Document gadx64NBXJjbo646ZZGJZxmZq

SPONSOR ElfAtochemS.A. Cours Michelet La Defense 10 92091 Paris-la-Defense CEDEX France TEST SUBSTANCE STUDY TITLE ACUTE ORAL TOXICITY IN RATS STUDY DIRECTOR Xavier Manciaux AR226-3141 CONFIDENTIAL IFM redierc'ie S M C. W CAflMl D S ii0 COC f S!EC SOClAt :.'.lBt^T - ;;005 VRU.l fffp.-^n ^< fl:"< r-'.-aciw STUDY COMPLETION DATE 13 December 1999 PERFORMING LABORATORY err Centre' International de Toxicologie BP 563 - 27005 Evreux - France LABORATORY STUDY NUMBER 18748 TAR Company S,^.0oe.< "" TSCACM. CENTRE INTERNATIONAL D TOXICOLOG.'E B. P. 563 2700S Evreux Cedex Tel.:+33 2 32 29 26 26 France CONTENTS STATEMENT OF THE STUDY DIRECTOR 4 OTHER SCIENTISTS INVOLVED IN THIS STUDY 4 STATEMENT OF QUALITY ASSURANCE UNIT 5 SUMMARY 6 RESUME 7 1. INTRODUCTION 8 2. MATERIALS AND METHODS 8 2.1 TEST SUBSTANCE 2.1.1 Identification 2.1.2 Formulation procedure 2.2 TEST SYSTEM 9 2.2.1 Animals 9 2.2.2 Environmental conditions 9 2.2.3 Food and water 9 2.3 TREATMENT 10 2.3.1 Fasting of the animals 10 2.3.2 Administration of the test substance 10 2.3.3 Chronology of the study 10 2.4 CLINICAL EXAMINATIONS 10 2.4.1 Clinical signs and mortality 10 2.4.2 Body weight 10 2.5 NECROPSY 11 2.6 DATA EVALUATION 11 - - 2.7 PROTOCOL ADHERENCE 11 2.8 ARCHIVING 11 3. RESULTS 12 3.1 CLINICAL EXAMINATIONS 12 3.1.1 Clinical signs and mortality (table 1) 12 3.1.2 Body weight (figures 1 and 2, tables 2 and 3) 12 . 3.2 PATHOLOGY (table 4) 4. CONCLUSION 12 contain TSCA}^ complysanifl2e.d-n-cs sal Lil/^tuay INO. is/48 i/udf^^^JU^lt Atocnem ii.A. 3 Figure 1: Body weight of treated rats 13 Figure 2: Body weight of CIT historical control rats 14 Table 1: Individual clinical signs and mortality 15 Table 2: Individual and mean body weight and weekly body weight change (g) 16 Table 3: Mean body weight and weekly body weight change of CIT historical control rats 17 Table 4: Individual macroscopic examinations at necropsy 18 APPENDICES 1. Test article description and analytical certificate 2. Diet formula 19 20 23 and 24 .^nTSC^Bl v.^.o^5^ Gon^23"" CIT/Study No. 18748 TARI^--------UF^' Atochem S.A. 4 STATEMENT OF THE STUDY DIRECTOR The study was performed in compliance with the principles of Good Laboratory Practice as described in: . OECD Principles on Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM (98) 17. . Decret N 90-206 du 7 mars 1990 concemant les Bonnes Pratiques de Laboratoire (Journal Officiel du 9 mars 1990), Ministere de 1'Industrie et de 1'Amenagement du Territoire. . Council Directive 87/18/EEC of 18 December 1986 on the harmonization of laws, regulations or administrative provisions relating to the application of the Principles of Good Laboratory Practice and the verification of their applications for tests on chemical substances (OJNo.L15ofl7.1-87). 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 CIT, Centre International de Toxicologie, BP 563, 27005 Evreux, France. Toxicology X. Manciaux Study Director Doctor of Pharmacy Date: 13 December 1999 OTHER SCIENTISTS INVOLVED IN THIS STUDY For Pharmacy: P.O. Guillaumat Doctor of Pharmacy For Toxicology: C. Pelcot Study Supervisor ^co^nTSC.cm Con^5^1- n ..'.-"ft "U^U<r."'