Document MoymK43J0Q63bypwRNK1V6dgz

AR226-3122 g a ttm ia * u v 5 a c E W j :i3 ^ g iw v s a ^ mr7BTn;p,'iiaBgT.gncnCTO T fa a y ^ j: CONFIDENTIAL DPT 429/984523/AC ACUTE ORAL TO XICITY TO THE RAT (FIXED DOSE M ETH O D) Sponsor DuPont Speciality Chemicals Jackson Laboratory Chambers Works Deepwater NJ 08023 USA Research Laboratory Huntingdon Life Sciences Ltd P.O. Box 2 Huntingdon Cambridgeshire PE18 6ES ENGLAND Report issued 6 May 1999 Page 1 of 17 Company Sanitized. Does not contain TSCA C-d CONTENTS COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS QUALITY ASSURANCE STATEMENT................. :.............................;........ RESPONSIBLE PERSONNEL............................................................... :........ SUMMARY........................................................................................................ INTRODUCTION.............................................................................................. TEST SUBSTANCE.......................................................................................... EXPERIMENTAL PROCEDURE................................................................... RESULTS.......................................................................................................... CONCLUSION.................................... ...........;................................................. TABLES 1. Mortality data................................. -- ........;..... " . '" `T""''......... 2. Sims of reaction to treatment - preliminary investigation .................... 3. Individual bodyweights & changes - preliminary investiga ons............ 4. Signs of reaction to treatment -m ain study............................................... 5. Individual and gfoup mean bodyweights.................................................. 6. Individual bodyweight changes................................................................. DPT 429/984523/AC Page 3 4 5 6 7 8 9 12 13 14 15 15 16 17 17 :2 : Company Sanitised. Dees not contain TGOA CB DPT 429/984523/AC COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS , irted in compliance with the following Good Laboratory Practice The study described in this report ^ co d t^ consider the data generated to be valid, standards and with the exception of that noted oeio The UK Good L abom ory Practice Regulations HOT (Statu,oty InstrumentNo 654). OECDP*KipleSo f G o o ^ ~ EC Council Directive 87/18/EEC '^ ^ M a r c h 1 9 (Official Journal from 1 May 1999 EC Commission Directive i y y y / n / ^ No L 77/8). Chemical an,ysis o f -- d tea. article fo, de-- n of ability, h o concenhahon was not undertaken for this study. Lewis A. McRae, M .I.Sc.T., C.Biol., M.I.Biol. Study Director, Huntingdon Life Sciences Ltd. csDate J !t ^ " \I :3 : Company SanltsEsd. Does not contain i SCACB) QUALITY ASSURANCE STATEMENT DPT 429/984523/AC The following have been inspected or audited in relation to this study Study Phases Inspected Standard Protocol Audit Process Based Inspections Husbandry . Housing/Environment Weighing of Animals Treatment Procedure Clinical Signs Post Mortem Records Audit Training Records Report Audit Date of Inspection 21 November 1997 14 September 1998 14 September 1998 14 September 1998 14 September 1998 14 September 1998 14 September 1998 14 September 1998 14 September 1998 4 January 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 17 September 1998 6 January 1999 Standard Protocol Audit: An audit of the standard protocol generated for this type o f study design was conducted and reported to Company