Document zQgen1qDZd7ZLkJOx352v78k3

AR226-3121 I *j ~Vl^l'K9^ Huntingdon . - .-._, f^_ ^^........^^.-^.-..s^.^^^^^^^^'^^^'^^^?^'^^'. . .. ,.,. .. .. --..>^/-t^^-^^j;^?^-;j^tfe^-^^^^^^^^^^.-a^ .^i^.%.g^^^^^Si^^^;5gi^^^^ipgfe^^,etS^^gS;^ -^.- ...,.... ^-'rv'''A:rrt"-'"" '" R\a ^^jj;'^h^^^^^0;;n^^^^"^5^-4'^ ;a,?,c.?^--.:^.^.;)^.fe^^^ff^%^iS^ " '-"^ ^^i^fe^s^^^^^^^^^^^^^^^^^^^^^^^^Ssj^^^^^^^i^^ ^s^jjigg^s^^a^s^^g^^^^^a^ga^^^^s^g_i^s.^,.i_^_^.i_^__ .,,_., -:.;.r^--^.?^-?;^^''1^^^^^^^3^^^s;S5^^^;:^^~^%^ '-'^-'^'^-^-^^^^S^'-g^'^^Sg^ffg^-^^n^^ --^:....-- -^----^--^-.^^-^ '^''^^-/^"^.-----^'-^^S.'^"^ . .' -- '-'------"""--------------------------------------------------------------------'----------------------------- ----- ------..gt - - Td \^Jy^*l ---.---------:;-:, --^^B, -. :- -'-k--.,-;. - ^-.."------ -----'-- -------------^----^^-^"-.^-.^..^:^-- -.g^-.:-^^--^--;...^^J~^i^ . ^-.-^^.'...i.:^^^- ._' -.^^^-^;.-^^c.--. ' .^^.- - ^'. ---^----^^^----:r^f^---^'^- -^"^--.J- ..';--^^.^-..-^-.-. ^^.- . - -- : - --,_-.: , . - .j - - - - --.-_ . . - ; . . -- - , .- . --- . - . . , ,, . .. - E,^ ^^^^ilil^^^^ .,^<FD^R''tc^ CONFIDENTIAL DPT 433/984485/AC ACUTE DERMAL TOXICITY TO THE RAT Sponsor DuPont Speciality Chemicals Jackson Laboratory Chambers Works Deepwater NJ 08023 USA Research Laboratory Huntingdon Life Sciences Ltd POBox2 Huntingdon Cambridgeshire PE186ES ENGLAND Report issued 5 May 1999 Page 1 of 16 Company Sanitized. Doss not cc'r.airs 7SCA C31* CONTENTS COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS QUALITY ASSURANCE STATEMENT. RESPONSIBLE PERSONNEL. SUMMARY. INTRODUCTION. TEST SUBSTANCE. EXPERIMENTAL PROCEDU-RE. RESULTS. CONCLUSION. TABLES 1. Dermal reaction...........................--........... 2. Individual bodyweight changes................ 3. Individual and group mean bodyweights. DPT 433/984485/AC Page 3 13 14 15 16 16 2 : Company Sanitised. Does no6 contain TSCA C: DPT433/984485/AC COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS The study described in this report was conducted in compliance with the following Good Laboratory Practice standards and I consider the-data generated to be valid. The UK Good Laboratory Practice Regulations. 1997 (Statutory Instrument No 654). OECD Principles of Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM(98)17. EC Council Directive 87/18/EEC of 18 December 1986 (Official Journal No L 15/29). Lewis A. McRae, M-I.Sc.T., C-BioL, MJ.Biol, Study Director, Huntingdon Life Sciences Ltd. o^i^Ulli Date ---- 3 : Company Sanitizes. Does ^ C3..ta;n TSCA C38 QUALITY ASSURANCE STATEMENT DPT 433/984485/AC The following have been inspected or audited in relation to this study Study Phases Inspected Standard Protocol Audit Process Based Inspections Housing/Environment Husbandry Weighing of Animals Treatment Procedure Scoring Records Audit Training Records Report Audit Date of Inspection 24 November 1997 14 September 1998 14 September 1998. 14 September T998 14 September 1998 14 September 1998 14 September 1998 14 September 1998 12 February 1999 Date of Reporting 24NoveniDer 1997 17 September 1998 17 September 1998 17 September 1998 17 September 1998 17 September 1998 17 September 1998 17 September 1998 16 February 1999 Standard Protocol Audit: An audit or the standard protocol generated for this type of study design was conducted and reported to Company Management as indicated above. Process based inspections: At or about the time this study was in progress inspections and audits of other routine and repetitive procedures employed on this type of study were carried out. These were promptly reported to appropriate Company Management Report Audit: This report has been audited by the Quality Assurance Department. This audit was conducted and reported to the Study Director and Company Management as indicated above. The methods, procedures and observations were found to be accurately described and tfae reported results to reflect the raw data. . - - '..k0-.'