Document 3ed556NwEoYqXKbZ6ZDz3qKza

Corning Hazleton Inc. P.O. Box 7545 Madison, WI 53707-7545 Deliveries: 3301 Kinsman Blvd., Madison, WI 53704 608.241.4471 608.241.7227 Fax A (tairOMi7 CO R N IN G Hazleton Sponsor: 3M St. Paul, Minnesota FINAL REPORT Study Title: Acute Dermal Toxicity Study of T -6342 in Rabbits (OECD Guidelines) Author: Steven M. Glaza Study Completion Date: October 24, 1995 Performing Laboratory: Corning Hazleton Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Project Identification: HWI 50800374 Page 1 of 32 000114 Page 2 of 32 HWI 50800374 COMPLIANCE STATEMENT Acute Dermal Toxicity Study of T-6342 in Rabbits (OECD Guidelines) This study was conducted in accordance with the Organization for Economic Cooperation and Development's Principles of Good Laboratory Practice, C(81)30(Final). Acute Studies Corning Hazleton Inc. ___________ R I V A S ' Date 000115 Page 3 of 32 HWI 50800374 QUALITY ASSURANCE STATEMENT This report has been reviewed by the Quality Assurance Unit of Hazleton Wisconsin, Inc., in accordance with the Organisation for Economic Cooperation and Development (OECD) Principles of Good Laboratory Practice, C (81)30(Final). The following inspections were conducted and findings reported to the Study Director and management. Written status reports of inspections and findings are issued to Hazleton management monthly according to standard operating procedures. Inspection Dates From To Phase Date Reported to Date to Study Director Management 08/31/95 08/31/95 Dose Administration 10/13/95 10/16/95 Data/Report Review 08/31/95 10/16/95 09/10/95 11/10/95 Repres Assurance Unit Date f o "2.*/- f r " 000116 Page 4 of 32 HWI 50800374 STUDY IDENTIFICATION Acute Dermal Toxicity Study of T-6342 in Rabbits (OECD Guidelines) Test Material Sponsor Sponsor's Representative Study Director Study Location Study Timetable Study Initiation Date Experimental (In-life) Start Date In-life End Date Experimental Termination Date Study Completion Date T-6342 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3220 Roger G. Perkins, PhD 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3220 (612) 733-3222 Steven M. Glaza Corning Hazleton Inc. P.O. Box 7545 Madison, HI 53707-7545 (608) 241-7292 Corning Hazleton Inc. 3301 Kinsman Boulevard Madison, HI 53704 August 25, 1995 August 31, 1995 September 14, 1995 October 24, 1995 October 24, 1995 (0117 Acute Studies Steven M. Glaza Study Director Manager Steven R. Sorenson Study Coordinator Patricia Padgham In-1ife Supervisor Rose M. Bridge Report Supervisor Toxicoloav SuDDort Kathy Myers Manager Calvin L. Horton Supervisor Oualitv Assurance Sherry R. W. Petsel Manager Page 5 of 32 HWI 50800374 KEY PERSONNEL Laboratory Animal Medicine Cindy J. Cary, DVM Dipl ornate, ACLAM Supervisor Anatomical Patholoav Thomas E. Palmer, PhD Anatomical Pathologist Deborah L. Pirkel/ Jack Serfort Supervisors Necropsy Anne Mosher Supervisor Pathology Data 000118 Page 6 of 32 CONTENTS Comp!i ance Statement Quality Assurance Statement Study Identification Key Personnel Summary Objective Test Material Test System Procedures Results Discussion Signature References Pathology Report Table 1 Mortality Summary 2 Individual and Mean Body Weights/BodyWeight Gains (g) 3 Individual Clinical Signs 4 Individual Dermal Irritation Observations 5 Individual Pathology Comments Appendix A Protocol Protocol Amendment No. Protocol Amendment No. 1 2 HWI 50800374 Page 2 3 4 5 7 7 7 8 9 10 11 11 11 12 13 14 15 16 18 19 20 31 32 000119 Page 7 of 32 HWI 50800374 SUMMARY The test material, T-6342, was evaluated for its acute dermal toxicity potential in male and female rabbits when administered as a single topical application at a level of 2,000 mg/kg of body weight. The estimated dermal LD,. for male and female rabbits was determined to be greater than 2,000 mg/kg. All animals appeared normal throughout the study. All animals exhibited body weight gain during the study with the exception of one male which exhibited a weight loss of 29 g during the first week. The test material produced slight to moderate dermal irritation. The gross necropsy at termination revealed no visible lesions. OBJECTIVE The objective of this study was to assess the systemic toxicity and relative skin irritancy of a test material when applied to the skin of