Document 1y580R4mwoa77652V1xvyqZB5

92091 SPONSOR ElfAtochemSA. Cours Michelet La Defense 10 Paris-la-Defense CEDEX Prance AR226-3098 STUDY TITLE ACUTE DERMAL IRRITATION IN RABBITS TEST SUBSTANCE STUDY DIRECTOR Xavier Manciaux STUDY COMPLETION DATE 20 May 1998 PERFORMING LABORATORY Centre International de Toxicologie (C.I.T.) Miserey - 27005 Evreux. - France LABORATORY STUDY NUMBER 16046TAL Company San?fzed. Do^ ""' <"'"fa;- CONTENTS STATEMENT OF THE STUDY DIRECTOR 4 OTHER SCIENTISTS INVOLVED IN THIS STUDY 4 STATEMENT OF QUALITY ASSURANCE UNIT SUMMARY RESUME 1. INTRODUCTION 2. MATERIALS AND METHODS 2.1 TEST SUBSTANCE 8 2.1.1 Identification 8 2.1.2 Formulation procedure 8 2.2 TEST SYSTEM 9 2.2.1 Animals 9 2.2.2 Environmental conditions 9 2.2.3 Food and water 9 2.3 TREATMENT 10 2.3.1 Preparation and selection of the animals 10 2.3.2 Study design 10 2.3.3 Application of the test substance 10 2.3.4 Date of treatment 10 2.4 CUTANEOUS EXAMINATIONS 10 2.5 DESCRIPTION AND EVALUATION OF CUTANEOUS REACTIONS 11 2.6 INTERPRETATION OF RESULTS AND CLASSIFICATION OF SUBSTANCES 11 ' 11 2.6.1 Interpretation of the results . 2.6.1.1 Criteria for irritation 11 2.6.1.2 Criteria for corrosion 11 2.6.2 Classification of the test substances 11 ' 2.6.2.1 Irritant substances 11 2.6.2.2 Corrosive substances 12 2.7 PROTOCOL ADHERENCE 12 2.8 ARCHIVING 12 3. RESULTS (table 1) 4. CONCLUSION __ 'KsfSBm'--S^^.--6-9'v^tea<?ret conta.in wTS&CApo- '13 pSBpany ^ Table 1: Individual cutaneous examinations and mean values of the scores recorded at each reading (24, 48 and 72 hours) for each animal 14 APPENDICES 1. Test article description 2. Diet formula 16 17 19 and 20 -e..r--mw-^^.nni^:^!^0160^' STATEMENT OF THE STUDY DIRECTOR The study was performed in compliance with the principles of Good Laboratory Practice as described in: . O.E.C.D. principles of Good Laboratory Practice, Decision Concerning Mutual Acceptance of Data in the Assessment of Chemicals, C(81)30(final) Annex 2. May 12,1981. . Decret N 90-206 du 7 mars 1990 concemant les Bonnes Pratiques de Laboratoire (Journal Officiel du 9 mars 1990), Ministere de 1'Industrie et de 1'Amenagement du Territoire. . Council Directive 87/18/E.E.C. of 18 December 1986 on the harmonization of laws, regulations or administrative provisions relating to the application of the Principles of Good Laboratory Practice and the verification of their applications for tests on chemical substances (O.J. No. L 15 of 17.1.87). I declare that this report constitutes a true and faithful record of the procedures undertaken and the results obtained during the performance of the study. This study was performed at the Centre International de Toxicologie (C.I.T.), Miserey, 27005 Evreux, France. Toxicology y" ^^o^-v\ --~i X. Manciaux Study Director Date: 20 May 1998 Doctor of Pharmacy OTHER SCIENTISTS INVOLVED IN THIS STUDY For Pharmacy: P. 0. Guillaumat Doctor of Pharmacy For Toxicology: C. Pelcot Study Supervisor 'fewanySanW..^r.Ot--oneoslcnoonilcainTSCACBl w STATEMENT OF QUALITY ASSURANCE UNIT Type of inspections Protocol Report Inspections 28 August 1997 11 May 1998 Dates Reported to Study Director (*) 2 September 1997 12 May 1998 Reported to Management (*) 2 September 1997 12 May 1998 At about the same time as the study described in this report, "process-based" and routine facility inspections of critical procedures relevant to this study type were made by the Quality Assurance Unit. The findings of these inspections were reported to the Study Director and to C.I.T. Management. The inspections were performed in compliance with C.I.T. Quality Assurance Unit procedures and the Good Laboratory Practice. The reported methods and procedures were found to describe those used and the results to constitute an accurate and complete reflection of the study raw data. ( ^ \ ^ < ^ ^ ^ 0 )V L. Valette-TaIbi Date: 