1、 - API TRYOL 93 m 0732290 O540622 212 m American Petroleurn institute 1220 L Street, Northwest Washington, D.C. 20005 Health and Environmental Sciences Department Toluene The Effect on Pregnancy of the Rat (Inhalation Exposure) JUNE 1993 TOXICOLOGY REPORT NUMBER TR401 CAIS ABSTRACT NUMBER 40-31 150
2、API TR4OL 93 = 0732290 O540623 II59 American Petroleum Institute Health and Environmental Sciences Department QUAL I TY AS SURAJCE / GL P COMFL I MJCE STATEMENT Study Title: The Effect of Toluene on Pregnancy of the Rat (Inhalation Exposure) Testing Facility: Huntingdon Research Centre Ltd. Testing
3、Facility Number: APT 1/91309 : APT 2/91279 This study was reviewed by API Quality Assurance personnel under the direction of API Management on the dates indicated below for compliance with EPA TSCA Good Laboratory Practice (GLP) regulations. This study was conducted in accordance with EPA TSCA GLP r
4、egulations, with the exception of the item listed on page 2 of this statement. Copies of reports by API Quality Assurance personnel are available upon written request to the Director of the Health and Environmental Sciences Department of the American Petroleum Institute or his designee. Date(s) of T
5、ype of Inspection/Review Inspection Date of Report To Manaqement 4/6/90 5/31/90 6/26/90 7/5/90 7/9/90 7/30/90 7/31/90 9/19-20/90 10/14-15/90 5/7-10/91 2/20/92 6/3/92 7/13/92 9/15/92 Proposal Review 4/6/:90 SOP Review 5/31/90 Prel. Protocol Review 6/26/90 Prel. Protocol Review 7/5/90 Report Format Re
6、view 7/9/90 Prel. Atmosphere Gen. Data Review 7/30/90 Prel. Protocol Review 7/31/90 Audit Preliminary Data 10/11/90 Site visit, in-life and Necropsy Inspection 10/17/90 Draft Report Audit 6/5/91 2cd Draft Report Audit 2/20/92 3rd Draft Report Audit 6/3/92 Preliminary Report Audit Final Report Accept
7、ance k2izzzL-h Christine Sexsmcth Quality Assurance Coordinator 7/13/92 9/15/92 API TR401 93 O732290 0590629 095 = FOREWORD MI PUBLICATIONS NECESSARILY ADDRESS PROBLEMS OF A GENERAL NATURE. WITH RESPECT TO PARTICULAR CIRCUMSTANCES, LOCAL, STATE, Ah?) FEDERAL LAWS Ah?) REGULATIONS SHOULD BE REVIEWED.
8、 FACTURERS, OR SUPPLIERS TO WARN AND PROPERLY TRAIN AND EQUIP THEIR EMPLOYEES, AND OTHERS EXPOSED, CONCERNING HEALTH AND TIONS UNDER LOCAL, STATE, OR FEDERAL LAWS. NOTHING CONTAINED IN ANY API PUBLICATION IS TO BE CONSTRUED AS GRANTING ANY RIGHT, BY IMPLICATION OR OTHERWISE, FOR THE MANU- FACTURE, S
9、ALE, OR USE OF hW METHOD, APPARATUS, OR PRODUCT COV- THE PUBLICATION BE CONSTRUED AS INSURING ANYONE AGAINST LIA- API IS NOT UNDERTAKING TO MEET THE DUTIES OF EMPLOYERS, MANU- SAFETY RISKS AND PRECAUTIONS, NOR UNDERTAKING THEIR OBLIGA- ERED BY LETTERS PATENT. NEITHER SHOULD ANYTHING CONTAINED IN BIL
10、RY FOR INFRINGEMENT OF LETIERS PATENT. The range-finding study conducted to determine dose levels for this study can be found in report TR400, “A Preliminary Study of the Effect of Toluene on Pregnancy of the Rat.“ Addressee: API TR40L 93 = 0732290 0540625 T2L W APT 2/91279 TOLUENE THE EFFECT ON PRE
11、GNANCY OF THE RAT (Inhalation exposure) Study completed: Regulations: EPA (TSCA) 40 CFR 798.4350 American Petroleum Institute, 1220 L Street, Northwest, Washington, D.C. 20005, U.S.A. Report issued: 10 September 1992 Authors : Amanda J. Brooker, Caroline Brennan, Department of Reproductive Toxicolog
12、y. David M. John, Department of Pathology. Pearce C. Kieran, Terence J. Kenny, Derek W. Coombs, Department of Inhalation Toxicology. Huntingdon Research Centre Ltd., P.O. Box 2, Huntingdon, Cambridgeshire, PE18 6ES, ENGLAND. Page 1 of 213 API TR4OL 93 = 0732290 0540b2b 968 APT 2/91279 CONFIDENTIALIT
13、Y STATEMENT This report contains the unpublished results of research sponsored by AMERICAN PETROLEUM INSTITUTE. part, or reviewed or quoted in any other publication without the prior authorisation of the Sponsor. These results may not be published, either wholly or in :2: API TR40L 93 0732290 0540b2
14、7 BTY COMPLIANCE WITH GOOD LABORATORY PRACTICE STANDARDS HRC Report No. APT 2/91279 To the best of my knowledge and belief the Study described in this Report was conducted in compliance with the following Good Laboratory Practice Standards. Good Laboratory Practice, The United Kingdom Compliance Pro
15、gramme, Department of Health 6r Social Security 1986 and Subsequent revision, Department of Health, 1989. United States Environmental Protection Agency, (TSCA), Title 40 Code of Federal Regulations Part 792, Federal Register, 29 November 1983 and subsequent amendment Federal Register 17 August 1989.