rabbits.1 All procedures used in this study were in compliance with the Animal Welfare Act Regulations. In the opinion of the Sponsor and study director, the study did not unnecessarily duplicate any previous work. TEST MATERIAL Identification The test material was identified as T-6342 and described as a clear, colorless liquid. Purity and Stability The Sponsor assumes responsibility for purity and stability determinations (including under test conditions). Storage and Retention The test material was stored at room temperature. Any unused test material will be returned to the Sponsor after issuance of the final report according to Corning Hazleton (CHW) Standard Operating Procedure (SOP). Safety Precautions The test material handling procedures were according to CHW SOPs and policies. 000120 Page 8 of 32 HWI 50800374 Test Animal TEST SYSTEM Adult albino rabbits of the Hra:(NZW)SPF strain were procured from HRP, Inc., Kalamazoo, Michigan on August 9, 1995. Housing After receipt, the animals were acclimated for a period of at least 7 days. During acclimation and throughout the study, the animals were individually housed in screen-bottom stainless steel cages. Environmental controls for the animal room were set to maintain a temperature of 19 to 23C, a relative humidity of 50% 20%, and a 12-hour 1ight/12-hour dark lighting cycle. In cases where variations from these conditions existed, they were documented and considered to have had no adverse effect on the study outcome. Animal Diet The animals were provided access to water ad libitum and a measured amount of Laboratory Rabbit Diet HF #5326, PMI Feeds, Inc. The feed is routinely analyzed by the manufacturer for nutritional components and environmental contaminants. Samples of the water are periodically analyzed by CHW. There were no known contaminants in the feed or water at levels that would have interfered with or affected the results of the study. Animal Selection and Preparation of Exposure Area Five male and five female healthy, acclimated rabbits, weighing from 2,386 to 2,675 g, were used for a single dose level of 2,000 mg/kg of body weight. The animals were identified by animal number and corresponding ear tag throughout the study. On the day before test material application, each rabbit's back was clipped free of hair with an electric clipper. The clipped area made up not less than 10% of the total body surface. The animals were clipped as needed throughout the study. Justification for Species Selection Historically, the New Zealand White albino rabbit has been the animal of choice because of the large amount of background information on this species. 000121 Page 9 of 32 HWI 50800374 PROCEDURES Preparation of Test Material The test material was administered as received. An individual dose was calculated and weighed out based on each animal's body weight on the day of test administration. Treatment The test material was applied to the intact skin on each animal's back at a dose level of 2,000 mg/kg of body weight. The test material was applied to the test site at a rate of approximately 0.03 g/cm2 in a thin and uniform layer. The area of application (approximately 200 cm2 )was covered with a 9.5-cm x 21.0-cm, 4-ply gauze patch secured with paper tape and overwrapped with Saran Wrap and Elastoplast tape to provide an occlusive dressing. Collars were used to restrain the test animals during the 24-hour exposure period. At the end of the 24-hour exposure period, the restraining collars and bandages were removed and the test sites were washed using tap water and disposable paper towels. Reason for Route of Administration Historically, the dermal route has been the route of choice based on the method of Draize.2 Observations Clinical observations and mortality checks were conducted at approximately 1, 2.5, and 4 hours after test material administration. Additional clinical observations and twice a day mortality checks (morning and afternoon) were conducted daily thereafter for 14 days. Body weights were determined before test material application (Day 0), at Day 7, and at termination of the in-life phase (Day 14). The initial dermal irritation reading was made approximately 30 minutes after removal of the test material according to the Draize technique (recorded as the Day 1 score). Subsequent readings of dermal irritation were made on Days 3, 7, 10, and 14. 