Document eE8GwQX0YRvdZvbqE4rNoe6y

AR226-3087 DuPont HLO-1998-00730 TRADE SECRET Study Title Acute Dermal Irritation/Corrosion Study with H-22482 in Rabbits FINAL REPORT Haskell Laboratory Project ID: HLO-1998-00730 Performing Laboratory Project ID: WIL-189161 Data Requirement Organization for Economic Cooperation and Development (OECD), Part 404 (1992) European Economic Community (EEC) 92/69 Annex V - Method B4 (1992) Author Tom G. Kem, B.S. Study Completed On January 15, 1998 Performing Laboratory WIL Research Laboratories, Inc. 1407 George Road Ashland, Ohio 44805-9281 for E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, P.O. Box 50 Newark, Delaware 19714 Page 1 of 38 Company Sanitized. Does not contain TSCA CBJ WIL RESEARCH LABORATORIES,INC. A Subsidiaiyof Great Lakes Chemical Corporation.Ashland, OH 44805-9281 (419) 289-8700 Project No.: WDL-189161 Sponsor: E. I. (hi Font de Nemours and Company DuPont HUM998-00730 Acuie Dennal IrriEttion/CoiTOBion Study with H-2-^.nP.^ SPECIFIC COUNTRY REQUOffiMENIS (This page was intentionally left blank; dieat will imert fippropmte statcmeot.) -2Cce? ncS cant?", TSCA CW 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 COMPLIANCE STATEMENT 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 of Good Laboratory Practice [C(81)30(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. Study Director: Submitter: E. I. du Pont de Nemours and Company Sponsor: E. I. du Pont de Nemours and Company . Tom G". Ki--em--. --B.--S. Supervisor - Acute Toxicology WIL Research Laboratories. Inc. Date ///^ Sponsoring Company Representative^ Registration Manager -3^cW^rsfi T~''^--,Vf--^.v r-v- Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 Substance Tested: Acute Dermal Irritation/Corrosion Study with H-22482 in Rabbits GENERAL INFORMATION Synonyms/Codes: Hasicell Number Composition: 22482 Known Impurities: CAS Registry Number: Stability: Sponsor: Study Initiated-Completed: In Life Phase Initiated-Completed: In the arsence of visible evidence to the contrary, the test substance was assumed to be stable under the conditions of administration. DuPont Specialty Chemicals E. I. du Pont de Nemours and Company Wilmington, Delaware 19898 June 19. 1997 - January 15, 1998 June 20. 1997 - July 1, 1997 All raw data and the final report will be stored in the archives of WIL Research Laboratories, Inc., Ashland. Ohio, in the archive of Haskell Laboratory for Toxicology and Industrial Medicine, E. I. du Pont de Nemours and Company, Newark, Delaware, or at Iron Mountain, 200 Todds Lane, Wilmington, Delaware (formerly DuPont Records Management Center, Wilmington, Delaware). -4i Does not eonfam TSCA CBI Project No.: 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 ^pacific Country Requirements Page Good Laboratory Practice Coirpliance Statement General Information Summary Personnel and Report Submission Quality Assurance Unit Statement Objective and Introduction Materials 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 OF TABLES 1. Individual Derma! Scores 19 2. Incidences of Dermal Response 20 3. Individual Body Weights (Grams) 21 INDEX OF APPENDICES A. Scoring Criteria for Dermal Reactions 22 B. Protocol 24 -5.U&fe& , jsi... i'h.ii corUfcf;"; .''SC.w CBf Project No.: WIL-189161 Sponsor: E. I. du Font de Nemours and Company DuPoni HLO-1998-00730 Acute Dermal Irritation/Corrosion Study with H.22482 in Rabbits SUMMARY: The primary dermal irritation and corrosion potentials of H-22482 were evaluated in this study with New Zealand White rabbits. The study consisted of single 0.5-gram doses of the test substance that were applied to the clipped, intact skin of three albino rabbits under semi-occlusive dressings. Initially, a single screen animal was dosed to evaluate the irritative/corrosive potential of the test substance. This single rabbit received three sequential applications of the test substance for three minutes, one hour, and four hours. No severe dermal irritation or 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 made in accordance with the method of Draize (Appendix A) and corrosion evaluations were made immediately following unwrap of the three-minute and one-hour exposure sites. The fourhour exposure sites were observed for dermal irritation at 30-60 minutes and 24,48, and 72 hours after patch removal and once daily through day 7 if irritation persisted. For the screen animal, there was no evidence of corrosion following the three-minute, one- hour. and four-hour exposures. There was no dermal irritation for the three-minute and o.'