Management as indicated above. Process based .perdons: The methods, procedures and observances were found to be accurately described and the reported results reflect the raw data. ..... ...................... Margaret Blows, Quality Assurance Group Leader, Department of Quality Assurance, Huntingdon Life Sciences Ltd. Date Company Sanitised. Dossi in T 'jC < r espo n sib le pe r so n n e l DPT 429/984523/AC Lewis A. McRae, M.I.Sc.T., C.Biol., M.I.Bxol., Study Director, Department of Acute Toxicology. :5 : Company Sanitized. Do not coats' 80A CBl SUMMARY DPT 429/984523/AC rhis study was perfro_rmedj tto_ aoscsoeessss tmhee a^cute oral toxicity o to the rat. The method followed was based on that descnb f , cltv AimeX to Directive 92/69/EEC (Official Journal No. t %t t S ' " ?"*(oral)-" ' T a i ,, Of Chemicals No , `Acute Ora, T oxicity-F ixed Dose Medrod' OECD Guideline for Testing of Adopted 17 July 1992. . A of ten f t * rats (five males * < * f ^ T ; i ^ ^ ^ s=,heKs,su,deiines . . iere were d aths in the main phase of the study in which a group of ten rats recewed a osa0e o a dose ^level ouf 5i 0^0 mulcg,/.k-gDb-ody"weig~ht. inical signs of reaction to treatment ^ 8 ^ " d dm livation and ungroomed appearance nota ' m i huIlche(] posture (males only) had resol S t - SfficS-) <* ^ 4'^ iDaU" y ay 8. Hrats were considered to have achieved satisfactory bodyweight gains during the study. abnormalities were observed in animals killed at study termmatron on Day 15. lo macroseoprc abnormalrtre J ^ ^ ^ w a s indicated 5 be 500mg(kg -he discriminating (non-letbal) oral dose to rats odyweight. not Dn the basis of findings in f J w equire labelling with the risk phrase R22 Hamtrul 1/FF.C.. ^ h Commission Directive :6 : Company Sanitized. Do s not contsln TSCA C3f INTRODUCTION DPT 429/984523/AC , - .. a single oral dose to the rat. The Tlie study was conducted in compliance with: i s s s s r - s " 1" " OECD Guideline for Testing o f Chemicals No.420 `Acute Oral Toxicity - Fixed Dose Method Adopted 17 July 1992. suitable model for this type of study and is the animal The rat was chosen as it has been shown to be a recommended in the test guidelines. help define the toxicity of the test substance and to be in The dose levels used in this study were chosen to compliance with the test guidelines. The p r o ,, was appsoved by = 17 July 1998 and by the Study Director on 27 August 1998. " 7 * * '" 8' * " ^ . The experimental phase of the study was conducted between the 2 September and 13 November 19 9 '. " 7 : Company Sanm^e-. Doss not conetslrs TSC; * 3 test substance d p t 429/984523/AC Two years from date of receipt 3 23 June 1998 Company Sarntsz---d. Does not cd'Ous.:n j a c k =- ex pe r im e n ta l pr o c e d u r e DPT 429/984523/AC ANIMAL m a n a g e m e n t England. M tmals in * . niain cffiScTiX S r,,! ------- . - ....... The rats were allocated without conscious b r a s f t o '^ s X S S b l e Rodent Cages - in gdfips of up to five rats of the same * m ma1 ^ ,, e re fitted with grid C S f o e t u t ' ^ o ^ o ^ S u n d e r ita y s.i cages were suspended in mobile stainless steel racks in Room 6 of Building R14. A standard l a b o r e d rcalen, die, tiS ; " ^ 7 r s " i n g The < - * * s^ Sciences Ltd. as quarterly summaries. * ^ -- --- Lift Thermostatic controls were set to maintain a j p e L r a " ^ 3 ^ w * racori,S controlled but was anticipated to be m ^ " ^M easurement of these parameters revealed that animal s s i i s . r 's i s = - ; i r ? : -- - -- 12* -rf artificial light (0700 - 1900 hours) in each 24-hour period. Director and Home Office licensee. TEST SUBSTANCE PREPARATION In the main concentration of iTF/o v/v in distilled water and administered at a dose vo in order to achieve the desired dosage. a 1 .. The test substance was prepared on the day o f dosing. 