??1^.'^^. te'.\."T'>............. Margaret Blows, Quality Assurance Group Leader, Department of Quality Assurance, Huntingdon Life Sciences Ltd. ....^..rte.-^.wn.. Date Company SsnS'zad. Does not cc-*a'r? TSGA C3! RESPONSIBLE PERSONNEL DPT433/984485/AC Lewis A. McRae, M.I.SC.T., C.Biol., M.I.Biol., Study Director, Department of Acute Toxicology. 5 : Company Ssnitised. Daas p.oi contain TSCA csi SUMMARY DPT 433/984485/AC A study was performed to assess the acute dermal toxicity ou^^^^HB|to the rat. The method followed was based on that described in: EEC Methods for the determination of toxicity. Annex to Directive 92/69/EEC (Official Journal No. L383A, 29.12.92), Part B, Method B;3. Acule'toxichy (deimal). - ....-..,,.. OECD Guideline for Testing of Chemicals No. 402 "Acute Dermal Toxicity". Adopted: 24 February 1987. - A group of ten rats (five males and five females) received a single topical application of the test substance administered at a dose level of 2QOQ.mg/kgbodyweight with dose volume increased to compensate for water content (75%). All dosages referenced hereafter are expressed as dose concentrations after adjustment for oqUBU^The the 75% water content main study dose level selection was supported by preliminary study findings and in compliance with the test guidelines. All animals were killed as scheduled at study termination (Day 15) and subjected to 'a macroscopic examination. There was no clinical signs of reaction to treatment observed in any animal throughout the study. Primarily transient slight to well-defined dermal irritation (erythema/oedema up to Grade 2) was notable in the majority of rats with a similar level of response more persistent in one female. These reactions were accompanied in one rat by desquamation over the treatment site. In the majority of rats, all reactions had resolved by Day 7 and in one rat by Day 12. Minor bodyweight fluctuations were noted in the occasion animal. However, the majority of animals were considered to have achieved satisfactory bodyweight gains throughout the study. No macroscopic abnormalities were observed for animals killed at study termination on Day 15. The acute lethal dermal dose to rats ofBlHIB!B!H|was demonstrated to be greater than 2000 mg/kg bodyweight On the basis of findings in this study and in accord with EU hazard classificatiorM^^^^^lHwill not require labelling with the risk phrase R21 "Harmful in contact with skin", in accordance with Commission Directive 93/2 I/EEC. :6 : Company Sanitized. Does n&t coriiain TSCA GSi INTRODUCTION DPT 433/984485/AC The study was designed to assess the toxicity oJM^JjBfollowing a single dermal administration to the rat The rats were dosed by topical application as the test substance may come in contact with the skin during handling or use. The study was conducted in compliance with the following guidelines: EEC Methods for the determination of toxicity. Annex to Directive 92/69/EEC (Official Journal No. L383A, 29.12.92), Part B, Method B.3. Acute toxicity (dermal). OECD Guideline for Testing of Chemicals No. 402 "Acute Dermal Toxicity". Adopted: 24 February 1987. ^ The rat was chosen as me test species as it has been shown to be a suitable model for this type of study and is the animal recommended by the test guidelines. The dose level (2000 mg/kg) for the study was chosen after review of preliminary study findings and in compliance with the test guidelines. The protocol was approved by Huntingdon Life Sciences Management on 7 July 1998,. by the Sponsor on 17 July 1998 and by the Study Director on 27 August 1998. The experimental phase of the study was conducted between the 7 September and 17 December 1998. : 7 c,,^^-00-'^""^061 Identity: Chemical name: Intended use: Appearance: Storage conditions: Lot number: Expiry: Purity: Sample received: TEST SUBSTANCE DPT 433/984485/AC Room temperature