000122 Page 10 of 32 HWI 50800374 Pathology At termination of the in-life phase, all animals were euthanized, subjected to an abbreviated gross necropsy examination, and any abnormalities were recorded. After necropsy, the animals were discarded and no tissues were saved. Statistical Analyses No statistical analyses were required by the protocol. Location of Raw Data. Records, and Final Report The raw data, records, and an original signed copy of the final report will be retained in the archives of CHW in accordance with CHW SOP. Mortality RESULTS A summary of the survival rate is in Table 1. No mortality was observed during the study. The estimated dermal LD50 for male and female rabbits was determined to be greater than 2,000 mg/kg of body weight. Body Weights Individual and mean body weights and body weight gains are in Table 2. All animals exhibited body weight gain throughout the study with the exception of one male which exhibited a weight loss of 29 g during the first week. Clinical Signs Individual clinical signs are in Table 3. All animals appeared normal throughout the study. Dermal Reactions Individual dermal irritation observations are in Table 4. Dermal irritation (based on the most severe score for each animal at any time point) consisted of slight to moderate erythema, edema, and atonia, and slight desquamation, coriaceousness, and fissuring. No other dermal irritation was observed. 000123 Page 11 of 32 HWI 50800374 Pathology Individual gross necropsy pathology findings are in Table 5. A summary report by the study pathologist is on Page 12. There were no visible lesions observed at necropsy. DISCUSSION The acute dermal toxicity of T-6342 was evaluated in male and female rabbits when administered as a single topical application at a level of 2,000 mg/kg of body weight. The estimated dermal LD50 for male and female rabbits was determined to be greater than 2,000 mg/kg. All animals appeared normal throughout the study. All animals exhibited body weight gain during the study with the exception of one male which exhibited a weight loss of 29 g during the first week. The test material produced slight to moderate dermal irritation. The gross necropsy at termination revealed no visible lesions. SIGNATURE Date REFERENCES 1. "Acute Dermal Toxicity," Organisation for Economic Cooperation and Development's Guidelines for Testing of Chemicals, Section 402 (adopted May 12, 1981). 2. Draize, J. H., "Acute Dermal Toxicity (Single Exposure)," In: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics - Dermal Toxicity, Association of Food and Drug Officials of the U.S., pp. 54-55 (1959). 000124 Page 12 of 32 HWI 50800374 PATHOLOGY REPORT There were 10 rabbits (five males and five females) euthanized and necropsied at the termination of the study. The dose level, day of death, and gross observations recorded for each animal are in the Individual Pathology Comments that follow this report. There were no visible lesions in any of the animals. (50800374.rpt) 092695 Date 000125 Page 13 of 32 HWI 50800374 Dose Level (rciq/ kq) . 2,000 2,000 Table 1 Mortality Summary Mortality Result Sex No. Died/ No. Dosed M 0/5 F 0/5 000126 Page 14 of 32 HWI 50800374 Table 2 Individual and Mean Body Weights/Body Weight Gains (g) Animal Number F56261 F56271 F56265 F56266 F56267 Mean Day 0 Weioht Dav 7 Weiaht Gain* Males (2,000 ma/ka) 2,553 2,481 2,391 2,526 2,529 2,524 2,562 2,393 2,577 2,613 -29 81 2 51 84 2,496 2,534 38 Dav 14 Weiaht Gain 2,642 2,582 2,480 2,707 2,714 2,625 89 101 89 181 185 129 F56268 F56269 F56264 F56270 F56274 Mean Females (2,000 ma/kol 2,534 2,448 2,386 2,675 2,487 2,685 2,484 2,512 2,679 2,521 151 36 126 4 34 2,506 2,576 70 2,770 2,555 2,536 2,684 2,582 2,625 236 107 150 9 95 119 * Gain from the Day 0 body weight. 