Document 1Qq2yE4xvp93VjQL6wmegO7BE

F MAM - -3-2T/ TRADE SECRET ftkUM -.32X1 DuPont-11238 Study Title H-25357: Approximate Lethal Dose (ALD) by Skin Absorption in Rabbits Laboratory Project ID: DuPont-11238 A u t h o r : Carol Finlay, B.A. S tu d y C o m p l e t e d o n : August 1,2002 P e r f o r m in g L a b o r a t o r y : E.I. du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 Page 1 of 13 Osmpm Sanitized. Do m noi contain TSCA CBI H-25357: Approximate Lethal Dose (ALP) by Skin Absorption in Rabbits DuPont-11238 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of the data obtained from this study. R eview ed by: Associate Scientist -Q ^ < 7^4 Issued by Study D irector: l i/ IfKi 1Finlay, Am Staff T oxicologist ^ lz- - 2 - Company Sanitized. Does not contain TSCA CB1 H-25357: Approximate Lethal Dose (ALD) by Skin Absorption in Rabbits STUDY INFORMATION Substance Tested: 3 Svnonvms/Codes: H-25357 Submitter's Notebook Numberis): Haskell Number: 25357 DuPont-11238 Known Impurities Physical Characteristics: Amber liquid Sponsor: E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study hhtiated/Completed: June 17, 2002 / (see report cover page) In-Life Initiated/Completed: July 2, 2002 / July 16,2002 - 3 - Company Sanitized. Does not contain TSCA CB1 H-25357: Approximate Lethal Dose (ALP) by Skin Absorption in Rabbits DuPont-11238 STUDY PERSONNEL Study Director: Carol Finlay, B.A. Management: Scott E. Loveless, Ph.D. Primary Technician: James C. Mackay II Toxicology Report Preparation: Sean M. Callaghan, B.A. Management: Nancy S. Selzer, M.S. Laboratory Veterinarian: William Singleton, D.V.M., A.C.L.A.M. -4Company Sanitized. Does not contain TSCA CBS H-25357: Approximate Lethal Dose (ALD) by Skin Absorption in Rabbits DuPont-11238 SUMMARY H-25357 was applied to the shaved, intact skin of 2 male New Zealand White rabbits at a dose of 5000 mg/kg. The application site was semi-occluded for approximately 24 hours after which the test substance was removed. The rabbits were observed for clinical signs o f toxicity, body weight effects, and dermal effects for 14 days following removal of the test substance (weekends excluded). No death occurred. The rabbits exhibited no clinical signs during the study. Body weight loss of approximately 2% of initial weight occurred in one o f the rabbits by the day after application of the test substance. No biologically important weight loss occurred in the other rabbit. The substance produced no dermal irritation in either rabbit. Under the conditions of this test, the ALD was greater than 5000 mg/kg of body weight. This substance is considered to be very low in toxicity (ALD greater than 5000 mg/kg) when administered as a single dermal dose to rabbits. /f**,**\ -5Corapany Sanitized. Does not contain TSCA CBS H-25357: Approximate Lethal Dose (ALP) by Skin Absorption in Rabbits DuPont-11238 INTRODUCTION The purpose of this study was to determine an approximate lethal dose (ALD) by skin absorption o f H-25357. The ALD was defined as the lowest dose o f the test substance administered that caused the death of a test animal within 14 days of exposure. If 2 animals are dosed with 5000 mg/kg of test substance and neither animal dies, then the dermal ALD will be reported to be greater than 5000 mg/kg. MATERIALS AND METHODS A. Test Substance The test substance, H-25357, was supplied by the sponsor as an amber liquid. The test substance was inverted to mix before each amount for dosing was removed. The test substance appeared to be stable under the conditions of the study. No evidence of instability, such as a change in color or physical state, was observed. B. Animal Husbandry Young adult male HM:(NZW)fBR New Zealand White rabbits were received from Covance Research Products, Denver, Pennsylvania. The rabbits were housed singly in stainless steel, wire-mesh cages suspended above cage boards. Each rabbit was assigned a unique identification number which was recorded on a card affixed to the cage. The last 3 digits o f the identification number were written on the inside of each rabbit's ear with a water insoluble marker. The rabbits were offered approximately 125 grams of PMI Nutrition International, Inc. Certified High Fiber Rabbit LabDiet 5325 daily during the study. Water was available ad libitum. 