?hour sites following unwrap. The test substance induced erythema (score of 1 or 2) on all three of 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 numerical scores obtained from the quantitative evaluation of each dermal response (erythema and edema) observed in the rabbits at the 24-, 48-, and 72-hour observations (excluding the three-minute and onehour exposures). These values are as follows: Animal Number 23581 23588 23591 Erythema 1.67 1.33 0.00 Edema 0.00 0.00 0.00 Based on the overall average of 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 of the European Communities (EEC Directive 93/21), H-22482 is not classified as an irritant. -6- Project No.: WIL-189161 Sponsor: E. I. du Font de Nemours and Company DuPont HLO-1998-00730 PERSONNEL AND REPORT SUBMISSION: Key Personnel: Jenny R. Randail Daniel W. Sved, Ph.D. Camey B. Jackson, D.V.M., D.A.C.V.P., D.A.C.V.P.M. Report Prepared Bv: Biologist n - Acute Toxicology Director of Metabolism and Analytical Chemistry Assistant Director of Pathology and Veterinary Medicine ^^ ^ W^vwk I Anita F. Wilmot, B.S. Biologist n/Report Writer II - Acute Toxicology 1^2. Date Reviewed Bv: ^{y) fe.^/iL^ I Jonathan M. Hurrtif^sxxjjBB.S. Sr^e\ni"io- r_ Bm iolo_g_i-s*t. - A* c--u--^t--e To--x.-i:cology ^ ^ ^ ^ / : ^ . ^ ^ ^aiyR. Kiplinger;B.S. ' Manager of General Toxicology Approved and Submitted Bv: i is m ' Date ^di<hT Date Study Monitor: Tom 6. Kem, B.S. Study Director ^MJH^ <\^.g/Z 0. ff Tracy A. Filliben Toxicology Associate ^ / n Date 3/1^ Date -7Company Sanitized. Does nei contain TSCA CBi Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company QUALITY ASSURANCE UNIT STATEMENT: DuPont HLO-1998-00730 Date(s) of tospecttonfs} 6/20/97 7/24/97 7/24/97 Phase Inspected Test Material Dispensation and Administration Study Records (1-1) Draft Report Datt(s) Findings Reported to Snidy Director 6/20/97 7/24/97 7/24/97 Date(s) Findings Reported 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 of WIL Research Laboratories, Inc., and the protocol and protocol amendment(s), if any. Quality Assurance findings, derived from the inspections) during the conduct of the study and from the inspections of the raw data and draft report, are documented and have been reported to the Study Director. 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 we Archives at WIL Research Laboratories, Inc., or another location specified by me Sponsor. O^^v^A^^C^. Deborah L. Little Manager of Quality Assurance ilf^h^ Date 'SlytTipg;^ Project No.: WIL-189161 Sponsor: E. I. du Pom de Nemours and Company DuPont HLO-1998-00730 OBJECTIVE AND INTRODUCTION: The objective of the study was to determine the irritative and corrosive potentials of the test substance following a single exposure to the skin of albino rabbits. The protocol was designed and the study was conducted in general compliance with the Organization for Economic Cooperation and Development (OECD) Guidelines for Testing of Chemicals, Section 404 and European Economic Communities (EEC) guidelines in the Official Journal of the European Communities, 92/69, B4 (1992). The study was conducted in compliance with the U.S. EPA FIFRA and TSCA Good Laboratory Practice Standards (40 CFR Part 160 and 792, respectively) and the OECD Principles of Good Laboratory Practice [C(81) 30 (Final) Annex 2]. Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-i 998-00730 MATLPTALS AND METHODS. Study Title: WIL Study No.: Experimental Start Date: Experimental Termination Date: Test Facility: Study Sponsor: Study Director: Sponsor Representative: Acute Derma! 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 HaskeU Laboratory for Toxicology and Industrial Medicine Elkton Road, P.O. Box 50 Newark. DE 19714 Tom G. Kem, B.S. Senior Group Supervisor - Acute Toxicology Tracy A. Filliben Toxicology Associate Test System: Species: Strain: Justification for Species Selection: Source: Number on Study: Body Weipht Range: Albino rabbit New Zealand White The New Zealand White rabbit is generally recognized as appropriate for primary skin irritation studies. Considerable historical control data have been accumulated with mis species for this type of study. Covance Research Products, Inc. Denver. PA One male and two females 3189 to 3464 grams at initiation of dosing -10Sartfeed. Doss not contain T8CA CB? Project No.: WIL-189161 Sponsor: E. I. du Pont dc Nemours and Company DuPont HLO-1998-00730 Test System (continued): Age at Start of Study: Young adult Method of Identification: Eartag Housing: Individual suspended wire-mesh cages. The animals were maintained by the animal husbandry staff of WIL Research Laboratories, Inc., in accordance with Standard Operating Procedures. Quarantine: The animals were acclimated to laboratory conditions for a minimum of seven days prior to initiation of dosing. Food and Water: PMI Feeds, Inc. Certified Rabbit LabDiet 5322 was provided at approximately 150 grams per day. Analysis of feed was performed and provided by the manufacturer. Reverse osmosis-treated municipal water was provided ad libitum. Water 