Document 6bMBZ9JRR8BLva6D8yK2833j4

AR226-3087 DuPont H LQ -1998-00730 TRADE SECRET Study Title Acute Dermal Irritation/Corrosion Study with H-22482 in R abbits FINAL REPORT Haskell Laboratory Project ID : H LQ -1998-00730 Perform ing Laboratory Project ID : W IL-189161 Data Requirement Organization for Economic Cooperation and Development (OECD), Part 404 (1992) European Economic Community (EEC) 92/69 Annex V - M ethod B4 (1992) A uthor Tom G. Kern, B.S. Study Completed On January 15, 1998 Performing Laboratory W IL Research Laboratories, Inc. 1407 George Road Ashland, Ohio 44805-9281 for E. I. du Pont de Nemours and Company H askell Laboratory for Toxicology and Industrial M edicine Elkton Road, P.O . Box 50 Newark, Delaware 19714 Page 1 o f 38 Company Sanitized. Does not contain TSCA CBI W IL R E SE A R C H LA B O R A T O R IES, INC. A Subsidiary of Great Lakes Chemical Corporation, Ashland, O H 44805-9281 (419) 289-8700 Project No.: W1L-189I61 Sponsor: E. I. du Pont de Nemours and Company DoPOat HUM998-0073Q Study with H-22482 In Rabbit i (This page was intentionally left blank; client will insert appropriate '`` Ir .-` w Project No.: WIL-189161 Sponsor: E. 1. du Pont de Nemours and Company DuPont HLO-1998-00730 Acute Dermal Irritation/Corrosion Study with H-22482 in Rabbits GOOD LABORATORY PRACTICE COM PLIANCE STATEM ENT This study was conducted in compliance with EPA FIFRA (40 CFR 160) and EPA TSCA (40 CFR 792) Good Laboratory Practice Standards, and OECD Principles o f Good Laboratory Practice [C(81)3Q(Final), Annex 2] with the following exception: Although H-22482 was characterized in a DuPont Laboratory, the test substance was not characterized according to Good laboratory Practice. This deviation did not affect the validity of the study. Submitter: E. I. du Pont de Nemours and Company Sponsor: E. I. du Pont de Nemours and Company D ate Study Director: / / Tom O '. Rem. B.S. roup Supervisor - Acute Toxicology W IL Research Laboratories. Inc. I Sponsoring Company Representative:____ Registration Manager -3- Project N o.: WIL-189161 Sponsor: E. . du Pont de Nemours and Company DuPont H LO -1998-00730 Substance Tested: Acute Dermal Irritation/Corrosion Study with H-22482 in Rabbits GENERAL INFORMATION Svnonvm s/C odes: Haskell Num ber: Com position: Known Im purities: CAS Registry Number: S ta b ility : In the absence of visible evidence to the contrary, the test substance was assumed to be stable under the conditions of adm inistration. Sponsor: DuPont Specialty Chemicals E. I. du Pont de Nemours and Company W ilmington, Delaware 19898 Study Initiated-Compieted: June 19, 1997 - January 15, 1998 In Life Phase Initiated-Com pleted: June 20, 1997 - July 1, 1997 All raw data and the final report will be stored in the archives o f WIL Research Laboratories, Inc., Ashland, O hio, in the archives o f Haskell Laboratory for Toxicology and Industrial M edicine, E. I. du Pont de Nemours and Company, Newark, Delaware, o r at Iron Mountain, 200 Todds Lane, Wilmington, Delaware (formerly DuPont Records Management Center, W ilmington, Delaware). -4ompany Sanitized. Does not contain TSCA CBI Project N o.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 Acute Dermal Irritation/Corrosion Study with H-22482 in Rabbits TABLE OF CONTENTS Specific Country Requirements Page Good Laboratory Practice Compliance Statement General Information Summary Personnel and Report Submission Quality Assurance Unit Statement Objective and Introduction M aterials and Methods Observations Data Analysis and Interpretation of Results Results Conclusions Page 2 3 4 6 7 8 9 10 13 14 16 17 INDEX O F TABLES 1. Individual Dermal Scores 2. Incidences o f Dermal Response 3. Individual Body Weights (Grams) 19 20 21 INDEX O F APPENDICES A. Scoring C riteria for Dermal Reactions B. Protocol . 22 24 -5VOflipalj ii'Ssii: /su: oru Project N o.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 SUMMARY: Acute Dermal Irritation/Corrosion Study with H -22482 in Rabbits The prim ary dermal irritation and corrosion potentials o f H-22482 were evaluated in this study with New Zealand White rabbits. The study consisted o f single 0.5-gram doses o f the test substance that were applied to the clipped, intact skin o f three albino rabbits under semi-occlusive dressings. Initially, a single screen animal was dosed to evaluate the irritative/corrosive potential o f the test substance. This single rabbit received three sequential applications o f the test substance for three m inutes, one hour, and four hours. No severe dermal irritation o r corrosion was observed following any exposure to the screen animal. Based on these results, two additional rabbits received single four-hour exposures of the test substance. Dermal observations were mad* in accordance with the method o f Draize (Appendix A) and corrosion evaluations were maHp immediately following unwrap of the three-minute ami one-hour exposure sites. The fourhour exposure sites were observed for dermal irritation at 30-60 m inutes and 2 4 ,4 8 , and 72 hours after patch removal and once daily through day 7 if irritation