~-13^ I ' CIT/Study No. 18748T Bit' Atochem S.A. STATEMENT OF QUALITY ASSURANCE UNIT Type of inspections Protocol Report Inspections 4 June 1999 10 November 1999 Dates Reported to Study Director (*) 4 June 1999 8 December 1999 Reported to Management (*) 4 June 1999 10 December 1999 In addition to the above-mentioned inspections, at about the same time as the study described in the present report, "process-based" and routine facility inspections of critical procedures relevant to this study type were also made by the Quality Assurance Unit. The findings of these inspections were reported to the Study Director and to CTT Management. The Inspectionswere performed in compliance with CIT Quality Assurance Unit procedures and the Good Laboratory Practice. The reported methods and procedures were found to describe those used and the results to constitute an accurate and complete reflection of the study raw data. (^o^r^ L. Valette-TaIbi Date: 13 December 1999 Doctor of Biochemistry Head of Quality Assurance Unit and Scientific Archives (*) The dates indicated correspond to the dates of signature of audit reports by Study Director and Management. G ^ ^ y -..-,,', sos^ct/-.-"'r1Aa'iO'^^ CIT/Study No. 18/48 1 Alf----^B^mcii mocnem O.A. SUMMARY substancqH^^BHIIH|m|HmHwas At the reoues^fElf Atochem S.A; Paris-la-Defense, France, the acute oral toxicity of the test evaluated in rats according to OECD (No. 401,24th February 1987) and EC'(92/69/EEC, B.I, 31st July 1992) guidelines. The study was conducted in compliance with the principles of Good Laboratory Practice Regulations. Methods The test substance was administered by oral route (gavage) to one group of ten fasted Sprague-Dawley rats (five males and five females). The test substance was administered undiluted at the dose of 2000 mg/kg, taking into consideration that its specific gravity was 1.1 g/ml. Clinical signs, mortality and body weight gain were checked for a period of up to 14 days following the single administration of the test substance. All animals were subjected to necropsy. Results No deaths occurred at 2000 mg/kg. The general behaviour and body weight gain of the animals were not affected by treatment with the test substance. No apparent abnormalities were observed at necropsy in all animals. Conclusion Under our experimental conditions, the oral LDo of the test . |UUB|s substance|fH^JJUH ""this dose. ^ equal to or higher than 2000 mg/kg in rats. No signs of toxicity were observed at According to the classification criteria laid down in Commission Directive 93/2 I/EEC (27th April 1993) adapting to technical progress for the eighteenth time Council Directive 67/548/EEC, the test substance does not present a significant acute toxic risk if swallowed. ;onte5"TSCACBl Comps"y eS.<-.^rw...T.-e^d.Soes^ dl/^tuay INO. is/48 l/u^^^^^H^^Hcn Alocnem ^./\. / RESUME ll^^^HBii^BHiBBlpP1'68 A la demande_de ElfLAtochem _S.A.. Paris-la-Defense, France, la toxicite aigue du produit administration unique par voie orale chez Ie Rat a ete evaluee conformement aux lignesairectrices de 1'OCDE (n 401, 24 fevrier 1987) et de la CEE (92/69/EEC, B.I, 31 juillet 1992). L'etude a ete realisee conformement aux regles de Bonnes Pratiques de Laboratoire. Mefhode Le produit a ete administre par voie orale (gavage) a un groupe de 10 rats Sprague-Dawley (5 males et 5 femelles) mis a la diete hydrique. L'administration a ete effectuee avec le produit non dilue a la dose de 2000 mg/kg, en tenant compte de sa densite (d - 1,1 g/ml). Les signes cliniques, la mortalite et 1'evolution ponderale des animaux ont ete suivis pendant une periode de 14 jours apres 1'administration unique du produit. Un examen anatomopathologique a ete effectue sur tous les animaux. Resultats La mortalite est nulle a la dose de 2000 mg/kg. Aucun signe clinique n'est observe pendant 1'etude. L'evolution ponderale des animaux n'est pas influencee par le traitement. L'autopsie des animaux ne met en evidence aucune anomalie apparente. Conclusion Dans nos conditions experimentales, la DL 0 orale du produifHIB est superieure ou egale a 2000 mg/kg chez le Rat. Aucun signe de toxicite n'est observe a cette dose. Selon les criteres de classification decrits dans la