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 o f the study: Water samples are analyzed for total bacterial counts, and the presence o f coliforms, lead, and other contaminants. Feed samples are analyzed for total bacterial, spore, and fungal counts. 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 o f key contaminants, including specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The -6Company Sanitized. Does nol contain TSCA CBI H-25357: Approximate Lethal Dose (ALD) by Skin Absorption in Rabbits DuPont-11238 presence of these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity o f 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 o f the study. Rabbits were weighed and observed for general health during the 7-day quarantine period. Animal rooms were maintained on a timer-controlled, 12-hour light/12-hour dark cycle. Environmental conditions of the rooms were targeted for a temperature o f 20C 1C and relative humidity of 50% 10%. Excursions outside these ranges were judged to have been of insufficient magnitude and/or duration to have adversely affected the validity of the study. C. Protocol On the day before dosing, the fur o f each rabbit was closely shaved to expose the back from the scapular to the lumbar region. The test substance was measured for each animal on the day o f treatment at a dose o f 5000 mg/kg o f body weight. The amount o f test substance designated for each animal was calculated based on the test substance density of 1.09 g/mL and the body weight collected prior to treatment. The rabbits weighed 2266 or 2164 grams on the day of treatment. The area to be treated (approximately 6" x 5" or 190 square centimeters*) was marked on the dorsal skin of each rabbit with water-insoluble marker. It was not feasible to apply the test substance directly to the skin. The aliquot of the test substance designated for an animal was spread onto a 2 ply, 6" x 5" gauze pad. The gauze pad was placed onto the test site. The rabbits were then wrapped with successive layers of stretch gauze bandage and adhesive bandage. The rabbits were fitted with plastic collars to prevent ingestion of the test substance or disruption of the wrappings. After wrapping, the rabbits were returned to their cages. The rabbits were observed for clinical signs of toxicity approximately 2 hours after dosing. Approximately 24 hours after treatment, the rabbits were removed from their cages and the wrappings and collars were removed. Excess test substance was washed from the animals' backs with Lander soap and warm water, and the skin dried with a paper towel. The rabbits were then returned to their cages. Approximately 1 hour after test substance removal, the animals were weighed and observed for clinical signs of toxicity and dermal irritation. Observations for dermal irritation were made according to the Draize Scale (Table 1). A glossary of dermal effects and abbreviations is presented as Table 2. Observations for clinical signs of toxicity and dermal irritation were made daily for 14 days after treatment (weekends excluded). Observations for mortality and signs of illness, injury, or abnormal behavior were made daily throughout the study. The animals were weighed on the day of treatment and on days 1, 7, and 14 following treatment. One hundred ninety square centimeters (or approximately 6" x 5") is approximately equal to 10 percent of the total body surface of rabbits in the 2-3 kg body weight range. -7 Company Sanitized. Does not contain TSCA CBI H-25357: Approximate Lethal Dose (ALP) by Skin Absorption in Rabbits RESULTS AND DISCUSSION DuPont-11238 A. Dose Information and Mortality No death occurred. The dose regimen and the mortality resulting over the 15-day test period are detailed below. Animal Number 35563 35561 Dose (mg/kg) 5000 5000 Amount (mL) 10.4 9.9 Initial Body Weight (g) 2266 2164 Mortality No No B. Clinical Observations, Body Weights, and Skin Responses The rabbits exhibited no clinical signs during the study. Body weight loss o f approximately 2% of initial weight occurred in one of the rabbits by the day after application of the test substance. No biologically important weight loss occurred in the other rabbit. The substance produced no dermal irritation in either rabbit. Individual body weights and individual clinical signs o f toxicity are presented in Table 3. Individual dermal responses are presented in Table 4. CONCLUSIONS Under the conditions of this test, the ALD was greater than 5000 mg/kg of body weight. This substance is considered to be very low in toxicity (ALD greater than 5000 mg/kg) when administered as a single dermal dose to rabbits. 