persisted. For the screen animal, there was no evidence o f corrosion following the three-m inute, onehour, and four-hour exposures. There was no dermal irritation for the three-m inute and o.tehour sites following unwrap. The test substance induced erythema (score o f 1 o r 2) on all three o f the four-hour exposure sites. There was no edema or other dermal findings. All irritation was reversible and completely subsided by day 7 or earlier. Mean values were calculated for each animal separately by using num erical scores obtained from the quantitative evaluation o f each dermal response (erythem a and edem a) observed in the rabbits at the 24-, 48-, and 72-hour observations (excluding the three-m inute and onehour exposures). These values are as follows: mal Number 23581 23588 23591 Ervthema 1.67 1.33 0.00 -sSSML 0 .0 0 0 .0 0 0.00 Based on the overall average o f the nean values for erythema and edema calculated according to EEC Commission Directive 93/21 relative to the general classification and labeling requirements for dangerous substances published in the Official Journal o f the European Communities (EEC Directive 93/21), H-22482 is not classified as an irritant. -6- Project N o.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 PERSONNEL AND REPORT SUBMISSION: Kev Personnel: Jenny R. Randall Daniel W. Sved, Ph.D. Carney B. Jackson, D .V .M ., D .A .C .V .P ., D .A .C.V .P.M . Report Prepared Bv: Biologist n - Acute Toxicology Director o f Metabolism and Analytical Chemistry Assistant D irector of Pathology and Veterinary Medicine J Anita F. W ilmot, B.S. Biologist n/R epoit Writer II - Acute Toxicology Reviewed Bv: ijjrji D ate `V , 1Jonathan M. H urlyjB .S. Snior Biologist - Acute Toxicology B.S. Manager of General Toxicology t f fo F D ate Tracy A. Filliben Toxicology Associate D ate -7Company Sanitized. Does not contain TSCA CBi Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 QUALITY ASSURANCE UNIT STATEMENT: Daie(s) o f 6/20/97 7/24/97 7/24/97 Phase tasgetd Test Material Dispensation and Administration Study Records (1-1) Draft Report Daie(s) Findings Reported to 6/20/97 7/24/97 7/24/97 Date(s) Findings Repented to 7/29/97 8/29/97 8/29/97 This study was conducted and inspected in accordance with the Good Laboratory Practice Regulations, the Standard Operating Procedures o f WIL Research Laboratories, In c., ami the protocol and protocol amendment(s), if any. Quality Assurance findings, derived from the inspection(s) during the conduct o f die study and from die inspections o f die raw data and draft report, are documented and have been reported to the Study D irector. A status report is submitted to management monthly. The raw data, the retention sample(s), if applicable, and the final report will be stored in die Archives at W IL Research Laboratories, Inc., or another location specified by die Sponsor. _____ ................................................................... Deborah L. Little M anager o f Quality Assurance ... D ate -8(gf.-mps'^ySsnFasd. i:c-v;.: OBJECTIVE AND INTRODUCTION The objective o f the study was to determine the irritative and corrosive potentials o f the test substance following a single exposure to the skin o f albino rabbits. The protocol was designed and the study was conducted in general compliance with the Organization for Economic Cooperation and Development (OECD) nufrfrtjres for Testing o f Chem icals, Section 404 and European Economic Communities (ETC) giiiripiifi in the Official Journal o f the European Communities, 92/69, B4 (1992). The study was conducted in compliance with the U .S. EPA FIERA and TSCA Good Laboratory Practice Standards (40 CFR Part 160 and 792, respectively) and the OECD Principles o f Good Laboratory Practice [C(81) 30 (Final) Annex 2J. ! -9- Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 MATLPfALS AND METHODS Studv Title: W IL Study No. : ExDerimentai Start Date: Experimental Termination Date: Test Facility: Studv Soonsor: Studv Director: Sponsor Representative: Acute Dermal Irritation/Corrosion Study with H-22482 in Rabbits WIL-189161 June 20. 1997 July 1. 1997 WIL Research Laboratories, Inc. Ashland, OH 44805-9281 E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, P.O . Box 50 Newark, DE 19714 Tom G. Kern, B.S. Senior Group Supervisor - Acute Toxicology Tracy A. Filliben Toxicology Associate Test System: Species: Strain: Justification for Species Selection: Source: Number on Studv: Body Weieht Ranee: Albino rabbit New Zealand White The New Zealand White rabbit is generally recognized as appropriate for pnm aiy skin irritation studies. Considerable historical control data have been accumulated with this species for this type of study. Covance Research Products, Inc. Denver, PA One male and two females 3189 to 3464 grams at initiation o f dosing -10Sw npanj, Sanflfe,. B oss oof c a n t* , tsca cm Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO -1998-00730 Test System (continued): Age at Start o f Study: Young adult Method o f Identification: Eartag H ousing: Individual suspended wire-mesh cages. The animals were maintained by