Document 0Jjd3bOj1L7GO6rm789ELoy9k

AR226-2977 FOR DU PONT USE ONLY Study Title Delayed Contact Hypersensitivity Test in Guinea Pigs with H-19173 Study Completed On July 17, 1992 Contracting Laboratory Pharmakon Research International, Inc. P.0. Box 313 Waverly, Pennsylvania 18471 Author Susan Armondi for E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, P. 0. Box 50 Newark, Delaware 19714 Study Monitor Scott E. Loveless Medical Research No. Laboratory Project ID Haskell Laboratory Report No. 161-92 Page 1 of 60 Company Sanitized. Does not contain TSCA CBI Amended Final Report Revised Du Pont HLO 161-92 COMPLIANCE STATEMENT This study was conducted in compliance with the Principles of Good Laboratory Practice (GLP) as promulgated by the following regulatory agencies: U.S. Food and Drug Administration, as stated in the Federal Register, 21 CFR, Part 58, Friday, September 4, 1987. U.S. Environmental Protection Agency as stated in the Federal Register, 40 CFR, Part 160, Thursday, August 17, 1989 U.S. Environmental Protection Agency as stated in the Federal Register, 40 CFR Part 792, Thursday, August 17, 1989. Organization for Economic Co-operation and Development Guidelines for Testing Chemicals (OECD), ISBN 92-64-12221-4, adopted by the council at its 535th meeting on 12th May, 1981. Study Number: PH 424-DU-012-91 "To the best of my knowledge, this study was conducted in accordance with applicable Good Laboratory Practice regulations; there were no deviations from these regulations that impacted on study conclusion." JlltjQ(U(c?- (m mnOmrl^ Study Director M l / f/A,9 , I 9 2 ^ Dat/ ^ ' ' 2 Company Sanitized. Does not contain TSCA CBI Material Tested: Medical Research No. Synonym: Haskell Test Code No Pharmakon Study No.: Physical Form: Purity: Composition: GENERAL INFORMATION Test Material Du Pont HLO 161-92 H-19173 PH 424-DU-012-91 Slightly yellow liquid dispersion 100% Synonym: Submitter's Notebook CAS Registry No.: Stability: None availble In the absence of visible evidence to the contrary, the test material was assumed to be stable under the conditions of administration. Material Tested: Purity: Synonyms : Other Codes: Positive Control Material N-Phenyl-1,4-Phenylenediamine 98% 1,4-Benzenediamine p-phenylenediamine (PPD) Aldrich Chemical Co., Lot #05012MK Aldrich Chemical Co., Catalog #24139-3 -3Company Sanitized. Does not contain TSCA CBI CAS Registry No . ; GENERAL INFORMATION (CONT.) 101-54-2 Du Pont HLO 161-92 Sponsor; Study Initiated - Completed; In-Life Phase Initiated - Completed; Pharmakon Notebook No.; In the absence of visible evidence to the contrary, the test material was assumed to be stable under the conditions of administration. Du Pont Chemicals E. I. du Pont de Nemours and Company Wilmington, Delaware 10-28-91 - 07-17-92 01-28-92 - 03-07-92 - 4 - Company Sanitized. Does not contain TSCA CBI Du Pont HLO 161-92 DU PONT CENTRAL RESEARCH AND DEVELOPMENT HASKELL LABORATORY FOR TOXICOLOGY AND INDUSTRIAL MEDICINE K. D. Dastur Du Pont Chemicals Brandywine Building July 17, 1992 Delayed Contact Hypersensitivity Test in Guinea Pigs with H-19173 Attached is a report from Pharmakon Research International, Inc. (PH 424-DU-012-91). Included in the report are test methods, results, and conclusions. H-19173 at topical doses of 100% and 10% (v/v) and an intradermal dose of 1.0% (v/v), was tested on the clipped, intact skin of male and female guinea pigs. p-Phenylenediamine at a topical dose of 35% and an intradermal dose of 1.0% (w/v) was used to demonstrate the ability of the test system to detect a skin sensitizer (positive control group). In addition, a group of male and female guinea pigs was treated with 0.9% saline at induction and distilled water at challenge (vehicle control group). At challenge, a group of previously untreated guinea pigs was treated with H-19173 at topical doses of 100% and 10% (v/v), distilled water, and 35% p-phenylenediamine on separate, intact fjcin sites (negative control group). One animal died during the course of the study. Necropsy of this animal revealed no visible lesions. Theideath of this animal was considered spurious in nature and not related to treatment. During the induction phase, no to slight erythema was observed in the majority of the test article-treated animals. Following the third intradermal injection, a single guinea pig presented with a mild erythema and after the fourth intradermal injection, moderate erythema was noted in another guinea pig. Slight erythema was observed in one vehicle control animal following the third induction. No to severe erythema with/without blanching was observed in the positive control animals. During the challenge phase, no signs of erythema were observed in the test, vehicle, or negative control animals. No to moderate erythema was observed in the positive control group treated with p-phenylenediamine at a 35% concentration. Under the conditions of this study, H-19173 did not produce delayed contact hypersensitivity in guinea pigs. Scott E. Loveless, Ph.D. Senior Research Toxicologist - 5 Company Sanitized. Does not contain TSCA CBI Amended Final Report Revised Du Pont HLO 161-92 Skin Sensitization Test with H-19,173 in Guinea Pigs PH- 424-DU-012-91 Reason for Amendment: Submitted to E.I. du Pont de Nemours & Company Newark, Delaware In order to include page number on compliance statement, footnote on page 23, and correct weight range of animals on page 47. 0 i W ( . ia Susan E. Armondi, LAT Study Director 9. M ' y ' 6 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 Skin Sensitization Test with H--19,173 in Guinea Pigs SUMMARY Test Article H-19,173 at topical doses of 100% and 10% iv/v) and an intradermal dose of 1.0% (v/v), was tested on the clipped ^J-act s k m of male and female guinea pigs. p-Phenylenediamine (PPD) at a topical dose of 35% and an intradermal dose of 1.0% (w/v) was used to demonstrate the ability of the test system to detect a skin sensitizer (positive control group). During the primary irritation phase, no signs of erythema were observed m the test, vehicle, or positive control animals. One animal died during the course of the study. Necropsy of this animal revealed no visible lesions. The death of this animal was considered spurious in nature and not related to treatment. During the induction phase, no to moderate erythema was observed in the test^article-treated animals. Slight erythema was observed in one vehicle control animal at 24 hours following the third induction. No to severe erythema with/without blanching was observed m the positive control animals. , During the challenge phase, no signs of erythema were observed m the test, vehicle, or negative control animals. No to moderate erythema was observed in the positive control group treated with o- phenylenediamine at a 35% concentration. P Under the conditions of this study, H-19,173 did not cause delayed contact hypersensitivity in guinea pigs. Study Director: -P/AfTYOYn Susan E. Armondi, LAT Reviewed and Approved for Issue: M C lrO in . 