20 May 1998 Doctor of Biochemistry Head of Quality Assurance Unit and Scientific Archives (*) The dates indicated correspond to the dates of signature of audit reports by Study Director and Management. -.tedfloe^-o^0"181"^^^1 -. SUMMARY At the request of Elf Atochem S.A., Paris-lg-Defense, France, the potential of the test substance HB^IHBHHHHHBBBBi10 i^uce skin irritation was evaluated in rabbits to^^^). ' according (No. 404, 17th July 1992) and E.C. (92/69/E.E.C., 84, 31st July 1992) guidelines. The study was conducted in compliance with the Principles of Good Laboratory Practice Regulations. Methods The study design was established according to available information on the test substance and the above guidelines. The test substance was applied for 4 hours to three male New Zealand White rabbits. A single dose of 0.5 ml of the undiluted test substance was applied to the closely-clipped skin of one flank. The test substance was held in contact with the skin by means of a semi-occlusive dressing. Cutaneous reactions were observed approximately 1 hour, 24, 48 and 72 hours after removal of the dressing and then daily until the end of the observation period. The mean values of the scores for erythema and oedema were calculated for each animal. Results Well-defined to severe cutaneous reactions were observed in all animals from day 1. These reactions persisted up to day 11 at the latest. Dryness of the skin was noted in all animals from day 5 or 6 up to day 13 or 15. Mean scores over 24, 48 and 72 hours for each animal were 3.0, 2.0 and 2.0 for erythema and 4.0, 2.0 and 2.7 for oedema. Conclusion Under our experimental conditions, the test substance is irritant when applied topically to rabbits. According to the classification criteria laid down in Commission Directive 93/21/E.E.C. (27th April 1993) adapting to technical progress for .the eighteenth time Council Directive 67/548/E.E.C., the test substance was considered irritating to skin. company S^Ooc^c.".^8"08' RESUME MmHmBBlHHHB!]3?1'^ ,A la demande deElfAtochem S.A., j^aris-la-Defense, France, les proprietes imtantes du produit I application cutanee ont etc evaluees chez Ie Lapin "selon les lignes directrices de 1'O.C.D.E. (No. 404, 17 juillet 1992) et de la C.E.E. (92/69/E.E.C., B4,31juilletl992). L'etude a ete realisee conformement aux regles de Bonnes Pratiques de Laboratoire. Methode L'etude a ete realisee selon les informations disponibles sur Ie produit et les lignes directrices mentionnees ci-dessus. Le produit a ete applique pendant 4 heures sur 3 lapins males New Zealand White. Une dose unique de 0,5 ml de produit tel quel a ete appliquee sur une surface de peau tondue d'un des flancs. Le produit a ete maintenu en contact avec la peau au moyen d'un pansement semi-occlusif. Les reactions cutanees ont etc observees environ 1 heure, 24, 48 et 72 heures apres 1'enlevement du pansement puis quotidiennement jusqu'a la fin de la periode d'observation. La moyenne des scores a etc calculee pour 1'erytheme et pour 1'oedeme pour chaque animal. Resultats Des reactions cutanees bien definies a severes sont observees chez tous les animaux a partir du jour 1. Ces reactions persistent jusqu'au jour 11 au plus. Une secheresse cutanee est notee chez tous les animaux du jour 5 ou 6 au jour 13 ou 15. Les scores moyens calcules pour chaque animal apres 24, 48 et 72 heures sont de 3,0; 2,0 et 2,0 pour 1'erytheme et de 4,0; 2,0 et 2,7 pour 1'oedeme. Conclusion Dans nos. conditions experimentales. le produit|| considere irritant par vote cutanee chez le Lapin. '" Selon les criteres de classification decrits dans la Directive 93/21/C.E.E (27 avdl 1993) portant dix-huitieme adaptation au progres technique de la Directive 67/548/C.E.E., le produit est considere irritant pour la peau. 