the animal husbandry staff o f WIL Research Laboratories, Inc., in accordance with Standard Operating Procedures. . Q u aran tin e: The animals were acclimated to laboratory conditions for a minimum o f seven days prior to initiation o f dosing. Food and W ater: PMI Feeds, Inc. Certified Rabbit LabDiet 5322 was provided at approximately 150 grams per day. Analysis o f feed was performed and provided by the manufacturer. Reverse osmosis-treated municipal water was provided ad libitum. W ater was analyzed in accordance with Standard Operating Procedures. Contaminants were not present in animal feed or water at levels expected to interfere with the objective o f this study. Results o f analyses are available upon Sponsor request. Environmental Conditions: Animal room with controlled temperature (65.867.1F), humidity (49.7-52.1% ), and light (12 hours light/12 hours dark). Test Substance Preparation: Individual 0.5-gram doses of the test substance were weighed into plastic weigh boats that were covered, labeled, and transported to the animal room for dosing. Sufficient deionized water was dispensed for moistening the test substance. Route and Rationale of Test Substance Administration: The route o f test substance administration was direct application to clipped, intact skin. This route of administration is standard for .assessment o f local dermal irritative/corrosive potential. -11- Project N o.: WIL-189J61 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 M ethod o f Test Substance Administration: On the day prior to dosing, the hair was removed from the backs and flanks o f the rabbits using an Oster small animal clipper. The clipped area on each animal constituted approximately 25% o f the total body surface. The 0.5-g test sites, delineated with waterinsoluble ink, were located lateral to the midline o f die back on each rabbit. Approximately 0.1 m illiliters o f deionized water were added to the three-minute site o f the screen anim al. The other exposure sites were not moistened with deionized water as per protocol. The test substance was characterized as a semi-solid after preparation for dosing and with grease-like consistency after bandage removal. M oistening was considered to be unnecessary as die test substance was observed to have made good contact with skin upon unwrap. Each dose was applied to an area o f skin approximately 6 cm2 under a secured two-ply, one-inch square gauze patch that was overwrapped with gauze bandaging and secured with Dermiform tape. The wrappings were used to retard evaporation and keep the test substance in close rantqrt with the skin. Collars were applied to each animal for die one- and four-hour exposure periods to prevent ingestion o f the test substance and/or wrappings. Only one test snhctanr was applied to each rabbit. Dosage Level(s)/Group(s)/Treatm ent Regimen: The dosage level was 0.5 g/site. There was one group o f three rabbits. Initially, a single animal was selected and treated with the test substance in order to m ininw a unnecessary use o f additional rabbits in case the test substance was corrosive or severely irritating. One screen animal received three separate semi-occluded applications o f the test substance for the three-m inute, one-hour, and four-hour exposure periods. Following the three-minute and one-hour exposures, the patches were removed and the sites were immediately evaluated for corrosion and dermal irritation (see Appendix A for scoring criteria). No other observations were conducted on the three-minute and one-hour exposure sites. The four-hour exposure site was evaluated for dermal irritation at 30-60 minutes after patch removal and further observed in accordance with the protocol. As no corrosion or severe dermal irritation was observed on these sites, two additional animals subsequently received a single, four-hour, semi-occluded exposure. At the end o f each exposure, the collars (if applicable) and bandages o f each rabbit were removed and the sites were wiped with disposable paper towels moistened with tepid tap water. -12- Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 OBSERVATIONS: M ortality: The rabbits were observed twice daily (morning and afternoon) for m ortality for the duration o f the study. Dermal Observations: The three-m inute and one-hour dose sites were observed once immediately after bandage removal for the presence or absence o f corrosion. Dermal irritation was also evaluated and graded in accordance with the method o f Draize (Appendix A). The four-hour application sites were observed for erythema, edema, and other dermal findings approximately 30-60 minutes and 24,48, and 72 hours after patch removal and once daily through day 7 if irritation persisted. Dermal irritation was graded in accordance with the method o f Draize (Appendix A). The adjacent areas o f untreated skin were used for comparison. In order to facilitate dermal observations, the areas o f application (four-hour site) o f two rabbits were clipped free o f hair after collecting die 72-hour scores. Clinical Observations: The rabbits were observed for