Document gb8j9M4b6pjJKa9g4nx5nMwD9

SPCW.SQJ? 92091 ElfAtochemS.A. Cours Michelet La Defense 10 Paris-la-Defense CEDEX France AR226-3068 >->^'lNjC-UL'JClN 1 Lrt-L STUDY TITLE ACUTE DERMAL IRRITATION IN RABBITS STUDY DIRECTOR Stephane de Jouffrey STUDY COMPLETION DATE 3 February 1997 PERFORMING LABORATORY Centre International de Toxicologie (C.I.T.) Miserey - 27005 Evreux - France LABORATORY STUDY NUMBER 14887 TAL Company Sanitized. Does not contain TS'CA CB? CONTENTS STATEMENT OF THE STUDY DIRECTOR OTHER SCIENTISTS INVOLVED EN TfflS STUDY. 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 Preparation 8 2.2. TEST SYSTEM 2.2.1 Animals 9 " 9 2.2.2 Environmental conditions 9 2.2.3 Food and water 9 2.3. TREATMENT 2.3.1 Preparation and selection of the animals 10 . 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 11 2.5. DESCRIPTION AND EVALUATION OF CUTANEOUS REACTIONS 11 2.6. INTERPRETATION OF RESULTS AND CLASSIFICATION OF SUBSTANCES 11 2.6.1 Interpretation of the results 11 2.6.1.1 Criteria for irritation ^ 11. 2.6.1.2 Criteria for corrosion 12 2.6.2 Classification of the test substance 12 2.6.2.1 Irritant substances 12 2.6.2.2 Corrosive substances ' 12 2.7. ARCHIVES 12 3. RESULTS (table 1) 13 4. CONCLUSION . 13 F Table 1: Mean values of the scores recorded at each reading (24, 48 and 72 hours) for all animals (4-hour exposure) three 14 ^any sa""^ed es no, contain TSCA CM APPENDICES 1. Test article description 2. Diet formula 15 16 18 and 19 ^"""----"----nrso^, STATEMENT OF THE STUDY DIRECTOR The study was performed in compliance with the following Principles of Good Laboratory Practice Regulations: . 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 conceraant les Bonnes Pratiques de Laboratoire (Journal Officiel du 9 mars 1990), Ministere de 1'Industrie et de 1'Amenagement du Territoire. 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 <S. de Jdufflrey Date: 3 February 1' Study-Director Doctor of Veterinary Medicine Head of Short-term and Environmental Toxicology OTHER SCIENTISTS INVOLVED IN THIS STUDY For Pharmacy: J. Richard Doctor of Pharmacy For Toxicology: C. Pelcot Study Supervisor Company Sanitized. Does not contain TSCA CBB STATEMENT OF QUALITY ASSURANCE UNIT 1. Specific study inspections Type of inspections Inspections Protocol Report 21 Oct. 96 20 Jan. 97 Dates Report to Study Director (*) 29 Oct. 96 21 Jan. 97 Report to Management (*) 29 Oct. 96 21 Jan. 97 2. Routine inspections performed on other studies of the same type according to a frequency defined in Q.A.U. procedures Inspected phase Inspections Dates Report toReport to Study Director (*) Management (*) Treatment/test substance Preparation/test substance 13 Aug. 96 25 Sept. 96 22 Aug. 96 25 Sept. 96 22 Aug. 96 25 Sept. 96 The inspections were performed in compliance with C.I.T. Quality Assurance Unit procedures and the Good Laboratory Practice Regulations. (*) The dates mentioned correspond to the dates of signature of audit reports by Study Director and Management. (^HjdtfT^ L. Valette-TaIbi Date: 3 February Doctor of Biochemistry Head of Quality Assurance Unit and Scientific Archives 1997 Company Sanitized. Does not contain TSCA CBf SUMMARY At the request of Elf Atochem S.A., Paris-la-Defense, France, the potential of the test substance according to O.E.C.D. (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 test substance in its original form was applied to the closelyclipped skin of the flank. The test substance was held in contact with the skin by means of a semi-occlusive dressing. Cutaneous reactions were observed approximately one hour, 24, 48 and 72 hours after removal of the dressing. The mean values of the scores for erythema and oedema were calculated for each animal. Results Very slight or well-defined erythema was noted in all