11OmCYiC* Susan E. Armondi, LAT Study Director 7 Company Sanitized. Does not contain TSCA CBI Amended Final Report Revised Du Pont HLO 161-92 QUALITY ASSURANCE UNIT STATEMENT Study No.: PH 424-DU-012-91 Study Director: Susan E . Armondi The Quality Assurance Unit conducted the inspections listed below and reported the results to the study director and to management on the dates indicated. The following inspections were performed: Interval Date In Life Phase January 30, 1992 Challenge Phase March 5, 1992 Reporting Phase March 30, 1992 June 12, 1992 Amended Final July 8, 1992 Date OAU Report Issued To Study Director To Management March 31, 1992 June 12, 1992 July 8, 1992 March 31, 1992 June 12, 1992 July 8, 1992 8 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 INTRODUCTION The purpose of this study was to evaluate the potential of H-19,173 to produce delayed hypersensitivity when applied to the skin of guinea pigs. Sensitization was defined as a significant score increase at challenge over the response observed after the primary application of the test material to the test guinea pigs, or the response observed in the negative controls. A significant score increase defined as a 2-or-more step increase (e.g., from 0 to 2, from 1 to 3, etc.) has been used at Haskell Laboratory for its ability to identify compounds that are sensitizers. The sensitivity of the test system and procedures to detect chemical sensitizers was evaluated with p-phenylenediamine. MATERIAL AND METHODS A. Animal Husbandry Young adult male and female Duncan Hartley albino guinea pigs (generally 8 to 10 weeks of age) were received from BuckberG Lab Animals, Tomkins Cove, New York. The guinea pigs were housed singly in suspended, stainless steel, %" wire-mesh cages. Each guinea pig was assigned an identification card on which a unique animal number, study number, dose level, and sex were recorded and affixed to the cage. Guinea pigs were ear tagged. Purina Certified Guinea Pig Chow1* #5026 and water were available ad libitum. Guinea pigs were acclimated and observed for general health during a period of approximately five days. 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 of 22 3C (66- 77F) and relative humidity of 30 70%. Any excursions outside these ranges were of small magnitude and/or brief duration and did not adversely affect the validity of the study. B. Protocol A copy of the signed original protocol is attached as Appendix A * A preliminary range-finding test was conducted to estimate the primary irritation potential of the test material. The results of the range-finding study were used to select the exposure concentrations for the main study. The main sensitization study consisted of 3 phases: a primary irritation phase, an induction phase, and a challenge phase; During each phase, skin responses were scored according to the system presented in Table I. During the study, body weights were recorded weekly. 9 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 The range-finding test was conducted on 1 male and 2 female guinea pigs ranging in weight from 375 to 417 grams. Aliquots (approximately 0.05 mL) of 1.0%, 10%, 50%, and 100% (v/v) solutions of the test material in distilled water were applied and lightly rubbed onto four separate test sites on the clipped, intact skin on the back of each animal. Irritation responses were scored approximately 24 and 48 hours after treatment. The primary irritation phase was conducted in 10 guinea pigs (5 males and 5 females), weighing from 428 to 512 grams, by applying and lightly rubbing in 1 drop (approximately 0.05 mL) of 100% and 10% (v/v) solutions of the test material in distilled water onto two separate sites of clipped, intact shoulder skin of each animal. In addition, 10 positive control guinea pigs (5 males and 5 females), weighing from 429 to 506 grams, were treated by applying and lightly rubbing in 1 drop of 35% (w/v) suspension of p-phenylenediamine in distilled water onto the clipped, intact left shoulder skin of each animal. Five vehicle control guinea pigs (3 males and 2 females), weighing from 424 to 522 grams, were treated with distilled water in the same manner at one site only. Dermal responses were scored approximately 24 and 48 hours after application of the test material. Two days after the primary irritation phase, the induction phase of the study was initiated using the same 10 test guinea pigs in which primary irritation had been evaluated. Induction consisted of a series of 4 sacral intradermal injections (1 each week) of 0.1 mL of 1.0% (v/v) solution of H-19,173 in 0.9% saline. A virgin site on the back of each animal was used for each injection. The same injection procedure was followed for the positive control guinea pigs using 0.1 mL of a 1.0% (w/v) suspension of p-phenylenediamine in 0.9% saline. The vehicle control animals were treated with 0.1 mL 0.9% saline. Skin responses were evaluated approximately 24 hours after each induction. One animal died during the course of the study. Necropsy of this animal revealed no visible lesions. The death of this animal was considered spurious in nature and not related to treatment. Two weeks after the last induction treatment, the test guinea pigs were challenged for sensitization by applying and lightly rubbing in 1 drop (approximately 0.05 mL) of 100% and 10% (v/v) solutions of the test material in distilled water onto two separate sites of clipped, intact shoulder skin. The positive control animals were challenged for sensitization by applying and lightly rubbing in 1 drop (approximately 0.05 mL) of 35% (w/v) solutions of pphenylenediamine in distilled water onto the clipped intact left shoulder skin. The sites used during challenge were the same used on each animal during the primary irritation phase, respectively. The vehicle control animals were treated with distilled water in the same manner. Also, 2 male and 3 female previously untreated guinea pigs, weighing 415 to 509 grams at study initiation, were treated by applying 1 drop each (approximately 0.05 mL) of 100% and 10 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 10% (v/v) suspensions of the test material in distilled water onto two separate sites of clipped, intact skin and served as negative control animals. These animals also received 0.05 mL of distilled water and 0.05 mL of p-phenylenediamine (35%) in distilled water on