clinical signs of toxicity at each dermal evaluation (excluding the three-m inute and one-hour exposures). Body W eights: Body weights were obtained and recorded on study day 0 (initiation) and at term ination from the study. Term ination: anim al's Upon term ination, the rabbits were euthanized by intravenous injection of sodium pentobarbital solution and discarded. -13Cksmpany Sanitized. Doe not contain TSCA CBS Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 DATA ANALYSIS AND INTERPRETATION OF RESULTS: A- EEC Classification The numerical values corresponding to each animal for erythema and edema were recorded at each observation period. Mean values for each dermal response (erythema mid edema) were calculated for each animal separately from numerical scores obtained at the 24-, 48-, and 72-hour observations (excluding the three-minute and one-hour exposures). The results were interpreted according to EEC Commission Directive 93/21 relative to the general classification and labeling requirements for dangerous substances. Interpretation according to Annex VI Skin Corrosion o r Irritation Criteria: 1. Corrosive The test substance will be considered to be corrosive if it produces full thickness destruction o f the skin tissue on at least one animal during die test for skin irritation. The substance will be classified as corrosive and assigned the symbol "C* and the inrfir.artrm o f danger "corrosive. " Risk phrases shall be assigned in accordance with the following criteria: R35 "CAUSES SEVERE BURNS" If, when applied to healthy intact animal skin, full thickness destruction o f skin tissue occurs as a result of up to three-minute exposure or if this result can be predicted. R34 "CAUSES BURNS" If, when applied to healthy intact animal skin, full thickness destruction o f skin tissue occurs as a result o f up to four hour exposure o r if this can be predicted. 2. Irritant The test substance will be classified as "IRRITANT" and will require the "Xi" symbol and the indication o f danger "IRRITANT" in accordance with the following criteria: R38 "IRRITATING TO SKIN" If, when applied to healthy intact animal skin for up to four hours, significant inflammation is caused and is present 24 hours o r more after the end o f the exposure period. Inflammation is significant if the mean values o f the scores for either erythema and eschar formation or edema formation corresponds to one or m ote o f the following mean values calculated for each animal separately and has been observed in two or more animals: -14'Soropar.y S Project N o.: W IL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 DATA ANALYSIS AND INTERPRETATION OF RESULTS (continued): | D erm al Lesion Erythema and eschar formation Edema M ean Value 2 .0 o r more 2.0 o r more f All scores at each of the reading times (24, 48, and 72 hours) for an effect should be used in calculating the respective mean values. Inflammation of the skin is also significant if it persists in at least two animals at the end of the observation time. Particular effects such as hyperplasia, scaling, discoloration, fissures, scabs, and alopecia should be taken into account. -15Company Sanitized. Sees o? esnfa'n TSCA CBf Project N o.: WL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 RESULTS: Mortality: No deaths occurred during the study. Dermal Observations (Tables 1 and 2V For the screen animal, there was no evidence "*f corrosion following any o f the three different exposure tim es (three-minute, one-hour, and four-hour exposures). There was no dermal irritation observed for die three-minute o r one-hour exposures. Erythema (score o f 1 or 2) was noted on all three four-hour exposure sites at the 30-60 minute observation. Erythema totally cleared for one animal each at 24 hours, day 6, and day 7. There was no edema or other dermal findings. ' Clinical Observations (Table 31: No clinical signs o f toxicity were observed during the study. Body W eiputs (Table 3): No rem arkable changes or differences were noted in body weights during die study. -16&CMcisl Project No.: WIL-189161 Sponsor: . I. du Pont de Nemours and Company DuPont HLO -1998-00730 CONCLUSIONS. . Under the conditions o f testing and according to the guidance provided in EEC Directive 93/21, H-22482 is not classified as an irritant. -17Company Sanitized. Does not contain TSCA CBS Project N o.: W IL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont H LO -1998-00730 Acute Dermal Irritation/C orrosion Study with H-22482 in Rabbits TABLES 1-3 -18- S am p g n y 3 gss?n e e o n a r T S Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company TABLE 1 DuPont HLO-1998-00730 ACUTE DERMAL IRRITAHON/CORROSION STUDY WITH H-22482 IN BABBITS INDIVIDUAL DERMAL SCORES E rythem a Animal Se* Site 1 hr 24 hr J S J L J I M . 4d 23581 M A lr 2r 2r l r lr 23588 F A lr 2r lr lr lr 23591 F A 2r Or Or Or 5d 6d 7d lr l r Or lr 0 Edema Animal f a _Sitg J J j r . 