES (215) AVE WT /SURV CONTROL (a0 25 mg) voi'-V^;^ SURVIVAL IN HIGHEST (^.o /^ )CONC NUMBER OF REPLICATES L NUMBER OF ORGANISM/REPLICATE NORMAL DISTRIBUTION? HOMOGENEOUS VARIANCE? NOEC METHOD NOEC SURVIVAL NOEC REPRODUCTION/GROWTH ----X^-. R^Au^/g-r^---------- -\V^. e-s-^^\'sV,<A Cof't-^-N \^I'VA ; oo 10 -70 'S. O'/. 10 ) ^ MO S+K 2- o.s o.Z I e-ApR-2001 TITLE: Test Date 01/04/2001 01/22/2001 01/31/2001 02/05/2001 02/12/2001 03/14/2001 Test Name NOEC 2 2 2 2 1.25 2.5 Mode Mode/0.5 Mode'0.5 2.00 2.00 2.00 2.00 2.00 4.00 4.00 4.00 4.00 4.00 1.00 1.00 1.00 1.00 1.00 105 OLVER LAEOP.ATORIES, INC Company Santtlzed. Does not contain TSCA CBI lo-Apr-2001 FATHEAD MINNOW CHRONIC SURVIVAL CONTROL CHART 8 7 a) -a ,6; i^ 0 55 E 3 '^4 03 ! '5)3 0 UJ -02 2 1 . --------------E--------------s--------------a------------------y------ ^^ 0 i 01/04/2001 01/22/2001 01/31/2001 02/05/2001 02/12/2001 03/14/2001 -m- NOEC ------ Mode ------ Mode/0.5 ------ Mode*0.5 Olver Laboratories, Inc 1 06 ^^ ^A -^mpany San^d. Does no, CB( 1 e-ApR-zoo i TITLE: FATHEAD MINNOW CHRONIC-GROWTH _________Reference Toxicant: Sodium Chloride (g/L) Test Date Test Name Mode Mode/O.S Mode'0.5 NO EC 01/04/2001 01/22/2001 01/31/2001 02/05/2001 2 2 2.00 2 200 5 2.00 4.00 4.00 4.00 1 00 1 00 1 00 02/12/2001 1.25 2.00 4.00 1.00 03/14/2001 1.25 2.00 4.00 1.00 107 'Company Sanitized. Does not conlain TSCA CBI OLVE.R LABORATORIES, INC 16-Apr-2001 FATHEAD MINNOW CHRONIC GROWTH CONTROL CHART (u -a c b i_ _o 5s E I4 w '5)3 o uj -02,, -B- z 01/04/2001 01/22/2001 NOEC 01/31/2001 Mode 02/05/2001 02/12/2001 0.3,,/,1,4,,/2,0.,0,,1 Mode/0.5 Mode*0.5 Olver Laboratories, Inc. 108 Company Sanitized. Does not contain TSCA CB1 I 6-APR-200 I TITLE: Test Date 12/01/2000 12/27/2000 01/03/2001 02/07/2001 03/15/2001 CER10DAPHN1A CHRONIC TEST_SURVIVAL Reference Toxicant: Sodium Chloride (g/L) Test Name Mode Mode/0 5 Mode'0 5 NOEC 2 1 2.00 2 2.00 1.5 2.00 2 2.00 4.00 4.00 4.00 4.00 1.00 1.00 1.00 1.00 109 Ol-VE:R LABORATORIES, INC 'Company "Sanitized. Does not contain TSCA CBI 16-Apr-2001 CERIODAPHNIA CHRONIC SURVIVAL CONTROL CHART 4 35 1 3 i o j 25 E 3 2 ^ m 0) ^ '0)1 5 ^x ^^^ 0 UJ 0 i 1 \^ Z "^ /Bi--^ 05 0 1 2 / 0 I1/2000 -- NO E I' 12/27/2000 C ------ M I01/03/2001 ode ---- ]' 02/07/2001 -- Mode/0.5 ---- '03/15 -- Mode*0.5 / 2 0 0 1 - Olver Laboratories, Inc no Company Sanitized. Does not contain TSCA CB1 I S-APR-200 I TITLE: CER10DAPHN1A CHRONIC TEST-Reproduction _________Reference Toxicant: Sodium Chloride (g/L-l_ Test Date Test Name Mode Mode/0.5 Mode'0.5 Nnpf 12/01/2000 025 12/27/2000 0.25 0.25 0.50 0.13 01/03/2001 02/07/2001 0.25 0.19 0.25 0.25 0.50 0.50 0.13 0.13 03/15/2001 0.5 0.25 0.50 0-13 111 OLVER LABORATORIES INC Comoany Sanitized. Docs nnt contain T8CA CB1 -lG-Apr-2001 CERIODAPHN1A CHRONIC REPRODUCTION CONTROL CHART 2 1.5 as ~a 1 1 u E 1 O5 CO ^ 'm -0 - 0 iu 0 z -0.5 i ----------HE -1 12/01/2000 ' 12/27/2000 B -----------------^- ^ ^ ^ Q1/03/2001 ' 02^07/2001 03/15/2001 -- NOEC ------ Mode ------ Mode/0.5 ------ Mode*0.5 - Olver Laboratories, Inc. 112 Company Sanitized. Does not contain TSCA CBl APPENDIX 10 DILUTION WATER QUALITY ANALYSES DATA 1i 1i 3 Company -> Sanitized. Does not contain TSCA CBI /^>Ji WCTD' Consulting Engineers Environmental Laboratories [^roRATORlES 1116 South Main Street Blacksburg, Virginia 24060 Client Sheet No.: Date: Client: Source: Shipping Information: 0004151 May 8,2000 Olver Laboratories Incorporated Job No.: Date Received: 30001 4/12/00 Blacksburg, Virginia Delivered to Olver Laboratories Incorporated by Olver Personnel Sample No.: Date Collected: Time Collected: Description: 123511 4/12/00 9 45 a m and 3:00 p m. EPA Moderately Hard Dilution Water pH (Laboratory) (EPA 150 1) Alkalinity as CaCOa (EPA 3101) Conductivity (EPA 120.1) Tofal Cyanide (EPA 335 2) Ammonia as N (SM 4500 NH^.F) Nitrate as N (ERA 352.1) Nitnteas N (EPA 354 1) Hardness as CaCC^EPA 200 7) Total Phenols (EPA 420 1) MDL = Minimum Detection Limit N/A = Not Applicable ND = Not Detectable Result 809 60 317 NO ND ND ND 100 ND mg/L ^mhos/cm mg/L MIU. N/A 10 1 0 0005 0.1 0 1 001 1. 