animals at the 1-hour reading. It persisted for 24 hours in one animal, and for 48 hours in the two other animals. No oedema was observed. Mean scores over 24, 48 and 72 hours for individual animal were 0.3, 1.0 and 0.7 for erythema and 0.0 for oedema. Conclusion Under our experimental conditions, the test substance^ was considered non-irritant when administered by cutaneous route in rabbits. Company Sanitized. Does not contain TSCA CBl RESUME ^HBI^BIIHBHHH9 A la demande de Elf Atochem S.A., Paris-la-Defense, France, 1'irritation cutanee poiivant etre induite par Ie produit ^ ete evaluee chez Ie Lapin conformement aux lignes directrices de 1'O.C.D.E. (No. 404, 17th July 1992) et de la C.E.E. (92/69/E.E.C., B^, 31st July 1992). L'etude a ete realisee conformement aux regles de Bonnes Pratiques de Laboratoire. Methodes 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 sur le flanc. Le produit a ete maintenu en contact avec la peau au moyen d'un pansement semi-occlusif, Les reactions cutanees ont ete observees environ 1 heure, 24, 48 et 72 heures apres 1'enlevement du pansement. La moyenne des scores a ete calculee pour chaque animal pour 1'erytheme et pour 1'oedeme. Results ts Un erytheme tres leger ou bien defini est observe chez tous les animaux a la lecture 1 heure. II persiste pendant 24 heures chez 1 animal, et pendant 48 heures chez les 2 autres. Aucun oedeme n'est note. La moyenne des scores sur les temps 24, 48 et 72 heures pour chaque animal est de 0,3 ; 1,0 et 0,7 pour 1'erytheme et de 0,0 pour 1'oedeme. Conclusion Dans nos conditions experimentales, le produit, non irritant par voie cutanee chez le Lapin. ' est considere Company Sanitized. Does not contain TSCACffi 1. INTRODUCTION The objective of this study was to evaluate the potential of the test substance to induce dermal irritation following a single administration in 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 existence of hazards to Man likely to arise from cutaneous exposure 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., By 31st July 1992. 2. MATERIALS AND METHODS 2.1. TEST SUBSTANCE 2.1.1 Identification The test substance ^^U_^_^_^_f^__ised in the study was supplied by Elf Atochem S.A. Documentation supplied by the Sponsor identified the test substance as follows: . name: ^^^^^^ . batch number: ____ . Elf Atochem filing number:^|^|IHr . description: dark brown liquid . container: one plastic flask . date of receipt: 11 October 1996 . storage conditions: at room temperature and protected from light. 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 beginning of the study,, the analytical certificate was not available. The pH of the test substance as mentioned in the safety data sheet was 8.5. 2.1.2 Preparation The test substance was applied in its original form. CompanySanitized. Ooes not contain TSCA CBi 2.2. TEST SYSTEM 2.2.1 Animals Sex, species, strain: male New Zealand White rabbits. Reason for this choice: species commonly requested by the international regulations 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 regulations and taking into account that a good correlation of results can be obtained with either three or six animals (i). 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 standard deviation of 2.2 0.2 kg. Acclimatization: at least five days before the beginning of the study. 2.2.2 Environmental conditions During the acclimatization period and during the main test, the environmental conditions in the animal room were set as follows: . temperature: 183C . relative humidity: 30 to 70% . light/dark cycle: 12 h/12 h . ventilation: approximately 12 cycles/hour of filtered, non-recycled air. The temperature and relative humidity were recorded continuously and records retained. The housing conditions (temperature, relative humidity and ventilation) were checked monthly. 