Directive 93/21/CEE (27 avril 1993) portant dix-huitieme adaptation au progres technique de la Directive 67/548/CEE, le produit ne presente pas de risque toxique aigii significatif en cas d'ingestion. ,.ccn^nTSCACBl . nQas^"1-- Company ^riulze-' CIT/Study No. 18748 TAEM|^B--------EIfAtochem S.A. 8 1. INTRODUCTION The objective of this study was to evaluate the toxicity of the test substance following a single oral administration in rats. { In the assessment of the toxic characteristics of a test substance, determination of acute oral toxicity is an initial step. It provides information on health hazards likely to arise following a short-term exposure by the oral route in humans. The study was conducted in compliance with: . OECD guideline No. 401, 24th February 1987, . EC Directive No. 92/69/EEC, B.I, 31st July 1992. 2. MATERIALS AND METHODS 2.1 TEST SUBSTANCE The test substancej||^U|HHused in the study was supplied by Elf Atochem S.A. The test substance was identified as follows: - protocol and labellingfHHBJk . Elf Atochem filing number: CAL 1392/99 . description: amber liquid . container: one smoked glass flask . date of receipt: 5 May 1999 . storage conditions: at room temperature and protected from light . expiry date: May 2000. Data relating to the characterization of the test substance are documented in a test article description and in analytical certificate (presented in appendix 1) provided by the Sponsor. 2.1.2 Formulation procedure The test substance was used undiluted. ,^onlainTSCACai CompanySa.n,--'1a--n--oe-s0r6' 1-""- CIT/Study No. 18748 T, Elf Atochem S.A. 2.2 TEST SYSTEM 2.2.1 Animals Species, strain: rat, Sprague-Dawley ICO: OFA-SD (IOPS Caw). Reason for this choice: rodent species generally accepted by regulatory authorities for this type of study. Breeder Ufa Credo, 69210 L'Arbresle, France. Number and sex: one group of ten animals (five males and five females). Age/weight: on the day of treatment, the animals were approximately 6 weeks old, and had a mean body weight standard deviation of 179 8 g for the males and 144 5 g for the females. Acclimatization: at least 5 days before the beginning of the study. Identification of the animals: the animals were identified individually by eamotches. 2.2.2 Environmental conditions During the acclimatization period and throughout the study, the conditions in the animal room were set as follows: .temperature: 212C . relative humidity: 30 to 70% . light/dark cycle: 12h/12h . ventilation: approximately 12 cycles/hour of filtered, non-recycled air. The temperature and relative humidity were under continuous control and recording. The records were checked daily and filed. In addition to these daily checks, the housing conditions and corresponding instrumentation and equipment are verified and calibrated at regular intervals. The animals were housed in polycarbonate cages (48 cm x 27 cm x 20 cm). Each cage contained one to seven animals of the same sex during the acclimatization period and five rats of the same sex during the treatment period. Each cage contained dust-free sawdust (SICSA, 94142 Alfortville, France). Bacteriological and chemical analyses of the sawdust, including the detection of possible contaminants (pesticides, heavy metals), are performed regularly by external laboratories. The results of these analyses are archived at CIT. 2.2.3 Food and water All the animals had free access to A04C pelleted diet (UAR, 91360 Villemoisson-sur-Orge, France), except as noted in "2.3.1 Fasting of the animals". Each batch of food "was analysed by the supplier for composition and contaminant levels. The diet formula is presented in appendix 2. Drinking water filtered by a FG Millipore