0005 mg/L pmhos/cm mg/L mg/L mg/L mg/L mg/L mg/L Date/Time Anal yzed 4/12/00, 4/20/00, 5/02/00, 4/20/00, 4/17/00. 4/14/00, 4/14/00. 4/18/00, 4/24/00, 7 45 p m. 11 00a.m 10 45am 8 15am 6 00 p m. 3 00 p m. 1 30 p m. 5 00pm 1 10pm. SAH ACC ACC DF SAH BCW SAH HT DF All tests according to Standard Methods for the Examination of Water and Wastewater. 18th Edition, Test Methods for Evaluating Solid Waste (Physical/Chemical). 2r>d and 3rd Editions, and Methods for the Chemical Analysis of Water and Wastes. EPA By ^BnAt R^it/cfhcey^^_(}_________ Laboratory Manager Unless otherwise indicated. Ihis report sets forth Ihe results of our analysis of samples delivered lo our laboratory and shall nol be construed to be a representation by Oluer Laboratones Incorporated as to the source or method of procuring such samples All reports are submitted as the confidential property of clients and authonzalion (or publication of any statements contained in our reports is reserved pending our wntten approval Blacksburg. VA (540) 552-6974 1 1 4 Page 1 of 6 N \OATA\LAB\3000nCLNTSH"nOOO-n51 Company Sanitized. Does not contain TSCA CBl OLVER LABORATORIES Client Sheet No Client Date 0004151 Olver Laboratories Incorporated May 8,2000 Sample No.: Date Collected: Time Collected: Description: 123511 4/12/00 9 45 a.m and 3:00 p m. EPA Moderately Hard Dilution Water Analysis Total Aluminum (EPA 202 2) Dissolved Aluminum (EPA 202 2) Total Arsenic (EPA 200 9) Dissolved Arsenic (EPA 200 9) Total Cadmium (EPA 200 9) Dissolved Cadmium (EPA 200 9) Total Chromium III (EPA 200 9-SM 3500 Cr.D) Dissolved Chromium 111(EPA 200.9-SM 3500 Cr.D) Total Hexavalent Chromium (SM 3500 Cr.D) Dissolved Hexavalent Chromium (SM 3500 Cr D) Total Cobalt (EPA 200 9) Dissolved Cobalt (EPA 200 9) Total Copper (EPA 200 9) Dissolved Copper (EPA 200 9) Total Iron (EPA 236 2) Dissolved Iron (EPA 236 2) Total Lead (EPA 200 9) Dissolved Lead (EPA 200.9) Total Mercury (EPA 245 1) Dissolved Mercury (EPA 245.1) Total Nickel (EPA 200 9) Dissolved Nickel (EPA 200.9) Total Selenium (EPA 200 9) Dissolved Selenium (EPA 200 9) Total Sliver (EPA 200.9) Dissolved Silver (EPA 200.9) Total Zinc (EPA 200 7) Dissolved Zinc-(EPA 200 7) Pnonty Pollutant Organics- Acid Compounds (EPA 625): 2-Cntoruphenol 2,4-Dichlorophenol 2,4-Dimethylphenol 4,6-Dinitro-o-cresoI 2,4-Dinitrophenol 2-Nltrophenol 4-Nitrophenol p-Chloro-m-cresol Pentachlorophenol Phenol 2,4,6-Tnchlorophenol Result 0 005 ND ND ND ND ND ND ND NO ND ND ND ND ND 0 006 0 006 NO ND ND ND ND ND ND ND ND ND ND ND mg/L mg/L mg/L ND ND ND ND ND ND ND ND ND ND ND ME1. 0003 0003 0001 0001 00001 00001 0001 0001 0005 0005 0001 0001 0001 0001 0001 0001 0001 0001 00002 00002 0001 0001 0002 0002 00002 00002 0005 0005 mo/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L 10 Aig/1- 10 figiL 10 Aig/L 10 ^g/L 50 ^g/L 10 /tg/L 50 ^9/L 20 A'g/L 50 ^g/L 10 A'9/L 10 ."g/L Date/Time Analyzed 5/02/00, 12.00 pm 5/02/00, 1200pm 4/25/00. 2:00 pm 4/25/00; 2-00 p m 4/20/00; 9.30 a m 4/20/00; 9 30 a m 4/20/00, 4 00 p m 4/20/00, 4 00 p m 4/13/00, t-3(Tpm 4/13/00,1 30pm 4/26/00, 3 00 p m 4/26/00, 3 00 p m 4/18/00,9 00pm 4/16/00:9 00pm 5/03/00:12 00 p m 5/03/00; 12 00 p m 5/01/00; 2 00 p m 5/01/00; 2 00pm 4/21/00; 9 00 a m 4/21/00; 9 00pm 4/16/00:7 30am 4/16/00; 7 30 am 4/17/00; 11.45 am 4/17/00. 