'SanIlWO^^con^nW^- 1. INTRODUCTION The objective of this study was to evaluate the potential of the test substanca| to induce skin irritation following a single topical application to rabbits. - In the assessment of the toxic characteristics of a test substance, determination of the irritant and/or corrosive effects on the skin of mammals is an important initial step. Information derived from this test serves to indicate the possible hazards likely to arise from exposure of the skin to the test substance. This study was conducted in compliance with: . O.E.C.D. guideline No. 404,17th July 1992. . E.C. Directive No. 92/69/E.E.C.. B4,31st July 1992. 2. MATERIALS AND METHODS 2.1 TEST SUBSTANCE The test substance|B^^----B!|HBJusien dthe study was supplied by ElfAtochem S.A. The test substance was identified as follows: . batch number: protocol and labeUinAg^(^JB^^^^^\ ^J^ - . Elf Atochem filing number^U------B1 . on the test article description----||------------r container: one plastic flask . date of receipt: 23 August 1997 . storage conditions: at room temperature and protected from light . expiry date: August 1998. Data relating to the characterization of the test substance are documented in a test article description (presented in appendix 1) provided by the Sponsor. At the finalization of the study report, an analytical certificate was not available. Characterization of the test substance, which appropriately defines the tested batch, is under the responsibility of the Sponsor. The pH of the test substance ^IIHBHHBJ 2.1.2 Formulation procedure The test substance was used undiluted. 'CompanySartTOzecl.Does ttol contain TSCA CT 2.2 TEST SYSTEM 2.2.1 Animals Sex, species, strain: male New Zealand White rabbits. Justification of the test system : species generally accepted by regulatory authorities for this type of study. Breeder: Elevage Cunicole de Val de Selle, 80160 Prouzel, France. Number of animals and identification: three animals were used, as recommended by the international guidelines and taking into account that a good correlation of results can be obtained with either three or six animals (1), The animals were identified individually with a metal tag in the ear. Weight: on the day of treatment, the animals had a mean body weight of 2.5 0.2 kg. Acclimatization: at least 5 days before the beginning of the study. 2.2.2 Environmental conditions The conditions in the animal room were set as follows: .temperature: 183C . relative humidity: 30 to 70% . light/dark cycle: 12h/12h . ventilation: approximately 12 cycles/hour of filtered, non-recycled air. The temperature and relative humidity woce under continuous control and recording. The records were checked daily and retained. In addition to these daily checks, the housing conditions and corresponding instrumentation and equipment were verified and calibrated at regular intervals. The animals were housed individually in polystyrene cages (35 cm x 55 cm x 32 cm or 48.2 cm x 58 cm x 36.5 cm). Each cage was equipped with a food container and a water bottle. 2.2.3 Food and water During the study, the animals had free access to 112C pelleted diet (U.A.R., Villemoisson-sur-Orge, France). Each batch of food was analysed by the supplier for composition and contaminant levels. The diet formula is presented in appendix 2. 91360 Drinking water filtered by a F.G. Millipore membrane (0.22 micron) was provided ad libitum. Bacteriological and chemical analysis of the water and diet, including the detection of possible contaminants (pesticides, heavy metals and nitrosamines), are performed regularly by external laboratories. The results of these analyses are archived at C.I.T. No contaminants are known to be present in the diet or drinking water at levels which may be expected to interfere with or prejudice the outcome of the study. (1) Hatoum, N.S.; Leach, C.L.; Talsma, D.M.; Gibbons, R.D.; Garvin, P.J.: A statistical basis for using fewer Rabbits in dermal irritation testing. Journal of the American College of Toxicology. 