24 hr 48 hr 72 hr 23581 M A 0 0 0 0 23588 F A 0 0 0 0 23591 F A 0 0 0 0 4d 0 0 - - _____ 2 1 00 0 00 - h r - H our d = Day M = Male F * Female r = Residual test m aterial present on the site - = Dermal iiritation previously subsided; animal was terminated from the study A = Four-hou;- sites EEC: Animal Number 23581 23588 23591 1.67 1.33 0.00 0.00 0.00 0.00 19 Company Sanitized', Does not contain TSCA CBI Project N o.: WIL-189161 Spon r: E. I. du Pont de Nemours and Company TABLE 2 DuPont HLO-1998-00730 ACUTE DERMAL 1RRITATION/CORROSION STUDY WITH H-22482 IN RABBITS INCIDENCES OF DERMAL RESPONSE (SCORES)* Score 0 1 2 Erythema*1 1 hr il& r 48 hr 7gM M 0/3 1/3 1/3 1/3 0/2b 0/2b 1/2C l / l d 2/3 0/3 1/3 2/3 2/2 2/2 1/2 0/1 1/3 2/3 1/3 0/3 0/2 0/2 0/2 0/1 Score 0 1 2 Edemab I h r 24 hr 4 8 r 72 hr J d . 3/3 3/3 3/3 3/3 2/2b 0/3 0/3 0/3 0/3 0/2 0/3 0/3 0/3 0/3 0/2 5d 2/2b 0/2 0/2 M. 2/2c 0/2 0/2 7d l/ld 0/1 0/1 "According to EPA test guidelines. bFour-hour sites evaluated. T w o rabbits were evaluated for dermal irritation since irritation for 01 animal had totally cleared. dOne remaining rabbit was evaluated for dermal irritation since dermal irritation bad com pletely subsided for the other two animals. hr = Hour d = Day 20 iiemaany Sanffteerf'. &e-ts no? cent?." 'i Cc a CM Project N o.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company TABLE 3 DuPont HLO-1998-00730 ACUTE DERMAL IRRITATION/CORROSION STUDY W ITH H-22482 IN RABBITS INDIVIDUAL BODY WEIGHTS (GRAMS) Group Initiation T erm ination Sex (Oav m ()Imnitceirvatl)f 0.5 g/site 23581 23588 23591 M F F 3464 3258 3189 3668 (day 7) 3279 (day 6) 3283 (72 bouts) M = Male F = Female CLINICAL OBSERVATIONS No clinical findings were noted for any animals rfnrin the study. 21 Company Sanitized. Does noi contain TSCA CBI  .7. - * 22 Company Sanitized. nei aonimn SC GB! itlTllmmmi i..... .... .......... 11!M inilirii^iiilii^Hli111 iiii iii iiiili ii iih'|-|iMiirifi^^tf:Mihiin ail Project No.: W IL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 Value APPENDIX A ACUTE DERMAL IRRITATION/CORROSiON STUDY WITH H-22482 IN RARBITS SCORING CRITERIA FOR DERMAL REACTIONS Evaluation of Dermal Reactions* Erythema and Eschar Formation 0 No erythema 1 Vety slight erythema (barely perceptible, edges o f area not well defined) 2 Mild erythema (pale red in color and edges definable) 3 Moderate to severe erythema (definite red in color and area well defined) 4 Severe erythema (beet or crimson red) to slight eschar formation (injuries in depth) Edema Formation 0 No edema 1 Very slight edema (barely perceptible, edges of area not well defined) 2 Mild edema (edges o f area well defined by definite raising) 3 Moderate edema (raised approximately 1 mm) 4 Severe edema (raised more than 1 mm and extending beyond area o f exposure) *Draize, J.H ., 1959. The Appraisal of the Safety of Chemicals in Foods, Drags and Cosmetics. Dermal Toxicity, pp. 46-59. Assoc, of Food and Drug Officials o f the U .S ., Topeka. Kansas. 23 Company Sanitized. Does not contain TSCA CBi I Project No.: W IL-189161 Sponsor: 1: I. du Pont dc Nemours and Company DuPont HLO-1998-00730 Acute Dermal Irritation/Corrosion Study with H-22482 in Rahhiis APPENDIX B Protocol - 2 4 - Ssfflpafi Sanftzs, S ees Ret easiffn TSCA CBS DuPont H LO -1998-00730 Study Number: WL- / 9 / / t w m MASTER PROTOCOL COVER SHEET Sponsor E I. du Pont de Nemours and Company Master Protoco! No.: E06 (Sponsor) Sponsor Proje No. A. Jills gf Study; Acute Demut] Itritation/Cotrosion Study in Albino Rabbits 1) n . BERSONNEL INVOLVED IN THE S T tm v A. Sponsor Representative Name: Tracy A. Fiiliben Title: Toxicology Associate 2) m . STUDY SCHEDHT.E DATA A. Proposed Experimental Stan Date: 3- Proposed Experimental Termination Date: C- Proposed Audited Draft Report Date: 3) IV. TEST MATERIA! . DATA A. Identification^) B- LotNumber^! c . Esinnr: D. Stability- Stable (Sponsor) (Sponsor) 4 7A 7 * A /< 5 7 (W E.) (W E.) (W E.) (Sponsor) (Sponsor) (Sponsor) (Sponsor) W IL RESEARCH LABORATORIES, INC. A Subsidiary of Great Lakes C h e tS " Corporation, Ashland, OH 44805-9281 (419) 289-8700 25 Company Santifeesl. Does not contain TSCA CB( *~ n-.- / & / ! o ra .) Masitr Protocol Cover Sheet Page Two________ cuti DuPont HLO -1998-00730 E. Physical Description- F. Storage ConditionsG. Personnel Safety Para- Ambient temperature Normal laboratory procedures. For complete available caf-ry H. CAS Number; information see attached M Snc fWTL) (Sponsor) (Sponsor) (Sponsor) C Reason for Protocol Modification: I ) Sponsor Representative is specified. 2) Proposed scheduling information is added to the protocol. 3) Test material data are added to the protocol. Approved By: Prepared By. r E.I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine WIL Research Laboratories, Inc. Ashland, OH 44805-9281 Elkton Road P.O. Box JO Newark. DE 19714 Spon^Sr Representative /a? /? ? Date- " 26 npgF SanlfeeoL vom noi cattiate t. s? DuPont H L O -1998-00730 PROTOCOL Acute Dermal Irritation/Corrosion Study in Albino Rabbits (OECD/EEC Guidelines) Master Protocol No.: E06 For E. I. du Pont de Nemours and Company Haskell Laboratories for Toxicology and Industrial Medicine Elkton Road P.O. Box 50 Newark. DE 19714 By WIL Research Laboratories. Inc. Ashland. OH 44805-9281 June 4. 