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 of this study. Results of 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 of test substance administration was direct application to clipped, intact skin. This route of administration is standard for .assessment of local dermal irritative/corrosive potential. CA CB. Project No.: WIL-189161 Sponsor: E. I. du Font tie Nemours and Company DuPont HLO-1998-00730 Method of Test Substance Administration: On the day prior to dosing, the hair was removed from the backs and flanks of the rabbits using an Oster small animal clipper. The clipped area on each animal constituted approximately 25% of the total body surface. The 0.5-g test sites, delineated v'ith waterinsoluble ink, were located lateral to the midline of the back on each rabbit. Approximately 0.1 milliliters of deionized water were added to the three-minute site of the screen animal. 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. Moistening was considered to be unnecessary as me test substance was observed to have made good contact with skin upon unwrap. Each dose was applied to an area of 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 contact with the skin. Collars were applied to each animal for me one- and four-hour exposure periods to prevent ingestion of the test substance and/or wrappings. Only one test substance was applied to each rabbit. Dosage Level(s)/Groupfs)/Treatment Regimen: The dosage level was 0.5 g/site. There was one group of three rabbits. Initially, a single animal was selected and treated with the tsst substance in order to minimize unnecessary use of additional rabbits in case the test substance was corrosive or severely irritating. One screen animal received three separate semi-occluded applications of the test substance for the three-minute, 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 of each exposure, the collars (if applicable) and bandages of each rabbit were removed and the sites were wiped with disposable paper towels moistened with tepid lap water. -12- Project No.: WIL-189161 Sponsor: E. I. du Font de Ncmours and Company DuPont HLO-1998-00730 OBSERVATIONS: Mortality: The rabbits were observed twice daily (morning and afternoon) for mortality for the duration of the study. Dermal Observations: The three-minute and one-hour dose sites were observed once immediately after bandage removal for the presence or absence of corrosion. Dermal irritation was also evaluated and graded in accordance with the method of 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 of Draize (Appendix A). The adjacem areas of untreated skin were used for comparison. In order to facilitate derma! observations, the areas of application (four-hour site) of two rabbits were clipped free of hair after collecting me 72-hour dermal scores. Clinical Observations: The rabbits were observed for clinical signs oftoxicity at each dermal evaluation (excluding the three-minute and one-hour exposures). Body Weights: Body weights were obtained and recorded on study day 0 (initiation) and at each animal's termination from the study. Termination: Upon termination, the rabbits were euthanized by intravenous injection of sodium pentobarbital solution and discarded. -13CorapanySaniBzed, Does ncifeonfam 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 and edema) were calculated for each animal separately from numerical scores obtained at die 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 or Irritation Criteria: 1- Corrosive The test substance will be considered to be corrosive if it produces full thickness destruction of the skin tissue on at least one animal during me test for skin irritation. The substance will be classified as corrosive and assigned me symbol "C" and the indication of 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 of 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 of skin tissue occurs as a result of up to four hour exposure or if this can be predicted. 2. Irritant The test substance will be classified as "IRRITANT" and will require the "Xi" symbol and the indication of 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 or more after me end of the exposure period. Inflammation is significant if the mean values of die scores for either erythema and eschar formation or edema formation corresponds to one or more of the following mean values calculated for each animal separately and has been observed in two or more animals: -14- r^-i'-^,,a.-',&-,e--p,,e--l;-^-- ''"'c-'-^T-''""-'-''? -UF^Siil^-^r ^^ "--'--'--' -'-- ' -'J''it,"<- 3in <.3w'A Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company DuPont HLO-1998-00730 DATA ANALYSIS AND INTERPRETATION OF RESULTS (continued): Dermal Lesion Erythema and eschar formation Edema Mean Value 2.0 or more 2.0 or more 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. loflammadon 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. -15eompany Sanitized. Does's sot eanfa'n TSCA CBf Project No.: WIL-189161 Sponsor: E. I. du Port de Neuuurs arid Company DuPont HLO-1998-00730 RESULTS: Mortality: No deaths occurred during the study. Dennal Observations fTaMes 1 and 2): For the screen animal, there was no evidence "f corrosion following assy of the three different exposure times (three-minute, one-hour, and four-hour exposures). There was no dermal irritation observed for we three-minute or one-hour exposures. Erythema (score of 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 H-able 3}: No clinical signs of toxicity were observed during me study. Body Weijiiits (Table 3): No remarkable changes or differences were noted in body weights during die study. -16- eci?ipaiiy oan.tiS-ea. 'ti<i:t .;.-'. ^>,.,'H'aifi tSCft CQi Project No.: WIL-189161 Sponsor: . I. du Pont de Nemours and Company DuPont HLO-1998-00730 CONCLUSIONS. Under the conditions of testing and according to the guidance provided in EEC Directive . 93/21, H-22482 is not classified as an irritant. /^Fom G/Kern. B.S. ^r Stmudvy Director /A/^ Date -17Company Sanitized. Does not confam T8CA CBi Project No.: WIL-189161 Sponsor: E. I. du Font de Nemours and Company DuPont HLO-1998-00730 Acute Dermal Irritation/Corrosion Study with H-22482 in Rabbits TABLES 1-3 -18- SaWtpsm' SsniSzed, ")Q'S'?. "w- 5o'"<?@??' T^r'A ":r;f Project No.: WIL-189161 Sponsor: E. I. du Pont de Nemours and Company TABLE 1 DuPont HLO-1998-00730 ACUTE DERMAL IRRITATION/CORROSION STUDY WITH H-22482 IN RABBITS Animal Sex Site 23581 23588 23591 M A PA F A INDIVIDUAL DERMAL SCORES Erythema 1 hr 24 hr 48 hr 72 hr 4d Ir 2r 2r Ir Ir lr 2r lr lr lr 2r Or Or Or - 5d 6d 7d lr Ir Or lr 0 - - , - Edema ^mnal Se Site 1 hr 24 hr 48 hr 72 hr 4d 5d fid 7d ^ {| 23581 23588 23591 MPp AAA OO0 OO0 0 0 0 0 0 0 0 0 - 00- 000 - - - hr = Hour d = Day M = Male F = Female r = Residual test material present on the site - = Dermal iiritation previously subsided; animal was terminated from the study A = Four-hour sites EEC: Animal Number 23581 23588 23591 JEnahema. 1.67 1.33 0.00 ^dana. 0.00 0.00 0.00 1 19 91, J- -' Company Sanitized. Does not contain TSCA CBI Project No.: WIL-189161 Spoil r: E. I. du Pont de Nemours and Company TABLE 2 DuPont HLO-1998-00730 ACUTE DERMAL IRRITATION/CORROSION STUDY WITH H-22482 IN RABBITS INCIDENCES OF DERMAL RESPONSE (SCORES)' core 0 1 2 I irythema1' 1 hr 24 hr 48 hr 72 hr -M. Ji Jd. JV. 0/3 1/3 1/3 1/3 0/2- 0/2" 1/2 l/l" 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 Edema1' gore 1 hr 24 hr 48j;r 72 hr Jd. Jd. M .21 0 3/3 3/3 3/3 3/3 2/211 2/21' 2/26 i/r1 1 0/3 0/3 0/3 0/3 0/2 0/2 0/2 0/1 2 0/3 0/3 0/3 0/3 0/2 0/2 0/2 0/1 'According to EPA test guidelines. "Four-hour sites evaluated. Two rabbits were evaluated for dermal irritation since irritation for one animal had totally cleared. ''One remaining rabbit was evaluated for dermal irritation since dermal irritation bad completely subsided for the other two animals. hr = d = Hour Day 20 Project No.: WIL-189161 Sponsor: E. I. du Font de Nemours and Company TABLES DuPont HLO-1998-00730 ACUTE DERMAL IRRITATION/CORROSION STUDY WITH H-22482 IN RABBITS INDIVIDUAL BODY WEIGHTS (GRAMS) Group 0.5 g/site Animal Sex 23581 M 23588 F 23591 F Initiation (DavO) 3464 3258 3189 Termination (tntsrvat) 3668 (day 7) 3279 (day 6) 3283 (72 homs) M Male F Female CLINICAL OBSERVATIONS No clinical findings were noted for any animals during the study. 21 "Company Sanitized. Ooes noi eoniarn TSCA CBI Project No.: WIL-189161 Sponsor: E. I. du Font de Nemours and Company APPENDIX A DuPoni HLO-1998-00730 ACUTE DERMAL IRRITATION/CORROSION STUDY WITH H-22482 IN RABBITS SCORING CRITERIA FOR DERMAL REACTIONS Evaluation of Dermal Reactions* Erythema and Eschar Formation 0 No erythema 1 Very slight erythema (barely perceptible, edges of area not well defined) 2 Mi.ld 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 No edema Very slight edema (barely perceptible, edges of area not well defined) Mild edema (edges of area well defined by definite raising) Moderate edema (raised approximately 1 mm) Severe edema (raised more than 1 nun and extending beyond area of exposure) wi *Draize, J.H., 1959. The Appraisal of the Safety of Chemicals in Foods. Drugs and Cosmetics. Dermal Toxicity. pp. 46-59. Assoc. of Food Drug Officials of the U.S., Topeka. Kansas. 23 Company Sanitized. Sees not eonta'm TSCA CBi Project No.: W11..-189161 .Sponsor: 1-;. 1. du Pent tic Ncmours and Company DuPoni HLO-1998-007:^0 Acute Dermal Irritation/Corrosion Study with H-22482 in Rabbits APPENDIX B Protocol '24- SarsiEizsc, ^oes nes (.^HtaEn TSCA CBS Snidy Number: OTL. /jS9// DuPont HLO-1998-00730 rani\ MASTER PROTOCOL COVER SHEET Sponsor E.I. du Font de Nernoun md Company Msster Protocol No.: E05 (Sponior) A. Title of StudyAcute DenMiImtation/Coirosion Study in Albino RtbbiB B. Protocol Moriifi^ffn') n. PERSONNH. INVOLVED IN THE STUDY A. Sponsor ReonKenranve Nttue: Tncy A. Filliben Tide: Toxicology Associate 2) m. STUDY 'CTBDULEpATA A. EB!BOse(LEn>erimental San DateB. Eroposei?Experimental Tennii|?tip" Ra": C. faooostaiAudiied Draft Renon Da(<- 3) IV.TESTMATEMALPATA ^VG^f ^ ^ At7/9'7 IS/IT/iS'7 .^ . T B. ^^n^TfllBHBilH ' [HIHI . D- Siabiiity- Stable Bj (Sponsor) (Sponsor) (Sponsor) cvm^\ fvm\ {. wn\, (Sponsor) (Spaasor) (Sponsor) (Sponsor) WIL RESEARCH LABORATORIES. INC. A Subsidiary of Great Lakes Chemical Corporation, Ashland, OH 44805-9281 (419)289-8700 25 Company SaniiE@a!. Ooes not contain TSCA CB( wn-- ^'//<s' / WIL) Masier Protocol Cover Shed Page Tu'0 DuPont HLO-1998-00730 01--------| ] E. Phvsica) Description: F. Storaj'e.Cpnditipns: Arnbicn! ismpcratm-c G. Personnel Safeiv Data: Normal laboratory procedures. CWTL) (Sponsor) For complete available safety H. CAS Number information see attached MSDS Cl^SU^ \ (Sponsor) (Sponsor) C. Reason for Protocol Modification: 1) Sponsor Representative :s specified. 2) Proposed scheduling information is added to the protocol. 3) Test material data are added to the protocol. Approved By: E.I. du Font de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkion Road P.O. Box JO Newark. DE 19714 s ^ ^ ^ . ^ ^ A ^ SponEw Representative ^1/3^7 Date Prepared By. WIL Research Laboratories, Inc. Ashland. OH 44805-9281 ^r Tom ff. Kern, B.S. Study Director ^/^/^ 7 7Date 26 DuPont HLO-1998-00730 PROTOCOL Acute Dermal Irritation/Corrosion Study in Albino Rabbits (OECD/EEC Guidelines) Master Protocol No.: E06 For E. I. du Poni de Nemours and Company Haskel) Laboratories for Toxicology and Industrial Medicine Elk-ton Road P.O. Box 50 Newark. DE 19714 By WIL Research Laboratories. Inc. Ashland. OH 44805-9281 June 4. 1997 WIL RESEARCH LABORATORIESIN, C. A Subsidiaryof Great Lakes Chemical Corporation,Ashland. OH 44805-9281 (419)289-8700 27 Csmpany anitiz@8. Does not eon8a?n TSCA CB( ACUTE DERMAL IRRITATION/CORROSION STUDY IN ALBINO RABHITS DuPoni HLO-1998-00730 Master Protocol No.: E06 J OBJECTIVE OF STUDY eTxopodseutreermtointehethsekinirroitfatailvbeinaondrabcboirtrso. sive potential of the test material following a single This protocol has been designed and the study will be conducted in genera! compliance the Organization for Economic Cooperation and Development (OECD) Guidelines for with Testing OofffCichiaelmJicoaulrsn,alSoecf ttihoen E4u0r4opaenadn European Economic Communities (EEC) guidelines in the Communities, 92/69. B4 (1992). The study will Laboratory be conducted in compliance with the U.S. EPA FIFRA and TSCA Good Principles ofPrGaocotidceLaSbtaonrdaatordrys P(4ra0ctCicFeR[CP(a8rIt)s 31060(Fainnadl)7A92n,nerxes2p]e. ctively) and the OECD H PERSONNEL rNVOLVED IN THE STUDY A. Sponsor Representative To be specified by Master Protocol Cover Sheet B. WIL Study Director Tom G. Kem, B.S. Group Supervisor - Acute Toxicology C. WIL Deputy Director Jonathan M. Hurley, B.S. Senior Biologist - Acute Toxicology D. Study Responsibilities 1. Tames L. Scfaardein, MS., A.T.S. Director of Research 2 Christopher P. Chengelis. Ph.D., D-A.B.T. Senior Toxicologist 3 JozefJ.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 Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 11. PERSONNEL INVOLVED IN THE STUDY (continued) 5. Ronald E. Wilwn, B.S Director of Informational Systems 6. Daniel W. Sved, Ph.D. Director of Metabolism and Analytical Ct-mistry 7. Sally A. Keeis, A.S. Manager of Vivarium 8. Deborah L. Little Manager of Quality Assurance 9. Kerin Clevidence, B.S. Group Supervisor of Gross Pathology and-Developmenta! Toxicology Laboratory 10. Chariene M. Lindsey, M.A. Manager of Technical Report Writing 11. Camey B. Jackson. D.V.M.. D.A.C.V.P., D.A.C.V.P.M. Assistant Director of Pathology and Veterinary Medicine 12. Jeany R. RandaH Biologist n - Acute Toxicology 111. STUDY SCHEDULE DATA A. Proposed Experimental Stan Date To be added by Master Protocol Cover Sheet B. Proposed Experimental Termination Date: To be added by Master Protocol Cover Sheet C. Proposed Audited Draft Repon Daie: To be added by Master Protocol Cover Sheet IV TEST SUBSTANCE DATA A. Identification: B. Lot Nuinbe." C. Purity: D Stability: To be added by Master Protocol Cover Sheet To be added by Master Protocol Cover Sheet Tu be added by Master Protocol Caver Sheet To be added by Mastel Protocol Cover Sheet Page 3 of 12 ?9 Company Sanitized. Does mi contarn TSCA CBS Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 IV. TEST SUBSTANCE DATA (continued) E. Physical Description: F. Storage Conditions: G. Personnel Safety Data: V. TEST SYSTEM To be documented by WIL Research Laboratories, added by Master Protocol Cover Sheet Inc. and To be added by Master Protocol Cover Sheei To be added by Master Protocol Cover Sheet A. Species: B. Strain: C. Source: D. Number on Study: E. Sex: F. Body Weight Range: G. Approximate Age: H. Identification System: L Justification for Selecrinn Albino rabbit New Zealand White Covance Research Products, Inc. Denver, PA or Kalanmoo, MI Three animals (except as noted in section Vin. C. of this protocol). Males and/or females 2.0 kg or greater at initiation ofuosing. 