11 45am 4/16/00; 5 00pm. 4/18/00; 5 00pm 4/22/00.11.-30 am 4/22/00; 11.30 am 4/26/00:7 56 am * Analyst HT HT HT HT BN BN HT HT SW SAH HT HT HT/BN HT/BN HT HT HT HT BN BN BN BN BN BN HT HT BN BN J5L Page 2 of 6 N \DATA\LAB\30001\CLNTSH'noOO't 151 115 Company Sanitized. Does not contain TSCA CBI OLVER LABORATORIES Client Sheet No Client Date 0004151 Olver Laboratories Incorporated May 8,2000 Sample No- Date Collected: Time Collected: Description: 123511 4/12/00 9 45 a m and 3 00 p m EPA Moderately Hard Dilution Water Analysis Priority Pollutant Orgarncs (continued) 'Base/Neutral Compounds (EPA 625) Acenaphthene Acenaphthylene Anthracene Benzidine . Benzo(a)anthracene Benzo(a)pyrene 3,4-Benzofluoranthene Ben2o(g,h.i)perylene Benzo(k)fluoranthene Bis(2-chloroethoxy)nnethane Bis(2-chloroethyl)ether Bis(2-chloroisopropyl)ether Bis(2-ethylhexyl)phthalate 4-Bromophenyl Phenyl Ether Butyl Benzyl Phthalate 2-Chloronaphthalene 4-Chlorophenyl Phenyl Ether Chrysene , Dibenzo(a,h)anthracene 1.2-Dichloroben2ene 1.3-DichlQi-Dbenzene 1.4-Dichlorobenzene 3,3'-Dichlorobenzidine Diethyl Phthalate Dirnethyl Phthalate Di-n-Butyl Phthalate 2,4-Dinitrotoluene 2,6-Dinitrotoluene Di-n-Octyl Pnthalate 1,2-Diphenylhydrazine Fluoranthene Fluarene Hexachlorobenzene Resutt ND NO NO ND HD ND ND NO ND ND ND ND ND NO ND ND ND ND ND ND NO ND MD NO ND ND ND NO NO N D NQ ND ND 10 .ug/L 10 /^gA. 10 ^g/L 10 ug/L 10 ^igft. 10 ^g/L 10 ^g/L 10 ,ug/L 10 ^ig/L 10 ^ig/L 10 pg/L 10 ^ig/L 10 ^g/l_ 10 ^tgA. 10 A'9/L 10 ^g/L 10 ^g/L 10 ^g/L 10 ^g/L 10 ^g/L 10 ^g/L 10 ^g/L 20 ^ig/L 10 A'g/L 10 Atg/L 10 Atg/L 10 ^g/L 10 ,ug/L 10 ^gA. 10 ^igA10 ^g/L 10 ^g/L 10 MIL Date/Time 4/28/00, 7 56 a m Page 3 of 6 N \DATA\LAB\3000HCLNTSHT\0004151 11 fi Company Sanitized. Does not contain TSCA CB1 OLVER LABORATORIES Client Sheet No Client Date 0004151 Olver Laboratories Incorporated May 8,2000 Sample No.: Date Collected: Time Collected: Description: 123511 4/12/00 9-45 a m. and 3:00 p m. EPA Moderately Hard Dilution Water Analysis Result Base/Neutral Compounds (EPA 625) (continued). . Hexachlorobutadiene NO Hexachlorocyclopentadiene ND Hexachloroethane ND lndeno(1,2,3-cd)pyrene NO Isophorone, ND Naphthalene NO Nitrobenzene ND N-Nrtrbso-dimethyfamine ND N-Nitroso-di-n-propylamine ND N-Nilroso-diphenylamine ND Phenanthrene ND Pyrene ND 1,2,4-Tnchlorobenzene ND MDL 10 HQl\- 10 ^g/L 10 A'g/L 10 A'9/L 10 ^g/L 10 ^g/L 10 fig/L 10 f^gfL 10 ^g/L 10 At9/L 10 ^g/L 10 ^g/L 10 A'g/L Date/Time Analyzed 4/28/00, 7 56 a m Page 4 of 6 N \OATA\Lft,B\30001 U:LNTSHT\0004151 1 17 Company Sanitized. Does not contain TSCA CBI OLVER LABORATORIES Client Sheet No Client Date 0004151 Olver Laboratories Incorporated May B, 2000 Sample No.: Date Collected: Time Collected: Description: 123511 4/12/00 9-45 am and 3 00 p.m. EPA Moderately Hard Dilution Water Analysis Volatile Compounds (EPA 624)Acrolein Acrylonitnie Benzene Bromodichltiromethane Bromoform Bromo methane Carbon Tetrachlonde Chlorobenzene Chloroethane 2-ChlDroethyt Vinyl Ether Chloroform Chloromethane Dibromochlorometha ne 1.1-Dichloroetnane 1.2-Dichloroethane 1.1-Dichloroetnene trans-1,2-DichloToetlTene 1.2-DiGhloropropane ci^-1,3-OichlorQpropene trans-1,3-Dichloropropene Ethylbenzene Methylene Chlonde sym-Tetrachloroethane Tetrachloroethena Toluene 1.1.1-Tnchloroethane 1.1.2-Tnchloroethane Tnchloroethene TnchlorDfluorometha ne Vinyl Chlonde Result ND ND ND ND ND ND ND ND ND ND ND ND NO ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND MDL 10 ug/L 10 ng!L 5 ^g/L 5 ^g/L 5 iiqlL 10 ^g/L tQ ^g/L 5 ^g/L 10 /^g/L 5 ^g/L 5 ^g/L 10 Aig/L 5 ^g/L 10 ^g/L 5 ug/L 10 /^g/L 10 Aig/L 5 /^g/L 5 ^g/L 5 ^g/L 5 pg/L 5 HQIL 5 ^g/L 5 pg/L 5 /2g/L 10 ^g/L 5 ^s/L 10 ^g/L 10 ^gA10 pg/L Date/Time Analyzed 4/13/00. 