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TRADE SECRET AR226-3182 DuPont-6711 Study Title Jff^ Acute Oral Toxicity Fixed Dose Method Laboratory Project ID: DuPont-6711 TEST GUIDELINE: OECD Guidelines for Testing of Chemicals Section 4: Health Effects, No. 420 (1992) AUTHOR: Carol Finlay, B.A. STUDY COMPLETED ON: September 17, 2001 PERFORMING LABORATORY: E. I. du Font de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 Company Sanitized. Doss nol contain TSCA CB1 Page 1 of 27 Acute Oral Toxicity - Fixed Dose Method_________ '_______DuPont-6711 PAGE RESERVED FOR SPECIFIC COUNTRY REQUIREMENTS Company Sanitized. Doas no? conlain TSCA CB1 -2- Acute Oral Toxicity - Fixed Dose Method DuPont-6711 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT This study was conducted in compliance with U.S. EPA TSCA (40 CFR part 792) Good Laboratory Practice Standards, which are consistent with the OECD Principles of Good Laboratory Practice (as revised in 1997) published in ENV/MC/CHEM(98) 17 except for the items documented below. The items listed do not impact the validity of the study. The test substance was characterized after initiation of this study. Although the characterization (ISO 9001) was not performed under Good Laboratory Practice Standards, the accuracy of the data is considered sufficient for the purposes of this study. The dosing preparations used in the study were not analyzed for stability, homogeneity, or accuracy of concentration. The procedures used by trained staff to prepare the dosing preparations ensured: the accuracy of concentration because the test substance was weighed on an analytical balance accurate to 3 decimal places and the vehicle in which it was suspended was accurately measured in a flask graduated in 1 mL increments, homogeneity because the mixtures were stirred prior to dosing and while portions were removed for dose administration, and stability because the time between dose preparation and administration was kept to a minimum (less than 1 hour). Applicant / Sponsor: Telomer Research Program Arlington, Virginia U.S.A. Study Director: ^^/Igf. ^L^_________ Cfarol Finlay ^ Staff Scientist {/} - ^U-t' ^ Date Company Sanitized. Does no? contain TSCA CB1 ~~- .cute Oral Toxicity - Fixed Dose Method QUALITY ASSURANCE STATEMENT Haskell Sample Number(s): 24691 Dates of Inspections: Conduct: May 17, 2001 Records, Reports: April 5, 2001; September 9, 10, 2001 Dates Findings Reported to: Study Director: September 10, 2001 Management: September 13, 2001 DuPont-6711 Reported by: ^^l^Qf0^ ( } ' -- c ^ r^--~ fJoseph C. Hamill Sr. Quality Assurance Auditor )~7- Jg/- 2t>tf' Date Company Sanitized. Does no} contain TSCA C' -4- Acute Oral Toxicity - Fixed Dose Method _______DuPont-6711 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. Pathology Evohiations Reported by: * > L^CL C^'\i \I ^ J-^-/y rU-isa J. Lewis LaWratwy Technician Pathology Evaluations Reviewed by; & ^J. ^W^y^ ^k ^\ ------^ \1AA^ V G. Tracy Makovec, D.V.M. Diplomate A.C.V.P. </^Jraf^rj^fe4^s Reviewed by: < C. Macleay n Associate Scientist Issued by Study Director: f {L/\J^ fiCV\.[.JLt^ Ca^lFmlay, B.A.Q Staff Scientist t^-Sgo -aoot Date ^-r<f s<'\ Date V\-^jUfsL^i Dafe fc) Company Sanitizsd. Dcss nc; contain TSCA CB1 5- Acute Oral Toxicity - Fixed Dose Method DuPont-6711 TABLE OF CONTENTS Page PAGE RESERVED FOR SPECIFIC COUNTRY REQUIREMENTS................................... 2 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT.................................. 3 QUALITY ASSURANCE STATEMENT...................................................................................4 CERTIFICATION ........................................................................................................................5 STUDY INFORMATION............................................................................................................. 8 STUDY PERSONNEL.................................................................................................................. 9 SUMMARY.................................................................................................................................. 10 INTRODUCTION....................................................................................................................... 11 MATERIALS AND METHODS ............................................................................................... 11 A. Test Guidelines.................................................................................................................. 