1997 WIL R E S E A R C H L A B O R A T O R IES, INC. A SuDsidiary of Great Lakes Chemical Corporation, Ashland. O H 44805-9281 (413) 289-8700 27 inpMry SanffzeS. Does not confarti TSCA CBI m ACUTE DERMAL IRRITATION/CORROSION STUDY IN ALBINO RABBITS DuPoni HLO-1998-00730 Master Protocol N o.: E06 1 OBJECTIVE O F S T im v to determine the irritative and corrosive potential of the test exposure to the skin of albino rabbits. ? " material following a smgle Principles of Good Laboraraiy Practice fC(Sl) 30 (Final) PERSONNEL INVOT.VFQ in TWF S T tm v A- Sponsor Representative To be specified by Master Protocol Cover Sheet M L Study Director Tom G. Kem, B.S. Group Supervisor - Acute Toxicology c M L Deputy Director Jonathan M. Hurley. B.S. Senior Biologist - Acute Toxicology D Study Responsibilities Tames L. Schardein, M S.. A.T.S Director of Research - Christopher P. Chengelis, Ph D., D.A.B.T. Senior Toxicologist 3 Jozef J.W.M Menens. Ph.D.. D.A.B.T. Staff Toxicologist 4 Gary R. Kiplinger. B.S Manager of General Toxicology Page 2 of 12 28 " * - TSCA Good y) " Kl 0ECD ?*50A Acute Dermal Imtation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 11 ^ S Q N N m j N yO LVEP IN THF STfm v (continued) 5 Ronald E. Wilcon, B.S Director of Informational Systems 6. Daniel W. Sved, Ph.D. Director of Metabolism and Analytical CKmistry 7. Sally A. Keets, A.S. Manager of Vivarium 8. Deborah L. Little Manager of Quality Assurance 9. Kerin Clevidence. B.S. Group Supervisor of Gross Pathology and-Developmental Toxicology Laboratory 10. Charlene M. Lindsey. M.A. Manager of Technical Report Writing 11. Carney B. Jackson. D.V.M.. D.A.C. V.P., D A C V P M Assistant Director of Pathology and Veterinary Medicine ' 12. Jenny R. Randall Biologist II - Acute Toxicology HI. STUDY SCHEDm F d a t a A' to w s e d frm im -r,,l S,,n p 1g To by MasIer p ^ , ^ ^ B f a p ^ m a l Tern,W inn D m : To be added by Master Protocol Cover Steel ' C ' & g * Bl nr>l* P,,pon n i ^ T <* dded by Master Protocol Cover Sheet IV TEST SUBSTANCE D a t a A. Identification B. Lot Numbs.C. Purity: D Stability- To be added by Master Protocol Cover Sheet To be added by Master Protocol Cover Sheet To be added by Master Protocol Cover Sheet To be added by Mastet Protocol Cover Sheet Page 3 of 12 29 Company Sanitized Does not contain TSC CB1 Acute Dermal Imtation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 IV lE -S T S U B S X m i DATA (continued) F Storage ConriirinncG- Personnel Safety rw-. v - TEST SV.STFM A. Species: B. Strain: C. Source: D. Number on Study E. Sex: F - Body Weight R anppG- Approximate Ap>H. Identification S y s te m - F Justification for Selection To be documented by WIL Research Laboratories, added by Master Protocol Cover Sheet Inc. and To be added by Master Protocol Cover Sheet To be added by Master Protocol Cover Sheet Albino rabbit New Zealand White Covance Research Products, Inc. Denver, PA or Kalamazoo, MI Three animals (except as noted in section Vffi C of this protocol). ` Males and/or females 2.0 leg or greater at initiation of dosing. Young adult Each animal will be uniquely identified by a plastic eartag displaying me animal number. Individual cage cards will be affixed to each cage and will display the animal number group and study number. .' This species and strain is generally recognized as appropriate ior primary skin irritation studies The experimental design uses the procedures and standards S l byth' " d prop0"d Page 4 of 12 30 iSomp-mf SsjiiVaKZtr tJC/B- Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-U073 VI SPECIFIC M AINTENANCE SCHFDII7 F A. Animal Housing The animais wl be housed individually in suspended wire-mesh cages in an environmentally controlled room. Animals will be housed in clean cages elevated above cage-board or other suitable material that will be changed at least three times each week. The facilities at WIL Research laboratories. Inc. are fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care International (AAALAC International). B. Environmental Conditions Controls wl be set to maintain temperature at 67 4F and relative humidity at approximately 30-70%, Fluorescent lighting controlled by light timers will provide illumination for a 12-hour light/dark photoperiod. Temperature and relative humidity will be recorded once day. ' C. Drinking Water Reverse osmosis treated municipal water will be available ad libitum. Filters servicing the automatic watering system are changed regularly according to Standard Operating Procedures. Water supplying the laboratory is analyzed for contaminants according to standard Operating Procedures. No contaminants known to be in the water are expected to interfere with the validity of the study. D. Basal Diet PMI Feeds, Inc. Certified Rabbit LabDiet 5322 will be offered at approximately 150 g/day during the study. Periodic analyses of the certified feed for the presence of heavy metals and pesticides are performed and provided by the manufacturer to ensure that none are present in concentrations that would be expected to affect the outcome of the study. v n . EXPERIM ENTAL DESIGN ` A- Animal