Eanifesd. Doas not ccr.Sa^n i SCA CBt Company EXPERIMENTAL PROCEDURE DPT 433/984485/AC :y\ ANIMAL MANAGEMENT The animals chosen for this study were selected from a stock supply of healthy male and female CD rats of Sprague-Dawley origin (Hsd:Sprague-Dawley (CD)) obtained from Harlan U.K. Ltd., Bicester, Oxon, England. ~"---~~~------ Animals in the main study were in the weight range of 221 to 308 g and approximately eight to eleven weeks of age prior to dosing (Day 1). All the rats in the main study were acclimatised to the experimental environment for a period of seven days prior to the start of the study. The rats were allocated without conscious bias to cages within the treatment groups. They were housed in groups of up to five rats of the same sex in metal cages during the acclimatisation period and on commencement of study individually housed in metal cages (RS Biotech Sub-Dividable Rodent Cages polished stainless steel (20cm high x 39cm wide x 39cm long)). The cages were fitted with grid floors to ensure rapid removal of waste material to undertrays. The cages were suspended in mobile stainless steel racks in Room 6 of Building R14. All animals were returned to group housing on Day 12 of the study (by which time all dermal reactions had resolved). A standard laboratory rodent diet (Special Diet Services RM1(E) SQC expanded pellet) and drinking water were provided ad libitum. The batch(es) of diet used for the study was analysed for certain nutrients, possible contaminants and micro-organisms. Results of routine physical and chemical examination of drinking water, as conducted by the supplier, are made available to Huntingdon Life Sciences Ltd. as quarterly summaries. Thermostatic controls were set to maintain a temperature of 22 3C. Relative humidity was not fully controlled but was anticipated to be in the range 30 - 70%. Temperature and humidity were recorded continuously using a seven day recorder. Actual measurement of these parameters revealed that animal room temperature was in the range 20 to 24C and relative humidity was in the range 30 - 60%. Permanent daily recordings of these parameters were made and these are archived with other Departmental raw data. Lighting was controlled by means of a time switch to provide 12 hours of artificial light (0700 - 1900 hours) in each 24-hour period. Each animal was identified by cage number and ear punching. Each cage was identified by a coloured label displaying the dose level, study schedule number, animal mark and the initials of the Study Director and Home Office licensee. TEST SUBSTANCE PREPARATION fl^^U^IHl^BB^ JI^Bj^HHwas administered as supplied a dose volume increased sufficiently to compensate for the water content I f bodyweight in the main study. ' : of the test material morder to achieve a dose level of 2000 me/kg The absorption and characterisation of the homogeneity, stability in vehicle and purity of the test substance was not undertaken in this study and remains the responsibility of the Sponsor. :9: . Company eSe---i^-ed"-Lb"1aasi^c^a^^0^ DPT 433/984485/AC TREATMENT PROCEDURE Preliminary study In the absence of precise dermal irritation information a preliminary study comprising of two rats (one male and one female) was dosed at 500 mg/kg bodyweight to assist in defining potential inritation/toxicity of the test substance and aid in selection of a suitable dosage for the main study. Main study ----------_-_- A group often rats (five males and five females) received a single dermal application of the test substance at a dose level of 2000 mg/kg bodyweight. The dose level was chosen after review of preliminary study results and in compliance with the test guidelines. The treatment regime and constitution of the group(s) are shown below: Phase of Study Preliminary Main , Dates dosed 07.09.98 26.11.98* 03.12.98 Dose (mg/kg) 500 2000 2000 Dose volume (ml/kg) 1.75 7 7 No. of rats M F 1 1 1 