4 06 p m >.nalv BP Page 5 of 6 N \DATA\LAB\30001\CI-NTSHTi0004151 1 18 Company Sanitized. Does not contain TSCACBI OLVER LABORATORIES Clieni Sheet No Client Date Sample No.: Date Collected: Time Collected: Description: Analysis Pesticides {EPA 608) . Aldnn alpha-BHC beta-BHC delta-BHC gamma-BHC Chlordane 4,4'-DDD' 4,4'-DDE 4.4'-DDT Dieldnn Endosulfan I Endosulfan II Endosulfan Sulfata Endnn Endnn Aldehyde Heptachlor Heptachlor Epoxide Toxaphene PCBs (EPA 608) PCS-1016 PCB-1221 PCB-1232 PCS-1242 PCB-1248 PCB-1254 PCB-1260 0004151 Olver Laboratories Incorporated May 8, 2000 123511 4/12/00 9:45 a.m and 3 00 p m. EPA Moderately Hard Dilution Water gesult ND ND ND ND NO ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND NO ND MDL 1 ngIL 1 ^g/L 1 A'g/L 1 ^g/L 1 ^g/L 1 ^ig/L 1 ^g/L 1 ^g/L 1 pg/L 1 figIL 1 WiL 1 ^g/L 1 pg/L 1 ^9/L 1 P8/L. 1 ^ig/L 1 ^g/L 1 pg/L 50 ^ig/L 50 pg/L 50 pg/L 50 ^ig/L 50 ^gA. 50 ^g/L 50 ^g/L Date/Time 4/21/00. 9 26 p m 4/21/00. 9 26 p m Page 6 of 6 1 19 N \DATA'>LAB\30001\CLNTSHT\00041 S 1 Company SgnHlzed. Ooes not contain TSCA CBI / , CHAIN OF CUSTODY RECORD ly^^ OLVER LABORATORIES Consulting Engineers and Environmental Laboratories { n6 soulh Main street E^ksburg, Virginia . 24060 Telephone: (5'10) 552-6974-FAX: (540) 552-1715 + Clicnl ihoutd complcle ill mfomialion leqilcilcd lojhe right and in llic unshaded ircu below. DO HOT WRITE IN THE SHADED AREAS * Sunplcs will be discarded or iclumcd In the client two wctlii tiler the inalyticil report hu been mailed,. un)c5i prior amncinc;ilj liavt been msdc '^a.wi.. J-liT.^l'IJI'l. . )!>// j ' Sample Location (ic, innucnI.ouirnllOOl.MWl, cic) , Dilc Colleclcd Time Cfllltllcd Client: Olver Laboratories. Incorporat Company Contact: Bioassnv Laboratory Address: Telephone:. .116 South Main Street. Black (540) 552-6974 Purchase Order No. M s ^, Sc.3 J Srnplc Conllncr Prcscrvallon .0 a Type Type n V3 0 k. 0 EU u t-- i3 0 =H: /?35f/-Moi ~ - a! ^'2. f5d5r ''3. ! -5fl ^^ I -Wl i,--^ -kJAZ. <-2 -K.OZ -- GPA Modcrtllcy Hcsd Walcr EPA Moderallcy Hard Wilcr EPA Modcrtllcy Haid Walcr Et'A Moderallcy Hajd Walcr CPA Moderallcy Hard Waler GPA Moderallcy llBrd Water EPA Moderallcy Htrd Water \ / - MC5 -- EPA Moderallcy Hard Walcr 4 i J M ^ ^ / .s.iin[ilf](;,l. (/Si)'ii.]i'.nc) i RerinqyiSiliedB^' (Signalftc) w N ^ - Re linquishedUy-(sgnalure) Relinquislieci By (Signature) %-L\00 '6->o0pr^ LEt Dlluiion Wller PE q^5cu^ mr DllBIIICT PE Wilo Ws Ljb Dfluiion PE 1 Orfn Wiitt ^^ 6Am Lib DIlulKKi W"ti GL Lil> Diluuon FE q^(^ ' Will ^^ Lib Dllullon Wuu GL 3p0 ^ ^n^rn \ / Lib DllllHOB PE W.IB Lib Dilutlfin PE Wlltr Due Tune w ^ Dilc Timt Received By: (Signature) F^Aj^jbc^^f Received By; (Sigrrature) Dale Time Received By (Signature) i ^NaOH 1I,SO,/ N,S.O, H,SO, UNO, HNO, None / rpHpuictllni ) w*ia^^ / Tolal Cyanid ( / Animonia 1 / 1 Tolal Phenol / To(alAI,Ai.C 1 Total Zn(lC / 1 Tolal Mercu ^ nfi)^5Wc8?l% 1 ll-.-ttafl^.jy,^,^ / 1 DIss Al, As Diss Zn(IC Special In Dale Tirnc 4.1ZtfB Dale T'linc Dale Time Temperatu ^il;ll|0^1 O <<,( 10 -i/n/p^u Coiliiiien ^.Ajt. TT-f CHAIN OF CUSTODY RECORD ^tSV 0 LVER LABORATORIES Consulting Engineers and Environmental Laboratories S^^^g. }} ^6 south Main street' Virginia 24060 Telephone: (540) 552-6974 FAX; (540) 552-1715 * Clicnl ihould complele ill mfDnmtion icqucslcd lo llie nghl uid in Ilic unshided >reai below DO NOT WRITE IN THE SHADED AREAS * Simple! will be discinled or relumed la Ihc dicnl Iwa weeks iflcr Ihc inilyliml report hus been nulled, unlcu pnor ananECfTlciils have been made Client;_______Qlver Laboratories. Incorporat Company