membrane (0.22 micron) was provided ad libitum. Bacteriological and chemical analyses of the water and diet, including the detection of possible contaminants (pesticides, heavy metals and nitrosamines), are performed regularly by external laboratories. The results of these analyses are archived at CIT. No contaminants were known to have been present in the diet, drinking water or bedding material at levels which may be expected to have interfered with or prejudiced the outcome of the study. .,-^,^-tcoBlal"TSCACBl Cowpa"^'st-"-' CIT/Study No. 1874 TA^UI^------hIr Atochem ^.A. iu 2J TREATMENT 2.3.1 Fasting of the animals The animals were fasted for an overnight period of approximately 18 hours before dosing, but had free access to water. Food was given back approximately 4 hours after administration of the test substance. 2.3.2 Administration of the test substance As the test substance was anticipated to be non-toxic at 2000 mg/kg, a limit test was performed by administering 2000 mg/kg of the test substance to one group of ten animals (five males and five females). The test substance was administered undiluted, taking into consideration its specific gravity (1.1 g/ml). The administration was performed in a single dose by oral route using a metal gavage tube fitted to a 1 ml plastic syringe (0.01 ml graduations). The volume administered to each animal was adjusted according to body weight determined on the day of treatment. 2.3.3 Chronology of the study The single administration was performed on 13 July 1999 in the morning (day 1) and was followed by a 14-day observation period until 27 July 1999 (day 15). 2.4 CLINICAL EXAMINATIONS 2.4.1 Clinical signs and mortality The animals were observed frequently during the hours following administration of the test substance, for detection of possible treatment-related clinical signs. Thereafter, observation of the animals was made at least once a day until day 15. Type, time of onset and duration of clinical signs were recorded for each animal individually. Time of death was recorded individually, in terms of the number of hours or days after dosing. 2.4.2 Body weight The animals were weighed individually just before administration of the test substance on day 1 and then on days 8 and 15. The body weight gain of the treated animals was compared to that of CIT control animals with the same initial body weight. Company eSa,.n-;.*--"."-- D""o'a"s' "ol contain TSCA CB1. 2.5 NECROPSY On day 15, all animals were killed by carbon dioxide asphyxiation and a macroscopic examination was performed. After opening the thoracic and abdominal cavities, a macroscopic examination of the main organs (digestive tract, heart, kidneys, liver, lungs, pancreas, spleen and any other organs with obvious abnormalities) was performed. In case of macroscopic lesions, organ samples were taken and preserved in 10% buffered formalin. No microscopic examination was performed. 2.6 DATA EVALUATION Evaluation of the toxicity of the test substance following a single oral administration in rats should include the relationship, if any. between the animals'exposure to the test substance and the incidence and severity of all abnormalities including behavioural and clinical abnormalities, macroscopic lesions, body weight changes, mortality and any other toxic effects. 2.7 PROTOCOL ADHERENCE The study was performed in accordance with Study Protocol No. 18748 TAR and subsequent amendments, with the following deviations from the agreed Study Protocol: . the temperature and relative humidity recorded in the animal room were sometimes outside of the target ranges specified in the protocol. These minor deviations were not considered to compromise the validity or integrity of the study. 