3 The absorption substance w as n oatnudndcehratraakcetneriinsatthioisnstudy and remains the respo nse iy formulated at a ml/kg bodyweight v ^" :9 : Company Sanitized, Does not contasn TSCA f DPT 429/984523/AC treatm ent procedure Preliminary investigations . In the absence o f precise toxicological information a preliminary study was conducted to help define tire toxic potential of the test substance and aid in selection/validation of the dosage selected -far use m fee main L d y . Initially, one female rat was treated wife the test sUbstance as s u ||l i j d ^ l |5 m |g g , but result of a marked toxic response thought to be associated with fee pH f H p V n(l " S r e s L o fa n im a l welfare, a fiirther female was treated at 125 mg/kg wife feeT est substance fonnulated " l i t e r and for further clarification of oral toxicity a female rat was dosed at 500 mg/kg" L bodyweight and finally one male and one female (per dosage) treated at 1000 and 2000 mg/kg. Main study A group often rats (five males and five females) received a single oral gavage dose of fee te^ u b rta n c e at f dose level of 500 mg/kg bodyweight. This dose level was chosen in compliance with the test guidelines. No control animals were included in this study. a d m in ist r a t io n o f t h e t e st su bsta nce The appropriate dose volume of fee test substance was administered to each rat by oral gavage usmg a syringe and cannula of fee appropriate gauge. W d The day of dosing was designated Day 1. OBSERVATIONS M ortality Cages of rats,were checked at least twice daily for mortalities. Clinical signs Animals were observed soon after dosing and at frequent intervals for fee remainder o f Day L On ^subsequent days, animals were observed once in fee morning and again at fee end of fee expemnental day (with fee exception o f fee last day of observation - morning only). The nature and seventy o f fee clinical signs and time were recorded at each observation. . Animals in the preliminary and main study were observed for up to 7 or 14 days respectively.after dosing. Bodyweight , The bodyweight o f each rat in the preliminary study was recorded on Days 1 (prior to dosing) and 8 (or at death) and in the main study on Days 1 (prior to dosing), 2, j , 4, 8 and 15. Company Sanifcsci. Does not contain TSCA C 10 ? DPT 429/984523/AC t e r m in a l stu d ies T erm ination T h . a to ra ls ( t o . s k iv e d tteaonea.) in the pmtimtaar, audy wen. killed on Day 8 (swdy nninaion) L a ftTmain smdy were killed o,, Day 15 by c a to n diox.de aaphyx.abon. M acroscopic pathology ..... to rS fid aC p i ^ S " c e r f S i e S 5 o ^ f w ^ r d e d . ' a r c h iv e s Huntingdon. Such cords wii, be rem ind for a minimum knowledge. DEVIATIONS FROM PROTOCOL T he were no deviations from t o pmrocol 4 a t w e consider^ <<> t a v . 6t d e to t a ^ to to.m egrim of tossm dy. ^ S a. 1000and S ^ ic b ^ " ^ w e also slightly ab^ve t o upper weigh. Ibn.. specified in t o study protocol. 