11 B. Test Substance................................................................................................................... 11 C. Animal Husbandry............................................................................................................ 11 D. Test Substance Preparation............................................................................................... 12 E. 12 Dosing............................................................................................................................... 1. Sighting 12 Study........................................................................................................................................ 2. Main Study ..........................................................;.......................,,.........................................................12 F. 13 Observations...................................................................................................................... 1. Sighting Study......................................................................................................................................13 2. Main Study............................................................................................................................................. 13 RESULTS AND DISCUSSION ................................................................................................. 14 In-life A. B. Toxicology......................................................................................................................... 14 Sighting 14 Study................................................................................................................... 1. Dose Information and Mortality Summary............................................................................................. 14 2. Body Weights........................................................................................................................................ 14 3. Clinical Signs ......................................................................................................................................... 14 Main Study........................................................................................................................ 14 1. Dose Information and Mortality Summary............................................................................................. 14 2. Body Weights.........................................................................................................................................15 3. Clinical Signs .........................................................................................................................................15 Pathology Evaluations................................................................................................................ 15 A. Gross Observations ........................................................................................................... 15 CONCLUSION............................................................................................................................ 15 RECORDS AND SAMPLE STORAGE ................................................................................... 15 Company Sanitized. Doe; not contain TSCA CBt -6- Acute Oral Toxicity - Fixed Dose Method DuPont-6711 ___ TABLE OF CONTENTS Page APPENDICES............................................................................................................................. 16 A. INDIVIDUAL BODY WEIGHTS.......................................................................................................................17 B. INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY RECORDS...............................................21 C. INDIVIDUAL ANIMAL GROSS OBSERVATIONS...................................................-...................................