2.8 ARCHIVING The study documentation and specimens generated during the course of the study are archived at CIT, 27005 Evreux, France, for 10 years after the end of the in vivo phase of the study. The archived study materials include: . protocol and possible amendments, . raw data, . correspondence, . final report and possible amendments. On completion of this period, the archived study materials will be returned to the Sponsor, or may be archived at CIT for a further period. Company Sanitized. Doss not contain TSCA CBt 3. RESULTS 3.1 CLINICAL EXAMINATIONS 3.1.1 Clinical signs and mortality (table 1) No clinical signs and no deaths were observed during the study. 3.1.2 Body weight (figures 1 and 2, tables 2 and 3) The body weight gain of the treated animals was similar to that of CIT historical control animals. 3.2 PATHOLOGY (table 4) Macroscopic examination of the main organs of the animals revealed no apparent abnormalities. 4. CONCLUSION Under our experimental conditions, the oral LDo of the test substanceHB^H^HBUB I^^B^^^ys equal to or higher than 2000 mg/kg in rats. No signs of toxicity were observed at Sdose*1 According to the classification criteria laid down in Commission Directive 93/2 I/EEC (27th April 1993) adapting to technical progress for the eighteenth time Council Directive 67/548/EEC, the test substance does not present a significant acute toxic risk if swallowed. ,-,.,,B^""ot"ai"Tsl;A<;& CcTOS"?38"' CIT/StudyNo. 18748T Figure 1: Body weight of treated rats ,lt Atoctiem Si.A. Cowpa"^8^3' CIT/Study No. 1874S lAKl^------------^ir Atocnem ;>.A. 14 Figure 2: Body weight ofCIT historical control rats e.g.: Crr Historical data of animals dosed by the oral route: results of control animals from October 1995 to December 1997. ,., ewpwr53"11'" ^^amwc^ CIT/Study No. 1S748 TAJ^BB|^Hft.it Atocnem S.A. 13 Table 1: Individual clinical signs and mortality Dose (mg/kg) Time Animals Males Females Mortality 2000 1 h 1 2h-4h } 01-02-03-04-05 06-07-08-09-10 No D2toD15 J min: minutes h : hour D : day Clinical signs None c^s^.------"--------^ CIT/Study No. lo74oTA ,11 Atocnem a.A. 10 Table 2: Individual and mean body weight and weekly body weight change (g) Dose mg/kg Volume Sex ml/kg Animals 1 Days (1) 8 (1) 2000 1.82 Male 01 194 84 278 72 02 179 74 253 58 03 176 76 252 54 04 175 73 248 62 05 173 68 241 61 M 179 75 254 61 SD 8 6 14 7 2000 1.82 Female 06 142 42 184 31 07 146 31 177 34 08 137 36 173 29 . 09 142 42 184 19 10 151 37 188 27 M 144 38 181 28 SD 5 5 6 6 (1) - Body weight gain M - Mean SD - Standard Deviation 15 350 311 306 310 302 316 19 215 211 202 203 215 209 6 nolccn.a.nTOC'1081 Con-pa"!^."^-0065 cn'/iStudy i^o. 18/48 i ,n Alocnem a.A. i / Table 3: Mean body weight and weekly body weight change of CIT historical control rats BODY WEIGHT OF CONTROL RATS (g) Days mg/kg ml/kg 1 8 0 10 Male M 182 261 SD 10 12 n 45 45 0 10 Female M 147 190 SD 10 15 n 45 45 M mean SD standard deviation . n number of animals 15 315 21 45 215 17 45 BODY WEIGHT CHANGE OF CONTROL RATS (g) mg/kg ml/kg 0 10 Male M SD - 0 10 Female M SD M : mean SD : standard deviation Days 1 to 8 79 7 43 8 8 to 15 54 13 25 7 e.g.: CIT Historical data of animals dosed by the oral route: results of control animals from October 1995 to December 1997. Aoonl^TSCACBl Sani^-0033"01 CofflpaW CIT/Study No. 18748T. it Atochem b.A. is Table 4: Individual macroscopic examinations at necropsy Dose mg/kg Time Males Animals Females 2000 D: day D 15 01-02-03-04-05 06-07-08-09-10 Macroscopic abnormalities No apparent abnormalities notconlatoTSCACBl Co^S^-0^ U 1/Stuay INO. 18 /46 1 AJ^^Bii^lHBlcir Aluc-ncin o./^.. * ' ^^ APPENDICES ^^^, ^^^SCAC^ CIT/Study No. 1S748 TAKUH^HfP|tlt Atoctiem S.A. 2U l.Test article description and analytical certificate .