1 4 4 In the above and following Tables, M: denotes male rats and F: denotes female rats. *: To ensure at the volume dosed the test substance would not prove to irritant initially only one male and one female were dosed- ADMINISTRATION OF TEST SUBSTANCE One day prior to treatment, hair was removed from the dorso-lumbar region of each rat with electric clippers -taking care to avoid damaging the skin, exposing an area equivalent to approximately 10% of the total body surface area. The test substance was applied by spreading it evenly over the prepared skin. The treatment area (approximately 50 mm x 50 mm) was covered with porous gauze held in place with a non irritating dressing, and further covered by a waterproof dressing encircled firmly around the trunk of the animal. Treatment in this manner was performed on Day 1 (day of dosing) of the study only. At the end of the 24 hours exposure period the dressings were carefully removed and the treated area of skin was washed with warm water (37C) to remove any residual test substance. The treated area was blotted dry with absorbent paper. No control animals were included in this study. 10 : con^ny S^ti^- Oo- -tcontain TSCACB1 OBSERVATIONS DPT 433/984485/AC Mortality Cages of rats were checked at least twice daily for mortalities. BK^/ Clinical signs JS^, Animals were observed soon after dosing and aT frequent intervals fbr tile remainder-of-Bay--h--Oir |^,'; subsequent days animals were observed on at least two occasions during the day (once in the morning and H^" ' sgai" at me en^ of the experimental day, with the exception of the day of study termination - morning only). Hf' -" The nature and severity of clinical signs and the time these were observed were recorded at each observation. Animals in the preliminary and main study were observed for 7 or 14 days respectively after dosing. Dermal responses Local dermal irritation at the treatment site was assessed daily using the following numerical scoring system: Erythema and eschar formation: No erythema 0 Slight erythema 1 Well-defined erythema 2 Moderate erythema 3 . Severe erythema (beet redness) to slight eschar formation (injuries in depth) 4 Oedema formation: No oedema 0 Slight oedema 1 Well-defined 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 the area of exposure) 4 Any other lesion not covered by this scoring system was as described. Bodyweight __Tlit; bodyweighl of each rat in the preliminary study was recorded on Days 1 (prior to dosing) and 8 and in the main study on Days 1 (prior to dosing) 8 and 15. TERMINAL STUDIES Termination The animals in the preliminary study were killed on Day 8 and all animals in the main study were killed on Day 15 by cervical dislocation. : 11 : Cornpsrsy Sanitized. Doss noi contain TSCA CBI DPT 433/984485/AC Macroscopic pathology All animals were subjected to a macroscopic examination which consisted of opening the thoracic and abdominal cavities, the cranial cavity was not examined as observations did not indicate any neurotoxicity. The macroscopic appearance of all examined organs was recorded. |T ARCHIVES _ ,_._....... S.^- All raw data and study related documents generated during the course of the study at Huntingdon Life .Sciences Ltd, together with a copy of the final report are lodged in the Huntingdon Life Sciences Ltd Archive, Huntingdon. Such records will be retained for a minimum period of five years from the date of issue of the final report. At the end of the five year retention period the Sponsor will be contacted and advice sought on the future requirements. Under no circumstances will any item be discarded without the Sponsor's knowledge. DEVIATIONS FROM THE PROTOCOL There were no deviations from the protocol. 12 . " Company.cS^^"----ed-D^no-^7^^ } ^ DPT433/984485/AC RESULTS r PRELIMINARY STUDY fe>-iA.- group of two rats (one male and one female) was dosed at 500 mg/kg bodyweight. There were no deaths. ^^.No dermal response was seen in either animal throughout-the.study._........