separate, intact skin sites. These animals also served as negative control animals for the vehicle and positive control materials. C. Records Retention All raw data and the final report will be stored in the archives of Pharmakon Research International, Inc., Waverly, PA. RESULTS AND CONCLUSIONS Weekly body weights are presented in Appendix B. There were no statistically or biologically significant differences observed in the group mean body weight data among the four dose groups. A copy of the original raw data is presented in Appendix C. One test article-treated animal died during the study. Necropsy of this animal revealed no visible lesions. The death of this animal was considered spurious in nature and not related to treatment. In the range-finding test, no signs of erythema were observed at any test site. Based on the results of the range-finding study, 100% and 10% concentrations were used for the main study. During the primary irritation phase, no signs of erythema were observed in any of the test, vehicle, or positive control animals. Individual animal data are presented in Table II. During the induction phase, no to moderate erythema was observed in the test article-treated animals. Slight erythema was observed in one vehicle control animal at 24 hours following the third induction. No to severe erythema with/without blanching was osberved in the positive control animals. Individual animal data are presented in Table III. Individual animal data are presented in Table IV. Dermal responses observed during the challenge phase are summarized in Table V. During the challenge phase, no signs of erythema were observed in the test, vehicle, or negative control animals. No to moderate erythema was observed in the positive control group treated with pphenylenediamine at a 35% concentration. Under the conditions of this study, H-19,173 did not cause delayed contact hypersensitivity in guinea pigs. 11 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 TABLE I SCORING SYSTEM USED TO EVALUATE SKIN RESPONSES Score 0 1 2 Skin Reaction No erythema or edema Slight erythema (barely perceptible, usually nonconfluent) Mild erythema (well defined, usually confluent) 3 Moderate erythema 4 Severe erythema (beet redness, with or without edema, eschar formation or necrosis) If edema, blanching, necrosis or eschar formation occurred, they were indicated using the following codes: ED = Edema N = Necrosis E = Eschar B = Blanching SN = Superficial Necrosis 12 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 GUINEA PIG NUMBER 1531 1532 1533 1534 1535 1536 1537 1538 1539 1540 TABLE II PRIMARY IRRITATION PHASE SKIN RESPONSES OBSERVED IN TEST GUINEA PIGS FOLLOWING TOPICAL EXPOSURE TO H-19.173 LEFT FRONT 100% 24 hr. 48 hr. RIGHT FRONT 10% 24 hr. 48 hr. 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 13 Company Sanitized. Does not contain T SCA CBI Amended Final Report Du Pont HLO 161-92 TABLE II (Cont'dl PRIMARY IRRITATION PHASE SKIN RESPONSES OBSERVED IN VEHICLE CONTROL GUINEA PIGS FOLLOWING TOPICAL EXPOSURE DISTILLED WATER GUINEA PIG NUMBER LEFT FRONT Distilled water 24 hr. 48 hr. 1551 00 1552 00 1553 00 1554 00 1555 00 14 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 TABLE II fCont'dl PRIMARY IRRITATION PHASE SKIN RESPONSES OBSERVED IN POSITIVE CONTROL GUINEA PIGS FOLLOWING TOPICAL EXPOSURE TO P-PHENYLENEDIAMINE GUINEA PIG NUMBER LEFT FRONT 35% 24 hr. 48 hr. 1541 1542 1543 00 00 00 1544 1545 00 00 1546 1547 1548 0 0 . .0 0 0 0 . 1549 00 . 1550 00 15 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 TABLE III INDUCTION PHASE SKIN RESPONSES OBSERVED IN TEST GUINEA PIGS 24 HOURS FOLLOWING INTRADERMAL (ID1 INJECTIONS OF 1.0% H-19.17^ GUINEA PIG NUMBER ID1 (LEFT) ID2 (RIGHT) ID3 (LEFT) ID4-- (RIGHT) 1531 1532 1533 1534 1535 1536 1537 1538 1539 1540 00 00 00 00 00 10 00 00 00 00 1 1 1 2 1 1 1 1 0. 1 1 1 1 1 1 1 1 1 3 1 16 Company Sanitized. Does not contain TSCA CBI Amended Final Report DU Pont HLO 161-92 TABLE III (continued1 INDUCTION PHASE SKIN RESPONSES OBSERVED IN VEHICLE CONTROL GUINEA PIGS 24 HOURS FOLLOWING INTRADERMAL fID) INJECTIONS OF 0.9% SALINE GUINEA PIG NUMBER IDI (LEFT) ID2 (RIGHT) ID3 (LEFT) ID4 (RIGHT) 1551 1552 1553 1554 1555 00 00 00 00 00 0 0 1 0 0 0 0 0 0 0 17 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 TABLE III INDUCTION PHASE SKIN RESPONSES OBSERVED IN POSITIVE CONTROL GUINEA PIGS 24 HOURS FOLLOWING INTRADERMAL fIDl INJECTIONS OF 1.0% p--PHENYLENEDIAMTNE GUINEA PIG NUMBER ID1 (LEFT) ID2 (RIGHT) ID3 (LEFT) ID4 (RIGHT) 1541 1542 1543 1544 1545 1546 1547 1548 1549 1550 01 3 0 1 .1 10 1 00 2 00 3 01 1 00 4B 00 2 00 2B 00 3 3 3 3B 4B 3 2 3B 2 4 3B B = Blanching 18 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 TABLE IV CHALLENGE PHASE SKIN RESPONSES OBSERVED IN TEST GUINEA PIGS FOLLOWING TOPICAL APPLICATION OF H-19.173 GUINEA PIG NUMBER LEFT FRONT 100% 24 hr. 48 hr. 1531 00 1532 dd 1533 00 1534 00 1535 00 1536 00 1537 00 1538 00 1539 00 1540 00 d = denotes animal dying on study. RIGHT FRONT 10% 24 hr. 48 hr 00 dd 00 00 00 00 00 00 00 00 19 Company Sanitized. Does not contain TSCA CBi Amended Final Report Du Pont HLO 161-92 TABLE IV (continued) CHALLENGE PHASE SKIN RESPONSES OBSERVED IN POSITIVE CONTROL GUINEA PIGS FOLLOWING TOPICAL APPLICATION OF P-PHENYLENEDIAMINE GUINEA PIG NUMBER 1541 1542 1543 1544 1545 1546 1547 . LEFT FRONT 35% 24 hr. 48 hr. 23 22 12 00 01 00 11 1548 1549 1550 11 12 01 20 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 TABLE IV fcontinued CHALLENGE PHASE SKIN RESPONSES OBSERVED IN VEHICLE CONTROL GUINEA PIGS FOLLOWING TOPICAL APPLICATION OF DISTILLED WATEP VEHICLE GUINEA PIG NUMBER LEFT FRONT Distilled Water 24 hr. 48 hr. 1551 00 1552 00 1553 00 1554 00 1555 00 21 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 TABLE IV (continued) CHALLENGE PHASE SKIN RESPONSES OBSERVED IN NEGATIVE CONTROL GUINEA PIGS FOLLOWING TOPICAL APPLICATION OF H-19,173 , P--PHENYLENEDIAMTNE AND DISTILLED WRTF.P GUINEA PIG NUMBER LEFT FRONT H-19.173 100% 24 hr. 48 hr. 1556 0 0 1557 0 0 1558 0 0 1559 0 0 1560 0 0 RIGHT FRONT H-19.173 10% 24 hr. 48 hr. LEFT REAR PPD 35% 24 hr. 48 hr. 000 0 0000 0000 0000 0000 RIGHT REAR Distilled Water 24 hr. 48 hr. 00 00 00 00 00 22 Company Sanitized. Does not contain TSCA CBI Amended Final Report Revised Du Pont HLO 161-92 Table V Summary of Skin Responses Challenge Phases! Positive Control Test Animals Responses No erythema Slight erythema Mild erythema Moderate erythema PPD 35%a 24 hr 48 hr 4/10 2/10 4/10 4/10 2/10 3/10 0/10 1/10 H --19,173 100%a ~ io%b 24 hr 48 hr 24 hr 9/9 9/9 9/9 9/9 0/9 0/9 0/9 0/9 0/9 0/9 0/9 0/9 0/9 0/9 0/9 0/9 Responses No erythema Negative Control H-19,173 PPD Vehicle Control 100%a 10%J 35%( Distilled