16、 Organisation for Economic Co-operation and Development, ISBN 92-64-12367-9, Paris 1982. Amanda J. Brooker, B.Sc. (Hons.), M.Sc., Date Study Director, Huntingdon Research Centre Ltd. Keith F. Rivett, B.Sc., IO. q.qz Date C.Biol., M.I.Biol., For Laboratory Management :3: API TR40L 93 m 0732290 054062
17、8 730 m APT 2/91279 We the undersigned, hereby declare that the work was performed under our supervision according to the procedures herein described, and that this report provides a correct and faithful record of the results obtained. AntLAOcad Amanda J. Brooker, B.Sc. (Hons.), M.Sc., Study Directo
18、r, Department of Reproductive Toxicology lon - animals with live young - group mean values Bodyweights - animals with live young - group mean values Bodyweight change - animals with live young - group mean values Liver weights - group mean values Litter data - group mean values Sex ratios and litter
19、 weight by sex - group mean values Malformations and anomalies - group incidences Malformations - incidence by type Visceral anomalies - incidence by type Skeletal anornalies - incidence by type Sternebra1 variants - group values APPEND I CES 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. Signs (post exposure) and
20、autopsy findings - individual incidence Water consumption - individual values Food consumption - individual values Bodyweights - individual values Liver weights - individual values Litter data - individual values Sex ratios and litter weight by sex - individual litter values Malformations and anomal
21、ies - individual incidence Malformations and anomalies - individual descriptions Skeletal variants - individual values 26 27 - 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 - 52 53 - 57 58 - 62 63 64 - 68 69 - 73 74 - 78 79 - 98 99 - 103 :a: API TRqOL 93 m 0732290 0540b33 OTB m APT 2/91279 Page ADENA
22、 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. Details of the inhalation exposure system, methodology and results Procedure for time-mating of animals Quality assurance aspects of nesting material Composition and quality assurance aspects of diet Quality assurance aspects of water Certificate of analysis of test c
23、ompound Post mortem findings of health check animals Foetal examinations - individual findings (includes those considered within normal limits) Historical control data for foetal changes - study incidences Summary of results from preliminary study 104 - 128 129 - 133 134 135 - 136 137 138 139 140 -
24、208 209 - 211 212 - 213 :9: APT 2/91279 SUMMARY 1. In this assessment of the effect of the solvent, Toluene, on pregnancy and in utero development of the rat, dosages of O, 250, 750, 1500 and 3000 ppm were administered by inhalation for a period of 6 hours a day from Days 6 to 15 of pregnancy inclus
25、ive. On Day 20 surviving females were sacrificed and subjected to post mortem examination. and foetuses subsequently examined for skeletal and visceral changes. Litter values were determined 2. Exposure to Toluene was associated with a dosage-related maternal response at 750 ppm and above. At 750 pp
26、m, reaction was confined to awareness to exposure and closed/half-closed eyelids; this is a non-specific response to exposure and is not considered to be biologically meaningful. Additional responses at 1500 ppm were uncoordinated gait, ataxia and hypersensitivity to knock on chamber wall. Bodyweigh
27、t gain was lower than controls during the first two days of treatment. At the highest dosage of 3000 ppm, similar signs were observed, as well as abnormal limb movements, lachrymation, increased respiration and, on isolated occasions, salivation and nystagmus of the eyeball. consumption reduced duri
28、ng the treatment period. was recorded during the first two days of treatment. Water consumption was increased and food Initial bodyweight loss 3. Treatment-related effects of maternal exposure on embryofoetal development were confined to 1500 and 3000 ppm and included an exposure-related, but minima
29、l reduction in litter and mean foetal weights, and an increase in foetuses with reduced or unossified sternebrae. Conclusion Within the context of this study into the effect of inhaled Toluene on pregnancy and in utero development of the rat, exposures of 1500 and 3000 ppm were associated with obvio
30、us signs of maternal and embryofoetal toxicity. The no-effect level for specific maternal toxicity and embryofoetal development (to Day 20) was considered to be 750 ppm. It is concluded that exposure to Toluene via the inhalation route does not show any selective effects on embryofoetal development.