--.-"--""' CIT/Study No. l874yi bit Atochem t>.A. /I TOXICOLOGY DEPARTMENT CONFIDENTIAL April 99 elf atochem s.a. La defense 10, cours Michelet 92091 Paris-la-Defense, France TEST ARTICLE DESCRIPTION IDENTITY Test article name Chemical name CAS number EINECS number Purity Origin and batch Batch Elf Atochem filing number PHYSICAL AND CHEMICAL PROPERTIES Appearance pH Density Boiling point Flash point Solubility amber liquid 51 HOOkg/m3 at20C 98-100 C no flash point water: insoluble soluble in alcohols, aromatic hydrocarbons TOXICOLOGICAL INFORMATIONS AND USE SAFETY See safety data sheet__ __ STORAGE AND DISPOSAL Storage Expiry date Disposal__ in dark and at room temperature may 2000 incineration .Company. Sanity. Does not cont^n TSCA CBl CIT/Study No. 18748T, eiF atochem ^ S i--\r--i'--^. :lf Atochem S.A. 22 Elf Atochem S.A. Usln d Vaiera-Soi'nt-Paul ZA Vinars Saini Paul Rteux B.P. 20 60870 Ricux (Fianco) TAI: 4<.74.<4.74 . Fax : 4.74.2.07 TtlBl: 140 2- ATO VSP ----------------------------------------y----------------.---- Villcn Suo( Pul. Ic 9 Novcnibre t'W -^-------------------------------------------- CERT1FICAT D'ANALYSE f ;t LotN Extrait Sec Tcrtio Butanol Acrylamidc Acidc acrylique Test d'extinction UDit6 % Rdsultat 30,3 Specification dc fabrication 29131 M6thodc d'amlysc LCU 622 % 0.95 Q\ 1 LCU653 % 0.07 OAO,1 LCU 658 % 0.1 Oi2 LCU 658 1 posilif positif. SA-t 021 Lc Chcfdc Scm'oe du Laboratoirc WWATURE: N0,^: Rnhert Gnippo Company Sanitized. Does not contain TSCACBI h"U''l--l,(..pita|3(;lU5norr.,or5 CIT/Study No. 18748 TAK|--------MB|^lt Atochem S.A. 23 2. Diet formula P^^co^TSCACft Company ,,S-a,,,^,,^,,",' Lii/anlay INO. io/'+o i/\. i^tl niu^iicin o.n. Ref: A04 COMPLETE DIET RAT AND MOUSE MAINTENANCE DIET Appearance: 15 nun diameter pellets or powder Conditioning: 25 kg double paper bag with aluminium on the outside Daily portion: Rat 18-25 g. Mouse 5-10 g, water ad libltum. FORMULA % Cereals and cereal byproducts...... 88 Vegetable protein (soya bean meal, yeast).................................. 7 Animal protein (fish).................... 2 Vitamin and mineral mixture....... 3 AVERAGE ANALYSIS % Calorific value (KCal/kg) Moisture........................... Proteins............................, Lipids................................ Carbohydrates (N.F.E.)..... Fibre.................................. Minerals (ash)................... 2900 12 17 3 58.7 4 5 AMINO ACID VALUES (calculated in mg/kg) MIr'IERALS (caIculated in nrig/kg) Nat CMV val. val. Total P ..... 5900 Ca 3300 K 6700 Na 300 Me 1900 Mn 50 Fe , 90 Cu 15 Zn 40 Co T I 0.3 0 5000 0 1600 100 40 150 15 45 1.5 0 5900 8300 6700 1900 2000 90 240 30 85 1.5 0.3 vrTAMINS (calculated pe-rkg) Nat CMV val. val. Total Arginine ..... Cystine ....... Lysine......... Methionine. Tryptophan. Glycine....... FATTY ACID VALUES (calculated in mg/kg) Palmitic acid..... Palmitoleic acid. Stearicacid......... Oleic acid........... Linoleic acid....... Linolenic acid..... 9800 2300 8500 3200 1900 8100 2600 Traces 500 8000 14500 Traces Vitamin A Vitamin D3 Vitamin Bl Vitamin B2 Vitamin B3 Vitamin B6 Vitamin B12 Vitamin E Vitamin K3 Vitamin PP Folic acid Biotin Choline Traces Traces 6mg 2mg 10 mg 1.3mg 0.01 mg 15 mg 0.25 mg 60 mg 0.5 mg 0.04 mg 1200mg 7500 IU 1500IU 1 mg 4.5 mg 6.5 mg 1.3mg O.P1 mg 15 mg 2.25 mg 15 mg Omg Omg 400 mg 7500 IU 1500IU 7mg 6.5 mg 16.5 mg 2.6 mg -0.02 mg 30 mg 2.5 mg 75 mg 0.5 mg 0.04 mg l600mg Available under quality "Control Ref.: A04 C " UAR,7rueGallieni,91360Villemoisson-Tei: 01.69.04.03.57 - Fax : 01.69.04.81.97 (Ref. Doc. UAR: 1992) Company San^."n"ossno