Receipt and Acclimation Each animal will be inspected by a qualified technician upon receipt. Rabbits judged to be in good health and suitable as test animals will be acclimated to laboratory' conditions for a minimum of seven days. All rabbits will be weighed initially and permanently identified with an eartag. During the acclimation period, each rabbit will be observed twice daity for changes in general appearance and behavior Prior to initiation of dosing, those animals judged to be suitable test subjects will be identified. Page 5 of 12 31 Company Sanitized. Does not contain TSCA CBI Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 XPERJMEN1 AL DESIGN (continued) . B- Baute and Rationale of Test Material A d m in i^ ti^ The route c f administration will be dermal in order to m e t . =*. corrosion. The detmal mote is , possible ^ o f ^ E H S E w C. Body Weighty ,. Ihe body weight of each animal v.ili be determined on study dav 0 and at " ^ ta>TM d ^ 7 *= ^ h m l a, i t i L e . VDI- BQ.^gE LEVELS AND TREATMENT REfiTMFM A- Treatment Levels The same dose level will be used for all three animals (0.5 ml or 0.5 g per test site). 11. Treatment Group *"* - -- - - S K S S E C. Initiation Procedure If it is anticipated that the test substance mav be severeiv . substance for three minutes, one hour, and four h l l E ' l L ' a E E i E 0" 11" of be dosed and observed for die presence of severe .mr.TM !! " [sure sue will 1 * unwrap. If no actions' 5 i= * - -~ i= r ^ V K a = If severe irritation (erythema, edema and/or e s c h a r!; nhew. _#v. jg * ESSfwSLT"T ^on the single aninud men. no forhe, teui request of the Sponsor. If corrosion is o h ^ '! ! f f d * " ^ SpCClflc ** immediately be terminated If neither a con-TM- ^ a"y exPosure* study will observed after the four-hour exposure the smn n f 3 Severe irritam effect is umnls. each ^ a ^ J e Z - b i o f e x p m L " '" ** Page 6 of 12 ' ) 32 Awfipaiij S snlfaee, Dl-s-;-;h,,- mi Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 IX. OBSERVATTONS (continued) Individual animals may be teimimted if no irritation is present at the 72- hour or any subseouent J bS ? J 0n' If " n " Uon f present at *" end seven days, observation will continue daily^ntil nnTM ndeS L `j ?5V*0US,y irreversible "P 10 14 ^ y s (additional cost). Hie study^eed not 5 ! aftar app,icsd" sptc" ca" !r " , d . X- GROSS PATHOI DPtY The animals will be euthanized by an intravenous injection of euthanasia solution unon r n t ^ u o n and discarded Collection of skin samples from the four-hour t m J m S L T S ?reserved m 10% " Otral-buffered formalin for histopatfaological examination at the specific wntten request of the Sponsor (additional cost). A gross necropsy e S S t i o i on major organ systems of the thoracic and visceral cavities will be conductedTM ail an,,,,,ic found dead or euthanized in extremis. animals XT PAIA ANALYSIS AND INTERPRETATION OP PFCttjjc A. EEC Classification The numerical values corresponding to each animal for erythema and edema will be recorded at each observation period. Mean group scores will be calculated ror each dermal reponse (eryAema and edema) by summing the scores obtained from all animals at the 24-, 4 ^ d -hour observations (excluding the 3 and 60 minute exposures). The three m ^nc corresponding to the three observation periods will be averaged and overall averages will be , o r c h ' t o- Th= a w u i i t a p * J S r . ' S J " lat'VE " "* g ra m l d a" i,iCSdTM " " ** - " `tonem s Interpretation according to Annex VI Skin Coirosion or Imtation Criteria: I. Corrosive . The test substance will be considered to be corrosive if it produces full if ue on al Iea* one animal during the test for skin S ta tio n . The substance will be classified as corrosive and assigned the symbol "C" and the foHtwhag c rite ria ^ ^ ^ te aSsigned ^ R35 "CAUSES SEVERE BURNS" If, when applied to healthy intact animal skin, full thickness destruction of skin p e e r e d 11" ^ 3 rCSUU f "P l thrCC m" Ute exPsures or if * is result can be Page 8 of 12 34 OB Acute Dermal irritation/Corrosion Srudy in Albino Rabbits DuPont HLO-1998-00730 a ' P ^ A A N A [ . V . ^ , m ^ T p ,, p r r T 1 T tn r , n[- r r | ,, ,, ( ............... R34 "CAUSES BURNS" If, when applied to healthy intact animat s|rin a.ii issue occurs as a result of up to four hour, e ^ o a u e 2. Irritant j. *Hic test substance will be classified ac "rpp nr a kit" * ... and the tadicetiou of ,, g e t -IRRITANT- in aceotdaTe Z R38 "IRRITATING TO SKIN" If, when applied to healthy intact animat h . tnr -, .. inflammation is caused and is p r e r e n t ^ I or S L ? " ? ! Si8nificant period.- Inflammation is significant if tl.e StSZSrJST*-- values of the <u-n the cxposurc fl Dermal Lesion I Erythema and eschar formation I Edema Mean Score 2.0 or more 2.0 or more | | Ail scores at each of the readme times (7d an __j . . _ used in calculating the respece.ve mean values ' UrS) " CffeCt wUI ** Inflammation of the skin is also significant if it Demists in > t . , end of the observatton . Particular S Lh L . ^ discoloration, fissures, scabs, and alopecia s h o u l d b e ^ J S T `` ` `" S' x n . REPORT summarizing main .a " " hT ' ' m nal">'' M X " W * a able draft report wl be provided approximately six w e d S ^ o l t o S ^ ' . 