____________...... C_._ iglinical signs were confined to increased sensitivity to touch seen in the female rat and notable for ^approximately 3 hours after dosing. There were no other signs of reaction and recovery was. complete by J?-. Day 2. I^Y fiddyweight changes were considered satisfactory for a study of this nature and duration. There were no macroscopic abnormalities noted in either animal at the terminal necropsy on Day 8. oqUUkHwas The results from the preliminary study indicated the acute lethal dermal dose to rats greater than 500 mg/kg bodyweight and on this basis no further preliminary study animals were used, as this result was considered adequate justification for progression to the main study at a dosage of 2000 mg/kg. MAIN STUDY There were no deaths and no evidence of a systemic response in any animal throughout the study oBHjBl|to following a single dermal application at a dose level of 2000 mg/kg bodyweight. a group often rats (five males and five females) DERMAL RESPONSES (Table 1) sk'-. Primarily transient slight to well-defined dermal irritation (erythema/oedema up to Grade 2) was notable in ^:; the majority of rats with a similar level of response more persistent in one female. These reactions were ^-.'.-accompanied in one rat by desquamation over the treatment site. In the majority of rats, all reactions had ^1:'--' "resolved by Day 7 and in one rat by Day 12. BODYWEIGHT (Tables 2 and 3) ;/ Minor bodyweight fluctuations were noted in the occasion animal. However, the majority of animals were considered to have achieved satisfactory bodyweight gains throughout the study. MACROSCOPIC EXAMINATION : No macroscopic abnormalities were observed for animals killed at study termination on Day 15. 1 3 " " .--ompany Seized. Does net c=:^TSCAC3) CONCLUSION DPT 433/984485/AC The acute lethal dermal dose to rats of| l^, bodyweight. |was demonstrated to be greater than 2000 mg/kg : 14 : Company SanSSszed. ~oes no? conts?" TSCA C3f TABLE 1 Dermal reactions DPT 433/984485/AC Dose (mg/kg) 2000 Animal No.& Sex 21 M 23 M 24 M 25 M 26 M 22 F 27 F 28 F 29 F 30 F - ... -,, :-JXys- after dosing 10 11 12 to 15 Z..-.-3.- 4^5.--.-.-6-.:-..7..-8-^9 E -i--o--o -. ^a.: 'o- " ^o" "6 '^"o 'o" o--0---- 0 1 .o o":o" o'^o-;.^^^":"0'"" 0 a E 0 0 ^^^^^^^^--^Q^^^;.^:^ 0 0 0- 0 "--Q ---0----6.'-^-0 v 0 ;0 -"0 . .0 E-- -.Q--^O--.-O^^O--,.,^- 0 0 0' 0 '0 0 ' i? - . '- 00 E 2 1..--0-0 o 0 i o o # :Q^Q^.... JL^0 - -. P -0- 0 -^0--'0'~-^ :,-0 ' '-'--"- --.- '-:"r^^ -"-- - ''o ^-^oyor:^- o ^- o o ^o^'tr".' o o^o'^ E Q o 0 0 0 0 0 ..0-:--0-,,--0--;--0. o 0 ooooooo If E 0 ,0 0 0 00 0 0100000000000000 E -1222221 0 1 1 1 ,1 1 110 E -f- 2100000 0 1210000 if E ~o~~ 6 oooooo 0 ooooooo a 1. -1.----1'- 0: -.00 ,.0 .00 0 -0 -(TO O . o O....P o ...o,....o;o. 0Ei? 0100000000000000.0Q,0, 0 00 Kev E Erythema 0 O.edema SS^S^characterised # by dryness, sloughing and /or scaling on treatment site c. (reported as desquamation) Not applicable-------------------------------------- : 15 : Company Seized. Cass no contain T3CACB? TABLE2 Individual and group mean bodyweights (g) DPT 433/984485/AC Phase of study Preliminary Main Dose (mg/kg) 50D 2000 Animal-No. Bodyweight (g) at Day & Sex I* 8 15 ....... 1M 284 ... .. ' 311 --^as^-----234-- 21M 238 287 .334 23 M 278 302 326 24 M 294 341 375 25 M 289 325 361 26 M 308 319 362 Mean 281 315 3:52 22F 241 248 255 27 F 227 235 249 28 F 221 239 249 29 F 239 253 254 30 F 233 235 233 Mean 232 242 248 *: Prior to dosing TABLES Individual and group mean bodyweight changes (g) Phase of study Preliminary Main Dose (mg/kg) 500 2000 Animal No. &Sex 1M 2F 21M 23 M. 24 M 25 M 26 M Mean 22F 27 F 28 F 29 F 30 F Mean Bodyweight gains (g) Day 1-8 27 Day 8-15 0 49 47 24 24 47 34 36 36 . 11 43 33 37 7 7 8 , 14 18 10 14 1 2 -2 10 6 16 Company Sanitized. Does no*, contain TSCA C51