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. Company Sanitized. Does not contain TSCA CBI H-25357: Approximate Lethal Dose (ALP) by Skin Absorption in Rabbits DuPont-1123 8 TABLES -9Company Sanitized. Does net contain TSCA CBI H-25357: Approximate Lethal Dose (ALD) by Skin Absorption in Rabbits DuPont-11238 TABLE 1 DRAIZE3 SCALE FOR SCORING SKIN IRRITATION Evaluation of Skin Reactions Score Erythema and eschar formation: No erythema........................................................................................... 0 Very slight erythema (barely perceptible)............................................ 1 (Slight) Well-defined erythema.................................................................................................. 2 (Mild) Moderate to severe erythema........................................................................................3 (Moderate) Severe erythema (beet redness) to slight eschar formation (injuries in depth)...... 4 (Severe) Edema formation: No edem a..................................................................................................................... 0 Very slight edema (barely perceptible)...................................................................... 1 (Slight) Slight edema (edges o f area well defined by definite raising)...................................2 (Mild) Moderate edema (raised approximately 1.0 m m )..................................................... 3 (Moderate) Severe edema (raised more than 1.0 mm extending beyond the area of exposure)..................................................................................................................4 (Severe) a Draize, J. H., "Dermal Toxicity." Appraisal of the Safety of Chemicals in Foods. Drugs and Cosmetics. The Editorial Committee of the Association of Food and Drug Officials of the United States, Austin, Texas, 1959, pp. 46-59. -10Company Sanitized. Does not contain TSCA CBI H-25357: Approximate Lethal Dose (ALP) by Skin Absorption in Rabbits DuPont-11238 TABLE 2 GLOSSARY OF DERMAL EFFECTS Blanching white appearance to skin Eschar scab on the skin that is more superficial than necrosis. This is considered to be severe irritation. Desquamation dry, flaking of the skin Fissuring a split or cleft in the top layer of skin without bleeding Fissuring with Bleeding a split or cleft in the skin with bleeding Hyperkeratosis thick, dry discoloration (usually but not limited to brown or white in color) of the top layer of skin. This is considered to be severe irritation. Sloughing peeling of the top layer o f skin, and epidermal scaling that has detached Necrosis area o f rough/hard/dry black or dark colored skin that may crater. This is considered to be corrosion and an irreversible effect. Epidermal Scaling platelike areas of the top layer of skin that have separated from but are still attached to viable skin. This may progress to sloughing. Thickening skin is firm and/or dense to the touch. Ulceration open sore representing loss of superficial tissue ABBREVIATIONS OF OTHER DERMAL EFFECTS - = No Effect A = Abraded B = Blanching C = Eschar D = Desquamation F = Fissuring G = Fissuring with Bleeding H = Hyperkeratosis I = Intact L = Sloughing N = Necrosis R = Raw Areas S = Epidermal Scaling T = Thickening X = Compound Adhered to Skin -11 - C om paq Sanitized. Doe* not contain TSCA CB9 H-25357: Approximate Lethal Dose (ALP) by Skin Absorption in Rabbits DuPont-11238 TABLE 3 INDIVIDUAL BODY WEIGHTS (g) RABBIT NUMBER 35563 35561 DOSE (mg/kg) 5000 5000 INITIAL WEIGHT 2266 2164 DAYS AFTER APPLICATION 1 7 14 2212 2562 2687 2143 2458 2562 CLINICAL SIGNS OF TOXICITY OBSERVED IN RABBITS No clinical signs of toxicity were observed in any o f the rabbits during the study. - 12- Ctjnyanf Saniiaed. Doss not contain TSCA CBI H-25357: Approximate Lethal Dose (ALP) by Skin Absorption in Rabbits DuPont-11238 RABBIT NUMBER 35563 35561 1 0 0 RABBIT NUMBER 35563 35561 1 0 0 TABLE 4 INDIVIDUAL DERMAL RESPONSES ERYTHEMA DAYS AF1PERAPP CATION 3 6 7 8 9 10 13 14 0 00 00 0 0 0 00000000 EDEMA DAYS AF1rER APP C A T IO N 3 6 7 8 9 10 13 14 0000000 0 000 0000 0 - 13 - c a'PMySsiifizei Daes nofcontain TSCACBI