Water** 24 hr 48 hr 24 hr 48 hr 24 hr 48 hr 24 hr 48 hr 5/5 5/5 5/5 5/5 5/5 5/5 5/5 5/5 Vehicle Control Distilled Water 24 hr 48 hr No erythema 5/5 5/5 a = left shoulder b = right shoulder c = left flank d = right flank 23 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 APPENDIX A PROTOCOL AND PROTOCOL AMENDMENTS a- 24 Company Sanitized. Does not contain TSCA CBI Amended Final Report 1 Du Pont HLO 161-92 PH ARM AKO N RESEARCH IN TERN A TIO N A L. WAVERLY, PENNSYLVANIA 18471 _. , 1 Protocol - 424D1 INC. PHONE (7171586-2411 Skin Sensitization Test in Guinea Pigs (7171586-3450 Sponsor: E.I. du.Pont de Nemours & Company P.0. Box 50 Elkton Road Newark, DE 19714 Testing Facility: Pharmakon Research International, Inc. Waverly, Pennsylvania 18471 Test Facility S.O.P. No.: PH-424 Study N o .: Purpose of the Study: To be assigned upon study initiation. This study is designed to evaluate the dermal sensitization potential of the test material in guinea pigs. Ownership of the Study: The sponsor owns the study. All raw data, analysis, and reports are the property of the sponsor. Study Monitor: Scott E. Loveless, Ph.D., E.I. du Pont de Nemours & Company Study Director: Susain Armondi, LAT, Pharmakon Research International, Inc. O.A.U. Responsible Personnel: Leslie J. Pinnell, M.S. Dates of Performance: Study will begin within one month of the receipt of the test chemical and authorized protocol. Good Laboratory Practices Statement: This study will be conducted in compliance with the U.S. Environmental Protection Agency Toxic Substances Control Act Good Laboratory Practice Standards (40 CFR Part 160) or Good Laboratory Practice Standards (40 CFR Part 792).. IACUC Statement: Protocol-424D'1' has been reviewed by the Institutional Animal Care and Use Committee (IACUC) and complies with acceptable standard animal welfare and humane care. 25 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 2 Protocol-42401 Delayed Contact Hypersensitivity in Guinea Pigs (Buehler) Tentative Dai- of Submission of Final Report- ; Records Maintained; Raw Data: Record Retentinn: Analytical Chemistry; trhneehilnn-?fephiafse*3of5otlhleowsitnugdyt.he completinyg of All raw data, protocol will Archives. final reports, be maintained documentation and in the Pharmakon Feed Lot Number Body weights, initial weekly and final Compound preparation Volume administration Observed signs Observed skin reactivity Maintained in a Standard Pharmakon Notebook All raw data and completed notebooks. taai " " .OM,uy Acy .m t:ne test article and test article/carrier mixture is the responsibility of the sponsor. If reouested Vnr IInncc. Sw5ilSl0&thrroPhuaghhn^itis0nsuRbecsoenatrrcahctIonrt,ercnoantdiuoctn-al,Y appropriate analytical analysis and will indicate the additional cost involved fol inw ^ ^TcaiPP a!*d evaluation of the appropriate analytical method. In the case where a 5 satisfactory method is not provided, Pharmakon Research International, inc. or its * subcontractor, at additional cost to the sponsor, will develop appropriate methods. Species: Strain: Supplier -(Sourcei: Sex: Agej. frt?c?0 /PnSr el(rcts to analyze the test article/carrier mixtures, Pharmakon Resea i"- "U 1 " U e c r S e S S e at appropriate designated intervals durinc the the s" pl*s TEST SYSTEM Guinea pig Hartley BuckerG or from Lab any Animals, other U. Tomkins Cove, New York S.D.A acceptable source. Male and Female (the sex used will be S o " ) ln the StUdy Young adult 053191 26 Company Sanitized. Does not contain TSCA CBI Amended Final Report ^ ,,,,. Du Pont HLO 161-92 Protocol-42401 3 Delayed Contact Hypersensitivity in Guinea Pigs (Buehler) Weight at. Initiatinn: No. on Studyr Method and Justification fnr Randomization ; 400 -- 650 grams uose'-Range-Finding: 3 studv1Mn?tr0li 5 (at the discretion of the S ' thlS group be excluded fron NenoVe control: 5 ( re-challenge is a second set of 5 naive, age- u s e d ^ negative control animals will be Test article: 10 PO ^ enntf0l: 10 (at the discretion of the Study Director after consultation with the fromythens t S ; o hiS 9rUP * * Y excluded Dositinn?nPS " P 1 b e .housed bY vertical cage cclliinniiccaall ssi?gTMns*2d*urianngimaaclcslismhaotwiionng weivlildebnece of fr" randomization. Randomization . rried out by use of a computer generated random number table. Before treatment commences, all guinea pigs will be Weighed, according to body weight and assigned to SummbbeSrsr !sogthuaUtlSeaUcshintgreaattmaebnlte gorfourpanwdiolml have we?hlarcd^Strib"tion according to body ' "lnea beyond the extremes of the ttrrefatamtenSthgtroruapnsg,e Wiilf1thneortebeisaasssitganetidsttoical!,, b^fSlflCant difference in mean body weight Acclimation Period : System of Identi fication ? c = S "S f Mil" ls "U 1 be "sea *> Minimum: five (5) days -- mui.jveu wicn an U I U >U group number tagged?3t*ment grUP' Guinea ^ are ear Research Facility Registration: Animal Rooms: HUSBANDRY U.S.D.A. Registration Animal Welfare Act 74 uauef m e SC 2131 et seg rT^i isolation by test system. Temperature/Humidity*?^ E v I ^ ' a t e m p ^ i u ^ ' -? s -" 3c` 053191 27 Company Sanitized. Does not contain TSCA CBI Amended Final Report Protocol-42401 Delayed Contact Housi ng- Du Pont HLO 161-92 4 Guinea pigs are housed individually +- cage xn stainless steel iire i e L tW per Sxze xn accordance with the " o H d f * cages- and Use of Laboi*at-rrr-^r for the Care Saniti 7:a-MnnFood: and f e e l e j r w L r i e s a n S S f e J e ^ w o ^ I e L Food Analysis: Water-: WaterAnalvsis: Rationale fnr Test System: = o ,, sid e rS ?\n i *""> belixfevveedaUrtoorbbeS wwixtthh!xnCaocncteapmtianbalnet liiemvietlss. are "Sn?rLn?hfSiSSnStiS"f f tobe capable of interferiSr with he t0 be .conduct of the study.? th th purpose or Fresh tap water, ad libitum. Water is monitored for ntervals Procedure according PH-018. to ^ ami^ants at periodic Standard Operating METHODS *' - i h f e l i j ` a i d1K e l o de M o " b tio ,, exposure to the t e S ,, , "S se fr" Rationale fnr Dose Selection: Justification fnr .Test System- Compound Preparation- y - S / o ^ S ^ o S o " ; r e S ? 5* - 2-00P . He? ^ f S f - * G u id e lin e s historically been used pig has chemicals for dermal sensitaSom f Liquids Liquids will be applied a s t-onr-; j when specific instructions r l ^ H t kit*?