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 for primary skin irritation Studies. The experimental design uses the procedures and standards required by the current and proposed international guidelines. Page 4 of 12 30 SofflpaiTf SgTiifesA Dc-Si- ,: Acme Dermal Irritation/Corrosion Study in Albino Rabbits DuPonI HLO-1998-U0730 VI SPECIRr MAINTENANCE SCHEDULE A. Animal Housing The animals will 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 thai 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 Inte-national (AAALAC International). B. Environmental Conditions Controls will be set to maintain temperature at 67 4F and relative humidity at approximately 30-70%. Fluorescent lighting conn-oiled by light timers will provide illumination for a 12-hour light/dark photoperiod. Temperature and relative humidity will be recorded once daily. C. Drinking Water Reverse osmosis treated municipal water will be available ad libinan. Fillers servicing die automatic watering system are changed regularly according to Standard Operating Procedures. Water supplying the laboratory is anaiyzcJ 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 LabDiet0 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 ui concentrations that would be expected to affect the outcome of the study. v". EXPERIMENTAL DESIGN A. Animal Receipt and Acclimation Each animal will be inspected by a qualified lectuurian 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 daily 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 HOK coniam TSCA CB( Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 VII. EXPERIMENTAL DESIGN (continued) B. Route and Rationale of Test Material Administration The and route cf administration will be dencal in older to lest the potential for dermal irritation corrosion. The dermal route is a possible route of exposure in humans. C. Body Weights The body weight of each animal v,ill be determined on study day 0 and at Rabbits remaining on study beyond day 7 will be weighed at least once a termination. week it' the study is extended. Vffl. DOSAGE LEVELS AND TREATMENT REGIMEN A. Treaaneo; Levels The same dose level will be used for all three animals (0.5 ml or 0.5 g per test site). ^- Treatment Groups Following stock the acclimation period, three rabbits will be arbitrarily selected from available based upon health and a body weight of 2.0 kg or greater. No separate control group will be utilized; each animal will serve; as its own control. All test sites will be left intact. C. Initiation Procedure If it is anticipated ihat the test substance may be severeJy irritating and/or saunbimstaalncsec.reeTnhrseteudsyitewsillonbethdisosseidng:loe eavnaimluaaltewtihlle riercrietiavteivein/cdoivrirdousaivl e poctoenrrtoiaslivoef, athseintgeslet substance for three minutes, one hour. and four hours. exposures of the test The mres-minute be dosed and observed for the presence of severe irritation/corrosion exposure site will immediately following . the unwrap. If no serious skin reaction is present on the tLree-minute site, exposure site will be dosed immediately following and observed .or 'he presence of severe tue one-hour irritation/corrosion the unwrap. If no serious skin reaction is present on the one-hour asfittee,r ttheset fsouubrs-thaonucer ex{ osure site will be dosed and observed t.;Droximately 30-60 minutes removal. If severe irritation (erythema, edema and/or eschar) is on the single animal screen, no furher testing will be obsa-.ed after any exposure conducted without the specific period request of the Sponsor. If corrosion is observed after any exposure, the written study will immediately be terminated. If neither a corrosive effect nor a severe irritant observed afrer the four-hour exposure, the study will be completed using two effect is additional animals, each receivii.