^ i) Company Sanitized. Dcas net contain TSCA CBT -7- Acute Oral Toxicity - Fixed Dose Method STUDY INFORMATION 9th Collective Nomenclature: Synonyms/Codes:, DuPont-6711 fc Haskell Number: 24691 sy CAS Registry Number: Composition:! Purity:^ Known Impurities: Physical Characteristics: White to yellow wax Stability: The test substance appeared to be stable under the conditions of the study; no evidence of instability was observed. Sponsor: Telomer Research Program 1200 South Hayes Street Arlington, Virginia 22202-5050 U.S.A. Study Initiated/Completed: April 2, 2001 / (see report cover page) In-Life Initiated/Completed: April 25, 2001 / May 17, 2001 Company Sanitised. Fcos nc? contain T3CA CBI -8- Acute Oral Toxicity - Fixed Dose Method STUDY PERSONNEL Study Director: Management: Primary Technician: Carol Finlay, B.A. Judith C. Stadler, Ph.D., D.A.B.T. James C. Mackay D Pathologist: Management: Pathology Report by: G. Tracy Makovec, D.V.M. Steven R. Frame, D.V.M., Ph.D. Lisa J. Lewis Toxicology Report Preparation: Wanda F. Diribokowilz Laboratory Veterinarian: William Singleton, D.V.M., A.C.L.A.M. DuPont-6711 Company San^od. 5---:3 n=? contain TSCA CW -9- Acute Oral Toxicity - Fixed Dose Method DuPont-6711 SUMMARY oJHIIBU|U|Bwere Single doses administered by intragastricmtubation to fasted ^l^^l|HHBBil male and femaleTats. In the preliminarysighting study, a single dose was administered to fasted'female rats at a dose of 500 or 2000 mg/kg. Smcenotoxicity was seen in these rats, the limit dose of 2000 mg/kg was selected for the main study. In the main study, the test substance was administered to a group of 5 fasted male rats and a group of 5 fasted female rats at a dose of 2000 mg/kg. The rats were observed for mortality, body weight effects, and clinical signs oftoxicity for 14 days after dosing. The rats were necropsied to detect grossly observable evidence of organ or tissue damage or dysfunction. No deaths occurred during the study. The rats exhibited no clinical signs oftoxicity. No significant body weight losses occurred after dosing. No gross lesions were present in the rats at necropsy. Under the conditions of this study, the minimum lethal dose was greater than 2000 mg/kg. According to the guidance provided in the OECD testing guideline^^^^^, compound which does not present a significant acute toxic risk if swallowed. <3- ra, Company Sanifec-cf -'-n---,--., n,,o,,?. contam TSCA CBI 10- Acute Oral Toxicity - Fixed Dose Method DuPont-6711 INTRODUCTION o f _ _ _ _ _ The purpose of this study was to assess the acute oral toxicity administered by oral gavage to male and female rats. The dose selected for the main study, 2000 mg/kg, was derived from a preliminary sighting study. In the sighting study, no toxicity was observed in rats dosed at 500 or 2000 mg/kg. MATERIALS AND METHODS A. Test Guideline The study design complies with the following testing guideline: Organisation for Economic Co-Operation and Development (OECD) (1992). 420 Acute Oral Toxicity - Fixed Dose Method. Guideline/or Testing of Chemicals. B. Test Substance The test substance------^--------HflHwas.subpypthlieesdponsor as a white to yellow wax. The test substanceappearedTobestableunder the conditions of the study. No evidence of instability, such as a change in color, was observed. C. Animal Husbandry Male and female Crl:CD(SD)IGS BR rats were received from Charles River Laboratories, Inc., Raleigh, North Carolina. Rats were housed singly in suspended, stainless steel, wire-mesh cages. Each rat was assigned a unique identification number which was recorded on a card affixed to the cage. The rats were tail-marked, using a water-insoluble marker, with the last 3 digits of the animal number. PMI Nutrition International, me. Certified Rodent LabDiet 5002 and water were available ad libitum except as noted in section E. Dosing. As specified in the Haskell Laboratory animal health and environmental monitoring program, the following procedures are performed periodically to ensure that contaminant levels are below those that would be expected to impact the scientific integrity of the study: Water samples are analyzed for total bacterial counts, and the presence ofcolifbrms, lead, and other contaminants. Feed samples are analyzed for total bacterial, spore and fungal counts. Company SaniSh-s;*. P-^s no? contain TSCA CBI -^_ .cute Oral Toxicity - Fixed Dose Method___________________DuPont-6711 Samples from freshly washed cages and cage racks are analyzed to ensure adequate sanitation by the cagewashers. Certified animal feed is used, guaranteed by the manufacturer to meet specified nutritional requirements and not to exceed stated maximum concentrations of key contaminants, including specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presence of these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity of the study. The animal health and environmental monitoring program is administered by the attending laboratory animal veterinarian. Evaluation of these data did not indicate any conditions that affected the validity of the study. Rats were quarantined, weighed, and observed for general health for 6 days. Animal rooms were maintainedon a timer-cohtrolled, 