Document ZBvqJ12arpGnojVv9X8Z5Bwd7

AR226-3121 CONFIDENTIAL DPT 433/984485/AC acute derm al to xicity to the rat Sponsor DuPont Speciality Chemicals Jackson Laboratory Chambers Works Deepwater NJ 08023 USA Research Laboratory Huntingdon Life Sciences Ltd PO Box 2 Huntingdon Cambridgeshire PE18 6ES ENGLAND Report issued 5 May 1999 Page 1 of 16 .. Company Sanitized. Does not contasn TSC CS CONTENTS COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS QUALITY ASSURANCE STATEMENT..................-.................................... RESPONSIBLE PERSONNEL....................................................................... SUMMARY...................................................................................................... INTRODUCTION............................................................................................ TEST SUBSTANCE........................................................................................ EXPERIMENTAL PROCEDURE.................................................................. RESULTS........................................................................................................ CONCLUSION......................................................................................... :..... TABLES 1. Dermal reaction......................................... -............................................... 2. Individual bodyweight changes........ -.................................................... 3. Individual and group mean bodyweights................................................. DPT 433/984485/AC Page 3 4 5 6 7 8 9 13 14 . 15 16 16 :2 : Company Sanitized. Doss not corda-n TSCA C DPT 433/984485/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), EhW/MC/CHEM(98) 17. EC Council Directive 87/18/EEC of 18 December 1986 (Official Journal No L 15/29). Lewis A. McRae, MJ.Sc.T., C.BioL, M.I.Biol., Study Director, Huntingdon Life Sciences Ltd. 3: 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 1998 14 September 1998 14 September 1998 14 September 1998 14 September 1998 12 February 1999 Date of Reporting 24 November 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 ondi, of the standard precoci generated for this type of study design as conducted and reported to Company Management as indicated above. Process based inspections: At or about other routine and repetitive procedures employed on this type ot study v e . , c promptly reported to appropriate Company Management Report Audit- This report has been audited by the Quality Assurance DepartmenL Tlns aud.t was S t t d reported^) the Study Director and Company Management as mdtcamd above. The methods, procedures and observations were found to be accurately desenbed and the report results to reflect the raw data. ............ Margaret Blows, Quality Assurance Group Leader, Department of Quality Assurance, Huntingdon Life Sciences Ltd. Date ri*'' t imiti!nWWl :4 : Company S; 3d. Does not co tain TSCA CSS responsible personnel Lewis A. McRae, MXSc.T., C.Biol., Study Director, Department of Acute Toxicology. DPT 433/984485/AC :5 : Company Sanitized. Dsas not contain TSCa o s ! SUMMARY DPT 433/984485/AC A study was performed to assess the acute dermal toxicity o was based on that described in: to the rat. The method followed EEC Methods for the derenninafion of toxieky, A * o D a t i v e 92/69/EEC (Official kmnta. No. L383A 29.12.92), Part B, Method B.3. Acute toxicity (dermatf. OECD Guideline for Testing of Chemicals No. 402 "Acute Dermal Toxicity . Adopted. 24 FebruaI^ 1987. , A group often rate (five malesand d ^ f l d ^ L c r e l l e t o compensate for water administered at a dose level 0 - ,,.pressed as dose concentrations after adjustment for content (75%). All the 75% water content 1ne ni" u ? J study findings and in compliance with the test guidelines. level se,action was supported by prelimrutuy A,, animals were killed as scheduled at sbtdy .emanation (Day 15) and s u b je c t to a macreatopic examination. There was no clinical signs o f reaction to treanuent obsereed in any animal throughout the study. Primarily nansien, slight to well-defined demtri " ^ " " f ^ e ^ " w e re " T re " treaonen, she. In dte m ^orhy of rah, al, reacfions had resolved by Day 7 and in one rat by Day 12. Minor bodyweight flucmations were noted in the occasion animal. H o - - . ^ majority of mimais were considered to have achieved satisfactory bodyweight gams throughout die study. No macroscopic abnonnalhies were observed for annuals killed a, study termination on Day 15. Tie acute lethal dermal dose to rate of lodyweight |was demonstrated to be greater than 2000 mg/kg ut Directive 93/21/EE. "n s f p ^ i rlmc.hS ' Company