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 All the animals had free access to 112 C pelleted diet (U.A.R., 91360 Villemoisson-sur-Orge, France). Each batch of food was analysed (composition and contaminants) by the supplier. The diet formula is presented in appendix 2. Drinking water filtered by a F.G. Millipore membrane (0.22 micron) was provided adiibitimi. Bacteriological and chemical analysis of the water and diet and detection of possible contaminants (pesticides, heavy metals and nitrosamines) are performed periodically. Results are archived at C.I.T. It was verifred that no contaminants in the diet'or water at levels likely to influence the outcome of the study were present. (l)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). Company Sanitized. Does not contain TSCA CB!) 10 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. The skin of each animal was examined in order to use only animals without any signs of cutaneous irritation. Animals showing signs of cutaneous irritation, cutaneous defects or pre-existing dermal injury were not used. 2.3.2 Study design The study design was established according to available information on the test substance and following the O.E.C.D. and E.C. guidelines. As no irritant effects were anticipated, the test substance was applied for 4 hours to three animals. 2.3.3 Application of the test substance The test substance was used in its original form. A single dose of 0.5 ml of the test substance was prepared 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 (Laboratoires 3M Sante, 92245 Malakoff, France). The untreated skin served as control. No residual test substance was noted at removal of the dressing. 2.3.4 Date of treatment Animal number 01 02 03 Date of treatment (day 1) 5 November 1996 5 November 1996 5 November 1996 End of the observation period 8 November 1996 8 November 1996 8 November 1996 C<wnpaySanitized. | 11 2.4. CUTANEOUS EXAMINATIONS The skin was examined approximately one hour, 24, 48 and 72 hours after removal of the dressing. Following the O.E.C.D. and E.C. guidelines: . when there is no evidence of dermal irritation after 72 hours, the study is ended. . when there is persistent cutaneous irritation after 72 hours, the observation period is 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 are observed, the animals are killed on humane ground. Any change in the animals' behaviour was noted. 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 taking into consideration the nature and the reversibility of i 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 skin if, when it is applied to healthy intact animal skin for up to 4 hours, significant inflammation is caused and which pcrs-ists for 24 hours or more after the end of the exposure period. CompanySanitized. Does not contain TSCA< 12 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 substance 2.6.2.1 Irritant substances - symbol Xi: indication of danger: "irritant", - phrase indicating the nature of special risks: R 38: "Irritating to skin" 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, fisures, 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 burns" If, when applied to healthy intact animal skin, full thickness destruction of tissue occurs as a result of up to 4 hours exposure, or if this result can be predicted. skin R 35: "Causes severe burns" 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. ARCHIVES The .study documentation and materials, namely: . protocol and possible amendments, . raw data, . correspondence, . final report and possible amendments, are stored in the archives of C.I.T., Miserey, 27005 Evreux, France, for five years after the end of the /"/; vivo phase of the study. At the end of this period, the study documentation will be returned to the Sponsor. ed Does not contain TSCACBS; 3. RESULTS (table 1) Very slight or well-defined erythema (grade 1 or 2) was noted in all animals at the 1-hour reading. It persisted for 24 hours in one animal, and for 48 hours in the two other animals. No oedema was observed. Mean scores over 24, 48 and 72 hours for individual animal were 0.3, 1.0 and 0.7 for erythema and 0.0 for oedema. 