12-hbur lighf/12-hour dark cycle. Environmental conditions of the rooms were targeted for a temperature of 23 1C and relative humidity of 50 10%. Excursions outside these ranges were of small magnitude and/or brief duration and did not adversely affect the validity of the study. D. Test Substance Preparation The test substance was melted and stirred in a water bath (approximately 75-80C) for IP\ approximately 30 minutes to assure uniformity before each suspension was prepared. ^ E. Dosing 1. Sighting Study A single oral dose ^I^^BH|^BB|B melted and suspended in 0.5% aqueous methylcellulose, was administered by intragastric intubation to a fasted female rat at a dose of 500 mg/kg. Since toxicity was not seen at this dose, another fasted female rat was dosed at 2000 mg/kg. The rats were fasted approximately 17 or 18 hours prior to dosing with food being returned to the rats approximately 3 hours after dosing. The rats were approximately 9 weeks old on the day of dosing. The individual dose volumes were calculated using the suspension concentration and the fasted body weight obtained prior to dosing. The rats were dosed at a volume of 10 mL per kg of body weight. The dosing suspensions were stirred prior to and throughout the dosing procedure. 2. Main Study fUBBH^^IIR1^6^^ Single oral doses an(^ suspended in 0.5% aqueous methylcellulose, were administered by intragastric intubation to a group of 5 fasted male rats and a group of 5 fasted female rats at a dose of 2000 mg/kg. The rats were fasted approximately 18 hours prior to dosing with food being returned to the rats approximately 3.5 hours after dosing. The rats were approximately 9 weeks old on the day of dosing. Individual dose volumes Company Sanifeec?. Decs -otc-ntain TSCA CB1 -^- Acute Oral Toxicity - Fixed Dose Method___________________DuPont-6711 were calculated using the suspension concentration and the fasted body weights obtained prior to dosing. The rats were dosed at a volume of 10 mL per kg of body weight. The dosing suspension was stirred prior to and throughout the dosing procedure. F. Observations 1. Sighting Study Observations for mortality and signs of illness, injury, or abnormal behavior were made daily throughout the study. The rats were observed for clinical signs oftoxicity twice on the day of dosing and once each day thereafter. The rats were weighed on test days -1, 0 (day of dosing), 1, and 7. On test day 7, the rats were euthanized by carbon dioxide asphyxiation. 2. Main Study Observations for mortality and signs of illness, injury, or abnormal behavior were made daily throughout the study. The rats were observed for clinical signs oftoxicity twice on the day of dosing and once each day thereafter. The rats were weighed on test days -1, 0 (day of dosing), 1, 2, 3, 7, and 14. On test day 14, the rats were euthanized and necropsied to detect grossly observable evidence of organ or tissue damage or dysfunction. The rats were euthanized by carbon dioxide asphyxiation followed by exsanguination. Company Samfiz.d. Does not -nta.n T8CA C- - 13- .cute Oral Toxicity - Fixed Dose Method DuPont-6711 RESULTS AND DISCUSSION In-life Toxicology A. Sighting Study 1. Dose Information and Mortality Summary No deaths occurred. The dose information and the mortality data are summarized in the table below. DOSE (mg/kg) FEMALE 500 2000 FASTED BODY WEIGHT (g) 217.4 213.7 SUSPENSION CONCENTRATION (mg/mL) DOSE VOLUME (mL) MORTALITY 50 2.2 No 200 2.1 No 2. Body Weights (Appendix A) The rats exhibited no body weight losses after dosing. 3. Clinical Signs (Appendix B) The rats exhibited no clinical signs oftoxicity. B. Main Study 1. Dose Information and Mortality Summary No deaths occurred. The dose information and the mortality data are summarized in the table below. Company Sanitized. Doss not contain TSCA CB\ - 14- DOSE (mg/kg) MALE 2000 FEMALE 2000 'Acute Oral Toxicity - Fixed Dose Method AVERAGE FASTED BODY WEIGHT (g) SUSPENSION CONCENTRATION (mg/mL) 249.<5 200 210.3 200 AVERAGE DOSE VOLUME (mL) 2.5 2.1 DuPont-6711 MORTALITY RATIO 0/5 0/5 2. Body Weights (Appendix A) The rats exhibited no significant body weight losses after dosing. 3. Clinical Signs (Appendix B) The rats exhibited no clinical signs oftoxicity. The end of the tail was missing from one male rat during the study. Pathology Evaluations A. Gross Observations No gross lesions were present in the rats. CONCLUSION Under the conditions of this study, the minimum lethal dose was gri According to the guidance provided in the OECD testing guideline} compound which does not present a significant acute toxic risk if swallowed. 