^ a single four-hour exposure. Page 6 of 12 32 ^QPfSKi 1'3C., Acme Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 V.ri. DOSAGE LEVELS AND TREATMENT REGIMEN (continued) D- Method of Administration Approximately24 hours prior to dermal applications, be clipped free will constitute of hair with an Osier small animal the back and flanks clipper. The clipped of each rabbit will abnormalities approximately 25% of the total body surface area on each animal of the or injuries will be excluded. back and delineated The test sites will be alorecaa.tedAlnatiemraalls with dermal with four dots made with to the midline approximately 2.5 centimeters apart (~6 cm2) arranged in a water-insoluble ink spaced square. Approximately0.5 ml of the undiluted test substance applied evenly solutions (if to each test site covering an area at or 0.5 g of solid approximately 6 or semi-solid will be prepared) will (additional cost). Solid be perfonned at the specific cm2. Analysis of dosing written request of the materials will be slightly moistened with water or Sponsor other suitable vehicle (amount recorded) to form a thick documented in'die study records. paste. The amount of vehicle used will be Each test site will be immediately gauze patch. The patches are covered with secured in place a wo-ply. approximately one-icsh square then be wrapped with gauze bandaging that with surgical porous tape. is The trunk will irritating tape. period.? Collars will secured with be applied to each animal for the several wrappings of non- study, to prevent ingesiion all test substance of the test material and/or onewrappings. and four-hour For the exposure Only ons test substance applied is ^sumed to be a-ailable for absorption by purpose of this will be applied to each ^aibit. me test system. IX. OBSERVATIONS After each exposure, the bandages will be off with skin). disposable towels moistened with removed water (as and the residual thoroughly test substance gently wiped Alternate moistening materials may be used (to be as possible documented without irritating the to remove residua! test substance upon approval by the Study Directori.n study records) in order Approximaiely-30-60minutes after test substance test sites will be examined Draize Technique and the degree of removal on four-hour exposure erythema and edema sites only, the (Appendix A). Additional examinations will be recorded performed according to the 24, 48 and 72 hours afte: patch continue daily up to seven removal. d<iys after dosing. will only be conducted immediately If irritation is present at 72 at approximately hours, Examination and recording of scoring erythema will exposure sites. The animals will upon bandage removal on the three-minute andand edema excluding the 3 and 60-minute also be observed for clinical Signs at each dermal one-hour mortality and signs of illness, injuerxyp, oosruraebsn)o. rmTahl ebeahnaimviaolrs. will be observed twice edvaaliulyatfioonr The areas of application will be clipped necessary and as needed during the study ro frfeaeciliotaftehair following the 72-hour evaluation, if accurate dermal observations. Page 7 of 12 33 Ssniffzed, Does not confarn TSCA CSt Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 IX. OBSERVATIONS (continued) Individual animals may be teiminiied if no irritation is present ac the 72- hour or any subsequent observation. If irritation is present at the end of seven days, observation will continue daily until it subsides or is obviously irreversible up to 14 days (additions? cost). The snidy need not normally exceed 14 days after application unless specifically requested and authorized by the Sponsor (additional cost). X. GROSS PATHOLOGY The animals will be euthanized by an intravenous injection of euthanasia solution upon termination and discarded. Collection of skin samples from the four-hour exposure sites may be collected and preserved in 10% neutral-buffered formalin for histopathological examination at the specific written request of the Sponsor (additional cost). A gross necropsy examination on major organ systems of the dioracic and visceral cavities will be conducted on all animals found dead or euthanized in extremis. XI. DATA ANALYSIS AND INTERPRETATION OF RESULTS 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 response (erythema and edema) by summing the scores obtained from all animals at the 24-, 48- and 72-hour observations (excluding the 3 and 60 minute exposures). The three means corresponding to the three observation periods will be averaged and overall averages will be obtained for each dermal lesion. The results will be 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 or Irritation Criteria: I. Corrosive The test substance will be considered to be corrosive if it produces full thickness destruction of the skin tissue on at least one animal during the test for skin irritation. The substance will be classified as corrosive and assigned the symbol "C" and die indication of danger "corrosive." Risk phrases shall be assigned in accordance with the following criteria: R35 "CAUSES SFVERE BURNS" If, when applied to healthy intact animal skin, full thickness destruction of skin tissue occurs as a result of up to three minute exposures or if this result can be predicted. Page 8 of 12 34 Acuie Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-1998-00730 XI. DATA ANALYSIS AND INTERPRETATION OF RESULTS fcontinun R34 "CAUSES BURNS" Irritant If, when tissue applied to healthy intact animal skin. full thickness desmiction of skin occurs as a result of up to four hours exposure or if this can be predicted. The test substance and the indication will be classified as "IRRITANT" and will require the "Xi" symbol of 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 or more after toe end of the period.- Inflammation erytheiaa and is significant if cLe mean values of the scores exposure for either the following eschar formation mean values: or edema formation corresponds to one or more of 1Dermal Lesion Erythema and eschar formation Edema Mean Score 2.0 or more 2.0 or more Ail scores used in at each of the reading tunes (24, 48. and 72 hours) for an effect will be calculating the respective mean values. XII. Inflammation end of the of (he skin is also significant if it persists in at least two animals at the observation tune Particular effects such as hyperplasia, scaling, discoloration, fissures, scabs, and alopecia should be taken into account. The final report will include the following receipt, methods, individual dermal summary, objective, test substance identification and summarizing mean values for observations, mortality, each individual body weights, a table classification of me test substance based rabbit, gross necropsy findings (if applicable), and draft report will be provided on dermal irritation properties. One copy of the audited copies of me report will be praopvpirdoexdimuaptoenlyfmsixaliwzaeteikosn.following completion of the study. Two Page 9 of 12 35 'tern TSCA CW 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 by LWabILoraRtoersieeasr,cIhnc.Laboratories. Inc. will be collected and maintained by WIL Research XIV. WORK PRODUCT Sponsor will have title to all documenrauon records, raw data, slides, specimens, or other work product generated during the perfonr.^nce 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, Inc. 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 will be 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. Quality The raw data and draft report will be audited by the WIL Assurance Unit prior to submission to the Sponsor to assure that the final report accurately describes the conduct and the findings of the study. XVI. PROTOCOL MODIFICATION This Master Protocol shall be used for multiple studies. A Masw Protocol Cover Sheet shall be prepared for each study which will use this design. The Master Protocol ami Master Protocol Cover Sheet shall constitute the protocol for an individual study. In addition, the date of Study Director signature on the Master Protocol Cover Sheet shall be considered me date of study initiation. Modification of the protocol may be accomplished during the course of this investigation. However, no changes will be made in the study design without the verbal or written permission of the Sponsor" In me event that the Sponsor verbally requests or approves a change in the protocol, such changes will be made by appropriate documentation in the form of a protocol amendment. All alterations of the protocol and reasons for the modifications) will be signed by the Study Director and the Sponsor Representative. Page 10 of 12 36 CSoiap&nyyansSizea. F3os^ ^.,;"^-u;ite!n'? Y'yCA OS Acute Dermal Irritation/Corrosion Study in Albino Rabbits DuPont HLO-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: 1. The Sponsor signature on this protocol documents for the Study Director the Sponsor's assurance that the study described in this protocol does not unnecessarily duplicate previous experiment:;. 2. Whenever possible, procedures used in this study have been designed to avoid or minimize idniswcorimttefonrtl,abdiosrtaretossryoSr tpaanidnartod aOnpimeraaltsin. gAPlrlomceedthuoredss. are described in this study protocol or 3. .Animals that experience severe or chronic pain or distress that cannot be relieved will be painlessly euthanized as deemed 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 confonnance with me above-referenced regulation. XVm. PRC;OCOL APPROVAL E. I. du Pont de Nemours and Company Haskell Laboratories for Toxicology and Industrial Medicine Elkton Road P.O. Box 50 Newark. DE 19714 i Mrt. ,/i U' Carol Finlayf B A. Sponsor Representative 1/t,/1^ Daic WIL Research Laboratories, Inc. Ashland, OH 44805-9281 ^'Tom G. Kem, B.S. Study Director _ ^ / ADate^ \2^y<g.^X^A^, Iffacy Filliben Sponsor Representative ^ / ^Date Page ll of 12 37 Company SarsESized. Does not eontsfn TSCA CB8 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 zrea 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 ^SS'-i^f'srsW,UBS"! ?^:?C-:?P?S??TgCQ C^