-Pt mxxxng. The use of a vehicle^on ^ ^ lublon or bep desotib,d in the study records ^ d "^ " 111 053191 28 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 Protocol-4 24D1 Delayed Contact Hypersensitivity in Guinea Pigs (Buehler) Solids The test material will be pulverized if appropriate. The test material may be moistened with distilled water or a suitable vehicle to ensure good contact with the skin When a vehicle is used, the influence of the' vehicle on penetration of the skin by the test substance will be considered. Absorption of the Test Material : Vehicl p.; Semi-Sol irig .. Extremely viscous liquids may be heated to approximateiy 50*c to dosing, with the approval of the study Monitor. The procedure used will STSSTrSoS? StUdyr"0rtS"a For the purpose of this study, all of the test ro Kriai applied to test sites is asIumSd to be absorbed by the test system. All calculations and the evaluation of effects will be based on the applied dose. J"L ^ehfcle varies, depending on the f the.test substance. Commonly used vehicles are distilled water, saline, ethanol:distilled water (80:20, v/v) , dimethvl phthalate, acetone:dimethyl phthalate (l-g Y r Propylene glycol. The vehicle u^ed fiial report?ented 10 the StUdy records'a.nd the Volume Administration : Route of Administration ; Rationale for Route of Administrate on ; Approximately 0.05 ml/dose " d control materials will be applied topically and/or injected intradermally, to the clipped, intact shoulder skin. Y . Skin contact is a possible worker and consumer exposure route. TM er ^-.and Description Dose-Range-Finding: 3 nOrf ASnmiinmaailsr-- per-- Dose Group: _1 . _ ^ Vehicie control: 5 (at the discretion of the Study Director, this group may be excluded from the study). Negative control: 5 (if re-challenge is necessary, a second set of 5 naive, age- matched negative control animals will be used.} Test article: 10 Positive controi: io (at the discretion of the Director after consultation with the fto"',t S ns S ; . r ' ls 9roup "*Y be excludsd 053191 29 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 Protocol-42401 6 '" w a " Method of Rt-n^y Performance. " *** ^ * - . Plg. (Bushler, igf?=E?ag^ rrior to initiation of range-finding study S i l ^ L das ^StUdy' a dse- of t0 S a i s nv m SbUdieS 33 SUbjeCted back and sides* Ihe ? S i YmCiip?ed over the concentrations `w i n b e ^ L e * of the test compound (soffrif 01\ bhe solubility the maximum soluble cincent?atioi be.tested at lower concentrations) t n and at 2 or 3 at 100% (as received or S ^ r '"i1 be testad Sponsor's instructions) andPfrPtred per the concentrations. Each cru?nf=> ^ r 3 lower with up to 4 d i f f e r e n t ^ TM Plg ? ay be dosed different sites n TOr, oncentrations at some cases where (tta 5 S ^ i tl0, ite> In common solvent, it will bf soluble in a . by topical application teSted as r e i v e d appropriate test site a ^ ^ v h K ^ 601117 on the the end of the dosing s v r i S S ^ 13 gently with of approximately 25 in COVer an area Observations for s i a ^ diameter. ssA srs 'J r * sxtes ,,in be soored ao=oSin5l?S'T2 L tifate<i M m a i r Trritatinn ' ' per a U " 2 the middle of the back onf -h shoulder and the highest slight to non-irrfi^-Wl11 receive concentration as deteminZrt J tatlng range-finding study. The se>^TO* tbe dose~ receive one-tenth of t-hpf ond site will site will receive l d r o ^ r*nCenbration Each 1) of the appropriate ~niapp5xilnate1}'' *05 gently i,,to S f '' TM " * i the dosing syringe to cover an S the nd Qf approximately 25 mm in d i a L f area control gninoo pigs, " / S f i h l 'S r hicle a *- 1 site as above usinrr +-k w*. Seated positive control guinea^ijl nly- The 1 site with the p o s i t i v e c n ^ ^ b treated at control guinea pigs W i11 ol Tbe negative the challenge phase. o b s e ^ ? unticated until dermal irritation w i n h ! T fc ns for signs of hoars after a p p U c i S n ? r'=rd>a 24 L d 053191 30 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 7 Protocol-42401 Delayed Contact Hypersensitivity in Guinea Pigs (Buehler) 161-92 -ucna-i-L-i-^d--LlO-n- - XUUUC.L1U11 } T"f flft induction application will be made ' *ftef.the 18_hour dermal observation is made for the primary irritation phase. If the test material is soluble in an appropriate vehicle ' ?*9% saline or dimethyl phthalate, ' induction will be performed by intradermal injection. If the test" material is insoluble in tne conuaonly used vehicles a topical induction procedure will be used. The hair at the appropriate application site will be clipped on the day prior to each application or, if a topical induction procedure is employed, at least weekly during induction. After clipping, the area to be a?aL b^ eline?ted for the convenience of and at the discretion of the study director or the experimenter. The hair will be clipped as needed during the study for evaluation of the dermal irritation. A total of 4 intradermal injections will be made, 1 per week at l week intervals, a virgin site will be used for each injection. A 0.1 ml aliquot of a 1% solution of the test material in an acceptable solvent (e.g., 0.9% saline dimethy! phthalate, e t c , wifl'be almlSsSered by intradermal injection to the clipped- sacral area of each test guinea pig. if distilled water is used as a vehicle in the primary irritation phase, 0.9% saline will be used for the intradermal injections. The vehicle control guinea pigs will be treated as above using the vehicle only. The positive control guinea pigs will be treated as above using the positive control material. - Topical Administratenn ` A total of 9 applications will be made to the sacral area, 3 applications per week for 3 weeks, one drop (approximately 0.05 ml) of a minimally irritating concentration of the test material in the acceptable solvent (e.g., dimethyl phthalate, 0.9% saline, etc. will be ^ ^ d -dir?fily to the skin *nd then gently rubbed m with the end of the syringe, to cover an area approximately 25 mm in diameter on each test guinea pig. The vehicle control guinea vphfrT111 Feceive similar treatments using the vehicle only. The positive control guinea piqs 1 receive similar treatments using the positive control article. 053191 31 Company Sanitized. Does not contain TSCA CBI Amended Final Report 8 Du Pont HLO 161-92 Protocol-42401 Delayed Contact Hypersensitivity in Guinea Pigs (Buehler) Challenge Phase ttrreeaattmmennt,tethhe 1c4huadl?lfengaeftterreathtemelnatswtililndbuection administered. The test material will be administered in the same manner as in the primary irritation phase, treating with the same concentrations at the same sites. Following the same procedure the negative control animals will 'be treated with the same f thf test material, and may be treated with the vehicle and positive control material (up to 4 separate test sites). The vehicle control animals will be treated with the vehicle only. The positive control animals w i n raGeive the positive control article, if the test results are conclusive and the test material has either clearly caused allergic sensitization or is not a sensitizer, the test is terminated and the guinea pigs are euthanized. If the reactions are less than conclusive, a second challenge (rechallenge) may be performed approximately l week after the challenge using the same test guinea pigs and additional naive control guinea pigs. The vehicle and positive control animals may not be rechallenged. 