0 mg/kg.. s a RECORDS AND SAMPLE STORAGE Specimens (if applicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at Iron Mountain Records Management, Wilmington, Delaware. Coffipany Samtized. Dcss not contain TSCA C'?.\ -15- Acute Oral Toxicity - Fixed Dose Method DuPont-6711 APPENDICES Company SanFtized. C!--;s n"? contain TSCA CBI 16- Acute Oral Toxicity - Fixed Dose Method___________________DuPont-6711 APPENDIX A Individual Body Weights Company Sanitized. Doss not contain TSCA CBI 17- Acute Oral Toxicity - Fixed Dose Method Animal Number 648018 INDIVIDUAL BODY WEIGHTS OF FEMALE RATS FROM SIGHTING STUDY DOSE: 500mg/kg Day -I3 236.7 Day O" 217.4 Day 1 227.6 Day 7 251.0 Animal Number 648020 Day -1 231.1 Day 013 213.7 DOSE: 2000mg/kg Day 1 Day 7 216.0 235.0 a Weight before fasting b Fasted body weight Company Sanitized. Do 18- \.cute Oral Toxicity - Fixed Dose Method Animal Number 648472 648473 648474 648475 648476 Day -1 274.0 276.0 257.0 268.0 278.0 INDIVIDUAL BODY WEIGHTS OF MALE RATS FROM MAIN STUDY D OSE: 2000 mg/:kg Day O" 254.1 254.8 235.6 247.7 255.7 Day 1 269.1 272.6 255.4 271.7 276.4 Day 2 295.0 292.9 272.3 287.8 297.0 Day 3 305.0 307.1 282.3 295.4 306.1 Day 7 Da 325.0 3 328.5 3 299.9 3 310.4 3 325.1 3 " Weight before fasting b Fasted body weight Company Sanitized. Doe ""'Sg-:*'''*^^.------^.*.]!-., -----1 19- ^' Acute Oral Toxicity - Fixed Dose Method Animal Number 648488 648489 648490 648491 648492 INDIVIDUAL BODY WEIGHTS OF FEMALE RATS FROM MAIN STUDY DOSE: 2000mg/kg Day -1" 230.0 230.0 230.0 222.0 229.0 Day O" 213.5 214.5 211.7 201.3 210.5 Day 1 226.6 226.4 226.9 217.1 221.6 Day 2 240.6 245.3 249.5 229.4 238.9 Day 3 248.9 240.7 252.4 236.3 242.3 Day 7 Da 261.3 2 246.9 2 251.2 2 247.3 2 244.8 2 3 Weight before fasting b Fasted body weight 20- Company Sanitized ""Dooe .cute Oral Toxicity - Fixed Dose Method DuPont-6711 APPENDIX B Individual Clinical Observations and Mortality Records Company Sanitized ^esnn otcon^TSCACB, -21 - Acute Oral Toxicity - Fixed Dose Method INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY RECORDS IN FEMALE RATS FROM SI DOSE: 500mg/kg Sex Animal Observation F 648018 General observation. No Abnormality Detected Sacrificed by design Days 0-7 7 DOSE: 2000mg/kg Sex Animal Observation F 648020 General observation. No Abnormality Detected Sacrificed by design Days 0-7 7 -22- Company Sanitized. Doe ^S) =: - JlBAcute Oral Toxicity - Fixed Dose Method - - INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY RECORDS IN MALE RATS FROM M DOSE: 2000mg/kg Sex Animal Observation M 648472 General observation. No Abnormality Detected Sacrificed by design M 648473 General observation. No Abnormality Detected Sacrificed by design M 648474 General observation. No Abnormality Detected End of tail missing Sacrificed by design M 648475 General observation. No Abnormality Detected Sacrificed by design M 648476 General observation. No Abnormality Detected Sacrificed by design Days -1-14 14 -1-14 14 -1-4 5-14 14 -1-14 14 -1-14 14 23- Company Sanitized. Do 'Acute Oral Toxicity - Fixed Dose Method INDIVIDUAL CLINICAL OBSERVATIONS AND MORTALITY RECORDS IN FEMALE RATS FROM DOSE: 2000mg/kg Sex Animal Observation 648488 648489 648490 648491 648492 General observation. No Abnormality Detected Sacrificed by design General observation. No Abnormality Detected Sacrificed by design General observation. No Abnormality Detected Sacrificed by design General observation. No Abnormality Detected Sacrificed by design General observation. No Abnormality Detected Sacrificed by design Days -1-14 14 -1-14 14 -1-14 14 -1-14 14 -1-14 14 .24- Company Sanitized. Doss n Acute Oral Toxicity - Fixed Dose Method___________________DuPont-6711 APPENDIX C Individual Animal Gross Observations Company Saniffzed. Doss not contain TSCA CBI .25- Acute Oral Toxicity - Fixed Dose Method INDIVIDUAL ANIMAL GROSS OBSERVATIONS IN MALE RATS FROM MAIN STUD LESIONS |LESION I | ANIMAL TREATMENT GENERAL ORGAN NO ABNORMALITY DETECTED Figures in parentheses is the number of animals grossly examined for this tissue The absence of a number indicates the finding specified was not identified .26- Company Sanitized. D Acute Oral Toxicity - Fixed Dose Method INDIVIDUAL ANIMAL GROSS OBSERVATIONS IN FEMALE RATS FROM MAIN STU LESIONS LESION I ANIMAL TREATMENT GENERAL ORGAN NO ABNORMALITY DETECTED Figures in parentheses is the number of animals grossly examined for this tissue The absence of a number indicates the finding specified was not identified .27- Company Sanitized. D