000127 Page 15 of 32 HWI 50800374 Animal Number Observation F5626I Appeared normal F56271 Appeared normal F56265 Appeared normal F56266 Appeared normal F56267 Appeared normal F56268 Appeared normal F56269 Appeared normal F56264 Appeared normal F56270 Appeared normal F56274 Appeared normal / Condition existed. Table 3 Individual Clinical Signs ______ Hour ____________________________ Day 1.0 L i 4.0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 Males (2.000 ma/ka) / / / / / / / / / / / / // / / / / / / / / / / // / / / / / / / / // / / / / / / / / / / / // / / / / / / Females (2.000 ma/ka) / / / // / / / / / / / / // / / / / / / / / / / / / // / / / / / / / / / / / / / // / / / / / / / / / / / // / / / / / / ooo1 2 s Page 16 of 32 HWI 50800374 Animal Number F56261 F56271 F56265 F56266 F56267 Table 4 Individual Dermal Irritation Observations Observation Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring Observation Period (Davi 1 3 7* 10 14* Males 11111 1000 0 10 0 0 0 000 10 000 0 0 00 10 0 100 00 100 0 0 00 0 0 0 00000 00 0 0 0 00 0 0 0 110 0 0 100 0 0 100 0 0 000 0 0 00000 00000 11211 100 0 0 100 0 0 000 10 00000 00 0 0 0 1000 0 0000 0 100 0 0 00000 00000 00000 * Animals shaved before dermal observation. 000129 Page 17 of 32 HWI 50800374 --i O O --I o O CM--I o I O O O O O O O O Table 4 (Continued) Individual Dermal Irritation Observations Animal Number F56268 F56269 F56264 F56270 F56274 Observation Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring Erythema Edema Atonia Desquamation Coriaceousness Fissuring _______ Observation Period (Day) 1 3 7* io Females 1122 1111 100 0 000 1 00 10 00 1 1 1222 122 2 2 10 0 000 1 00 10 000 1 1110 100 0 0000 0000 000 0 0000 12 2 2 122 1 000 0 000 1 00 10 000 1 112 1 0111 0000 000 1 0010 0000 * Animals shaved before dermal observation. r-1 1--i O >--i o o oooooo 000130 Page 18 of 32 HWI 50800374 Table 5 Individual Pathology Comments Dose Level: 2,000 mg/kg of Body Weight Animal Test Dav Number Sex Died Sacrificed NecroDsv Observation F56261 M - 14 No visible lesions. F56271 M - 14 No visible lesions. F56265 M - 14 No visible lesions. F56266 M - 14 No visible lesions. F56267 M - 14 No visible lesions. F56268 F - 14 No visible lesions. F56269 F - 14 No visible lesions. F56264 F - 14 No visible lesions. F56270 F - 14 No visible lesions. F56274 F . 14 No visible lesions. - Not applicable. 000131 Page 19 of 32 APPENDIX A Protocol Protocol Amendment No. 1 Protocol Amendment No. 2 HWI 50800374 000132 Page 20 of 32 Sample Submittal Form This form is to be used when submitting samples for routine acute testing. Special testing needs can be easily arranged by contacting the Acute Toxicology Department at (608) 'N 241-4471, Ext. 7292, or the Client Service Center at (608) 241-7210.______________________________________ Submitted by:. HWI Study No. >ToSo_o37*f Enclose with samples and send to: Hazleton Wisconsin 3301 Kinsman Boulevard Madison, Wisconsin 53704 Date Sample Sent: kiz/fr Company:------- --------------------------------------------------- ;-------------- Invoice t o : _______ P.O. Number: Number of Reports Required: 3. Full GLP Compliance:__ -Y es _____No .FDA (21 CFR 58) .EPA (TSCA--40 CFR 792) ____ EPA (FIFRA)---40 CFR 160) _ X _ OECD Sample Nam e:. T - ( o 3 4 3 . Physical Description: _ Special Handling Precautions: 'c r fTl S 0 5 > b l ' aW Storage Requirements: Room Temperature ____ Refrigerated ____ O th er_____________ Disposal: Return to submitter _____ Dispose of according to Hazleton SOPs Tests Acute Oral Toxicity in Rats 1__ TP8084 Up and down LD50 procedure ------TP3206 FHSA screen; 5M -5F at 5.0 g/kg ___ Conduct defined study if death occurs at 5.0 g/kg ___ TP3013 EPA screen; 5M-5F at 5.0 g/kg ___ Conduct defined study if death occurs at 5.0 g/kg --A T P 2 0 6 9 OECD screen; 5M -5F at 5.0 g/kg ___ Conduct defined study if death occurs at 5.0 g/kg Special instructions:__________________________________ Acute Dermal Toxicity in Rabbits ------TP3207 FHSA screen; 5M -5F at 2.0 g/kg ------TP3016 EPA screen; 5M-5F at 2.0 g/kg v -- Conduct defined study if death occurs at 2.0 g/kg -X -T P 2 0 7 0 OECD screen; 5M -5F at 2.0 g/kg _ --- Conduct defined study if death occurs at 2.0 g/kg Special instructions:__________________________________ Primary Skin Irritation ___ TP3208 FHSA; 6 rabb'its-1 abraded, 1 intact site/rabbit ___ TP3014 EPA; 6 rabbits-1 intact site/rabbit _ y ,T P 2 0 7 1 OECD; 3 rabbits-1 intact site/rabbit ___ TP4206 DOT corrosivity; 6 rabbits-1 intact site/rabbit ------TP7145 Phototoxicity; 6 rabbits-2 intact sites/rabbit (one site with UVA exposure) Special instructions:__________________________________ Primary Eye Irritation ___ TP6360 Low-volume procedure; 6 rabbits unwashed ___ TP3209 FHSA; 6 rabbits unwashed ------TP2012 1978 EPA; 6 rabbits unwashed, 3 washed ___ TP3015 1982 EPA; 6 rabbits unwashed -X -T P 2 0 7 2 OECD; 3 rabbits unwashed ___ 3 rabbits washed at 4 seconds ------3 rabbits washed at 30 seconds Special instructions:_________________________________ I For Hazleton Use Only Protocol Issue D ate:________ ^ tudy Director:. White copy-- Hazleton Yellow copy-- Submitter Guinea Pig Sensitization ___ TP2017 EPA Magnusson-Kligman maximization -TP6164 OECD Magnussun-Kligman maximization ------TP2008 Buehler sensitization -- TP6289 Photoallergenic contact dermatitis (Armstrong) Special instructions:.