Contact: Bioassav Laboratory Address: 1116 South Main Street. Plack Telephone:. (540^ 552-6974 Purchase Order No. Sample Location (i c , influent, ouinill 001, MW1, etc) s f^> 1 U a Dile Time U Sxnipic Container Prescrvalton 'S ^3 5 Collcclcd Collcclcil Type Type o rt ca Q. U U E 0 0 U % EPA Motlcrmlcy Hard Walcr EPAMoilcrBllcyllMdWalcr EPAModcralleyIlariIWaler %Z 00 a^ 1 ojm Lib Dtlulion Wllei GL Lib Dilullon OL ^- ^7- WlItT Lib Dilulian Wiiti GL None None HC1 / ( Dissolved M / L 1'tloiny Pol / Z. P,! ill; Ml Relinquished By. (Signature) Due Time Received By: (Signature) LLlLhs^L ^S Dale Time RecervedBy.(SignalOre) DaicTime I Received By: (Signature) '-] > l.ll l Dale Tune j 4,l&00|44^ . 1' I- " -fflAA. Dl;e rime | Mmlioil 0\\ Dale Time (.';;lillll'.'l APPENDIX 11 CHEMICAL ANALYSES OF BRINE SHRIMP AND YCT FOOD MIXTURES 122 Company Sanitized. Dees not contain TSCA CB1 OLVER LABORATORIES 1300 Blue Spruce Drive, Suite C Fort Collins, Colorado 80524 Toll Free: 800/331-5916 Tel: 970/484-5091 Fax:970/484-2514 SALMON STARTER #1 38-4763-91-50 PROCESS DATE: 07/26/00 GUARANTEED ANALYSIS Crude Protein, Min Crude Fat, Mm....... Crude Fiber, Max.... 55 0% .15.0% ...2 0% Toxic Metals afld Pesticide Analysis (Note: This analysis is on Lot 3/16/00) Performed by Energy Labs, Billings, MT Aluminum Arsenic Cadmium Chromium Cobalt Copper Iron Lead Mercury Nickel Silver Zinc Total Metak Detection Limit "S/g 5 5 1 a 5 3 3 5 1 5 5 5 Result "E/E 20 <5 <1 <5 <S 9 347 <S <1 <5 <5 103 EPA Methods Compound* SCSI: Oi-ganoctilonne Pesticides & 8082. Result Compound* "e/s PCB's Analysis Repon RESU LT ug/g Aldrin alpha-BHC beta-BHC delta-BHC gamma-BHC (Lindane) alpha-Chlordane gamma-ChIordane 4,4'-DDD 4.4'-DDE 4,4' -DDT Dieldnn Endosulfan I Endosulfan 11 Endosulfan sul-Tate Endnn Endrin aldehyde <0.10 <0.10 <0.10 <0.10 <0.10 <0.10 <0.10 <0.20 <0.20 <0.20 <0.20 <0.10 <0.20 <0.20 0.20 <0.20 Endrin ketone Heptachlor Heptachlor eponde Methozychlor Chlordane (technical) Toxaphene Aroclor-1016 Aroclor-1221 Aroclor-1232 Aroclor-1242 Aroclor-1248 Aroclor-1254 Aroclor-1260 Aroclor-1262 Aroclor-1268 <0.20 <0.10 <0.10 <1.0 <l.O <10 <2.0 <4.0 <2.0 <2.0 <2.0 <2.0 <2.0 <Z.O <2.0 'Compound analyzed for but not detected. INGREDIENTS Fish Meal, Soy Flour, Com Gluten Meal, Blood Meal, Fish Oil, Liquid Fish Solubles, Torula Dried Yeast, Brewers Yeast, Soy Lecithin, Lignin Sulfonate, L-Ascorbyl-2-Polyphospbate (Vitamin C), Choline Chloride, Ethoxyqum (preservative), Canthaxanthin (pigrnenling agent). Vitamin B12 Supplement, Niacin, Calciurn Pantothenate, R-iboflavin, Pyndoxine Hydrochlonde, Vitarmn A Acetate, D-Activated Animal Sterol (souice of Vitamin D3), Thiainine Morionicrale, Folic Acid, Biolm, dI-Alpha Tocophero! Acetate (source of Vitamin E), Menadione (Vitamin K), Zinc Sulfate. Copper Sulfate, Majiganous Sulfale, Ethylencdiamuie Dihydroiodide, Cobalt Carbonate, Ferrous Sulfate, Calcium Carbonate MANLTFACTUTUED BY: ZEIGLER BROS., INC. P.O. BOX 95, GARDNERS, PA 17324 Aquatic BioSystems, Inc Quality Research Organisms - 123 Company Sanitized. Does not contain TSCA CB1 1300 Blue Spruce Drive, Suite C Fort Collins, Colorado 80524 Toll Free: 800/331-5916 Tel: 970/484-5091 Fax:970/484-2514 YTC TOTAL SOLIDS MEASUREMENT (Method from EPA/5 05/8-89-002 a) YTC Process Date: Average Total Solids 12/4/00: Best if used by 04/30/01 1760 mg/1 Ingredient T'"t Numbers 'Pines International Wheat Grass WG98S00090; Zeigler Salmon Starter#l: 38-4763-9150, Fleischmanns Yeast- G-3 Analyzed Metals Aluminum Arsenic ; Cadmium Chromium Copper Iron Lead Mercury Nickel Silver Zinc EPA Required Toxic Metals Report Limits Results (me/L) 0.1 <0.1 0.005 <0.005 0.001 0.001 0.01 <0.01 0.01 0.01 0.03 0.18 0.01 <0.01 0.001 O.OOl 0.01 0.005 0.01 0.01 0.005 0.42 Compounds Aldrin alpha-BHC beta-BHC delta-BHC Eamma-BHC fLmdane) alpha-Chlordane gamma-Chlordane 4,4"-DDD 4.4' - DDE 4,4' -DDT nieldrin Endosulfan I EndosuUan n Eodosulfan sulfate Endrin Endrin aldehyde Endrin ketone Heptachlor Heptachlor epoxide Melhozychlor Chlordane (technical) Toxaphene Aroclor-1016 Aroclor-1221 Aroclor-1232 Aroclor-1242 Arocior-1248 Aroclor-1254 Aroclor-1260 Aroclor-1262 Aroclor-1268 Results (ug/L) <2.0 <2.0 <2.0 <2.0 <2.0 <2.0 <2.0 <4.0 <4.0 <4.0 <4.0 <2.0 <4.0 <-1.0 <4.0 <4.0 <4.Q <2.0 <2.0 <20 <20 <200 <40 <80 <40 <40 <40 <40 <40 <40 <40. U - Indicates compound was analyzed for but not detected '"Testing performed by Energy Labs, Billings, Montana Qualilier U - U U U U U U U U U U U U U U U U U U U U U U U U U U U U U U Aquatic BioSystems, Inc Quality Research Organisms Company SaAiJ'Aed.Does not contain TSCA Cfil LABORATORIES Consulting Engineers Environmental Laboratories 1116 South Main Street Blacksburg, Virginia 24060 Client Sheet No.: Date: Client: Source: Shipping Information: 0005275 July 11,2000 Job No.: Date Received: 60001 5/25/00 Olver Laboratories Incorporated Blacksburg, Virginia Delivered to Olver Laboratories Incorporated by Olver Personnel Sample No.: Time Collected: i Date Collected: Description: Analysis Total Total Total Total Total Total Total Total Total Total Total Total Aluminum (EPA 202.2) Arsenic (EPA 206 2) Cadmium (EPA 213 2) Chromium (EPA 218.2) Cobalt (EPA 219.2) Cqaper (EPA 220 2) Iron (EPA 200.7) Lead (EPA 238 2) Mercury (EPA 245.1) Nickel (EPA 200 9) Silver (EPA 272 2) Zinc (EPA 200.7) 124686 3:45 p m. 5/25/00 Tetramin Flakes (Lot No. 121599) Result MU [EEL Date/"rime Anal'yzed 350 NO 0.0125 1 05 0.425 13.3 905 0275 ND 13 0.055 823 mg/kg as received ** mg/kg as received mg/kg as received mg/kg as received mg/kg as received mg/kg as received mg/kg as received mg/kg as received mg/kg as received' mg/kg as received 312 0.15 0005 0.15 0.05 05 035 0.05 0.25 0.05 001 05 mg/kg as received" mg/kg as received mg/kg as received mg/kg as received mg/kg as received mg/kg as received* mg/kg as received mg/kg as received* mg/kg as received mg/kg as received mg/kg as received mg/kg as received 6/27/00; 7/01/00: 6/19/00; 7/10/00; 6/29/00; 6/14/00; 6/22/00; 6/23/00; 7/05/00; 8 00 a m. 10.00 a-m. 11.30a.m. 1 30 p.m. 1.00 p m. 3.00 p m. 2.00 p m. 2.00 p m. 2:00 p m 7/05/00; 2:00 p.m 6/27/00; 3-00 p m. 6/27/00; 8:00 a m Analyst AMT BN ' AMT BN HT HT HT HT HT HT/BN HT AMT MDL = Minimum Detection Limit ND = Not Detectable * Sample required dilution; MDL was raised accordingly. "* Due to low spike recovery, the Method of Standard Additions was used to analyze this sample. All tests according to Standard Methods for the Examination of Water and Wastewater. 18th Edition, Test Methods for Evaluating Solid Waste (Physical/Chemical). 2nd and 3rd Editions, and Methods for the Chemical Analysis of Water and Wastes. EPA. By f^l^hlL. Bnt Ritchey /T Laboratory Manager Unless otherwise indicated, this report sets forth the results of our analysis of samples delivered to our laboratory and shall nol be construed to be a representation by Otver Laboratories Incorporated as to the source or method of procuring such samples. All reports are submitted as the confidential property of dienis and authorization for publication of any statements contained in our reports is r&served pending ourwntten approval Blacksburg, VA (540) 552-6974 pa9e 1 or'4 135 P\DATAIB10\60Q01\CLNTSHTUX)05275 Company Sanitized. Does not contain TSCA CBI OLVER LABORATORIES Client Sheet No . Client Date Sample No.: Time Collected: Date Collected: Description: Analysis Pesticides (EPA 608)- Aldnn alpha-BHC beta-BHC delta-BHC gamma-BHC ^ Chlordana 4,4'-DDD 4,4'-DDE 4,4'-DDT Dieldnn Endosulfan I Endosulfan II Endosulfan Sulfate Endnn Endnn Aldehyde Heptachlor . Hepiacfator Epoxide Toxaphene PCBs (EPA S08):. PCB-1016 PCB-1221 PCB-1232 PCB-1242 PCS-1248 PCB-1254 PCS-1260 0005275 Olver Laboratories Incorporated July 11,2000 124686 3:45 p.m. 5/25/00 Tetramin Flakes (Lot No. 121599) Result ND ND ND NO ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND MQL 5 /^g/kg as received 5 ^g/kg as received 5 /^g/kg as received 5 ^g/kg as received 5 ^g/kg as received 50 /2g/kg as received 5 ^g/kg as received 5 ^g/kg as received 5 Aig/kg as received 5 /.g/kg as received 5 Afl/kg as received 5 ^g/kg as received 5 ^g/kg as received 5 Atg/kg as received 5 ^g/kg as received 5 ^g/kg as received 5 ^g/kg as received 50 ^g/kg as received 50 ^g/kg as received 50 ^g/kg as received 50 ^g/kg as received 50 /^g/kg as received 50 ^g/kg as received 50 /^g/kg as received 50 ^g/kg as received Date/Time Analyzed 6/06/00; 5.56 a m. 6/06/00, 5:56 a.m Analyst ASM/PMW ASM/PMW Page 2 of 4 126 P \OATAaiC60001\CLNTSHT\OOOS275 Company Sanitized. Does not contain TSCA CBl OLVER LABORATORIES Client Sheet No Client Date 0005275 Olver Laboratories Incorporated July 11, 2000 Sample No.: Time Collected: Date Collected: Description: Analysis Total Aluminum (EPA 202.2) Total Arsenic (EPA 206.2) Total Cadmium (EPA 213.2) Total Chromium (EPA 200.9) Total Cobalt (EPA 219.2) Total Copper (EPA 220-2) Total Iron (EPA 200.7) Total Lead (EPA 239.2) Total Mercury (EPA 245.1) Total Nickel (EPA 200 9) Total Sliver (EPA 272 2) Total Zinc (EPA 200.7) Pesticides (EPA 608): Aldnn " alpha-BHC beta-BHC delta-BHC ((anaHia-BHC Chlordane 4,4'-DDD 4,4'-DDE 4,4'-ODT Dieldnn Endosulfan I Endosulfan II Endosulfan Sulfate Endnn Endnn Aldehyde Heptachlor Heptachlor Epoxide Toxaphene PCBs (EPA 608): PCB-1016 PCS-1221 PCB-1232 PCB-1242 PCB-1248 PCB-1254 PCB-1260 124687 3:45 p.m. 5/25/00 TYC (Lot No. 51900) Result 0.118 0.22 NO 0.004 ND 0 054 0.372 0.012 ND 0.004 ND 0.521 mg/L mg/L*' mg/L mg/L mg/L mg/L mg/L " mg/L ND ND ND ND tiD ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND Ml3J. Date/Time Analysari Analyst 0025 0.15 0.0001 0001 0.001 0.001 0.007 0.1 0.25 0.001 0.0002 0005 mg/L' mg/L' mg/L mg/L mg/L mg/L mg/L mg/L* mg/L mg/L mg/L mg/L 6/27/00; 8:00 a.m AMT 7/01/00; 10:00 am. BN 6/19/00; 11:30 am. AMT 7/07/00; 11.30 am. BN 6/29/00. 1:00 p.m. HT 6/14/00; 3:00 p.m. - HT . 6/22/00; 2:00 p-m. HT 6/23/00; 2:00 p m. HT 6/26/00; 10.00am. 7/05/00: 2:00 p m. 6/27/00; 3.00 p m. 6/27/00; 8:00 a m. AMT HT/BN HT AMT 6/06/00; 6.24 a m. ASM/PMW 5 yg/kg as received 5 ^9/kg as received 5 ^g/kg as received 5 /ig/kg as received 5 /^g/kg as received 50 /'0/kg as received 5 yg/kg as received 5 ^g/kg as received 5 ^g/kg as received 5 ^9/kg as received 5 ^g/kg as received 5 ^g/kg as received 5 ^g/kg as received 5 M/kg as received 5 Aig/kg as received 5 ^g/kg as received 5 ^g/kg as received 50 ^g/kg as received 6/06/00; 6.24 a m. ASM/PMW 50 ^g/kg as received 50 ^g/kg as received 50 ^g/kg as received 50 ^g/kg as received 50 ^g/kg as received 50 Aig/kg as received "- 50 A's/kg as received * Sample required dilution; MDL was raised accordingly. " Due to low spike recovery, the Method of Standard Additions was used to analyze this sample Page 3 of 4 127 P \OATAIB)0\60001\CLMTSHT\000527S Company Sanitized. Does not contain TSCACBI OLVER LABORATORIES Client Sheet No Client Date 0005275 Olver Laboratories Incorporated July 11,2000 Sample No.: Time Collected: Date Collected: Description: 124688 5:00 p.m. 5/25/00 Brine Shrimp Nauplii (Lot No. 4400) Analysis "Total Aluminum (EPA 202.2) Total Arsenic (EPA 206.2) Total Cadmium (EPA 213 2) Total Chromium (EPA 200-9) Total Cobalt [EP/42W.2} Total Copper (EPA 220 2) Total Iron (EPA 200.7) Total Lead (EPA 239 2) Total Mercury (EPA 2451) Total Nickel (EPA 200.9) Total Sliver (EPA 272.2) Total Zinc (EPA 200.7) Pesticides (EPA 608): Aldnn alpha-BHC beta-BHC - deftaWlC gamma-BHC Chlordane 4,4'-ODD 4,4'-DDE 4.4'-DDT Dieldnn Endosulfan I Endosulfan li Endosutfan Sulfate Endnn Endnn Aldehyde Heptachlor Heptachlor Epoxide Toxaphene PCBs (EPA 608). PCB-1016 PCS-1221 PCB-1232 PCB-1242 PCS-1248 PCB-1254 PCB-1260 Result 0.007 0.08 ND 0 001 ND 0015 1.04 ND ND 0.003 ND 0 069 mg/L mg/L' mg/L mg/L mg/L mg/L " mg/L ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND NO MDL Date/Time Analyzed ' Analyst 0.005 mg/L ' 0.15 mg/L' 0.0001 mg/L 0.001 mg/L 0.001 mg/L 0001 mg/L 0007 mg/L 0.1 mg/L' 025 mg/L 0.001 mg/L 0.0002 mg/L 0005 mg/L 5 f^g/kg as received 5 /^g/kg as received 5 ^g/kg as received 5 Aig/kg as received 5 ^g/kg as received 50 ^ig/kg as received 5 ^g/kg as received 5 /^g/kg as received 5 .ug/kg as received 5 Aig/kg as received 5 ^g/kg as received 5 ^ig/kg as received 5 ^g/kg as received 5 Aig/kg as received 5 A;g/kg as received 5 ^g/kg as received 5 ^ig/kg as received 50 fxg/kg as received 50 f^g/kg as received 50 ^ig/kg as received 50 ^ig/kg as received 50 ^g/kg as received 50 ^ig/kg as received 50 ^g/kg as received 50 /^g/kg as received 6/27/00:8 00 a.m. 7/01/00; 10.00 a.m 6/19/00, 11:30 a.m. 7/07/00: 11:30 a.m. 6/29/00, 1.00p.m. 6/14/00.3 00p.m. 6/22/00,2:00 p.m. 6/23/00,2.00 p m 6/26/00; 10:00 a m 7/05/00,2-00 p m 6/27/00.3 00 p.m 6/27/00; 8:00 a m 6/06/00; 6:53 a m. 6/06/00, 6 53 a.m ;., AMT BN AMT BN HT HT HT HT AMT HT/BN HT AMT ASM/PMW ASM/PMW * Sample required dilution; MDL was raised accordingly. " Due to low spike recover/, the Method of Standard Additions was used to analyze this sample. Page 4 of 4 P M)ATA\BICA6000nCLNTSH'tM)Otl5275 128 Company Sanitized. Does not contain 7SCA CB1 CHAIN OF CUSTODY RECORD ^^? OLVER LABORATORIES Consulting Engineers nnd nnviionmenta! Laboratories 1116 s0"111 Mai" strect' S'a^sburg, Virginia 24060 Telephone: (540) 552-6974 PAX: f540) 552-1715 * Clicnl l]uuM cuniplcic ill infoniialion tcqucilcd lo llie iiehl mil In Ilic unshidcil ircgi below DONOTWRIKIH niCSIIADr.D AHUAS * Sini)ilci mil be ilnciiJcil m rciumci) la ilic clicni iwo wcch incr Itic anilylicil repon his txcn niailrj, unicis p(lor irrjilgcnictlis liivc been niadc CSiCCOS-Z-T- 5 Jubt to0t10\ Olver Sample Pll 11) H S;ini|ilc Locnilon (i c, innucnl, nuirallOOl, MW1, CIC) Dale Callcclcd '1 Illlt Collccliil Client;--O^cr Labor^+or^s T^ J ^ " Company Contact: Bio^squ Lo^rft-WM- fer . Address:___' .__________________ Telephone:. Purchase Order No. L. s U g a J *^ Sample Container PrcicrvallDn 'K ^ Type "1 ypc M a 0 L. 0 0. U U 5 0 0 U % ^ ,^ul f2^(,^ AfOl, T<'t:r<^mio P)ckc5- l~ot 'ZlSiq './ ^^0 ^pn Fopi So " 'i urd. P1''*-CSCtH A ^Oru, ^ t ^ 1 i^' 1 -Ddd-J&^ Hi, AS.) cr F^i-hu'd ^ [Z^^^ -^/l ^0 iz4t,^2 , ^ -^P TY<^ Food M|\4<Ar<.~ Lot 5;1oo 51-2-5/00 p^ -2'A'O 0 '5>r\m. S^TnVip Mo-upl;/-' Lo-f- Woo \f 5(75/00 Soo pto- /P ^c. A^ A&- MA ^on*. ^A Mcxv- v^' ! L-^ 1 I^hL^'^ ft'i A5,Cr P'^-Hl-tA t/-- l 1^' 1 Tt)^\ t^t.+ Al, A5,C P&ih'u' Sumplei(s) (Signnlnre) .All^Nl ^ KCjiiiquis icj By (Si.v,iw{\\iv.) - ! ^ ^ Z A^ l<.c1inc|iii.slicdDy\(Si a^ninc) l^eliiniiiislicd Dy: (Sigmilinc) . Dale IniiL. ^ i ^ D^lc TII^C Rcccived Dy (Signntiirc) ^^f}^,'^'^ r^(- ^<a-^^--. Ilccqi/cd Dy: (Si^iiiiturc) ^/ Pile Iline Received Dy: (SignnHire) Special lu ^<i. 'vIViJ Uali: I line ^ / 3 ^ Dale Tniic Tcinpcrnliir Samples tra Melliod o n^ic Coiiuncul