31、 : 10 : API TR40L 93 m 0732290 O540635 970 m APT 2/91279 INTRODUCTION This report describes an experiment performed to assess the effect of the solvent, Toluene, on the pregnancy of the rat and in utero development of offspring when administered by inhalation, from Days 6 to 15 of pregnancy inclusiv
32、e. The inhalation route of administration was chosen by the Sponsor as the most likely route of exposure in manufacture and use. based on the results of a preliminary study (AFT/l-R) performed in these laboratories and reported separately. The exposure levels were Toluene was supplied from BDH (Brit
33、ish Drug Houses Ltd, Poole, Dorset, U.K.) in sixty 2.5 litre bottles as a clear colourless liquid, (purity 99.9%, batch no. 2212770L, expiry date 1st August 1995). It was received in these laboratories on 16 July 1990 and was stored in the dark under ambient temperature. Key dates of the study were
34、as follows: Protocol approval by: Study Director: HRC Management: Sponsor : Arrival of animals: Mating - first mating set: - last mating set: Commencement of treatment Last day of treatment - first mating set: - last mating set: Terminal sacrifice - first mating set: - last mating set: * Written pro
35、tocol approval: 24 September 1990. 24 September 1990. 25 September 1990. (Sponsor confirmed protocol details by telephone call)*. 13 September 1990. 24 September 1990. 28 September 1990. 1 October 1990. 14 October 1990. 15 October 1990. 19 October 1990. 19 October 1990. : 11 : API TR401 93 0732290 0
36、540636 807 APT 2/91279 METHODS Animal management and non-exposure accommodation Sexually mature (9 - 10 weeks old, weight range 170 - 9 g) Specific Pathogen Free female rats (Crl: CD (SD) BR VAF/Plus strain) which were time-mated to identified males of the same strain in the Department of Reproducti
37、ve Toxicology Facility (details are presented in Addendum 21, were forwarded to the Inhalation Toxicology Facility. appearance of sperm in the vaginal smear or by the presence of a vaginal plug, was considered as Day O of pregnancy. The day of mating, 8s judged by the Time-mated animals were, within
38、 each batch, assigned to five groups on Day O of pregnancy taking into account, where possible, the distribution of males to which the females were mated and the bodyweight of the animals. allocation was used for animals from the second day of mating. allocation, the animals were ear-tattooed to giv
39、e individual identification, Prior to the co,mencement of treatment, all animals were inspected by a veterinary officer. A random Following Animal room controls for temperature and relative humidity were set at 21OC and 55% respectively. temperature and relative humidity ranged from 20C 2 2OC and 62
40、% 2 20% respectively. Lighting was controlled to give 12 hours light (8 am to 8 pm) and 12 hours dark per 24 hours. During the course of the study recorded values for The animals were housed individually in suspended stainless steel cages, equipped with solid and mesh sides and mesh floors. During e
41、xposure the animals were housed individually in suspended stainless steel mesh cages. cages constituting each treatment group were held on separate batteries, each in a separate ventilated cabinet, in order to minimise the possibility of inhalation of test substance vapour from the fur of, or exhale
42、d by, rats in other test groups. The Throughout the study each cage was identified by a label coloured according to the group an9 recording the study schedule number, animal numbers, details of treatment and the name of the Study Supervisor and Director. All animals were given free access to Biosure
43、 Laboratory Animal Diet No. 1 and to tap water, other than during inhalation exposure when food and water was withheld. Addenda 4 and 5) (Quality Assurance aspects for food and water are presented in : 12 : API TR401 93 0732290 O540637 743 = Group/ colour code APT 2/91279 Exposure level Toluene (PPr
44、n) Experimental design The experimental design was as follows: 1:White 2:Yellow 3 :Blue 4:Green 5 :Red Control 250 750 1500 3000 ?umber of rats ? 25 25 25 25 25 Animal numbers in batch I II III IV v 1-5 6-10 11-15 16-20 21-25 26-31 32-36 37-40 41-45 46-50 51-55 56-60 61-65 66-71 72-75 76-81 82-86 87
45、-90 91-96 97-100 101-105 106-110 111-115 116-121 122-125 Exposure of rats to the test substance Rats were exposed in whole-body exposure chambers for 6 hours per day from Days 6 to 15 of pregnancy inclusive. exposure system, methodology and results are presented in Addendum 1. All details relating t
46、o the inhalation Observation The following observations were made during the study: 1. Parent animals (a) Signs All animals were regularly handled and observed daily for obvious changes or signs of reaction to treatment. period, signs were recorded prior to exposure and post exposure for all animals
47、. intervals during exposure. to tapping on the inhalation chamber walls, were reported. and duration of observations were recorded. During the treatment A maximum of 8 animals were observed, at half-hourly Any adverse signs together with responses The times : 13 : API TR4OL 93 = 0732290 0540638 b8T
48、2. APT 2/91279 Mortalities All animals that died were weighed and subjected to post mortem examination. Pregnancy status was assessed. Food and water consumption Food consumption was measured from weighday to weighday commencing on Day O of pregnancy. Water consumption was measured daily, by weight,
49、 from Day O of pregnancy through to termination. Bodyweights All animals were weighed initially (=Day O of pregnancy) and on Days 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20. Bodyweights on Day 20 corrected for gravid uterine weights are also reported. Litter data and foetal examinations n Day 20 of pregnancy the animals were killed by CO, asphyxiation, dissected and examined for congenital abnormalities and macroscopic pathological changes in maternal organs. weighed and preserved in formalin. for all, but one animal in error. immediately to determine: The livers of al