9: 49-60 (1990). . _ Dtfes Hoi TSCA CB( c" 2.3 TREATMENT 2.3.1 Preparation and selection of the animals The day before treatment, the flanks of each animal were clipped using electric clippers and the skin of each animal was examined. Only animals with healthy intact skin were used. 2.3.2 Study design The study design was established according to available information on the test substance and the O.E.C.D. (No. 404) and B.C. (92/69/E.E.C., B4) guidelines. The test substance was applied for 4 hours to three animals. 2.3.3 Application of the test substance The test substance was used undiluted. A single dose of 0.5 ml of the test substance was placed on a 6 cm2 dry gauze pad (Cooperative Pharmaceutique Francaise, 77000 Melun, France), which was then applied to the right flank of the animals for 4 hours. The test substance and the gauze pad were held in contact with the skin by means of an adhesive hypoallergenic aerated semi-occlusive dressing (Laboratoires de Pansements et d'Hygiene. 21300 Chenove, France) and a restraining bandage (Coheban9), Laboratoires 3M Sante, 92245 Malakoff, France). The untreated skin served as control. No residual test substance was observed on removal of the dressing. 2.3.4 Date of treatment Aninlal number 01 02 03 Date of treatment (day 1) 7 October 1997 7 October 1997 .7 October 1997 End of the observation period 20 October 1997 20 October 1997 21 October 1997 2.4 CUTANEOUS EXAMINATIONS The skin was examined approximately 1 hour, 24, 48 and 72 hours after removal of the dressing. Following the O.E.C.D. and B.C. guidelines: . when there was no evidence of dermal irritation after 72 hours, the study was ended. . when there was persistent cutaneous imitation after 72 hours, the observation period was extended to a maximum .of 14 days (until day 15) in order to determine the progress of the lesions and their reversibility. . when severe irritant effects were observed, the animals were killed on humane grounds. Any change in the animals' behaviour was noted. Company Sanitized. Does not contain TSCA CM 2.5 DESCRIPTION AND EVALUATION OF CUTANEOUS REACTIONS Dermal irritation was evaluated for each animal according to the following scoring scale: Erythema and eschar formation: . no erythema........................................................................................................... 0 . very slight erythema (barely perceptible).............................................................. 1 . well-defined erythema........................................................................................... 2 . moderate to severe erythema................................................................................. 3 . severe erythema (beet redness) to slight eschar formation (injuries in depth)...... 4 Oedema formation . no oedema.............................................................................................................. 0 . very slight oedema (barely perceptible)................................................................. 1 . slight oedema (edges of area well-defined by definite raising)..........................-... 2 . moderate oedema (raised approximately 1 millimetre)......................................... 3 . severe oedema (raised more than 1 millimetre and extending beyond area of exposure)........................................................................................................... 4 Any other lesions were noted. 2.6 INTERPRETATION OF RESULTS AND CLASSIFICATION OF SUBSTANCES The results obtained were evaluated in conjunction with the nature and the reversibility of the findings observed. 