4. CONCLUSION Under our experimental conditions, the test substance FORAFAC 1203 (batch No. 25-28) was considered non-irritant when administered by cutaneous route in rabbits. Company Sanitized. Does not contain TSCA CBB 14 Table 1: Individual cutaneous examinations and mean values of the scores recorded at each reading (24, 48 and 72 hours) for each animal (4-hour exposure) Rabbit number Dermal Irritation Ih Dl Scores 24h 48h D2 D3 01 Erythema Oedema Other 02 Erythema Oedema Other 03 Erythema Oedema Other (1) niKin of scores on days 2. 3 and 4 li == hour D =day (+) = (-) = * = irril.'ml according to E.E.C. criteria iioii-irril;uit according to E.E.C. criteria None 1 1 0 0* 0* 0* 2 2 1 0* 0* 0* 2 1 1 0 0* 0* Mean Interpretation irritation (+) 72h score (1) (-) D4 0 0,3 (-) 0 0,0 (-) * 0 1,0 (-) 0 0,0 (-) . * 0 0,7 (-) 0 0,0 (-) * S9mpmfswll^^.,^ fSCAOH APPENDICES ,^^WM'a"u 16 1. Test article description company Sanitized. Does not contain TSCA Cffi 17 TOXICOLOGY DEPARTMENT CONFIDENTIAL 10 October 1996 elf atochem s.a. La defense 10, cedex42 92091 Paris-la-Defense, France TEST ARTICLE DESCRIPTION PHYSICAL AND CHEMICAL PROPERTIES Appearance Melting point Boiling point Flash point Solubility brownish liquid -22C 95C 50C water TOXICOLOGICAL INFORMATIONS AND USE SAFETY See safety data sheet 'STORAGE AND DISPOSAL Storage Expiry.date Disposal ____ in dark and at room temperature December 1997 incineration Company Sanitized. Does not contain TSCA CM 18 2. Diet formula C">P.nys.n,.,^.ooe,no,c.n,,,nTscACM 19 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 libitum. hUKMULA % Cereals ........................... Grain biproducts and legumes .......................... Vegetable proteins (soya bean meal, yeast) ................... Vitamin and mineral mix ture .. AVERAGE ANALYSIS % 43.8 49 4.2 3 Calorific value (KCal/kg) Moisture ........................ Proteins ........................... Lipids ................................... Carbohydrates (N.F.E.) ..-.-.- Fibre ...................................... Minerals (ash) ........................ AMINO ACID VALUES (calculated in mg/kg) 2200 10 13 2.7 49.3 17 8 Arginine ... Cystine ..... Lysine ....... Methionine Tryptophan Glycinc ..... FATTY ACID. VALUES (calculated in mg/kg) 6800 2100 4600 1600 1400 5200 Palmitic acid...... Palmitoleic acid. Stcaric acid......... Olcic acid........... Linolcic acid...... Linolenic acid.... 6400 0 600 6400 12100 2400 MINERALS (calculated iri mg/kg) Nat. CMV val. val. Total P.......... ... K ......... ... Na ....... ... M" ... Mn ...... FP Cu ........ Zn ........ Co ........ I ........... Cl 3500 4snn 11600 400 2100 40 160 12 30 0.1 0 500 3500 4500 0 1600 100 40 140 15 45 1.5 0 3000 7000 9000 11600 2000 2200 80 300 27 75 1.6 0 3500 vnFAMINS (ccalculated perkg) Nat. CMV val. val: Total Vitamin A Vitamin D3 Vitamin B 1 Vitamin B2 Vitamin B3 Vitamin B6 Vitamin B 12 Vitamin E Vitamin K3 Vitamin PP Folic acid Biotin Choline Meso-lnositol 2850 IU 30 IU 4.3 mg 3.8 mg 16 mg 1 mg Omg 16 mg 6mg 55 mg Omg Omg 850 mg Omg 6500 IU 1000 IU Omg 0 mg Omg Img Omg 10 mg 1 mg 5mg Omg Omg 200 mg Omg 9350IU 1030IU 4.3 mg 3.8 mg 16 mg 2mg Omg 26 mg 7mg 60 mg 0 mg Omg 1050mg Omg Available under quality "Control Ref.: 112 C" U.A.R-. 7njcGallieni,91360Villemoisson - Tel: 69.04.03.57 - Fax : 69.04.81.97 (Ref. Doc. UAR: 1992) Company Sanitized. Does not contain TSCACSi