Sanitized. Does not conttaa:in 7SCA CBt INTRODUCTION DPT 433/984485/AC The study was designed to assess the toxicity o ^ M ^ > U w m g a single demul a t a i n t o h o n to the rat The rats were dosed by topical application as die test substance may come in con wi during handling or use. The study was conducted in compliance with the following guidelines: EEC Mediods 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). i. OECD Guideline for Testing of Chemicals No. 402 "Acute Dermal Toxicity" Adopted: 24 February : 1987. $ The rat was chosen as the test species as it has been shown to be a suitable model for this type o f 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 :. noes not contain TSCA CB*. Company Sanrt-- Identity: Chemical name: Intended use: Appearance: Storage conditions: Lot number: Expiry: Purity: " Sample received: TEST SUBSTANCE DPT 433/984485/AC Room temperature t*) Two years from date of receipt 23 June 1998 :8 : Compur.y jifegd. Does not c . ;nta:n TsSCf :El. EXPERIMENTAL PROCEDURE DPT 433/984485/AC a n im a l m a n a g e m e n t 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 w^eks 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 o f 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 'as administered as to compensate for the water content bodyweight in the main study. 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 Se: Loss tiwi treatm ent procedure Preliminary study DPT 433/984485/AC test substance and aid in selection of a suitable dosage for the main study. Main study A a dose ------------------- ~ - - 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 MF 11 11 44 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 surface area. J d further covered by a waterproof dressing encircled firmly around the trunk of the anima . Treatment in this manner was performed on Day 1 (day of dosing) of the study only. with absorbent paper. No control animals were included in this study. 10 cgnitfced- Sees not contalo TSGA OB' Vinmsanv DPT 433/984485/AC OBSERVATIONS M ortality | Cages o f rats were checked at least twice daily for mortalities. '.'C linical signs ' ''limais were observed soon after dosing and at Sequent intervals for tire remainder rrf-Bay 1. Om - subsequent days animals were observed on at least two occasions during the day (once m the momin an the end of the experimental day, with the exception of the day of study termination - morning only). "i,e nature and severity o f clinical signs and the time these were observed were recorded at each observation. ;. Animals in the preliminary and main study were obsetved 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 Slight erythema : r Well-defined erythema 0 j .2 3 SevCT^e^nhema(beet redness) to slight eschar formation (injuries in depth) 4 Oedema formation: No oedema " --------- WeU-defomd oedema (edges of area well-defined by definite raising) Moderate oedema (raised approximately 1millimetre) Severe oedema (raised more than 1 millimetre and extending beyond the area of exposure) Any other lesion not covered by this scoring system was as described. 0 1 2 4 . Bodyweight Tim bodyweight of cad, rat in the prelimimny 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. ?' 5a r TERMINAL STUDIES 1: 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 Company Sanitized. Does not 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. ARCHIVES ----------------- 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 not contain TSCA C J;ed. 0033 RESULTS DPT 433/984485/AC PRELIMINARY STUDY A 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......... .................... :-------------- -- -- - Clinical si"ns 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 Day 2. Bodyweight 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. 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 systeqyc response in any animal throughout the study following a single dermal application a group of ten rats (five males and five females) at a dose level of 2000 mg/kg bodyweight. - DERMAL RESPONSES (Table 1) Primarilv 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. BODYWEIGHT (Tables 2 and 3) .v . 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. 13 : S a n d e d . D oss net contain TSCAC31 Company CONCLUSION DPT 433/984485/AC The acute lethal dermal dose to rats o f ^ ^ ^ ^ ^ w a s demonstrated to be greater than 2000 mg/kg bodyweight. : 14 Comps?!'/ S ;afl'dze ;ocs no? contai IS C A C3f TABLE 1 Dermal reactions DPT 433/984485/AC : 15 : Company Sanstszed. C ,g nc? contain T3 CA CB d p t 433/984485/AC TABLE 2 Individual and group mean bodyweights (g) TABLE 3 Individual and group mean bodyweigbt changes (g)