2.6.1 Interpretation of the results 2.6.1.1 Criteria for irritation A substance or a preparation is considered to be irritating to the skin if, when it is applied to healthy intact animal skin for up to 4 hours, significant inflammation is caused and which persists for 24 hours or more after the end of the exposure period. 2.6.1.2 Criteria for corrosion A substance or a preparation is considered to be corrosive if, when it is applied to healthy intact animal skin, it produces full thickness destruction of skin tissue on at least one animal during the test for skin irritation, or if the result can be predicted, for example: from strongly acid or alkaline reactions.' 2.6.2 Classification of the test substances 2.6.2.1 Irritant substances - symbol Xi, indication of danger "irritant", - phrases indicating the nature of special risks: R 38: "Irritating to skin" W^^^''1^' Inflammation is significant if: . the mean value of the scores is two or more for either erythema and eschar formation or oedema formation. The same will be the case where the test has been completed using three animals if the score for either erythema and eschar formation or oedema formation observed in two or more animals is equivalent to the value of two or more, . it persists in at least two animals at the end of the observation period. Specific effects such as hyperplasia, desquamation, discolouration, fissures, eschar and alopecia should be taken into account. All scores obtained at each reading time (24, 48 and 72 hours) for an effect are used by calculating the respective mean values. 2.6.2.2 Corrosive substances - symbol C, indication of danger: "corrosive", - phrases indicating the nature of special risks: - R 34: "Causes bums" If, when applied to healthy intact animal skin, full thickness destruction of skin tissue occurs as a result of up to 4 hours exposure, or if this result can be predicted. - R 35: "Causes severe bums" If, when applied to healthy intact animal skin, full thickness destruction of skin tissue occurs as a result of up to 3 minutes exposure, or if this result can be predicted. 2.7 PROTOCOL ADHERENCE The study was performed in accordance with Study Protocol No. 16046 TAL and subsequent amendments, with the following deviation from the agreed Study Protocol: - the relative humidity recorded in the animal room was sometimes outside of the target ranges specified in the protocol. This minor deviation was not considered to compromise the validity or integrity of the study. 2.8 ARCHIVING The study documentation and specimens generated during the course of the study are archived at C.I.T., 27005 Miserey, Evreux, France, for 10 years after the end of the in vivo phase of the study. The archived study materials include: . protocol and possible amendments, . raw data, . correspondence, . final report and possible amendments. On completion of this period, the archived study materials will be returned to the Sponsor, or may be archived at C.I.T. for a further period. l^^iflySanWzeir. Boesirolsonlain WA cw 3. RESULTS (table 1) Well-defined to severe cutaneous reactions (well-defined or moderate erythema, grade 2 or 3, and slight or severe oedema, grade 2 or 4) were observed in all animals from day 1. These reactions persisted up to day 11 at the latest. Dryness of the skin was noted in all animals from day 5 or 6 up to day 13 or 15. Mean scores over 24, 48 and 72 hours for each animal were 3.0, 2.0 and 2.0 for erythema and 4.0,2.0 and 2.7 for oedema. 