________________________________ _ -------------------------------- -------------------------------------------------- -- -- 000133 HAZLSTOfM WISCONSIN OSI OFFICE BOX 7545 ADISON WISCONSIN 53707-7545 Page 21 of 32 a C O R N IN O l orator* Services Campar* Sponsor: 3M St. Paul, Minnesota PROTOCOL TP2070 Study Title: Acute Dermal Toxicity Study in Rabbits (OECD Guidelines) Date: June 1, 1993 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Project Identification: HWI , S ^ S ' o o J 7 V P h o rt : \ *A A t I l> f l V F R * 3 30 1 KINSMAN BOULEVARD MAH SON WISCONSIN 000134 Page 22 of 32 TP2070 Page 2 STUDY IDENTIFICATION Acute Dermal Toxicity Study in Rabbits (OECD Guidelines) HWI No. Test Material Sponsor Sponsor's Representative Study Director Study Location Proposed Study Timetable Experimental Start Date Experimental Termination Date Final Report Date ,fo 8 o c a 3 ? Y (See sample submittal form) 3M Toxicology Services 220-2E-02 3M Center St. Paul, MN 55144 John L. Butenhoff, PhD 3M Toxicology Services 220-2E-02 3M Center St. Paul, MN 55144 (612) 733-1962 Steven M. Glaza Hazleton Wisconsin, Inc. P.0. Box 7545 Madison, WI 53707-7545 (608) 241-7292 Hazleton Wisconsin, Inc. Building No. 3 3802 Packers Avenue Madison, WI 53704 Week of S-.2&* 9 ^ Week of 9- i1-9S'' Week of/0 ' Page 23 of 32 TP2070 Page 3 1. Study Acute Dermal Toxicity Study in Rabbits (OECD Guidelines) 2. Purpose To assess the systemic toxicity and relative skin irritancy of a test material when applied to the skin of rabbits 3. Regulatory Compliance This study will be conducted in accordance with the following Good Laboratory Practice Regulations/Standards/Guidelines: [ ] Conduct as a Nonregulated Study [ ] 21 CFR 58 (FDA) [ ] 40 CFR 160 (EPA-FIFRA) [ ] 40 CFR 792 (EPA-TSCA) nC (81)30 (Final) (OECD) Notification No. 3850, August 10, 1984 (Japanese MAFF) [ ] Notification No. 313, March 31, 1982, and as amended by Notification No. 870, October 5, 1988 (Japanese MOHW) All procedures in this protocol are in compliance with the Animal Welfare Act Regulations. In the opinion of the Sponsor and study director, the study does not unnecessarily duplicate any previous work. 4. Quality Assurance For regulated studies, the protocol, study conduct, and the final report will be audited by the Quality Assurance Unit in accordance with Hazleton Wisconsin (HWI) Standard Operating Procedures (SOPs) and policies. 5. Test Material A. Identification (See sample submittal form) B. Physical Description (See sample submittal form) C. Purity and Stability The Sponsor assumes responsibility for purity and stability determinations (including under test conditions). Samples of test material/vehicle mixture(s) (if applicable) for concentration, solubility, homogeneity, and stability analyses will be taken before administration if requested by the Sponsor. These samples (if taken) will be sent to the Sponsor after experimental termination for possible analysis. 000136 Page 24 of 32 TP2070 Page 4 0. Storage (See sample submittal form) E. Reserve Samples Studies of less than 4 weeks in experimental duration will not have reserve samples retained. Reserve sample(s) of each batch/lot of test material will be taken if this study is more than 4 weeks in experimental duration. The test material reserve sample will be stored at HWI in a freezer set to maintain a temperature of below 0*C for 10 years per HWI SOP. The Sponsor will be contacted after 10 years for disposition in accordance with the appropriate regulatory Good Laboratory Practices. F. Retention Any unused test material will be discarded after issuance of the final report, unless directed otherwise by the Sponsor. G. Safety Precautions As required by HWI SOPs and policies 6. Experimental Design A. Animals (1) Species Rabbit (2) Strain/Source Hra:(N2W)SPF/Hazleton Research Products, Inc. (3) Aae at Initiation Adult (4) Weight at Initiation 2.0 to 3.0 kg (5) Number and Sex 5 males and 5 females for the initial dose level 5 males and/or 5 females for any additional dose levels (if required) (6) Identification Individual numbered ear tag 000137 Page 25 of 32 TP2070 Page 5 (7) Husbandry (a) Housing Individually, in screen-bottom stainless steel cages (heavy gauge) (b) Food A measured amount of High Fiber Rabbit Chow* #5326 (Purina Mills, Inc.)