0nCpy of the audited copies of die report will be provided upon final^uon. ^ comP ^ of the study. Two Page 9 of 12 35 Company Sanitized. Dees nof eontafo TSC cm Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 Xm. RECORDS . All original raw data records (as defined by the applicable GLPs and WIL SOPs) generated bv W1L Research Laboratories, Inc. will be collected and maintained by WIL Research Laboratories, Inc. " XTV. WORK PRODUCT Sponsor will have title to all documentation records, raw data, slides, specimens, or other woric product generated during the performance of the study. All work product including raw paper data, magnetically encoded records and specimens will be retained at no charge for a period of six months following issuance of the final report in the Archives at WIL Research Laboratories lac. Thereafter, WIL Research Laboratories will return all work product to Haskell Laboratory for storage All work product wi'l be stored in compliance with regulatory requirements. XV. QUALITY ASSURANCE The study wiUbe audited by the WIL Quality Assurance Unit while in progress to assure compliance with Good Laboratory Practice regulations and adherence to the protocol and to WIL Standard Operating Procedures. The raw data and draft report will be audited by the WIL Quality Assurance Unit pnor to submission to the Sponsor to assure that the final report accurately describes the conduct and the findings of the study. ^n XVI. PROTOCOL MODTFTrATTOM This Master Protocol shall be used for multiple studies. A Master Protocol Cover s w shall ta pidpared for each audy which will esc ihis design. I t e M ^ ttr I h o h S i i S S S Cover Sheet shall constitute the protocol for an individual study. In addition, the date of Studv Du-ector signature on the Master Protocol Cover Sheet shall be considered the date of studv initiation. * Modification of the protocol may be accomplished during the course of this investigation n chan.ges_wliI be made in the study design without the verbal or written permission of the Sponsor. In the event that the Sponsor verbally requests or approves a rhJnl*. ^ protocol, such changes will be made by appropriate documentation in the form of a protocol amendment. AH alterations of the protocol and reasons for the modifications) will be ign-H bv the Study Director and the Sponsor Representative. * Page 10 of 12 35 Goisip&ny Uanifaect u;iwhf YsSCA CBl Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont H LO -1998-00730 XVn- ANIMAL WELFARE ACT COMPLIANCE This study will comply with all applicable sections of the Final Rules of the Animal Welfare Act regulations (9 CFR). The Sponsor should make particular note of the following: L The Sponsor signature on this protocol documents for the Study Director the Sponsor's perim 6 t ' 3t ** StUdy described m *** Protocol does not unnecessarily duplicate previous 2. Whenever possible, procedures used in this study have been designed to avoid or minimiTM. discomfort, distress or pain to animals. All methods are described in this study protocol or in written laboratory Standard Operating Procedures. p 3. Animals that experience severe or chronic pain or distress that cannot be relieved will be painlessly euthanized as darned appropriate by the veterinary staff and Study Director. The Sponsor will be advised by the Study Director of all circumstances which could lead to this action in as timely a manner as possible. 4. Methods of euthanasia used during this study are in conformance with the above-referenced regulation. x v m . pro ; ocol apprdvat E. I. du Pont de Nemours and Company Haskell Laboratories for Toxicology and Industrial Medicine Elkton Road P.O. Box 50 Newark. DE 19714 WIL Research Laboratories, Inc. Ashland, OH 44805-9281 ----------%l.f\ [dut Carol Finlay/ B A. (j Sponsor Representative Date (L y Filliben Sponsor Representative __________ Date Date Page 11 of 12 37 Company Sanitized. Does not contain TSCA CBI Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 Value 0 1 2 3 4 APPENDIX A SCORING CRITERIA FOR DERMAL REACTIONS* Evaluation of Dermal Reactions* Erythema and Eschar Formation No erythema Very slight erythema (barely perceptible, edges or area not well defined) Mild erythema (pale red in color and edges definable) Moderate to severe erythema (definite red in color and area well defined) Severe erythema (beet or crimson red) to slight eschar formation (injuries in depth) Edema Formation 0 No edema 1 Very slight edema (barely perceptible, edges of area not well defined) 2 Mild edema (edges of area well defined by definite raising) 3 Moderate edema (raised approximately 1 mm) 4 Severe edema (raised more than 1 mm and extending beyond area of exposure) Draize, J.H., 1959. The Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics Dermal Toxicity, pp. 46-59. Assoc, of Food and Drug Officials of the U.S., Topeka, Kansas. Page 12 of 12 38 SferfJ1 fees' Umn ?mi c-ypfs^ TeA C8 F