4. CONCLUSION Under our experimental conditions, the test substance is irritant when applied topically to rabbits. According to the classification criteria laid down in Commission Directive 93/21/E.E.C. (27th April 1993) adapting to technical progress for the eighteenth time Council Directive 67/548/E.E.C., the test substance was considered irritating to skin. Cowoa-n? '$w6zeS. 1 Table 1: Individual cutaneous e:xaminaitions an<1 mean values of the scores recorded at each reading (24, 48 and 72 hours) i"or each;mimal Rabbit number Dermal Irritation ih Dl Scores 24h 48h D2 D3 Mean I nterpretation irritation (+) 72h score (1) (-) D4 01 Erythema 2 3 3 3 3.0 (+) Oedema 4 4 4 4 4.0 (+) Other * * * * 02 Erythema 2 2 2 2 2.0 (+) Oedema 4 2 2 2 2.0 (+) Other * * * * 03 Erythema 2 2 2 2 2.0 (+) Oedema 4 4 2 2 2.7 (+) Other * * * * (1) mean of scores on days 2, 3 and 4 h = hour D =day (+) = irritant according to E.E.C. criteria --' X^Wpirnyahmze<f. Does not contain Tgca ^ Table 1 (continued) ^--J Rabbit number Dermal Irritation Scores D5 D6 D7 D8 D9 DIG DI1 D12 D13 D14 D15 4200000000- 321 1000000- 01 Erythema Oedema Other *SSSSSSSS*. 02 Erythema 1 1 1 1 0 0 0 0 0 0 Oedema 0000000000 Other SSSSSSSSS* 03 Erythema 22222210000 Oedema 20000000000 Other *SSSSSSSSSS D =day * '=- None S = Dryness of the skin Cutaneous examination not performed .%apanySanilizea. D@es rot eeniain TSCA 661 ,,,,-.,,, .,,. ,,-.- -.-^_------_^-. APPENDICES CssspainySanitized. Does noi eonlain TSft^ CBt -"""^ ""'"" " ""1:----------J~ 1. Test article description CompanySanRizeA Does not contain TSCA CBf TOXICOLOGY DEPARTMENT CONFIDENTIAL II August 1997 elfatochem s.a. La defense 10, Cours Michelet 92091 Paris-la-Defense cedex, France TEST ARTICLE DESCRIPTION IDENTITY Test article name CAS number EINECS number Purity Origin Batch ElfAtochem filing number ElfAtochem, CERDATO, Serquigny PHYSICAL AND CHEMICAL PROPERTIES Appearance Specific gravity Boiling point Vapor pressure Flash point Solubility TOXICOLOGICAL INFORMATIONS AND USE SAFETY see fiche de donnees de securite STORAGE AND DISPOSAL Storage Expiry date Disposal in dark and at room temperature August 1998 - incineration ^^rnySanTOzeA Does not contain TSC^ W 2. Diet formula no1 contain TSCA C'Bt Ref: 112 COMPLETE DIET RABBIT MAINTENANCE DIET Appearance: 4.5 mm diameter granules Conditioning: bags of 25 kgs Daily portion: in accordance with race and body weight. Rabbits 100-150 g, water ad libltum. FORMULA % Cereals ..................................... Grain biproducts and legumes. Vegetable protein (soya bean meal, yeast) ............................. Vitamin and mineral mixture... AVERAGE ANALYSIS % Calorific value (Kcal/kg). Moisture........................... Proteins............................ Lipids............................... Carbohydrates (N.F.E.) Fibre................................. Minerals (ash) .................. AMINO ACID VALUES (calculated in mg/kg) Arginine..... Cystine....... Lysine ........ Methionine. Tryptophan., Glycine........ PATTY ACID VALUES (calculated in mg/kg) Palmitic acid..... Palmitoleic acid, Stearic acid......... Oleic acid........... Linoleic acid...... Linolenic acid.... 43.8 49 4.2 3 2200 10 13 2.7 49.3 17 MINIERALS (calcu Nat. P.................... .. Ca ................. . K ................... . Na ................. . Mg ................. . Mn ................ Fe................... Cu ................. Zn ................. Co ................. I Cl.................... .................. val. 3500 4500 11600 4002100 40 160 12 30 0.1 0' 500 [ated in mg/k^g) CMV val. 3500 4500 Total 7000 9000 0 1600 100 40 11600 2000 2200 80 140 300 15 27 45 75 1.5 1.6 0 0 3000 3500 8 6800 2100 4600 1600 1400 5200 6400 0 600 6400 vmAMINS (calculated per kg) Nat. CMV val. val. Total Vitamin A 2850 IU 6500IU 9350 IU Vitamin D3 30 IU 1000IU 1030IU Vitamin Bl 4.3 mg Omg 4.3 mg Vitamin B2 Vitamin B3 3.8 mg 16 mg Omg Omg 3.8 mg 16 mg Vitamin B6 Vitamin B 12 Img Omg 1 mg Omg 2mg Omg Vitamin E 16 mg 10 mg 26 mg Vitamin K3 Vitamin PP 6mg 55 mg 1 mg 5mg 7mg 60 mg Folic acid Omg Omg Omg Biotin Omg Omg Omg Choline 850 mg 200 mg 1050 mg ^leso-Inositol Omg Omg Omg 12100 2400 Available under quality "Control Ref.: 112 C" U.A.R., 7 rue Galliem, 91360 Villemoisson - Tel: 01.69.04.03.57 - Fax : 01.69.04.81.97 (Ref. Doc. UAR: 1992) S^l^nySiahWzea.OoesWeonlaintCSBC^ A