- The food is routinely analyzed by the manufacturer for nutritional components and environmental contaminants. (c) Water Ad libitum from an automatic system. Samples of the water are analyzed by HWI for total dissolved solids, hardness, and specified microbiological content and for selected elements, heavy metals, organophosphates, and chlorinated hydrocarbons. (d) Contaminants There are no known contaminants 1n the food or water that would interfere with this study. (e) Environment Environmental controls for the animal room will be set to maintain a temperature of 19 to 23'C, a relative humidity of 50% +20%, and a 12-hour light/12-hour dark cycle. (f) Acclimation At least 7 days (8) Selection of Test Animals Based on health and body weight according to HWI SOPs. An adequate number of extra animals will be purchased so that no animal in obviously poor health is placed on test. (9) Justification for Species Selection Historically, the New Zealand White albino rabbit has been the animal of choice because of the large amount of background information on this species. B. Dose Administration (1) Dose Level A single dose of 2,000 mg/kg of body weight will be administered to the intact skin of five males and five females. If no test material-related mortality is produced at this level, no further testing will be required. If any mortality occurs at the 2,000 mg/kg Page 26 of 32 TP2070 Page 6 level, additional dose levels may be added at the direction of the study director in order to meet the objectives of the study. (2) Preparation of Exposure Area On the day before test material application, the back of each rabbit will be clipped free of hair with an electric clipper. Not less than 10% of the total body surface area will be clipped. The animals will be clipped as needed throughout the study. (3) Dose Administration All animals will receive a single administration of test material. The dose will be based upon the animal's body weight just before administration of the test material. The area of application will be covered with as thin and uniform a layer as possible. If a liquid, the test material will be applied undiluted. If a solid, the test material will be moistened with 0.9% saline. Aerosol materials will be discharged Into a beaker and administered as a liquid. The area of application will be wrapped with a 10.0-cm x 10.0-cm gauze bandage secured with paper tape around all edges, overwrapped with Saran Wrap*, and secured with Elastoplast* tape to provide an occlusive dressing. The rabbits will be collared during the 24-hour application period. (4) Reason for Route of Administration Historically, the dermal route has been the route of choice based on the method of Draize. (5) Removal of Test Material Approximately 24 hours after test material application, the bandages and collars will be removed, and the residual test material will be removed using water or an appropriate solvent, if necessary. C. Observation of Animals (1) Clinical Observations At approximately 1, 2.5, and 4 hours after test material administration for clinical signs and mortality, daily thereafter for clinical signs, and twice daily (a.m. and p.m.) for mortality for at least 14 days. Observations may be extended when directed by the study director. (2) Reading of Dermal Irritation Approximately 30 minutes after bandage removal, the initial dermal irritation reading will be made and 000139 Page 27 of 32 TP2070 Page 7 recorded as the Day 1 reading (Attachment 1). Additional dermal irritation readings will be made on Study Days 3, 7, 10, and 14. Individual dermal irritation records will be maintained for each animal. (3) Body Weights Just before test material application, on Days 7 and 14, and at death (when survival exceeds 1 day) D. Pathology At termination of the experimental phase, surviving animals will be euthanized. All animals, whether dying during the study or euthanized, will be subjected to an abbreviated gross necropsy examination and all abnormalities will be recorded. After necropsy, the animals will be discarded and no tissues will be saved. E. Statistical Analyses Other than LD50 calculations (when applicable) no statistical analyses are required. 