1 Method Validation/ Positive Control ; The procedures and test system will be evaluated with a known dermal sensitizer to ` validity.of the test. Evaluation of the test system with a known dermalsensitizer may not be conducted concurrentlv 2 TM ? Very testing with a known dermal sensitizer will be performed periodically. Evaluation of Dermal Response; Dermal evaluations will be made between 23-25 ?r 47-49 hours following treatment but the data will be reported as the 24- or 48our evaluations. . Intervals; a. Primary Irritation Test - Dermal Wil1 t made approximately applicaiion?UrS test .b. Induction - Dermal evaluations will be made approximately 24 hours after each induction treatment. c. Challenge and Rechallenge (if reauirPdi Dermal evaluations will be made ^ } approximately 24 and 48 hours after the test material applications. 053191 32 Company Sanitized. Does not contain TSCA CBI Amended Final Report 9 Protocol-42401 Delayed Contact Hypersensitivity in Guinea Du Pont HLO Pigs (Buehler) 161-92 Interpretation nf Results: All test sites will to 4 as follows: be graded on a scale of 0. TABLE I Score 0 1 Skin Reaction No erythema or edema Slight erythema (barely perceptible, usually nonconfluent) 2 Mild erythema (well defined, usually confluent) y Score 3 4 Skin Reaction Moderate erythema Severe erythema (beet redness, with or without edema, eschar formation or necrosis) CUCI.CI' oJ-ancnmg, necrosis, superficial necrosis _or _eschar formation occur, they will r*u =beEdlenmdalcated using the following code: N = Necrosis E = Eschar B = Blanching SN = Superficial Necrosis Sensitization Ratina: sensitization is defined as a 2-or-more step fram 0 to 2/ from 1 to 3, etc ) " J 1 irritation scores in the test animals challenge6 5 T rV?* the ^gative c o n t r l t challenge. When the test and negative control animals have similar dermal irritation scores the test material will not be considered a ' " r * ThS rating of a sensitizer is as Ratio of Responsers 1 to 2/10 3 to 4/10 Ratio of Responsers 5 to 6/10 7 and above Decree of Sensiti Weak Sensitizer Mild Sensitizer Degree of Sensjtizaf-ipm Moderate Sensitizer Strong Sensitizer 053191 33 Company Sanitized. Does not contain TSCA CBI Amended Final Report 10 Du Pont HLO Protocol-424D1 Delayed Contact Hypersensitivity in Guinea Pigs (Buehler) 161-92 Necropsy; Euthanasia: Statistical Analysis: Recha1lenae: >* Positive Control Article: Supplier (Source): Special Handling Instructions : Analysis of Puritv: How Supplied: Rationale for Positive Control Article: This rating scale is not fixed rigidly. S<^ ? niific valuati?n f the data and judgement will be used to decide whether a material will be classified as a weak, mild, etc. sensitizer. Pathological evaluations of test animals will not ordinarily be performed. Exceptions will be noted in the study records and final report. Animals that survive the test period will be euthanized according to the Testing Facility's Euthanasia SOP and incinerated. Statistical analyses of data will not be performed, unless requested by the sponsor. ." -- w a aiijLiaaxs uiat are considered sensitized. Verbal instructions for rechallenge may be given by the sponsor, followed by written confirmation. All test a$ticle anfmalsf along with an additional group of four naive guinea pigs, can be rechallenged six (6) days after primary challenge, but not before. The negative and positive control groups will not be rechallenged. POSITIVE CONTROL ARTICLE N-Phenyl-1,4 phenylenediamine (PPD) Aldrich Chemical Company, Milwaukee, Wisconsin Standard precautions The purity is the responsibility of the manufacturer. 100 gm bottle As per sponsor's request. ACT/PT3 053191 34 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 11 Protocol-42401 Delayed Contact Hypersensitivity in Guinea Pigs (Buehler) Test Article (Name or Godei : u hni Chemical Abstract No. or Code N o .: Analysis of Puritv/Stabilitv: Carrier Mixtures: Analysis of the purity and stability of the test article is the responsibility of the sponsor. Analysis for stability, uniformity and correctness of concentration of the test article in the carrier is the responsibility of the sponsor. ___ Return Test Article Carrier Mixtures to the Sponsor V* Dispose of Test Article Carrier Mixtures Person to whom carrier mixtures should be sent: Shipping Instructions: Government Agency Submission:_____FDA % TSCA FIFRA _____ EEC _____ OECD ____ MHW _________ _______________Other AMENDMENTS Date Date lopmi APPROVAL OF PROTOCOL Study Monitor Study Director 053191 35 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 12 Protocol-42401 Delayed Contact Hypersensitivity in Guinea Pigs (Buehler) APPENDIX A I Identification: Test Article Information Test Article (Name or Code) : B iCU~13_____________ _ Lot or Sample No. :__________________________ _________ Physical Description: 5\rckH, Purity :______________ lool? * u Expiration Date:_______________ _________________ DeasSTy/Specific Gravity: Solubility (check one) : Water Pijp^Ue- Acetone_______ Ethanol_____ Corn Oil r DMSO__________ Other (please specify)_________________ Chemical Classification: Flammable____ Corrosive_____ Other_______ ___ H Storage Information: Material Storage (check one): Room Temperature XI ; Refrigerator _________ Freezer _______ ; Other (specify)_______________ III Handling Information: Known Hazards: { H o ( L - v ,ryt Precautions: Routine use of protective clothing includes laboratory coats, latex gloves, dust masks, and safety glasses. Other recommended precautions________________________ ___ In Case of Emergency Related to this substance, contact: Sot* (person) IV Disposition: _________ o f C&xrQ (company/division) at ^OZ- 4-T\ -foU (phone number) All materials will be returned to the sponsor three months following submission of the final report to the sponsor. Person and address to whom test articles are to be returned. Name:_______ Address: 36 Company Sanitized. Does not contain TSCA CBI Amended Final Report Protocol Amendments Skin Sensitization Test in Guinea Pigs PH 424-DU-012-91 Test Article: H-19,173 Du Pont HLO 161-92 Records Maintained: (page 2 of protocol, line 8) Original Statement Body weights, initial weekly and final Corrected Statement Body weights, initial, weekly and final (including means and standard deviations) Reason for Amendment: As per sponsor's request. Evaluation of Dermal Response: (page 8 of protocol) Original Statement Dermal evaluations will be made between 23-35 hours and/or 47-49 hours following treatment, but the data will be reported as the 24- or 48- our evaluations. Corrected Statement Dermal evaluations will be made between 23-25 hours and/or 47-49 hours following treatment, but the data will be reported as the 24- or 48-hour evaluation. Reason for Amendment : Typographical error. Compound Preparation: (page 5 of protocol) Original Statement Semi-Solids Extremely viscous liquids may be heated to approximately 50C to dosing, with the approval of the Study Monitor. The procedure used will be documented in the study records and included in the final report. 37 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 Protocol Amendments Skin Sensitization Test in Guinea Pigs PH 424-DU--012--91 Test Article: H-19,173 Reason for Amendment: Method and Justification for Randomization: (page 3 of protocol) Corrected Statement Semi-Solids Extremely viscous liquids may be heated to approximately 50C to facilitate dosing, with the approval of the Study Monitor. The procedure used will be documented in the study records and included in the final report. Typographical error. Original Statement Treat groups will be housed by vertical cage positioning. Reason for Amendment: Corrected Statement Treatment groups will be housed by vertical cage positioning. Typographical error Susan E. Armondi, LAT Study Director Pharmakon Research International, Inc. 38 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 APPENDIX B WEEKLY BODY WEIGHTS fcM 39 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 WEEKLY BODY WEIGHTS ifl) Animal Number Sex Initial Test Group Week II Week III Week IV Week V Week VI Final 1531 M 452 1532 M 512 1533 M 482 1534 M 483 1535 M 507 481 572 573 558 555 539 579 612 670 680 648 719 783 d 748* 638 621 717 788 797 612 675 719 692 732 613 658 702 757 766 Mean S .D . N 487 24 5 1536 F 446 1537 F 466 1538 F 428 1539 F 496 1540 F 452 548 38 5 506 509 448 548 489 610 650 707 727 744 . 43 53 61 55 50 5 5 5 44 537 572 625 654 683 558 574 634 648 668 492 534 558 583 579 593 610 669 710 713 509 549 588 620 641 * & I 2 CO Mean .D. 458 500 25 36 5 '5 538 568 615 643 657 40 29 43 47 51 5 5 5 55 = denotes animal dying on study = weight upon death, data not included in statistical analysis 40 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 WEEKLY BODY WEIGHTS fGi Animal Number Sex Initial Week IT Vehicle Control Group Week III Week IV Week V Week VI Fi nal 1551 M 519 1552 M 461 1553 M 522 576 518 593 642 710 772 825 840 581 613 668 712 739 668 713 780 830 845 Mean S.D. N 501 562 34 39 33 630 679 740 789 808 45 57 62 67 60 3 3 3 33 1554 1555 F 424 F 465 457 525 506 524 557 597 568 605 608 624 666 694 Mean S.D. N 445 491 29 48 22 532 561 596 636 651 36 52 40 43 61 2 2 2 22 41 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 WEEKLY BODY WEIGHTS G1 Animal Number Sex Initial Week II Positive Control Group Week III Week IV Week V Week VI Final 1541 M 506 1542 M 503 1543 M 494 1544 M 464 1545 M 499 557 532 544 511 548 616 666 714 746 776 584 617 668 691 710 588 637 677 721 746 571 611 674 710 732 602 647 653 683 697 Mean S.D. N 493 538 17 18 55 592 636 677 710 732 17 22 23 25 31 5 5 5 55 1546 F 429 1547 F 462 1548 F 471 1549 F 457 1550 F 499 460 533 534 488 542 522 526 591 594 633 558 592 627 657 679 590 618 694 744 753 534 536 585 608 599 589 642 683 723 744 Mean S.D. N 464 511 25 36 55 559 583 636 665 682 31 51 .51 67 67 5 5 5 55 42 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 WEEKLY BODY WEIGHTS (G Animal Number Sex Initial Week IT Negative Control Group Week ITT Week IV Week V Week VT Fi nfi1 1556 M 482 1557 M 509 546 575 619 663 742 801 812 636 679 761 826 839 Mean S D N 496 561 19 21 22 628 671 752 814 826 12 11 13 18 19 2 2 2 22 1558 F 415 1559 F 486 1560 F 447 454 548 468 485 497 530 556 571 592 650 664 725 754 525 526 587 563 608 Mean S.D. N 449 490 36 51 33 534 558 594 615 644 54 81 67 96 97 3 3 3 33 43 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 APPENDIX C COPY OF RAW DATA 44 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 PHARMAKON RESEARCH INTERNATIONAL, INC. Sponsor: f ~r Skin Sensitization Test in Guinea Pigs Dose-Range-Finding Study ` AJesmarts a. a?. P u r p o s e - ^ ^ A. Study No.: P / / V Z * / ~ b u -Q /-f/ _____________ Method: "Aq 77z&Tce6_ *3^2) * ______ Date of Initiation: / A W ? ^ Date of Termination = lllh/fa Test Article^ ' -- /?, /*73 ----- ~~ Description: - ! ' v/*.--^ Amount Received:---- ($ , i Z y (&io*s) Date submitted: /o/zjjA,__________ V e h i c l e ^ c * . -------------- Dose Levels: /->, ^ 2, Route of Administration: % ) / _______ Animal P.0. #: ^0 fl~ 0(03*}^ /3 W. Food Lot #: /OJL3 Ci IA Strain : Identification: d'y e /Uh3aC /Up. No. of Animals on Study:__.3 __________ SSeexx:: MMaallee / Female ^ Scale#: 5 ) / ^ d ) 3,,c>w,. Light Cycle Checked: / / / ^ . ^ Animals Clipped: ///ifcl.3^_____________ ATABL3(1) /juAA'jf} Qjwnrhrl^Study Director A/6> ?,<? Daittee jJlkillL. Date ` 45 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 PHARMAKON RESEARCH INTERNATIONAL, INC. fim \N.B. 8 Skin Sensitization Test in Guinea Pigs Page , / P-H-s-^-~Ran9e-Fi ^ i ng Study cAfsgC & / M f f 2 J S & J In this test, both sides of the animal are shaved anoTfexposed to'four concentrations of the test material. The different concentrations are applied concurrently to four different test sites. A given concentration is applied to a different test site on each guinea pig so as to minimize variation in response due to dfferent skin locations. The format for application of the test article is described below: Site 1 = /, O?t> Site 2 = /O 70 Site 3 = 5 - 0 yo Site 4 =/cr2?yo Animal Test article No. Sex and dose / 6* y / ' / ? / 7 3 _ 3 - 1 1 /-C? />. A*>ct /era7,, 24 Hour Scores Initial Time of Site Site Site Wt. {gmsl Dose 1 2 3 Site Final 4 Wt. (gfes) 3 7.iT 0o o o y/7 \/ 0, 0 o | O d o(/>Pm 0 , 0 0 10 ' / n }.. o jo 0 io 0 10 /r 48 Hour Scores o a sW 0 o 73t> 0 0 7/Z ____ ATABL3(2 Study Director Date ////>A v Date " 46 Company Sanitized. Does not contain TSCA CBI Amended Final Report Revised Du Pont HLO 161-92 PHARMAKON RESEARCH INTERNATIONAL, INC. Skin Sensitization Test in Guinea Pigs Page Title; Sponsor; , 1 . q U ihfrn-- tS>Ajfr mJLn Qo^ j T U ( ma *\i \ 1. + P c iy o _____________ _ _ purP s e d n ^ M Ji stM S i . study no.; P H 4^.4- "hn-oia--0!I i ^ k k 'd o ^ Q f1! jg i;m.i*PjT Method=R e l x A 4" Psi.-O'-Ho&H /H.D1 Technical irn;itiation; //S-Z/X^ Test Article; (H ~ l^.l ~7 3 Technical .. Termination: /p?a/f.o / f a Positive Control;w.pi^u-/ --------------- --- Lot Number; fjp j ( f y ^ 'ffiJptU Lot Number: ero /*,K OiJn.cl n,~.cL tj . Description: jU<y.,4lu ultimo J-i^ixc c^UgrpiT^e^cription: ^ Vehicle t 'bjsT, l l * J l^.iTca Amount Submitted; /%"!*. /J? Vehicle; 7xs~ .U e J u,rc* Receipt Date; //ag'/^A Route of Administration: rttcsrmnJ,, yeuhyi4iwl Animal p.o. #; 5011-0/03,93 (L.______ Toxicology L a b . N o .; ~vT____________ sP6 c ie s: j P x - f t ______________ Strain; Urt .i _________________ _ ,, . ,,, _ Weight Ranged ,-/'/Wt6Jrrbmi\c,-^e*X7*y W?*.j Food Lot #; Q/OlRS- !& i))019.2 /A No. of Animal's/on Study: ^ 0 Sex: Male / S & Female / S '$ Scale #: (_) 2 ) Light Cycle Checked i/Az/fzJZ, Animals Clipped: ^Wd!<UJf ' Ciil 7 M Format for Primary Irritation, Induction and Challenge Phases z-loiie.ni-iuA' c Itn o A v m J . C U b { , ^^h^nadum^ U \ (rmimiv^cyjj d&htL U f a / 1 1 - 2 Primary Irritation and Challenge 3-6 Sensitization Phase (Induction) , .rfhJCl ^^fives tigatoi: / U /f d 1-t/i/l.U )A a i u ^ r j . Study Director -3.J 2 I4 ,-7 Date 'hhyy Date 47 Company Sanitized. Does not contain TSCA CBI Du P o n t HLO 1 6 1 -9 2 Amended Final Report Noteboi Page PHARMAKON RESEARCH INTERNATIONAL, INC. Skin Sensitization Test in Guinea Pigs GENERAL OBSERVATIONS Study No. = PHysuj-bU-O/Zi - ? / Test Article ; A/ -/ 5^/73 Sponsor; 3T. di, / W f Date AN ** (appears normal) di Initials "1 j /m Jj k J ^ s T u t -l o / *g;2No/a. sr ~'..^ &>. * & & & ,, ? * .-Xf> / / / ? . / / 2 X - a-------------------------------------------- - ------- J - o T ^ d /SZP-/ ...... rfp / a / y 5. s2 M y / zzzzzzzzzzzT Z Z Z ZZzzzzz / ATABL3 (7) 48 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLo % i -92|| PHARMAKON RESEARCH INTERNATIONAL, INC. Skin Sensitization Test in Guines Pigs Test Article Preparation Study No.; Pl-Ma/J- O 13 Test Article: }-4 ~ ify"7 3 I C.X. d u P ctjF et Sponsor;_________ / U c w t U A S +- OtS. Primary Irritation Phase {Week I) :{>/froZ ^ L ; ?PT> P 3S~Z $ /. 7 5 ^ ^ , ??2> a * 'xae- a //>i/n.J /^C^r^crrt-- 'r{^ ~ q -- ~Jor-+-I*---Mam---1-- r J^U. r.srrrj). / ft- u Jlx.mn 7o -21^ -- ^ Induction (Week I) x( 2 />?,, ~ > oJ-*L 7 +- .^ L ^ C O ' Y 7^ /?.* -- --------------:-----;---------------------------------- 7---------------------------- ....... ....................... ^V3^?(P A o T o /sO/Hq 967js, m . y/sa/j^T^ y A Induction (Week II) *~Jfc 4X^-^_P- , :t- m ^ P . y p o 0 / - Q ?g-= ^ ^g a ?^/>-____'/ sCZLa - 12223- Induction (Week III) = ( g / / f l /*^*'/*K / M / Qsf/lfJP f ?. ?a J J h j i L i u J b U - ^ d . ^ m c a j ? - m J u ^ f m & d />/>/)/ Q }.D % - 7 / (yO, / yy%4 o<-d^ f f f i t-3. J /tf\l)/PH*l\Zh /ftrxD) t{u.uftoA._'-hhCfWr*>^J.lMiwwA^Jd I\//aa/jmmaa adjjfow o %if J /^/l Q . d t b i f o f x A t b t w L h & J L ^ u v , / m L J ^ CQ. ATABL3(3) 49 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 PHARMAKON RESEARCH INTERNATIONAL, INC. *-*rfc-*-**vcSlUJL^<tt.J.On GENERAL OBSERVATIONS Study No. : rH V a bu -O/Xi -~?7 Sponsor ; Z~. r/,, -A __ TDDaeatsteet Article; Z - / 9 /-7 ,? TT!-- r ------ -- ------PlsWUTlA.'i.i. *- a-rnjoa.^. AN = (appears normal) T n U jai ^ -- "3" ?/t?s 2 ^ 1 w ^ r ew , r ~aAo/a b/.-- j .s.z ^ v ~ . y JA 7>a -- y / / / 2 / j 9 p ' -1_ f - J7 >-/ / / , .i <*A^3mS!ms ~^q / ^ ^ s7^^/ 7 ~?7*7 z-Z - Z zzZ zzz zz i f / ATABL3(7) 50 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 PHARMAKON RESEARCH INTERNATIONAL, INC, N.B. Page Skin Sensitization Test in Guinea Pigs Test Article Preparation (Continued) Study_No,; Test Article ; M- - [ *7 3 Induction (Week IV) : (9 /,Q 7o /.t. elu PeiAjt oU_ -g?-nsor:____ X W m i / v s + Otr. O. U L ~ iZZ -- -- ------------------- 7i r>'b(D -- -------- ~** PPh SQ~< ^ /-UP >4_____ <g-- ---5^ a^iyJL^>. Challenge (Week VI) (p-S % Q.p~Uda^ZZ&L- -h -Y-Cnl' PcxUZZ^e. 3- S ^ u ^ ??j S o ,, i . - * C + c A . *J Z i ^ P7>h >.*<-% ~a> , 4n\Z~i L- ;^ U L ^ - , Rechallenge (Week VII): Y ^ . j/s'J-ii f? ATABL3(4) 51 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161-92 PHARMAKON RESEARCH INTERNAIONAL, INC. Skin Sensitization Test in Guinea Pi r,e 52 Company Sanitized. Does not contain TSCA CBI Amended Final Report Du Pont HLO 161 tyDteboi Page # PHARMAKON RESEARCH INTERNAIONAL, INC. Skin Sensitization Test in Guinea Pine Body Heights (grams) TestSArtJi^ ^ study Lot Number: Initial dumber Sex Week I Week II Week III / w U 6L L ttc z 'y - V V SSV 3 4 6*~oG <? Vo 3 r 567 l/>)0 rt sn /.w lP f(p r r Y . s n V f? V *? f/, 51! IsOcK i^ 7 v o x S3 66? / -- s-m*r--/-r-- /V 9 *h ? y s" 7 ftf &d 637 /S ~5~0 $ Y ff z/M/fl Js~i 6 rt > -/>6 /&b> Week IV Week V Week VI >00 0 /7 ir t (ff(o% 760 09? 776 I/O 037 0// 677 674 77! 7/ Y7 S7U0? 1 s7y7~7Y 0 6 7 LS3 / A 3 Y0^7s 7/ 6^0 5 7 / 07R7 S?3 O /Y 03.7 6?y 067 767 0^ 7yy? 767? 6& 0Y2 Z/ff 76 073 Yzs'ftv- (U4! o<% YS W7 * 703 TW J-g -te li % A )e ^ u c h n i ( Y . T rW ? /si r Y S ^ - > /3>S~7 " V o ? 6 6 6 O rt 030 /52f /537 7s& t $ Y/S^ Y Y 776 e> + Y%C> 6 6 * o /- YY7 //* 6/ii. i7/*7 Y? 626 'h-' 3'K- ---- 063 o7f 74g. 7 bl Vf7 6 .30 0 6 0(t>4 30 S 7 - / -?s> Ca7 70 i <Qf/76s7K 9> -Y WYA .? 666 -X7/_ 7 6 /7o7 V7 5 0 3 ILCtd-sQY -S'/Q 37m Y S c& ATABL3 (5) 'i'UC J & Y Y u V ^estiqator V / / ^/rV-> .lMl.A&. L\j\f)VrTn.''6 d ' h h y Study Director D'ate --- 53 Company Sanitized. Does not contain TSCA CBI li' /. PHARMAKON RESEARCH INTERNATIONAL, INC. Individual Animal Scores During Primary Irritation Phase CQ a < CD p-8 fj HC-V) P0)<OU 1-0 ro5S Oi-i'rEe rt>> race E oo o 3rt a t0* H a\ H C1O to SI PHARMAKON RESEARCH INTERNATIONAL, INC. Individual Animal Scores During Primary Irritation Phase U1 U1 D C *0 O 3rt atr1 o H CTl H I U3 W PHARMAKON RESEARCH INTERNATIONAL, INC. ^ Individual Animal Scores Dring Induction Sponsor :_.Tpti, jlA f ffo Lj Q,W! E T M T - A s t u d v Number: P K 4 ^^/-~U - 12. - i ( Test Article:___ f4 ~ l`l j ' 7 3 ________ Lot Number; fj Q~ ln G\ A T A B L 3 (10) CO O < fern (De o| CUo *H[CUIJ-1m(O1) in (DN S OKtcB rtOT > cro cl E oo O rt Ir* 0 M cn I--1 1> to Ol 3>5EW?5W> 1 PHARMAKON RESEARCH INTERNATIONAL, INC. CO o < JroPc Eg 0.8 hjO P(DOO) f-Q ,Q*J HO c ri" taCo ooE o 3r+ K tO3 Ol H I OO IO PHARMAKON RESE-- !H INTERNATIONAL^ INC. SPnsor_d !r. cL , Prtcf 4 l fJtm ttto* W V Test Article:_____ /-/ - f -7 3 --- ^naiier.^w Study Mumber; V l j l h 4 - h / l - a >p _a, Ul CD A T A B L 3 (10) sYvJy 3, hjO f*-U*mO 1--0 W S oH:rea ci>ca, ooE d w o 3 c+ W 0W H CPl M U1J to aaoday; x b u t j papuauiY Company Sanitized. Does not contain TSCA CB1 SponsorA Test Article: ---- PHARMAKON RESE.-- 'H INTERNATIONAL, INC. Individual Animal Scores after Challenge Studv Number: U~D / ~ 9 / Lot Number, _ / ) p f c f l Pnlolxj Notebook # en V0 o T) O 3 rt i f0 H cn H* i1n tu S Amended Final Report Du Pont HLO 161-92 PHARMAKON r e s e a r c h i n t e r n a t i o n a l . inc WAVEflU.PENNSYLVANIA IB47I Individual Necropsy sheet , m 'SBfi-3 60 Company Sanitized. Does not contain TSCA CBI