Document EdyekKNJYZBbaxJKoMQbejj2j

Coming Hazleton Inc. -f.0.go)@-7545 Madison, Wl 53707-7545 Deliveries:3301 Kinsman Blvd.,Madison, WI 608.241.4471 608.241.7227 Fax S3704 Sponsor: 3M St. Paul, Minnesota FINAL REPORT CORNING Hazleton OC ToCoEE lIV1,9,9-5 OCT 3 0 1995 3 IV4@- Study Title: Acute Dermal Toxicity Study of T-6342 in Rabbits (OECD Guidelines) Author: Steven M. Glaza Study Completion Date: October 24, 1995 Performina Laboratory: Corning Hazleton Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 LaboratoryProjectIdentification: HWI 50800374 Page I of 32 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). Steven M. Glaza Date Study Director Acute Studies Corning Hazleton Inc. Page 3 of 32 HWI 50800374 QUALITY ASSURANCE STATEMENT This report has been reviewed by the Quality Assurance Unit of Hazleton Wisconsin, Inc., in accordancewith the Organisation for Economic Cooperation and Development (OECD) Principles of Good Laboratory Practice, C(81)30(Final).The following inspectionswere conducted and findings reported to the Study Director and management. Written status reports of inspectionsand 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 Representatipe, Quality Assurance Unit Date 2-@(-'f 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 InitiationDate 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, WI 53707-7545 (608) 241-7292 Corning Hazleton Inc. 3301 Kinsman Boulevard Madison, WI 53704 August 25, 1995 August 31, 1995 September 14, 1995 October 24, 1995 October 24, 1995 Acute Studies Steven M. Glaza Study Director Manager Steven R. Sorenson Study Coordinator Patricia Padgham In-life Supervisor Rose M. Bridge Report Supervisor Toxicology Support Kathy Myers Manager Calvin L. Horton Supervisor Ouality Assurance Sherry R. W. Petsel Manager Page 5 of 32 HWI 50800374 KEY PERSONNEL Laboratory Animal Medicine Cindy J. Cary, DVM Diplomate, ACLAM Supervisor Anatomical Pathology Thomas E. Palmer, PhD Anatomical Pathologist Deborah L. Pirkel/ Jack Serfort Supervisors Necropsy Anne Mosher Supervisor Pathology Data Page 6 of 32 CONTENTS Compliance Statement Quality Assurance Statement Study Identification Key Personnel Summary Objective Test Material Test System Procedures Results Discussion Signature References Pathology Report Table I Mortality Summary 2 Individual and Mean Body Weights/Body Weight Gains (g) 3 IndividualClinical Signs 4 Individual Dermal IrritationObservations 5 Individual Pathology Comments Appendix A Protocol Protocol Amendment Protocol Amendment No. 1 No. 2 HWI 50800374 Paqe 2 3 4 5 7 7 7 8 9 10 11 11 11 12 13 14 15 16 18 19 20 31 32 Page 7 of 32 HWI 50800374 SUMMARY The test material, T-6342,'was evaluatedfor 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,bbO 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 irritancyof a test material when appliedto 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 responsibilityfor purity and stability determinations (includingunder test conditions). Storage and Retention The test material was stored at room temperature. Any unused test material will be returned to the Sponsor after issuanceof 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. Page 8 of 32 HWI 50800374 TEST SYSTEM Test Animal 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 acclimatedfor a period of at least 7 days. During acclimation and throughout the study, the animals were individually housed in screen-bottom stainless steel cages. Environmentalcontrols for the animal room were set to maintain a temperature of 190 to 230C, a relative humidity of 50% 20%, and a 12-hour light/12-hourdark 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 interferedwith 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. Justificationfor 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. 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 approximately0.03 g/c:2m in a thin and uniform layer. The area of application (approximately200 cm2 )was covered with a 9.5-cm x 21.0-cm, 4-ply gauze patch secured with paper tape and overwrapped with Saran WrapO and Elastoplast'Otape 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 terminationof the in-life phase (Day 14). The initial dermal irritation reading was made approximately30 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. 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 LD,, 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 Individualdermal irritationobservationsare 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. 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 administeredas 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 mg7kg. 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 terminationrevealed no visible lesions. SIGNATURE Steven M. Glaza Date Study Director Acute Studies 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. 54-56 (1959). 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. Th-oh@aE@- Pal Pathologist (50800374.rpt) 092695 02 Date Page 13 of 32 HWI 50800374 Dose Level (mg/kq) . 2,000 2,000 Table I Mortality Summary Mortality Result Sex No. Died/ No. Dosed m 0/5 F 0/5 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 Weight 2,553 2,481 2,391 2,526 2,529 2,496 Day 7 Weight Gain! Males (2,000 mg/kq) 2,524 -29 2,562 81 2,393 2 2,577 51 2,613 84 2,534 38 Day 14 Weight Gain* 2,642 89 2,582 101 2,480 89 2,707 181 2,714 185 2,625 129 Females (2,000 mg/kq) F56268 2,534 2,685 151 2,770 236 F56269 2,448 2,484 36 2,555 107 F56264 2,386 2,512 126 2,536 150 F56270 2,675 2,679 4 2,684 9 F56274 2,487 2,521 34 2,582 95 Mean 2,506 2,576 70 2,625 119 Gain from the Day 0 body weight. Animal Number Observation F56261 F56271 F56265 F56266 F56267 Appeared normal Appeared normal Appeared normal Appeared normal Appeared normal F56268 F56269 F56264 F56270 F56274 Appeared normal Appeared normal Appeared normal Appeared normal Appeared normal Condition existed. Page 15 of 32 HWI 50800374 Table 3 IndividualClinical Signs Hour 1.0 2.5 4.0 1 Day 5 6 7 8 9 10 11 12 13 14 Males (2,000 mg/kq) 1( ( 4( ( *1 0( / ( 0( / 1( 4( el 11 0( 1( 4( 4( 4( 0( 1( 11 .1 (/ / ( 1( ( 0( ( / 91 / / #I 1( 1( #1 / 1( / Femles (2,000 mg/kq) 1( 1( 1( 0( 01 4( 1( 0( 1( 4( 1( 0( 0( 0( 0( If 0( 0( 4( Page 16 of 32 HWI 50800374 Table 4 IndividualDemal IrritationObservations Animal Number Observation F56261 F56271 F56265 F56266 F56267 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 1 Males 1 1 1 0 0 0 1 1 0 0 0 0 I 1 1 0 0 0 1 1 1 0 0 0 1 0 1 0 0 0 Observation Period (Day) 3 7* 10 14* 1 1 1 1 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 2 1 1 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Animals shaved before dermal observation. Page 17 of 32 HWI 50800374 Table 4 (Continued) Individual Dermal IrritationObservations 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* 10 14* Females 1 1 2 2 1 1 1 1 1 0 1 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 1 1 0 1 2 2 2 2 1 2 2 2 1 2 1 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 1 1 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 2 2 2 1 1 2 2 1 1 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 1 0 1 1 2 1 0 0 1 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0 0 Animals shaved before dermal observation. Page 18 of 32 HWI 50800374 Table 5 Individual Pathology Comments Dose Level: 2,000 mg/kg of Body Weight Animal Test Day Number Sex Died Sacrificed, NecrODSY 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 visiblelesions. F56270 F - 14 No visible lesions. F56274 F - 14 No visible lesions. Not applicable. Page 19 of 32 APPENDIX A Protocol Protocol Amendment No. 1 Protocol Amendment No. 2 HWI 50800374 Page 20 of 32 Sample Submittal Form Thisformistobe usedwhensubmininsgamplesforroutine acutetestinSgpeciatlestinegedscanbe easilayrranged by contactitnhgeAcuteToxicoloDgeypartmenatt(608) 241-4471E,%l 7292,ortheClienStervicCeenteart(6oa) 241-7210. HWIStudyNo. Enclosewithsamples and send to: HazletonWisconsin 3301 Kinsman Boulevard Madison,Wisconsin 53704 S ubm hted by:- tz i< Company: P.O. Number: FullGLP Compliance: Yes -FDA (21CFR 58) -No -EPA(TSCA--40CFR792) Sample Name: T-(o 3qcq Date Sample Sent: Invoiceto: Number ofReportsRequired: EPA (FIFRA)-40CFR 160) OECD PhysicalDescription: SpecialHandlingPrecautions: F F (h s C)s StorageRequirements: Room Temperature Refrigerated Other Disposal: Ftetumtosubmftter Dispose ofaccordingto HazletonSOPs Test 4LcuteOral Toxicityin Rats -LTP8084 Up and down LD50 procedure -TP3206 FHSA screen;5M-5F at5.0g/kg Conduct definedstudyifdeathoccursat5.0g/kg -TP3013 EPA screen;SM-5F at5.0g/kg Conduct definedstudyifdeathoccursat5.0glkg -ITP-2069 OECD screen;5M-5F at5.0gtkg Conduct definedstudyitdeathoccursat5.0gtkg Specialinstructions: Primary Skin Irritation TP3208 FHSA; 6 rabbgs-Iabraded,I intacstite/rabbit -TP3014 EPA; 6 rabbks-Iintacstfte/rabbh -44TP2071 OECD: 3 rabbits-i1ntacstite/rabbit -TP4206 DOT corrosivh6y;rabbits-iIntacstite/rabbft -TP7145 Phatoloxici6tyr;abbks-2intacsthes/rabbft (onesitewithUVA exposure) Specialinstructions: Primary Eye Irritation Acute Dermal Toxicityin Rabbits -TP3207 FHSA screen;SM-SF at2.0g/kg -TP3016 EPA screen;5M-5F at2.0g/kg _)L7- Conductdefinedstudyd deathoccursat2.0gtkg P2070 OECD screen;5M-5F at2.0gtkg Conduct definedstudyifdeathoccursat2.0gtkg Specialinstructions: -TP6360 Low-volumeprocedure-6,rabbitusnwashed -TP3209 FHSA; 6 rabbitusnwashed TP2012 1978 EPA; 6 rabbitsunwashed, 3 washed -TP3015 1982 EPA; 6 rabbitusnwashed -.)LTP2072 OECD; 3 rabbitusnwashed - 3 rabbitwsashed at4 seconds 3 rabbitwsashed at30 seconds Spec-ialinstructions: ForHazletonUse Only ProtocolIsstuoerDate: Cls- 'turdyoEtDoicroelIescscuteor: Whitecopy-Hazleton Yellowcopy-Submftter Guinea Pig Sensitization -TP2017 EPA Magnusson-Kligmamnaximization -TP6164 OECD Magnussun-Kligmanmaximization -TP2008 Buehlersensitization -TP6289 Pholoallergenciocntactdermatit(iAsrmstrong) Specialinstructions: '"HAZLP.'tC:N W WI SCON S IN POSI O;FICE BOX 0 7$45 MADISON WISCONSO 537 7 7545 Page 21 of 32 a COFtNW4G LaMOtWV SeNCeSCOMWPV Sponsor: 3M St. Paul, Minnesota PROTOCOL TP2070 Study Title: Acute Dermal Toxicity Study in Rabbits (OECD Guidelines) June 1, 1993 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 LaboratoryProject Identification: HWI SbRIQ>C:@3?,Y p 1,n @0 R 4 1 4A I I f 11,441 1-1 'AAII 1)1 1 .V R V 330 1 K IN kiM A @f 9 0 IJL E VA R 1) M-1@ "()ts (iU N 4 1 122 1 -JYISC()tisiti 53 704 Page 22 of 32 STUDY IDENTIFICATION Acute Dermal Toxicity Study in Rabbits (OECD Guidelines) TP2070 Page 2 HWI No. Test Material Sponsor Sponsor's Representative Study Director Study Location Proposed Study Timetable Experimental Start Date Experimental Termination Date Final Report Date (See sample submittalform) 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.O. Box 7545 Madison, Wl 53707-7545 (608) 241-7292 Hazleton Wisconsin, Inc. BuildingNo. 3 3802 Packers Avenue Madison, Wl 53704 Week of 3-21? Week of "t.t.IWeek of tO - ;23-c),I- Page 23 of 32 TP2070 Page 3 I. Study Acute Dermal Toxicity Study in Rabbits (OECD Guidelines) 2. Purpose To assess the systemictoxicityand relativeskin irritancyof a test materialwhen appliedto the skin of rabbits 3. Regulatory Compliance This study will be conducted in accordancewith 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) C(81)30(Final)(OECD) NotificationNo. 3850, August 10, 1984 (JapaneseMAFF) 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 compliancewith the Animal Welfare Act Regulations. In the opinion of the Sponsor and study director,the study does not unnecessarilyduplicateany previous work. 4. Ouality Assurance For regulatedstudies, the protocol,study conduct,and the final report will be audited by the Quality AssuranceUnit 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. Purityand Stability The Sponsor assumes responsibilityfor purity and stability determinations(includingunder test conditions). Samples of testmaterial/vehiclmeixture(s)(if applicablef)or concentration,solubility,homogeneity,and stabilityanalyses will be taken before administrationif requestedby the Sponsor. These samples (if taken) will be sent to the Sponsor after experimentalterminationfor possibleanalysis. Page 24 of 32 TP2070 Page 4 0. Storage (See sample submittalform) E. Reserve Samples Studiesof less than 4 weeks in experimentaldurationwill not have reserve samples retained. Reservesample(s)of each batch/lotof test materialwill be taken if this study is more than 4 weeks in experimental duration. The test material reserve sample will be stored at NWI in a freezer set to maintain a temperatureof below O*C for 10 years per HWI SOP. The Sponsor will be contacted after 10 years for dispositionin accordancewith the appropriateregulatory Good LaboratoryPractices. F. Retention Any unused test material will be discarded after issuanceof the final report,unless directedotherwiseby the Sponsor. G. Safety Precautions As required by NWI SOPs and policies 6. ExperimentalDesign A. Animals (1) Species Rabbit (2) Strain/Source Hra:(NZW)SPF/HazletonResearch Products, Inc. (3) Age at Initiation Adult (4) Weight at Initiation 2.0 to 3.0 kg (5) Number and Sex 5 males and 5 femalesfor the initialdose level 5 males and/or 5 females for any additionaldose levels (ifrequired) (6) Identification Individual numbered ear tag Page 25 of 32 TP2070 Page 5 (7) Husbandry (a) Housing Individually,in screen-bottomstainlesssteel cages (heavy gauge) (b) Food A measured amount of High Fiber Rabbit Chow* #5326 (PurinaMills, Inc.). The food is routinelyanalyzed 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 specifiedmicrobiologicalcontent and for selected elements,heavy metals, organophosphates,and chlorinatedhydrocarbons. (d) Contaminants There are no known contaminants in the food or water that would interferewith this study. (e) Environment Environmentalcontrols for the animal room will be set to maintain a temperatureof 19 to 230C, a relative humidity of 50% +20%, and a 12-hour light/12-hourdark 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) Justificationfor SpeciesSelection Historically,the New Zealand White albino rabbit has been the animal of choice because of the large amount of background informationon this species. B. Dose Administration (1) Dose Level A single dose of 2,000 mg/kg of body weight will be administeredto the intact skin of five males and five females. If no test material-relatemdortalityis producedat this level,no furthertestingwill be required. If any mortalityoccurs at the 2,000 mg/kg Page 26 of 32 TP2070 Page 6 level, additionaldose levels may be added at the directionof the study director in order to meet the objectivesof the study. (2) Preparation of Exposure Area On the day before test materialapplication,the back of each rabbitwill be clipped free of hair with an electric clipper. Not less than 10% of the total body surface area will be clipped. The animalswill be clipped as needed throughoutthe study. (3) Dose Administration All animalswill receivea single administrationof test material. The dose will be based upon the animal's body weight just before administrationof the test material. The area of applicationwill be coveredwith as thin and uniforma layer as possible.If a liquid,the test materialwill be appliedundiluted. If a solid,the test materialwill be moistenedwith 0.9% saline. Aerosol materialswill be discharged into a beaker and administeredas a liquid. The area of applicationwill 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 occlusivedressing. The rabbitswi*ll be collaredduring the 24-hour applicationperiod. (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 Approximately24 hours after test material application, the bandages and collars will be removed, and the residual test material will be removed using water or an appropriatesolvent, if necessary. Observation of Animals (1) Clinical Observations At approximately1, 2.5, and 4 hours after test material administrationfor clinicalsigns and mortality,daily thereafterfor clinicalsigns,and twicedaily (a.m. and p.m.) for mortalityfor 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 initialdermal irritationreadingwill be made and Page 27 of 32 TP2070 Page 7 recordedas the Day I reading(Attachment1). Additional dermal irritationreadingswill be made on StudyDays 3, 7, 10, and 14. Individualdermalirritationrecordswill be maintainedfor each animal. (3)Body Weights Just beforetest materialapplication,on Days 7 and 14, and at death (whensurvivalexceedsI day) D. Pathology At terminationof the experimentalphase, survivinganimals will be euthanized.All animals,whetherdyingduringthe study or euthanized,will be subjectedto an abbreviatedgross necropsyexaminationand all abnormalitieswill be recorded. After necropsy,the animalswill be discardedand no tissues will be saved. E. StatisticalAnalyses Other than LD,,calculations(when applicable)no statistical analysesare required. 7. Report A final reportincludingthoseitems listedbelowwill be submitted: Descriptionof the test material Descriptionof the test system Procedures Dates of experimentalinitiationand termination Tabulationof mortalitydata by sex and dose level Descriptionof any toxiceffects/dermailrritation Tabulationof mean body weightsby sex and dose level LD . values by sex with 95% confidenceintervals lwhen applicable) Gross pathologyfindings/grosspathologyreport (whenapplicable) 8. Locationof Raw Data, Records,and Final Report Originaldata, or copiesthereof,will.beavailableat HWI to facilitateauditingthe studyduringits progressand before acceptanceof the finalreport. When the finalreport is completed, all originalpaperdata, includingthoseitemslistedbelowwill be retained in the archivesof HWI accordingto HWI SOP. Page 28 of 32 TP2070 Page 8 Protocol and protocol amendments Dose preparation records In-liferecords Body weights Dose administration Observations Anatomical pathology records Study correspondence Final report (originalsignedcopy) The followingsupportingrecordswill be retained at HWI but will not be archivedwith the study data. Animal receipt/acclimationrecords Water analysis records Animal room temperature and humidity records Refrigerator and freezer temperature records Instrument calibration and maintenance records Page 29 of 32 PROTOCOL APPROVAL John L. Butenhoff, PhD Date Sponsor's Representative 3M Steven M. Olaza Date Study Dire@tor Acute Toxicology Hazleton Wisconsin, Inc. Representative Date Quality Asturance Unit Hazleton Wisconsin, Inc. (TP207O,3M) TP2070 Page 9 Page 30 of 32 TP2070 Page 10 Attachment I Scoring Scale for Acute Dermal Reactions Erythema 0 - None I - Slight 2 - Moderate 3 - Severe Edema 0 - None 1 - Slight(barelyperceptibleto well definedby definiteraising) 2 - Moderate (raisedapproximatelyI mm) 3 - Severe(raisedmore than I mm) Atonia 0 - None I - Slight(slightimpairmenotf elasticity) 2 - Moderate(slowreturnto normal) 3 - Marked(noelasticity) Desquamation 0 - None I - Slight(slightscaling) 2 - Moderate(scalesand flakes) 3 - Marked (pronouncedflakingwith denuded areas) Coriaceousness 0 - None I - Slight(decreasein pliability) 2 - Moderate(leatherytexture) 3 - Marked(toughand brittle) Fissuring 0 - None I - Slight(definitecracksin epidermis) 2 - Moderate(cracksin dermis) 3 - Marked (crackswith bleeding) Page 31 of 32 HWI No. PROTOCOL AMENDMENTS SbSc@i C,@iZ37'@ Amendment No. Effective- 4-u6%JJT f4)5.@5- Portion of Protocol Being Modified: Page 2. Study Location Reason for Modification:To identify the location-wherethe study will be conducted, replace this sectionwith the followingchange: Modification: Study Location Hazleton Wisconsin. Inc. 3301 Kinsman Boulevard. Madison, Wl 53704 (G21/01-07-91) Study Director Approval: )ru.@L@ ol lhxt= Page 32 of 32 HWI No. ---50800374 PROTOCOL AMENDMENTS Amendment No. 2 Effective August 28, 1995 Portion of Protocol Being Modified: Page 6. 6. Experimental Design: B. Dose Administration:(3) Dose Administration. Reason for Modification: To correctlyidentifi-Ythe size of the gauze bandage used to wrar the area of application. Modification: Replace the seventh sentenceof this section with the following change: The area of arplicationwill be wrapped with a 4-ply gauze bandage (approximately9.5-cm x 21-cm) secured with paper tape around all edges, overwrapped with Saran Wrap*. and secured with ElastoDlastO)tape to provide an occlusive dressing. 4 Study Director Approval: -A -q!S- (G21/01-07-91)