7. Report A final report including those items listed below will be submitted: Description of the test material Description of the test system Procedures Dates of experimental initiation and termination Tabulation of mortality data by sex and dose level Description of any toxic effects/dermal irritation Tabulation of mean body weights by sex and dose level LD,0 values by sex with 95% confidence intervals (when applicable) Gross pathology findings/gross pathology report (when applicable) 8. Location of Raw Data. Records, and Final Report Original data, or copies thereof, will.be available at HWI to facilitate auditing the study during its progress and before acceptance of the final report. When the final report is completed, all original paper data, including those items listed below will be retained in the archives of HWI according to HWI SOP. credo Page 28 of 32 TP2070 Page 8 Protocol and protocol amendments Dose preparation records In-life records Body weights Dose administration Observations Anatomical pathology records Study correspondence Final report (original signed copy) The following supporting records will be retained at HWI but will not be archived with the study data. Animal receipt/acclimation records Water analysis records Animal room temperature and humidity records Refrigerator and freezer temperature records Instrument calibration and maintenance records 000141 Page 29 of 32 PROTOCOL APPROVAL TP2070 Page 9 w t, John L. Butenhoff, PhD Sponsor's Representative 3M Steven H. Claza Study Director Acute Toxicology Hazleton Wisconsin) Inc. 0 fiefacm. -X .y faiZ L Repreessentative Quality Assurance Unit Hazleton Wisconsin, Inc. (TP2070.3M) Date z\t, / m 1 -fa=v-r* Date 000142 Page 30 of 32 TP2070 Page 10 Attachment 1 Scoring Scale for Acute Dermal Reactions Erythema 0 - None 1 - Slight 2 - Moderate 3 - Severe Edema 0 - None 1 - Slight (barely perceptible to well defined by definite raising) 2 - Moderate (raised approximately 1 mm) 3 - Severe (raised more than 1 mm) Atonia 0 - None 1 - Slight (slight Impairment of elasticity) 2 - Moderate (slow return to normal) 3 - Marked (no elasticity) Desouamatlon 0 - None 1 - Slight (slight scaling) 2 - Moderate (scales and flakes) 3 - Marked (pronounced flaking with denuded areas) Coriaceousness 0 - None 1 - Slight (decrease in pliability) 2 - Moderate (leathery texture) 3 - Marked (tough and brittle) Flssurina 0 - None 1 - Slight (definite cracks In epidermis) 2 - Moderate (cracks in dermis) 3 - Marked (cracks with bleeding) 000143 Page 31 of 32 HWI No. PROTOCOL AMENDMENTS 3~QSO>37V Amendment No. / Effective /vIq &o j t 3 ^ fC)c)-S'~~ Portion of Protocol Being Modified: Page 2. Study Location Reason for Modification: To identify the location where the study will be conducted. replace this section with the following c h a n g e : _________________________ Modification: Study Location____________ Hazleton Wisconsin. Inc.____________________ 3301 Kinsman Boulevard. _____________________________ Madison. WI 53704_______________________________________ (G21/01-07-91) Study Director Approval: 000144 Page 32 of 32 HWI No. PROTOCOL AMENDMENTS 50800374 Amendment No. Effective August 28. 1995 Portion of Protocol Being Modified: Page 6. 6. Experimental Design: B. Dose Administration: (31 Dose Administration. Reason for Modification: To correctly identifiy the size of the gauze bandage used to wrap the area of application._________________________________________________ Modification: Replace the seventh sentence of this section with the following change: The area of application will be wrapped with a 4-olv gauze bandage fapproximately 9.5-cm x 21-cm) secured with paper tape around all edges, overwrapoed with Saran Wrap, and secured with Elastoolast18 tape to provide an occlusive dressing._________________ Study Director Approval: (621/01-07-91) 000145