1 1 - Company Sanft&ed. Does not contain TSCA Cl results DPT 429/984523/AC PRELIMINARY STOPY (Tables 1 ,2 and 3) n /aOI Two females r e c e iv e c H H H I I l|B llr^ was treated at 500 mg/kg and fm y one female treated at 2000 m j* * (Day8)' ^ terminatio" ^jrnti/mvrteadv aait and dark colouring to d0SaeS` . . v, r mitc for a study of this nature and duration. Bodyweight gains were cons,dered to be within a p ffled ,, ^ termination revealed ,, 1 C examination no abnonnalitiw. Examination o f ^ o n e ^ ^ ^^ enlarged S/\IS>tUSUUvWe1dw- i--n d-ie lader was fluid/food n a t i o n . U- u cp n f the study demonstrated that administration o: Findings from frus phase 1000 and 2000 mg/kg resulted m -5 it dosages of MAIN STUDY There were no deaths administration o ,,f ten mis (five males and five finales) following a single Oml a dose level of 500 mg/kg bodyweight. CLINICAL SIGNS (Table 4) ^ ed in all rats within seven Piloerection increased s a l i v a t i o n ^ ^ I ' S S S i w 5 - ^ " 2 , " n e 'T " ? '* 4 and piloerection resolving in all instances y l) 12 : Company Sanitized. Does not conissn i Su CB* DPT 429/984523/AC BODYWEIGHT (Tables 5 and 6) All rats were considered to have achieved satisfactory bodyweigh. gains during the study. MACROSCOPIC e x a m in a t io n Mo macroscopic abnormalities were observed for annuals hilled at shrdy tenninahon on Day 15. CONCLUSION lull tn rats ( 1 ^ 1 1 ^ demonstrated to be greater than 500 mg/kg The acute lethal oral dose to rats bodyweight. : 13 : Company Sanitized. Dog? not contain TSCA CSf table 1 Mortality data DPT 429/984523/AC administered as supplied Not applicable << : 14 : Company Sanitized. D b net contain TSC& CSt d pt 429/984523/AC TABLE 2 treatment - preliminary investigations Signs of reaction to 1 -------- No. of rats ingroup ol i 1XJ L showing signs Signs of reaction Piloerection Hunched posture Waddling/unsteady gait Increased salivation ^ Ungroomed appearance Thinappearance Lethargy .t Abnormal respiration Pallid extremities Walking on toes Abnormal faeces Protruding eyes Blue/cold extremities Dark colouring to eyes Increased lacrimation Body tremors Prostration______.-- . Dose (mg/kg) 125'+ 125 500 "lOOCK "2P" 1 I .1 MF M 1# 1# . 1 0 1 1 1 1# 1 1 1 1 1 l 0 0 0 1 1 1 0 0 0 0 1 0 0 0 0 0 1 11 1 1 0 0 11 1 11 1 01 1 0 0 0 0 0 0 0 11 11 00 00 00 00 11 1 0 1 1 1 1 1 0 0 11 1 00 0 0 0 0 0 0 0 0 11 00 1 001 00 1 1 0 0 1. Preliminary study. 2. Main Study +: . administered as supplied as< #: a: b: sCharaScterised b:y so=ft to liqu^id faecers/fewT , = = r - ' ' " " " ' " " c: tables and gains ( g ) - preliminary investigations Individual bodyweights 1L . I1! : 15 : Company Sanitized. Does note antain TSCA CBt TABLE 4 Signs of reaction to treatm ent - main study DPT 429/984523/AC Signs of reaction Piloerection Hunched posture Waddling/unsteady gait Increased salivation Ungroomed appearance" Thin appearance No. of rats in group of 5 per sex showing signs Dose ( 500 mg/kg) TST 55 55 50 55 5 1 a: Characterised by soiled/stained fur around the face/muzzle or ano/genital region or wet fur M: Male rats F: Female rats 1 i ..y*!__ j f'St DPT 429/984523/AC TABLES Individual and group mean bodyweights (g) - main study Dose 500 Animal No. & Sex 11 M 12 M 13 M 14 M 15 M Mean 16 F 17 F 18F 19 F 20 F Mean *: Prior to dosing Bodyweight (g) at Day 1* 2 3 4 8 15 86 86 88 98 127 187 89 '94 102 rnr par 78 87 94 99 122 174 86 85 95 91 103 101 113 112 146 210 144 204 85 91 98 108 136 194 95 109 118 117 139 154 88 99 108 117 135 156 84 93 102 109 124 145 97 108 120 123 142 172 94 100 112 118 136 160 92 102" 112 117 l3o 157 TABLE6 Individual and group mean bodyweight changes (g) - mam study Dose (mg/kg) 500 Animal No. & Sex 11 M 12 M 13 M 14 M 15 M Mean 16 F 17F 18F 19 F 20 F Mean Bodvweight'gains (g) Day 1-8 Day 8-15 41 60 53 51 44 52 60 64 59 60 51 57 44 15 48 20 40 21 45 30 42 24 44 22 Company Sanitised. Does not cer tain T3CA CB: