Document vBNYaND6xQExqN4K1daV6wDXb

MIN 313/023622 PERFLUOROOCTANESULFONYL FLUORIDE (POSF; T-7661.4) TOXICITY STUDY BY INHALATION ADMINISTRATION TO CD RATS FOR 13 WEEKS FOLLOWED BY A 4 WEEK RECOVERY PERIOD Volume 1 Sponsor 3M Center, 3M Corporate Toxicology, Building 220-2E-02, St Paul, MN 55133-3220, USA. Research Laboratory Huntingdon Life Sciences Ltd., Woolley Road, Alconbury, Huntingdon, Cambridgeshire, PE28 4HS, ENGLAND. Report issued: 28 September 2005 Page 1 of 740 MIN 313/023622 CONTENTS Page CONTENTS.................................................................................................................................... 2 COMPLIANCE WITH GOOD LABORATORY PRACTICE.................................................... 4 QUALITY ASSURANCE STATEMENT..................................................................................... 5 CONTRIBUTING SCIENTISTS................................................................................................... 6 SUMMARY.................................................................................................................................... 7 INTRODUCTION........................................................................................................................... 9 EXPERIMENTAL PROCEDURE................................................................................................. STUDY SCHEDULE AND STRUCTURE............................................................................... TEST SUBSTANCE AND FORMULATION........................................................................... ANIMAL MANAGEMENT...................................................................................................... SERIAL OBSERVATIONS....................................................................................................... NECROPSY AND HISTOLOGY.............................................................................................. PATHOLOGY............................................................................................................................ DATA TREATMENT................................................................................................................. QUALITY ASSURANCE AND ARCHIVING PROCEDURES............................................. 11 11 12 13 15 19 22 22 26 RESULTS........................................................................................................................................ 27 DISCUSSION................................................................................................................................. 35 REFERENCES................................................................................................................................ 36 FIGURES 1. Bodyweights - group mean values....................................................................................... 37 :2 : iVHN 313/023622 TABLES Page 1. Bodyweights - group mean values.......................................................................................... 38 2. Food consumption - group mean values................................................................................ 40 3. Haematology - group mean values......................................................................................... 42 4. Biochemistry - group mean values......................................................................................... 50 5. Urinalysis - group mean values.............................................................................................. 66 6. Organ weights - group mean values....................................................................................... 71 7. Macroscopic pathology - group distribution.......................................................................... 83 8. Microscopic pathology - group distribution.,,............................................. 87 APPENDICES 1. Weekly physical examination - individual findings.............................................................. 101 2. Daily post-dose observations - individual findings................................................................ 118 3. Bodyweights - individual values............................................................................................ 128 4. Food consumption - individual values................................................................................... 135 5. Haematology - individual values................................................ 138 6. Biochemistry - individual values........................................................................................... 150 7. Urinalysis - individual values............................................................................................. 178 8. Absolute organ weights - individual values........................................................................... 202 9. Individual pathological findings............................................................................................ 212 10. pH values for terminal urine.................................................................................................. 293 11. Cell proliferation findings...................................................................................................... 296 ADMINISTRATION OF POSF BY INHALATION TO RATS.................................................... 298 SCANNING ELECTRON MICROSCOPE EXAMINATION AND X-RAY MICROANALYSIS.......................................................................................................................... 369 ANALYTICAL PHASE REPORT.................................................................................................. 518 HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENTS......................... 735 EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENTS............................................ 738 Volume 1........................................................................................................................................... 1 Volume 2 ........................................................................................................................................... 366 :3 : MIN 313/023622 COMPLIANCE WITH GOOD LABORATORY PRACTICE PERFLUOROOCTANESULFONYL FLUORIDE (POSF; T-7661.4) TOXICITY STUDY BY INHALATION ADMINISTRATION TO CD RATS FOR 13 WEEKS FOLLOWED BY A 4 WEEK RECOVERY PERIOD 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 1999 (Statutory Instrument No 3106) as amended by Statutory Instrument 2004 No. 994. OECD Principles of Good Laboratory Practice (as revised in 1997), ENV/MC/CHEM(98)17. EC Commission Directive 1999/11/EC of 8 March 1999 (Official Journal No. L 77/8), as amended by EC Commission Directive 2004/10/EC of 11 February 2004 (Official Journal No. L 50/44). These principles of Good Laboratory Practice are accepted by the regulatory authorities of the United States of America and Japan on the basis o f intergovernmental agreements. An expiry date for the test substance was not given. The material was assumed to be stable for the duration of the study. Scanning Electron Microscopy and X-ray analysis was conducted by a facility which is not part of the GLP monitoring programme. No claim o f compliance with GLP is made in respect of these phases of the study. Terence J. Kenny, B.Sc. (Hons.), Study Director, Huntingdon Life Sciences Ltd. Date 4 MIN 313/023622 QUALITY ASSURANCE STATEMENT PERFLUOROOCTANESULFONYL FLUORIDE (POSF; T-7661.4) TOXICITY STUDY BY INHALATION ADMINISTRATION TO CD RATS FOR 13 WEEKS FOLLOWED BY A 4 WEEK RECOVERY PERIOD The following inspections and audits have been carried out in relation to this study: Study Phase Date of Inspection Date of Reporting Protocol Audit 10 December 2001 11 December 2001 Study Based Inspections Study preparation Test Item control and disposition Exposure and generation of test atmosphere Atmosphere sampling Clinical signs (post-dose) Clinical signs (weekly) Fresh urine collection for SEM analysis. Necropsy 20 December 2001 20 December 2001 20 December 2001 20 December 2001 20 December 2001 8 January 2002 14 January 2002 21 March 2002 20 December 2001 20 December 2001 20 December 2001 20 December 2001 20 December 2002 8 January 2002 14 January 2002 26 March 2002 Report Audit 9 June 2004 - 22 June 2004 19 July and 25 July 2005 22 September 2005 24 June 2004 25 July 2005 22 September 2005 Protocol Audit: An audit of the protocol for this study was conducted and reported to the Study Director and Company Management as indicated above. Study based inspections: Inspections of phases of this study were conducted and reported to the Study Director and Company Management as indicated above. Process based inspections: At or about the time this study was in progress inspections 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 the reported results to reflect the raw data. Details of the Quality Assurance inspections and audits pertaining to the Analytical Phase determining the level of POSF, PFOS and PFOA in liver and serum samples are presented in the Principal Investigator's report included as an addendum to this main report. Tracy Scaijfe, F.R.Q.A., Group Manager, Department of Quality Assurance. Huntingdon Life Sciences Ltd. :5: CONTRIBUTING SCIENTISTS MIN 313/023622 STUDY MANAGEMENT Terence J. Kenny, B.Sc. (Hons.), Study Director. Rhiannon Davies, B.Sc. (Hons.), Senior Study Supervisor. Derek W. Coombs, B.Sc., MSc., Senior Toxicologist. AEROSOL TECHNOLOGY AND ANALYSIS Ian S. Gilkison, M.A., Ph.D., Section Head, Aerosol Technology and Analysis. CLINICAL PATHOLOGY P. Travis, B.Sc., M.Sc., Head of Central Laboratory Services and Clinical Analytical Service Manager, CentraLabs Clinical Research. PATHOLOGY Samuel McCormick, M.V.B., M.R.C.V.S., Ph.D., F.R.C.Path., Director of Pathology. STATISTICS Graham F. Healey, B.Sc. (Hons.), M.Sc., A.R.C.S., Head, Department of Statistics. BIOANALYSIS Richard A. Grazzini, Exygen Research, 3058 Research Drive, State College, PA 16800, USA. :6: SUMMARY MIN 313/023622 The systemic toxic potential of the test substance, POSF, to Crl:CD (SD)IGS BR rats by inhalation administration was assessed over a period of 13 weeks, followed by a 4 week recovery period. Three groups of ten male and ten female rats received POSF by snout only inhalation at exposure levels of 26, 104 and 299 ppm by volume for 13 weeks. A similarly constituted Control group received air. A further 5 males and 5 females were allocated as satellite animals to Group 1 and 4 only and were dosed for 4 weeks only. Ten male and ten female rats in Groups 1-4 were exposed for 13 weeks, after which 5 males and 5 females per group were sacrificed. The remaining 5 male and 5 female rats in each group were allocated as recovery group animals and were allowed to recover for four weeks following the 13 weeks exposure. During the study, clinical condition, bodyweight, food consumption, haematology, blood chemistry, urinalysis, organ weight, macroscopic and microscopic pathology investigations were undertaken. Principal findings are summarised below: Results Mortality and clinical signs There were no treatment-related effects during the 13 weeks of the treatment period or in any of the recovery animals during the 4 week recovery period. Bodyweight Following 13 weeks of treatment with POSF there was a dose-related reduction in bodyweight gain. Food consumption A reduction in food consumption was seen in Group 4 (High dose) animals, with statistical significance attained for males, compared with Control following 13 weeks of treatment. Following 4 weeks of recovery female food consumption was still reduced in treated groups. Haematology There were no findings considered to be o f toxicological importance. Blood chemistry Alkaline phosphatase levels were higher for Group 3 (Intermediate dose) and Group 4 (High dose) males compared with Control and remained higher following 4 weeks of recovery. Alanine amino-transferase levels were higher in all treated male groups compared with Control, and remained higher following 4 weeks of recovery. Cholesterol levels were lower in all treated male groups compared with Control and remained lower following 4 weeks of recovery. Creatinine and triglyceride levels were reduced for all treated groups compared with Control, following 4 weeks of recovery the levels were comparable with Control. :7: MIN 313/023622 Urinalysis There were no findings considered to be of toxicological importance. Organ weights Absolute liver weights were greater than controls in both sexes of the Intermediate and High dose groups. The differences were statistically significant when adjusted for bodyweight. Following a 4 week recoveiy period the recovery animal's liver weights, when adjusted for bodyweight, remained statistically significantly higher for Group 3 (Inter dose) males and in both sexes of Group 4 (High dose). In satellite rats after 4 weeks exposure at the High dose level the liver weights were statistically significantly greater than control weights. Lung and bronchi weights (absolute and bodyweight adjusted) in High dose rats of both sexes were statistically significantly higher than controls after 13 weeks of exposure. After 4 weeks of recovery the lung and bronchi weights in male High dose rats remained statistically significantly higher than controls, lung and bronchi weights in females were similar to controls after 4 weeks of recovery. In Satellite animals after 4 weeks of expsoure at the High dose, lung and bronchi weights were statistically significantly higher than controls. Kidney weights, when adjusted for bodyweight, in male rats from the Intermediate and High dose were statistically significantly higher than controls after 13 weeks of exposure. There were no such differences in females. After 4 weeks of recovery the bodyweight adjusted kidney weights of High dose males were statistically significantly greater than controls. There were no histopathology findings which accounted for the higher bodyweight adjusted kidney weights noted in males. In satellite rats there were no differences in kidney weight after 4 weeks of exposure. Macroscopic pathology Enlargement of the liver was observed in 4/5 male rats treated with 300 ppm for 4 weeks compared with none in the Control male rats and in 3/5 male rats and 2/5 female rats treated with 300 ppm for 13 weeks compared with none in the Control rats. This finding was not apparent following 4 weeks of recovery. Microscopic pathology The following findings were related to the administration of 300 ppm of the compound: larynx - necrosis of ventral cartilage, with no evidence of recovery in males and with incomplete recovery in females lungs - foamy alveolar macrophages/septal thickening/scattered alveolar macrophages in both sexes with incomplete recovery in males and complete recovery in females liver - centrilobular hepatocyte hypertrophy extending to the midzonal area with males more notably affected, prominent centrilobular hepatocyte pigment in females, and incomplete recovery of these findings in both sexes. The administration of 30 ppm or 100 ppm was associated with centrilobular hepatocyte hypertrophy extending to the midzonal area or centrilobular hepatocyte hypertrophy in males, with incomplete recovery at these dose levels. Conclusion The administration of 30 ppm or 100 ppm was associated with centrilobular hepatocyte hypertrophy extending to the midzonal area or centrilobular hepatocyte hypertrophy extending to the midzonal area or centrilobular hepatocyte hypertrophy in males, with incomplete recovery at these dose levels. It was not possible to identify a no effect level in this study. :8: INTRODUCTION MIN 313/023622 Objective The objective of this study was to assess the systemic toxic potential of perfluorooctanesulfonyl flouride (POSF), when administered by snout only inhalation to rats for 13 weeks. Recovery from any effects was assessed during a 4-week recovery period. Satellite rats were allocated to Groups 1 and 4 and dosed for 4 weeks. Regulatory compliance The study was designed to meet the requirements of: Guidelines for the Testing of Chemicals - 413 European Economic Communities. The study was conducted in accordance with the requirements of current, internationally recognised Good Laboratory Practice Standards, and the applicable sections of the United Kingdom Animals (Scientific Procedures) Act 1986. Test system The rat was chosen as the test species because of its acceptance as a predictor of toxic change in man and the requirement for a rodent species by regulatory agencies. The Crl.CD (SD)IGS BR strain was used because of the historical control data available in this laboratory. Route of administration The inhalation route of administration was chosen to simulate the conditions of potential human exposure. Treatment groups and dosages The dosages used in this study (0, 30, 100 and 300 ppm) were selected in conjunction with the Sponsor with reference to previous work with this compound performed in these laboratories. (Huntingdon Life Sciences Report Number: MIN 312/014272). In that study rats were exposed to a target concentration of 300 ppm where transient clinical effects were seen. Study location The test system was maintained at the following laboratory: Huntingdon Life Sciences Ltd., Huntingdon Research Centre, Woolley Road, Alconbury, Huntingdon, Cambridgeshire, PE28 4HS, ENGLAND. Histology was performed at: Huntingdon Life Sciences Ltd., Eye Research Centre, Eye, Suffolk, IP23 7PX, ENGLAND. :9: Scanning electronic microscopy and X-ray analysis was performed at: Plymouth Electron Microscope Unit, University of Plymouth, Drake Circus, Plymouth, PL4 8AA. The bioanalytical phase was performed at: Exygen Research, 3058 Research Drive, State College, PA 16801, USA. MIN 313/023622 10 : EXPERIMENTAL PROCEDURE MIN 313/023622 STUDY SCHEDULE AND STRUCTURE Duration of treatment The test substance, POSF, was administered for 6 hours a day, 5 days a week over a period of 13 consecutive weeks and for satellite rats a period of 4 weeks. The necropsy procedures were completed in 1 day, during which time treatment continued up to the day prior to necropsy, and serial observations were recorded at appropriate intervals. The duration of treatment is reported as 13 weeks. Animals assigned to the recovery phase completed a further four weeks without treatment. Time schedule Study initiation: (Protocol signed by Study Director) Experimental start date: Treatment commenced: Necropsy completed: Satellite Study Main Study Recovery Study Experimental completion date: Study completion: 6 December 2001 6 December 2001 20 December 2001 15 January 2002 21 March 2002 19 April 2002 29 October 2002 28 September 2005 Identity of treatment groups The study consisted of one Control and three treated groups of rats, identified as follows: Group 1 2 3 4 Treatment Control POSF POSF POSF Target exposure level (ppm) 0 30 100 300 Main study (13 weeks) No. of animals Animal numbers Male Female Male Female 5 5 1-5 21-25 5 5 6-10 26-30 5 5 11-15 31-35 5 5 16-20 36-40 Cage numbers Male Female 15 26 37 48 11 MIN 313/023622 Group 1 2 3 4 Group 1 4 Treatment Control POSF POSF POSF Treatment Control POSF Target exposure level (ppm) 0 30 100 300 Target exposure level (ppm) 0 300 Recovery phase (13 weeks + 4 weeks recovery) No. of animals Animal numbers Cage numbers Male Female Male Female Male Female 5 5 41-45 61-65 9 13 5 5 46-50 66-70 10 14 5 5 51-55 71-75 11 15 5 5 56-60 76-80 12 16 No. of animals Male Female 55 55 Satellite study (4 weeks) Animal numbers Male Female 81-85 91-95 86-90 96-100 Cage numbers Male Female 17 19 18 20 TEST SUBSTANCE AND FORMULATION Test substance Information supplied by the Sponsor regarding the test substance is contained in the test substance data sheet, which is retained in the study records. The following information is given in summaiy: Identification: POSF, T7661.4 Description: Liquid Storage conditions: Room temperature Supplier: Sponsor Batch number: 040227 Date of receipt: 14 June 2001 Quantity received: 4 x 20 kg Stability: Assumed stable for duration of the study Purity: > 99.5 % A small sample (1 ml) was sealed in a suitable container and stored in Archives at an appropriate temperature. The Sponsor was responsible for the characterisation of the test substance and the documentation of the methods of synthesis, fabrication or derivation and stability. : 12 : MIN 313/023622 ANIMAL MANAGEMENT Animal supply, acclimatisation and allocation A total of 55 male and 55 female CrlrCD (SD)IGS BR rats were received from Charles River (UK) Ltd, Margate, Kent, England. The rats were ordered at 40-44 days of age and within a weight range of 11 g for each sex. On arrival, the animals were removed from the transit boxes and allocated to study cages. Using the sequence of cages in the battery, one animal at a time was placed in each cage with the procedure being repeated until each cage held the appropriate number of animals. Each sex was allocated separately. The latest Health Screen Report published by the animal supplier was provided to HLS. In addition, the additional consignment of animals included a health screen relating to the current status of the breeding colony. These documents were sent to Huntingdon Life Sciences Veterinary Service immediately upon receipt for review and subsequent archiving. The cages constituting each group were blocked together by sex and the groups were dispersed in batteries so that possible environmental influences arising from their spatial distribution were equilibrated, as far as was practicable. Additionally, batteries of cages were rotated around the room at weekly intervals to further minimise possible spatial variations. Each animal was assigned a number and uniquely identified within the study by a tail tattoo. Each cage label was colour-coded according to group and was uniquely numbered with cage and study number, as well as the identity of the occupants. The animals were allowed to acclimatise to the conditions described below for 14 days before treatment commenced. For those animals selected for this study, their age at the start of treatment was 55 to 58 days and their bodyweights were in the range of 239 g to 292 g for males and 170 g to 209 g for females. The spare animals were removed from the study room after treatment commenced. Animal housing, diet and water supply Animals were housed inside a restricted entry rodent facility (Building Y14, Room 007). The facility was designed and operated to minimise the entry of external biological and chemical agents and to minimise the transference of such agents between rooms. Before the study the room was cleaned and disinfected with a bactericide. Each animal room was kept at positive pressure with respect to the outside by its own supply of filtered fresh air, which was passed to atmosphere and not re-circulated. The temperature and relative humidity controls were maintained within the range of 19 to 23C and 40 to 70%, respectively. Periodic checks were made on the number of air changes in the animal rooms. Temperature and humidity were monitored continuously. Ranges occasionally deviated from target and the actual ranges recorded were 17 to 21 C and 29 to 74%. Deviations were minor and had no impact on the study outcome. Artificial lighting was controlled to give a cycle of 12 hours continuous light and 12 hours continuous dark per 24 hours. : 13 : MIN 313/023622 Alarms were activated if there was any failure of the ventilation system, or temperature limits were exceeded. A stand-by electricity supply was available to be automatically brought into operation should the public supply fail. The animals were housed five of one sex per cage, unless this number was reduced by mortality or isolation. The cages were made of a stainless steel body with a stainless steel mesh lid and floor, and were suspended above absorbent paper which was changed at appropriate intervals. Cages, cage-trays, food hoppers and water bottles were changed at appropriate intervals. The animals were allowed free access to a standard rodent diet (Rat and Mouse No. 1 Maintenance Diet from Special Diets Services Ltd., Witham, Essex, England), except during the 6 hours exposure or when urine was being collected and overnight before routine blood sampling. This diet contained no added antibiotic or other chemotherapeutic or prophylactic agent. Water taken from the public supply was freely available via polycarbonate bottles fitted with sipper tubes, except during the 6 hour exposure or when urine was being collected. Each batch of diet was analysed routinely by the supplier for various nutritional components and chemical and microbiological contaminants. Supplier's analytical certificates were scrutinised and approved before any batch of diet was released for use. The quality of the water supply is governed by regulations published by the Department for Environment, Food and Rural Affairs (formerly known as the Department of the Environment). Certificates of analysis were received routinely from the supplier. Since the results of these various analyses did not provide evidence of contamination that might have prejudiced the study, they are not presented. No other specific contaminants that were likely to have been present in the diet or water were analysed, as none that may have interfered with or prejudiced the outcome of the study was known. Administration The vapour-generated from the test substance was administered for 6 hours a day, for 5 days a week for 13 weeks, Control rats received air only consecutive days. The test material was delivered to an all glass vapouriser and generated as a vapour atmosphere, into a stream of air for administration to the rats by inhalation from snout only exposure chambers. The test substance was metered to the vaporiser from an infusion pump. The vapour/air mixture passed directly into the exposure chamber. The target chamber concentration was achieved by using different liquid feed rates controlled by the infusion pump and using syringes of an appropriate value. The target concentrations for treated rats were 30, 100 and 300 ppm. Control rats received air only. The rats were exposed to the control/test atmosphere using ADG snout-only exposure chambers (ADG Developments Ltd, Hitchin, Hertfordshire, England). All animals (including reserves) were subjected to restraint procedures and exposed to air only (Sham dosing), for 5 consecutive days prior to the start of exposures, in order to accustom animals to the restraining procedure. The rats were restrained once per day, increasing the `Sham dosing' period progressively (0.5,1,2,4 and 6 hours for Days -5 to -1 respectively). Details of administration and analysis of the test atmospheres together with the results obtained are presented in ADMINISTRATION OF POSF BY INHALATION TO RATS appended to this report. : 14 : MIN 313/023622 SERIAL OBSERVATIONS Dated and signed records of all activities relating to the day by day running and maintenance of the study within the animal unit as well as to the group observations and examinations outlined in this procedure were recorded in the Study Day Book. In addition, observations relating to individual animals made throughout the day were recorded. Serial observations performed on the animals from the Satellite groups were confined to clinical observations, bodyweights and food consumption. All observations described below were performed in cage number sequence except where otherwise indicated. Clinical observations Animals were inspected visually at least twice daily for evidence of ill-health or reaction to treatment. Cages and cage-trays were inspected daily for evidence of ill-health amongst the occupant(s), such as loose faeces. Any deviation from normal was recorded at the time in respect of nature and severity, date and time of onset, duration and progress of the observed condition, as appropriate. In addition, detailed observations were recorded daily, on the days of exposure as follows: Pre-exposure observation During exposure [however, observation was severely restricted due to the tube restraint] As each animal was returned to its home cage As late as possible in the working day In addition, a more detailed weekly physical examination was performed on each animal to monitor general health. During the acclimatisation and recoveiy periods, observations of the animals and their cages were recorded at least once per day. Bodyweight The weight of each rat was recorded one week before treatment commenced (Week -1), on the day that treatment commenced (Week 0), weekly throughout the treatment and recovery periods, and before necropsy. Food consumption The weight of food supplied to each cage, that remaining and an estimate of any spilled was recorded for the week before treatment started (Week -1), and each week throughout the treatment and recoveiy periods. From these records the mean weekly consumption per animal (g/rat/week) was calculated for each cage. : 15 : MIN 313/023622 Haematology, peripheral blood During Week 13 of treatment (before dosing) and in Week 5 of recovery, blood samples were obtained from all animals after overnight starvation. Animals were held under light general anaesthesia induced by isoflurane and blood samples were withdrawn from the retro-orbital sinus. Blood samples (nominally 0.5 ml) were collected into EDTA as anticoagulant and examined for the following characteristics: The following were measured using a Bayer-Technicon HIE haematology analyser: Haematocrit (Hct) Haemoglobin (Hb) Red blood cell count (RBC) Reticulocytes (Retie) Mean cell haemoglobin (MCH) Mean cell haemoglobin concentration (MCHC) Mean cell volume (MCV) Total white cell count (WBC) Differential WBC count Neutrophils (N) Lymphocytes (L) Eosinophils (E) Basophils (B) Monocytes (M) Large unstained cells (LUC) Platelet count (Pit) Reticulocyte count (Retie) - brilliant cresyl blue stain, examined by light microscopy for reticulocytes and abnormalities using a Sysmex R3000 Reticulocyte Counter. Blood film - Romanowsky stain, examined by light microscopy for abnormal morphology and unusual cell types, including normoblasts. The most common morphological changes, anisocytosis, micro/macrocytosis, hypo/hyperchromasia were recorded as follows: = += ++ = no abnormalities detected slight moderate Additional blood samples (nominally 0.5 ml) were taken into citrate anticoagulant and examined in respect of: Prothrombin time (PT) using an ACL 3000 Plus analyser and IL PT-Fibrinogen reagent Activated partial thromboplastin time (APTT) using an ACL 3000 Plus Analyser and IL APTT reagent : 16 : MIN 313/023622 Blood chemistry At the same time and using the same animals as for peripheral haematology, further blood samples (nominally 0.7 ml) were collected into lithium heparin as anticoagulant. All tubes were mechanically agitated for at least five minutes and the sample subsequently centrifuged in order to separate the plasma. After separation, the plasma was examined in respect of: Using a Hitachi 917 Clinical Chemistry Analyser: Alkaline phosphatase (ALP) Alanine aminotransferase (ALT) Aspartate aminotransferase (AST) Lactate dehydrogenase (LDH Total) and iso enzymes Creatine Phosphokinase (CPK Total) Sorbitol dehydrogenase (SDH) Total Bilirubin (Bili) Urea Creatinine (Creat) Glucose (Glue) Total cholesterol (Choi) Triglycerides (Trig) Sodium (Na) Potassium (K) Chloride (Cl) Calcium (Ca) Inorganic phosphorus (Phos) Total protein (Total Prot) Electrophoretic protein fractions; albumin (Alb), a l globulin (al), a2 globulin (a2), 3 globulin (Beta), y globulin (Gamma) were analysed with agarose gel, using a Beckman test kit, staining with Ponceau-S and scanning with a suitable densitometer. Albumin/globulin ratio (A/G Ratio) was calculated from total protein concentration and analysed albumin concentration. 17 MIN 313/023622 Urinalysis During Weeks 13 of treatment and Week 5 of recovery, overnight urine samples were collected from all animals. Animals were deprived of water from approximately 16:30/16:00 hours and placed in an individual metabolism cage without food or water; urine was collected until approximately 07:30/07:45 hours respectively the following day. The individual samples were examined for the following characteristics: Appearance (App.) Volume (Vol) pH Specific gravity (SG) Protein (Prot) Sodium (U-Na), potassium (U-K) and chloride (U-Cl) Glucose (Glue), ketones (Keto), bile pigments (Bili), haem pigments (Blood) by Multistix. Multistix are diagnostic reagents obtained from Bayer pic, Newbury, England and were used as qualitative indicators of analyte concentration. Results for haem pigments are reported as positive or negative only whilst results for glucose, ketones and bile pigments are reported according to the following convention: 0 TR += negative trace `small amount' of analyte A microscopic examination of the urine sediment was performed. An aliquot of the urine sample was centrifuged and the resulting deposit spread on a microscope slide. The deposit was examined for the presence of the following: Crystals Epithelial cells Leucocytes Erythrocytes Casts Spermatozoa and precursors Other abnormal components (Cryst) (Epi) (Leuc) (RBC) (Casts) (Sperm) (Abn.) The grading of cell frequency in the centrifuged deposit is as follows: 0 = None found in any field examined 1 = Few in some fields examined 2 = Few in all fields examined The sample residues were despatched to Exygen Research for test substance/metabolite analyses. : 18 : MIN 313/023622 Urine SEM x-ray analysis Individual urine samples were collected from main study animals during Week 13 of exposure, recovery group animals during the last week of recovery and from satellite animals during Week 4 of exposure. The samples were in addition to those collected overnight for urinalysis and metabolite assays. The samples were collected from the animals within 2 hours following lights on in the animal holding room where possible. For animals failing to produce a specimen during this interval, urine was collected at necropsy. The urine samples were immediately assayed for pH using a microelectrode, and prepared SEM stubs despatched to Plymouth University for calculi and crystal analysis by SEM X-ray element identification. NECROPSY AND HISTOLOGY Method of kill Animals killed during the study and those surviving until the end of the scheduled treatment or recovery period were killed by intraperitoneal injection of sodium pentobarbitone, followed by exsanguination. The sequence in which the animals were killed after completion of treatment or recovery period was selected to allow satisfactory inter-group comparison. Test substance/metabolite analyses Samples of blood (for serum) were obtained at necropsy by cardiac/aorta puncture while the rats were held under terminal sodium pentobarbitone anaesthesia. The blood samples (up to 4 ml) were run into tubes, allowed to clot at room temperature and the serum separated and frozen in liquid nitrogen prior to despatch to Exygen Research. Residual samples of urine following urinalysis parameters assay and urine samples collected from Satellite rats during Weeks 4 were frozen and despatched to Exygen Research. During necropsy samples of liver were collected from all animals. The samples were frozen liquid nitrogen and despatched to Exygen Research. Macroscopic pathology All animals were subject to a detailed necropsy. After a review of the history of each animal, a full macroscopic examination of the tissues was performed. All external features and orifices were examined visually. The cranial roof was removed to allow observation of the brain, pituitary gland and cranial nerves. After ventral mid-line incision, the neck and associated tissues and the thoracic, abdominal and pelvic cavities and their viscera were exposed and examined in situ. Any abnormal position, morphology or interaction was recorded. The requisite organs were weighed and external and cut surfaces of the organs and tissues were examined as appropriate. Any abnormality in the appearance or size of any organ and tissue was recorded and the required tissue samples preserved in appropriate fixative. Any photographs of unusual findings were taken at the discretion of the necropsy supervisor. : 19 : MIN 313/023622 The retained tissues were checked before disposal of the carcass. Organ weights The following organs, taken from each animal killed after 13 weeks of treatment or 4 weeks of recovery, were dissected free of adjacent fat and other contiguous tissue and the weights recorded: Adrenals Brain Heart Kidneys Liver Lungs with mainstem bronchi (Lungs & br) Bilateral organs were weighed together. Fixation Testes and epididymides were fixed in Bouin's solution, prior to transfer of these tissues to 70% industrial methylated spirit. Eyes were fixed in Davidson's fluid. Urinary bladder was fixed in Bouin's solution Samples (or the whole) of the other tissues listed below from all animals were preserved in 10% neutral buffered formalin. Adrenals Aorta - thoracic Brain Caecum Colon Duodenum Epididymides Eyes Femur and joint+ Harderian glands Head Heart * Ileum Jejunum Kidneys *a Lachrymal glands Larynx* Liver* Lungs * Lymph nodes - mandibular - mesenteric - tracheo-bronchial Mammary area - caudal Nasal turinates Oesophagus Optic nerves Ovaries Pancreas Pituitary Prostate Rectum Salivary glands+ Sciatic nerves+ Seminal vesicles Skeletal muscle - thighs+ Skin Spinal cord Spleen Sternum Stomach Testes Thymus Thyroid with parathyroids Tongue Trachea* Urinary bladder* Uterus and cervix Vagina + Only one processed for examination 3Sections included the pelvic epithelium : 20 : MIN 313/023622 Samples of any abnormal tissues were also retained and processed for examination. In those cases where a lesion was not clearly delineated, contiguous tissue was fixed with the grossly affected region and sectioned as appropriate. This extensive list of tissues preserved is intended to satisfy any possible future requirement for further examination of tissues. Samples of the head (including nasal cavity, paranasal sinuses and nasopharynx) and the remaining femur, salivary gland, sciatic nerve and skeletal muscle (thigh) were not examined histologically, but are retained against any future requirement for microscopic examination. Histology For those animals specified in the Pathology section, the relevant tissues with an asterisk (*) were subject to histological processing. Tissue samples were dehydrated, embedded in paraffin wax, sectioned at approximately four to five micron thickness and stained with haematoxylin and eosin, except the testes which were stained using a standard periodic acid/Schiff (PAS) method. Those tissues subjected to histological processing included the following regions: Heart - included auricular and ventricular regions Liver - section from all main lobes Lungs - section from two major lobes, to include bronchi Trachea - included bifurcation Urinary bladder - sections taken sagitally (using one half for SEM analysis and the other half for light microscopy and cell proliferation index) For bilateral organs, sections of both organs were prepared. A single section was prepared from each of the remaining tissues required for microscopic pathology. : 21 MIN 313/023622 PATHOLOGY Light microscopy Microscopic examination was performed as follows: All tissues preserved for examination were examined for all animals of Groups 1 (Control) and 4 (300 ppm) sacrificed on completion of the scheduled treatment period, and for all animals killed or dying during the study. Tissues reported at macroscopic examination as being grossly abnormal were examined for all Main study and Recovery animals. The following tissues, which were considered to exhibit a reaction to treatment at the high dosage, were examined for all Main study and Recovery animals; larynx, liver and lungs. Findings were either reported as "present" or assigned a severity grade. In the latter case one of the following five grades was used - minimal, slight, moderate, marked or severe. A reviewing pathologist undertook a peer review of the microscopic findings. PCNA Staining for cell proliferation Sections of urinary bladder were taken from all animals for PCNA immunostaining in order to assess the degree of cell proliferation present. A total of 3,000 cells were counted from 3 separate sections (1,000 cells/section) in order to determine the cell proliferation index. Positive PCNA staining cells were counted, and examined by the pathologist to assess the sections and count S-phase positive cells, if practicable. In addition, sections of the duodenum from each animal were taken and stained to act as positive controls for the immunostaining methodology. DATA TREATMENT This report contains serial observations pertaining to all weeks of treatment and recovery completed, together with signs data collected during the necropsy period. The only serial observations relating to the acclimatisation period included in this report relate to the Week-1 bodyweights and food consumption. Summary statistics (e.g. means and standard deviations) presented in this report were calculated from computer-stored individual raw data. The summary statistics and the individual data were stored in the computer to a certain number of decimal places, different for each parameter. For presentation purposes, however, they were usually rounded to fewer places. It is, therefore, not generally possible to reproduce the presented means and standard deviations exactly using the presented individual data. Throughout the report the following abbreviations are used: R N or n SD or sd Recovery Number of animals examined. Standard deviation. : 22 : MIN 313/023622 Definition of "Week" The first week of treatment started at midnight prior to treatment commencing and ended at midnight on the seventh day following. Subsequent experimental weeks of treatment and recovery were of the same duration. Signs A detailed history of individual animals that showed signs is presented in Appendix 1, as the weeks in which the sign was observed. Only animals with findings are presented. Observations during the recovery phase are recorded on a study week basis. Weeks of study relate to weeks of recovery as follows: Week of recovery 1R 2R 3R 4R 5R Week of Study 14 15 16 17 18 Death code key 7 = terminal kill 8 = recovery kill H = humane kill F = found dead Appendix 2 presents the individual observations of signs related to dose administration. Bodyweight Group mean weight changes were calculated from the weight changes of individual animals. Food consumption Values presented for the amount of food consumed in each cage in each experimental week allow for any animal that died or was killed during the week. Weekly group mean food consumptions and standard deviations were derived from unrounded cage values, which were weighted to allow for any deaths as follows: S[ad.fc] x w = 2(ad) S [a d .(fc -x w )2] Jsd 2 ( a d ) - l Where: x w = weighted mean, ad = animal days, fc = cage value and sd = standard deviation. Blood chemistry Albumin to globulin (A/G) ratios were calculated as: Albumin concentration A/G =----------------------------------------- Total protein - albumin concentration : 23 : MIN 313/023622 Urinalysis The abbreviations used in Appendix 7 have the following meanings: MY Medium yellow appearance TR Trace detected. PY Pale yellow appearance CPY Cloudy, pale yellow appearance CMY Cloudy, medium yellow appearance Group means and standard deviations are presented for volume, pH, specific gravity, protein, and electrolytes only. Organ weights The adjusted organ weights presented in Table 6 and Appendix 8 were those from the analysis of covariance. This analysis took the transformed absolute organ weights as the response variable and the original, untransformed, terminal bodyweights as the covariate. Linear regression lines were fitted to each group, assuming the lines for all groups were parallel to one another. The individual organ weights (and therefore the group mean organ weights) were adjusted towards the overall mean terminal bodyweight, thus removing the effect of bodyweight. The standard deviations were based on the variability after allowing the covariate, rather than the usual unadjusted standard deviations. Pathology Tissues which could not be examined are specified in the appendix. The absence of a comment for a tissue scheduled for examination therefore indicates that the tissue was examined and found to be normal. Tissues recorded as abnormal macroscopically but found to be normal microscopically are described as `No significant lesion' in the microscopic pathology appendix. In all tabular presentations of data the tissues specified in the protocol for histopathological examination precede other tissues. 24 : MIN 313/023622 Statistical analysis All statistical analyses were carried out separately for males and females. Data relating to food consumption was analysed on a cage basis. For all other parameters, the analyses were carried out using the individual animal as the basic experimental unit. The following data types were analysed at each timepoint separately:- . Bodyweight, using gains over appropriate study periods Food consumption, over appropriate study periods, using cumulative cage totals Blood chemistry, haematology and urinalysis Organ weights, both absolute and adjusted for terminal bodyweight Pathological findings, for the number of animals with and without each finding. For categorical data, including pathological findings, the proportion of animals was analysed using Fisher's Exact test (Fisher 1973) for each treated group versus the control. For continuous data, Bartlett's test (Bartlett 1937) was first applied to test the homogeneity of variance between the groups. Using tests dependent on the outcome of Bartlett's test, treated groups were then compared with the Control group, incorporating adjustment for multiple comparisons where necessary. The following sequence of statistical tests was used for bodyweight, food consumption, organ weight and clinical pathology data If 75% of the data (across all groups) were the same value, for example c, then a frequency analysis was applied. Treatment groups were compared using a Mantel test for a trend in proportions (Mantel 1963) and also pairwise Fisher's Exact tests (Fisher 1973) for each dose group against the control both for i) values <c versus values >=c, and for ii) values <=c versus values >c, as applicable. If Bartlett's test for variance homogeneity (Bartlett 1937) was not significant at the 1% level, then parametric analysis was applied. If the FI test for monotonicity of dose-response (Healey 1999) was not significant at the 1% level, Williams' test for a monotonic trend (Williams 1971, 1972) was applied. If the FI test was significant, suggesting that the doseresponse was not monotone, Dunnett's test (Dunnett 1955, 1964) was performed instead. If Bartlett's test was significant at the 1% level, then logarithmic and square-root transformations were tried. If Bartlett's test was still significant, then non-parametric tests were applied. If the HI test for monotonicity of dose-response (Healey 1999) was not significant at the 1% level, Shirley's test for a monotonic trend (Shirley 1977) was applied. If the HI test was significant, suggesting that the dose-response was not monotone, Steel's test (Steel 1959) was performed instead. Where appropriate, analysis of covariance was used in place of analysis of variance in the above sequence. For organ weight data, analysis of variance was performed using terminal bodyweight as covariate when the within group relationship between organ weight and bodyweight was significant at the 10% level (Angervall and Carlstrom, 1963), in an attempt to allow for differences in bodyweight which might influence the organ weights. : 25 : MIN 313/023622 Significant differences between Control and treated groups were expressed at the 5% (p<0.05), 1% (p<0.01) or 0.1% (p<0.001) level. Williams test is denoted by **'; Dunnett's test is donated by **', Shirley's test is donated by ` ' and t tests are denoted by '+ ' QUALITY ASSURANCE AND ARCHIVING PROCEDURES Quality Assurance Details of the Quality Assurance inspections and audits are presented on the Quality Assurance Statement. Archives Following completion of this study all raw data, specimens and samples, except those generated or used during any Sponsor's or supplier's analysis, were stored in the archives o f Huntingdon Life Sciences. Types of sample and specimen which are unsuitable, by reason of instability, for long term retention and archiving may be disposed of after the periods stated in Huntingdon Life Sciences Standard Operating Procedures. All data generated by Exygen Research and Plymouth University will be archived by Huntingdon Life Sciences. A copy of the final report and all Quality Assurance inspection records will be retained indefinitely. All other appropriate specimens and 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 above requirements. Under no circumstances will any item be discarded without the Sponsor's knowledge. 26 RESULTS MIN 313/023622 CHAMBER ATMOSPHERE CONDITIONS Chamber analysed concentration of POSF The data are presented in ADMINISTRATION OF POSF TO RATS BY INHALATION appended to this report. The mean chamber concentrations (ppm) are summarized below: Group 23 4 Mean chamber concentration (ppm) 26 104 299 The achieved concentrations were close to target. Signs and mortality (Appendix 1 and 2) Animal No. 63, Group 1 (Recovery Control) female was killed for humane reasons during Week 3 of the study. This animal had signs of head leaning to the right, displacement of hindlimbs, reduced body temperature, poor righting reflexes and abnormal gait. Animal No. 23, Group 2 (Low dose) died during the bleed for haematology and biochemistry during Week 13. Neither of the 2 deaths were considered to be related to exposure to POSF. There were no treatment-related effects during the 13 weeks of the treatment period or in any of the recovery animals during the 4 week recovery period. Bodyweight (Figure 1; Table 1; Appendix 3) Following 4 weeks of treatment there was a reduction in bodyweight for treated satellite animals compared with Control. Following 13 weeks of treatment there was a dosage-related reduction in bodyweight for treated rats, with statistical significance being attained for all treated males and Group 3 (Inter dose) and Group 4 (High dose) females, compared with Control. Following 4 weeks recovery Group 4 (High dose) animals bodyweights recovered, with bodyweights comparable or greater to Control. Food consumption (Table 2; Appendix 4) Throughout the 13-week exposure period food consumption by High-dose rats of both sexes was lower than that of the controls. To a lesser degree and with a few exceptions consumption by the Intermediate and Low dose groups was also lower than control consumption. Over the course of the 13 weeks of exposure the effect was small and achieved statistical significance in male High dose rats only, however, this was in contrast to pre-dose consumption where the 3 treated groups consumed more than the controls. During the 4 week recovery period consumption by all treated male groups increased while male control consumption remained constant such that consumption by all groups was similar. In females consumption by High dose, Low dose and controls increased. Consumption by the female Intermediate dose was similar to that seen during the dosing period. In all treated female groups consumption was lower than that o f controls over the recovery period. In satellite animals the High dose consumption was lower than controls in both sexes over the 4 weeks of exposure. : 27 : MIN 313/023622 Haematology (Table 3; Appendix 5) Following 13 weeks of treatment group mean reticulocyte counts were statistically significantly higher in Group 4 (High dose) males compared with Control, however no such effect was present in females and the difference is considered to be of no toxicological importance. Group mean neutrophils and monocyte counts were reduced in Week 13 for Group 3 (Intermediate dose) and Group 4 (High dose) males with statistically significance being attained for Group 4 (High dose), however there were considered to be no effect on these counts following 4 weeks of recovery. In Week 13 large unstained cells (LUC) counts were statistically significantly lower in treated males compared with Control and these remained lower following 4 weeks of recovery. These differences were not dose related and were independent of sex, they are considered unrelated to treatment. Following 4 weeks of recovery there was a reduced white blood cell count (WBC) in females, with reduced leucocyte (L) basophil (B) and monocyte (M) counts, with statistical significance being attained for Group 3 (Intermediate dose) and Group 4 (High dose) females. A longer prothrombin time (PT) was also evident in all treated groups following 4 weeks of recovery, with statistical significance being attained for all males and Group 3 (Intermediate dose) and Group 4 (High dose) females. These differences were not seen following 13 weeks of treatment with POSF and are therefore considered not to be of any toxicological significance. Other inter-group differences in the haematological parameters investigated in Week 13 or following a 4 week recovery period that attained statistical significance were small, not dose-related and inconsistent between the sexes and considered incidental and of no toxicological significance. Blood chemistry (Table 4; Appendix 6) Following 13 weeks of treatment group mean alkaline phosphatase (ALP) levels were statistically significantly higher for Group 3 (Intermediate dose) and Group 4 (High dose) males. The values for this parameter remained higher following 4 weeks of recovery with statistical significance being attained for Group 4 (High dose) compared with Control. Following 13 weeks of treatment alanine amino-transferase (ALT) levels were higher in treated males compared with Control, and remained higher following 4 weeks of recovery with statistical significance in Group 4 (High dose) only. Group mean lactate dehydrogenase (LDH) levels were lower in all treated groups of both sexes following 13 weeks of treatment compared with Control. However, following 4 weeks of recovery control values were lower and considered to be reasonably similar to those of test groups. The difference were not dose-related and statistical significance was attained for females values only they are considered to be of no toxicological importance. Cholesterol levels were lower in treated males following 13 weeks of treatment compared with Control, with statistical significance being attained for Group 3 (Intermediate dose) and Group 4 (High dose) males. Following 4 weeks of recovery the level was still lower than controls in treated males with statistical significance being attained in all treated groups. Creatinine (Creat) levels were reduced for all treated groups compared to Control, with statistical significance being attained for Group 2 (Low dose) females and Group 3 (Intermediate dose) and Group 4 (High dose) animals of both sexes following 13 weeks of treatment. Following 4 weeks recovery the levels in treated groups were comparable with Control, : 28 : MIN 313/023622 Triglyceride levels were reduced in treated groups with statistical significance attained for females only, following a 4 week recovery period there were no significant differences between the groups. Other inter-group differences in the biochemical parameters investigated in Week 13 or following a 4 week recovery period were small, not dose-related and inconsistent between the sexes and considered incidental and of no toxicological significance. Urinalysis (Table 5; Appendix 7) Inter-group differences in the parameters investigated in Week 13 and following 4 weeks of recovery were small, not dose-related and inconsistent between the sexes. None are considered to be of toxicological significance. Urinary pH (Appendix 10) pH values of treated and Control animals were similar. Organ weights (Table 6; Appendix 8) Absolute liver weights were greater than controls in both sexes of the Intermediate and High dose groups. The differences were statistically significant w'hen adjusted for bodyweight. Following a 4 week recovery period the recoveiy animal's liver weights, when adjusted for bodyweight, remained statistically significantly higher for Group 3 (Intermediate dose) males and Group 4 (High dose). In satellite rats after 4 weeks exposure at the High dose level the liver weights were statistically significantly greater than control weights. Lung and bronchi weights (absolute and bodyweight adjusted) in High dose rats of both sexes were statistically significantly higher than controls after 13 weeks of exposure. After 4 weeks of recovery the lung and bronchi weights in male High dose rats remained statistically significantly higher than controls, lung and bronchi weights in females were similar to controls after 4 weeks of recovery. In Satellite animals after 4 weeks of exposure at the High dose, lung and bronchi weights were statistically significantly higher than controls. Kidney weights, when adjusted for bodyweight, in male rats from the Intermediate and High dose were statistically significantly higher than controls after 13 weeks of exposure. There were no such differences in females. After 4 weeks of recovery the bodyweight adjusted kidney weights of High dose males were statistically significantly greater than controls. There were no histopathology findings which accounted for the higher bodyweight adjusted kidney weights noted in males. In satellite rats there were no differences in kidney weight after 4 weeks of exposure. Other inter-group differences were small, not dose-related and inconsistent between the sexes. Macropathology (Table 7; Appendix 9) The macroscopic examination performed at termination revealed the following change: Liver Enlargement was observed in 4/5 satellite male rats treated with 300 ppm for 4 weeks compared with none in the Control male macropathological rats. An enlargement of the liver was observed in 3/5 male rats and 2/5 female rats treated with 300 ppm for 13 weeks compared with none in the Control rats. Following 4 weeks of recoveiy no enlargement of the liver was seen amongst recovery animals or any other treatment-related macropathological findings. The incidence and distribution of all the findings were considered to fall within the background range of macroscopic changes. : 29 : MIN 313/023622 Micropathology (Table 8; Appendix 9) Treatment-related findings Larynx Necrosis of the ventral cartilage was associated with the administration of 300 ppm for 13 weeks. There were no findings which were considered to be related to treatment in rats receiving 30 ppm or 100 ppm. Dosage (ppm) Necrosis of ventral cartilage Total 0 0 Males 30 100 00 300 4a 0 0 Females 30 100 00 300 5b Number of larynx examined 5 5 5 5 5 5* 5 5 a - K0.05, b-/K0.01 with Fisher's Exact Test * Includes one sporadic animal Following the 4 week recovery period, there was no evidence of recovery from necrosis of the ventral cartilage in males previously receiving 300 ppm. The incidence of this finding in females previously receiving 300 ppm was only marginally reduced compared to that noted in the main part of the study. Males Females Dosage (ppm) 0 30 100 300 0 30 100 300 Necrosis of ventral cartilage Total 0 0 0 4a 0 0 0 3 Number of larynx examined 5 5 5 5 5* 5 5 5 a - ><0.05 with Fisher's Exact Test * Includes one sporadic animal Lungs Both the incidence and degree of foamy alveolar macrophages/septal thickening/scattered alveolar macrophages, were increased in rats receiving 300 ppm for 13 weeks when compared with controls. This finding correlated with the higher bodyweight adjusted lung and bronchi weights in rats receiving 300 ppm when compared with controls. Although foamy alveolar macrophages/septal thickening/scattered macrophages was noted as minimal in a female receiving 100 ppm, this level of occurrence is not unusual in controls and therefore may be ignored. There were no findings in the lungs which were related to the administration of the compound in rats receiving 30 ppm or 100 ppm. : 30 : MIN 313/023622 Dosage (ppm) Foamy alveolar macrophages/ septal thickening/scattered alveolar macrophages Minimal Slight Moderate Total 0 0 0 0 0 Males 30 100 300 0 0030 00 10 00 10 0 0 5b 0 Females 30 100 300 0 12 002 000 0 1 4a Number of lungs examined 5 5 5 5 5 5* 5 5 a -f?<0.05, b - /?<0.01 with Fisher's Exact Test * Includes one sporadic animal Following the 4 week recovery period there was good evidence of reversibility of the finding in females previously receiving 300 ppm but less convincing recovery in males previously receiving 300 ppm. This correlated with the bodyweight adjusted lung and bronchi weights, which remained high for males but not females, compared with their contemporaneous recovery Control groups. Dosage (ppm) Foamy alveolar macrophages/ septal thickening/scattered alveolar macrophages Minimal Slight Total 0 1 0 1 Males 30 100 300 0 112 0 0030 1 1 5a 0 Females 30 100 300 102 000 102 Number of lungs examined 5 5 5 5 5* 5 5 5 a - /K0.05 with Fisher's Exact Test * Includes one sporadic animal Liver Centrilobular hepatocyte hypertrophy extending to the midzonal area and centrilobular hypertrophy were associated with the administration o f 30 ppm, 100 ppm, or 300 ppm for 13 weeks in males with a dosage-relationship. In females these findings was less pronounced and were associated only with the administration of 300 ppm. Prominent centrilobular hepatocyte pigment was noted occasionally in females receiving 300 ppm and this was likely to reflect increased hepatocyte metabolic activity. Hepatocyte hypertrophy correlated positively with the enlargement of the liver noted at necropsy in 3/5 males and 2/5 females receiving 300 ppm, compared with none in controls. There was correlation between hepatocyte hypertrophy and the higher level of ALT. Bodyweight adjusted liver weights were statistically higher in rats receiving 100 ppm or 300 ppm compared with controls. : 31 : MIN 313/023622 Males Females Dosage (ppm) 0 30 100 300 0 30 100 300 Centrilobular hepatocyte hypertrophy extending to midzonal area Slight 0 4 0 0 0 0 0 3 Moderate 0 0 5 5 0 0 0 2 Total 0 4a 5b 5b 0 0 0 5b Centrilobular hepatocyte hypertrophy Slight 0 1 0 0 0 0 0 0 Total 0 1 0 0 0 0 0 0 Prominent centrilobular hepatocyte pigment Minimal 0 0 0 0 0 0 0 1 Total 0 0 0 0 0 0 0 1 Number of livers examined 5 5 5 5 5 5* 5 5 a - p<0.05, b - /?<0.01 with Fisher's Exact Test * Includes one sporadic animal Following the 4 week recovery period there was good evidence of reversibility of hepatocyte hypertrophy in all affected groups, particularly in females previously receiving 300 ppm, where only the centrilobular area was affected. The histological findings correlated with the statistically higher bodyweight adjusted liver weights remaining in males previously receiving 100 ppm or 300 ppm compared with their contemporaneous control group, and also with the increased ALT in all treated male groups. Dosage (ppm) Centrilobular hepatocyte hypertrophy extending into midzonal area Slight Total Centrilobular hepatocyte hypertrophy Minimal Slight Total Prominent centrilobular hepatocyte pigment Minimal Total Number of livers examined 0 0 0 0 0 0 0 0 5 Males 30 100 300 0 Females 30 100 300 02 50000 0 2 5b 0 0 0 0 13 0 0 0 04 20000 00 3 3 0 0 0 0 4a 00000 02 00 00 0 02 5 5 5 5* 5 5 5 a -p<0.05, b - </><0.01 with Fisher's Exact Test * Includes one sporadic animal : 32 : MIN 313/023622 Other findings Moderate centrilobular hepatocyte necrosis/inflammatory cell infiltration and sinusoidal congestion was noted in the liver of animal 56 (male previously receiving 300 ppm). Since this was a recovery rat and there was only a single incidence of this finding it is unlikely to be associated with treatment. In the laiynx arytenoid epithelial hyperplasia, hyperkeratosis/parakeratosis and subepithelial inflammatory cell infiltration; and ventral pouch epithelial hyperplasia and inflammatory cell infiltration; and ventrolateral epithelial hyperplasia and squamous metaplasia were noted in occasional animals. These findings were considered likely to be related to the presence of an irritant (probably foreign body food material), rather than to the administration of the compound. Incidental findings All other findings were considered to be part of the usual background pathology to be expected in this age and strain of rat and as such to be of no toxicological importance. In particular, there were no findings which accounted for the higher bodyweight adjusted kidney weight noted in males receiving 300 ppm compared with Controls which remained following the recovery period. Conclusion The following findings were related to the administration of 300 ppm of the compound: larynx - necrosis of ventral cartilage, with no evidence of recovery in males and with incomplete recoveiy in females lungs - foamy alveolar macrophages/septal thickening/scattered alveolar macrophages in both sexes with incomplete recovery in males and complete recovery in females liver - centrilobular hepatocyte hypertrophy extending to the midzonal area with males more notably affected, prominent centrilobular hepatocyte pigment in females, and incomplete recovery of these findings in both sexes. The administration of 30 ppm or 100 ppm was associated with centrilobular hepatocyte hypertrophy extending to the midzonal area or centrilobular hepatocyte hypertrophy in males, with incomplete recovery at these dose levels. It was not possible to identify a no effect level in this study. Cell proliferation (Appendix 11) There were no treatment-related findings. SEM and X-ray analysis of urine and bladder tissue The results of examination together with SEM photomicrographs are presented in the Scanning Electron Microscope Examination and X-Ray Microanalysis report appended to this Main report. : 33 : MIN 313/023622 Test substance and metabolite analysis PFOS in the rat liver samples ranged from non-detected levels to 535,000ng/g. PFOA in the rat liver samples ranged from non-detected levels to 8270 ng/g. There was no POSF detected in any of the rat liver samples. PFOS in the rat serum samples ranged from non-detected levels to 158,000 ng/mL, PFOA in the rat serum samples ranged from non-detected levels to 12,100 ng/mL. There was no POSF detected in any of the rat serum samples. PFOS in the rat urine samples ranged from nondetected levels to 4710 ng/mL. PFOA in the rat urine samples ranged from non-detected levels to 18,600 ng/mL. There was no POSF detected in any of the rat urine samples. The average percent recoveries standard deviations for PFOS in rat liver, serum, and urine samples were 96% 11%, 102% 14%, and 87%i 11%, respectively. The average percent recoveries standard deviations for PFOA in rat liver, serum, and urine samples were 99% 9%, 105% 14%, and 110% 17%, respectively. The average percent recoveries standard deviations for POSF in rat liver, serum, and urine samples were 99% 18%, 92% 18%, and 99% 18%, respectively. : 34 DISCUSSION MIN 313/023622 Administration of POSF to rats at dosages of 30, 100 and 300 ppm a day over 13 weeks produced no mortalities associated with treatment and no clinical signs that were considered to be treatment related. There was a reduction in food consumption in Group 4 (High dose) animals compared with Control following 13 weeks of treatment which was still apparent following 4 weeks recovery for females only. Liver weights were statistically significantly higher in treated animals and remained higher following 4 weeks of recovery. Lungs and bronchi weights were higher in treated males and females respectively compared with Control and following 4 weeks of recovery still remained high for males but not females previously exposed to the high concentration. Enlargement of the liver was observed in 4/5 male rats treated with 300 ppm for 4 weeks compared with none in the Control male rats and in 3/5 male rats and 2/5 female rats treated with 300 ppm for 13 weeks compared with none in the Control rats. This finding was not apparent following 4 weeks of recovery. Histopathological findings related to the administration of 300 ppm of the compound were seen. In the larynx necrosis of ventral cartilage, with no evidence of recovery in males and with incomplete recovery' in females. In the lungs foamy alveolar macrophages/septal thickening/scattered alveolar macrophages in both sexes with incomplete recovery in males and complete recovery in females. Centrilobular hepatocyte hypertrophy extending to the midzonal area and centrilobular hypertrophy were associated with the administration of 30, 100, or 300 ppm for 13 weeks in males. There was some evidence of a dosage-relationship in terms of incidence and degree and the findings correlated with the higher levels of ALT seen in treated males. In females centrilobular hepatocyte hypertrophy extending to the midzonal area only was seen in animals treated with 300 ppm. Prominent centrilobular hepatocyte pigment was noted occasionally in females receiving 300 ppm and this was likely to reflect increased hepatocyte metabolic activity. In both sexes the hepatocyte hypertrophy correlated positively with the enlargement of the liver and higher bodyweight adjusted liver weights noted at necropsy. Following the 4 week recovery period there was good evidence of reversibility of hepatocyte hypertrophy in all affected groups, particularly in females previously receiving 300 ppm, where only the centrilobular area was affected. The histological findings correlated with the statistically higher bodyweight adjusted liver weights remaining in males previously receiving 100 ppm or 300 ppm and also with the increased ALT in all treated male groups. It was not possible to identify a no effect level in this study. The Lowest Adverse Effect Level (LOAEL) was considered to be 30 ppm. : 35 : REFERENCES MIN 313/023622 Haematology PROCTOR, R.R. and RAPAPORT, S.I. (1961). The Partial Thromboplastin Time with Kaolin. Am J. Clin. Path., 36, 212 - 219. QUICK, A.J. (1942). In: The Haemorrhage Diseases and the Physiology o f Haemostasis, ch III, p 24 - 40. Statistics ANGERVALL, L and CARLSTROM, E (1963) Theoretical criteria for the use of relative organ weights and similar ratios in biology, Jnl. Theoret. Biol. 4:254-259. BARTLETT, M.S. (1937), Properties of sufficiency and statistical test, Proc. Roy. Soc A 160: 268 -282. DUNNETT, C.W. (1955) A multiple comparison procedure for comparing several treatments with a control. J. Am. Stat. Assoc., 50, 1096. DUNNETT, C.W. (1964) New tables for multiple comparisons with a control. Biometrics, 20, 482. FISHER, R.A. (1973) in: (Eds). Statistical Methodsfo r Research Workers. Oliver and Boyd, Edinburgh. HEALEY, G.F. (1999) The FI approximate parameter test for monotonicity. Statistical Specification ST9725, Department of Statistics, Huntingdon Life Sciences. HEALEY, G.F. (1999) The HI approximate non-parameter test for monotonicity. Statistical Specification ST9726, Department of Statistics, Huntingdon Life Sciences. MANTEL, N. (1963) Ch-square tests with one degree of freedom : Extentions of the Mantel Haenszel procedure. J.Amer. Statist. Ass., 58, 690-700. SHIRLEY, E. (1977), A non-parametric equivalent of Williams' test for contrasting increasing dose levels of a treatment, Biometrics, 33: 386 - 389. STEEL, R. G. D., (1959), A multiple comparison rank sum test: treatments versus control, Biometrics, 14:560-572. WILLIAMS, D.A. (1971), A test for differences between treatment means when several dose levels are compared with a zero dose control, Biometrics, 27: 103 - 117. WILLIAMS, D.A. (1972), The comparison of several dose levels with a zero dose control, Biometrics, 28: 519-531. : 36 : FIGURE 1 Bodyweight - group mean values (g) 0 1 2 3 4 5 6 7 8 9 10 11 12 13 R1 R2 R3 R4 Week MIN 313/023622 TABLE 1 BODYWEIGHT - GROUP MEAN VALUES (G) Main study and recovery phase animals GROUP :12 3 4 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL (PPM) : 0 30 100 300 SEX: --------------------- MALE--------------------- GROUP: 1 2 3 4 WEEK -------------------- FEMALE 12 3 -i 0 1 CO oo 2 3 4 5 6 7 8 9 10 11 12 13 14a 15a 16a 17a 233 235 236 234 178 175 177 261 267 264 258 191 187 189 286 290 284 274 202 201 201 310 310 303 296 213 208 206 333 333 324 316 220 219 214 356 352 346 335 232 224 224 376 367 362 346 238 234 228 393 383 379 360 246 241 236 409 399 392 369 255 246 240 421 412 406 379 258 251 245 433 423 416 382 264 255 248 444 430 422 390 270 259 251 454 438 433 395 277 263 252 463 444 442 399 277 264 257 462 445 441 395 274 260 252 479 451 455 425 291 295 257 492 462 470 437 303 301 264 506 470 476 454 305 306 265 517 481 485 465 305 308 269 Gain * ** Week 0-13 202 179 177 137 % of Control - 89 88 69 Weeks 13-17 50 44 47 52 % of Control - 88 94 104 83 73 - 88 27 28 - 104 * 63 76 20 74 Level of significance - all comparisons made with control * p 0.05, ** p <. 0.01 (Williams' test) a Recovery animals only Print Mo: 0001 Printed: 24.-JUN-02 Xybion protocol number: MIN 313 4 179 190 203 206 213 217 225 231 235 237 241 244 246 247 240 253 263 268 274 * 50 60 33 122 MIN 313/023622 TABLE 1 (Boclywcight - continued) Satellite study animals GROUP :i 2 34 COMPOUND : CONTROL -- POSF--- EXPOSURE LEVEL(PPM): 0 30 100 300 VLOO SEX: ----------MALE--------- --------FEMALE- GROUP : 1 4 1 4 WEEK -i 237 237 180 176 0 263 272 190 187 1 290 292 204 193 2 316 313 209 196 3 341 332 223 204 Gain Weeks 0-3 78 60 33 16 No differences of statistical significance (p> 0.05) Print No: 0002 Printed: 24-JUN-02 Xybion protocol number: MIN 313A 313/023622 TABLE 2 FOOD CONSUMPTION - GROUP MEAN VALUES (G/RATAVEEK) Main siudy and recovery phase animais GROUP :1 2 3 4 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL (PPM) : 0 30 100 300 Print No: 0002 Printed: 24-JUN-02 Xybion protocol number: MIN 313 SEX: --------------------- MALE--------------------- GROUP : 1 2 3 4 WEEK -------------------- FEMALE 12 3 4 -i 182 194 191 186 137 141 137 142 i 171 171 172 154 133 137 132 132 2 186 184 178 172 139 137 143 129 3 195 194 192 179 144 145 141 133 4 205 199 197 188 150 147 144 131 5 206 199 196 183 150 147 145 135 6 204 199 204 190 150 147 145 136 7 203 203 203 186 149 145 144 134 8 205 202 205 191 147 147 145 133 9 203 200 201 183 147 145 142 134 10 203 202 202 187 147 141 139 131 11 206 201 202 182 147 142 140 129 12 208 200 201 181 146 142 142 130 13 194 193 190 170 139 130 130 116 14* 198 197 205 190 152 156 138 135 15* 197 197 205 196 158 160 142 141 16* 201 206 206 199 190 136 140 137 17* 203 205 205 200 147 151 138 140 Average Weeks 1-13 199 196 196 180 145 143 141 131 % of Control - 98 98 90 - 99 97 90 Weeks 14-17 200 201 205 196 162 151 140 138 % of Control - 101 103 98 - 93 86 85 Level of significance - all comparisons made with control * p<, 0.05 (Williams' test) Recovery animals only MIN 313/023622 Satellite study phase GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 TABLE 2 (Food consumption - continued) 34 100 300 GROUP: WEEK 1 2 34i 2 1 169 2 185 3 197 161 138 177 145 188 149 3 Printed: 24-JUN-02 Xybion protocol number: MIN 313A 4 122 131 136 MIN 313/023622 TABLE 3 Haematology - group mean values Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 -- POSF-- 30 100 300 Group / sex Met Hb RBC Retie L/L g/dL xIO'TL % 1M Mean SD n 0.432 0.0129 3 15.4 0.10 3 8.24 0.263 3 2.78 0.567 3 2M Mean SD n 0.441 0.0127 5 15.7 0.30 5 8.18 0.278 5 2.02 0.262 5 3M Mean SD n 0.447 0.0202 5 15.8 0.74 5 8.34 0.255 5 2.00 0.430 5 4M Mean SD n 0.430 0.0319 5 15.2 1.63 5 7.83 0.816 5 3.44 4.050 5 Level of significance - all comparisons made with control p < , 0.05 (Shirley's test) *p<, 0.05 (Williams' test) MCH Pg 18.7 0.53 3 19.1 0.59 5 19.0 0.58 5 19.5 0.32 5 MCHC g/dL 35.6 0.84 3 35.5 0.62 5 35.5 0.38 5 35.4 1.39 5 MCV fL 52.4 0.23 3 53.9 1.60 5 53.6 1.47 5 55.1 2.18 5 WBC x109/L 12.22 4.073 3 13.47 4.667 5 10.98 0.766 5 11.71 1.002 5 N x109/L 2.64 2.018 3 3.25 2.585 5 1.59 0.614 5 * 0.90 0.219 5 MIN 313/023622 TABLE 3 (Haematology - continued) Day 91 Group Compound Exposure level (ppm) i Control 0 234 --POSF----- 30 100 300 Group /sex L xl09/L E x109/L B xio'/L M x109/L 1M Mean SD n 8.83 1.814 3 0.18 0.124 3 0.04 0.017 3 0.36 0.142 3 2M Mean SD n 9.61 2.079 5 0.18 0.037 5 0.04 0.031 5 0.30 0.146 5 3M Mean SD n 8.92 1.216 5 0.20 0.076 5 0.03 0.013 5 0.20 0.027 5 * 4M Mean 10.46 0.11 0.04 0.15 SD 1.098 0.033 0.011 0.111 n5 5 5 5 Level of significance - all comparisons made with control *p<, 0.05, **/>^0.01 (Williams' test) *p< 0.05 (Dunnett's test) LUC xl09/L 0.17 0.086 3 * 0.09 0.047 5 ** 0.05 0.013 5 ** 0.06 0.031 5 Pit xl07L 806 233.8 3 818 343.4 5 851 183.1 5 1067 227.2 5 PT sec 13.0 2 13.9 0.36 3 * 14.2 0.46 5 13.2 0.57 5 APTT sec 19.0 2 16.6 0.58 3 19.0 2.98 5 18.9 3.11 5 MIN 313/023622 TABLE 3 (Haematology - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 -- POSF-- 30 100 300 Group / sex Hct Hb RBC Retie L/L g/dL x10I2/L % IF Mean 0.423 15.2 7.60 2.31 SD 0.0126 0.38 0.305 0.500 n3333 2F Mean 0.425 15.1 7.62 2.09 SD 0.0112 0.44 0.214 0.409 n5 5 55 3F Mean 0.421 15.1 7.55 2.53 SD 0.0168 0.71 0.256 0.971 n5 5 5 5 4F Mean 0.431 15.6 7.95 1.74 SD 0.0109 0.29 0.333 0.372 n5 5 5 5 No differences of statistical significance (p> 0.05) MCH Pg 19.9 0.44 3 19.8 0.47 5 20.0 0.78 5 19.6 0.70 5 MCHC g/dL 35.8 0.55 3 35.6 0.23 5 35.8 0.61 5 36.1 0.31 5 MCV fL 55.7 0.61 3 55.8 1.40 5 55.9 1.53 5 54.4 2.20 5 WBC x109/L 8.31 2.996 3 9.07 1.461 5 9.48 2.893 5 9.52 1.461 5 N x109/L 1.29 0.719 3 1.56 0.823 5 1.63 1.058 5 1.25 0.493 5 MIN 313/023622 TABLE 3 (Haematology - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 -- POSF-- 30 100 300 Group / sex L x109/L E x109/L B x109/L M x109/L IF Mean 6.58 0.15 0.02 0.20 SD 2.189 0.026 0.010 0.050 n3 3 3 3 2F Mean 6.91 0.20 0.02 0.28 SD 0.972 0.104 0.004 0.090 n5 555 3F Mean 7.38 0.16 0.03 0.22 SD 1.748 0.090 0.019 0.100 n5555 4F Mean 7.86 0.16 0.02 0.17 SD 1.041 0.075 0.008 0.103 n55 55 Level of significance - all comparisons made with control * p<, 0.05 (Williams' test) LUC x109/L 0.06 0.032 3 0.10 0.058 5 0.06 0.015 5 0.06 0.023 5 Pit x109/L 1058 111.2 3 935 48.3 5 892 58.2 5 1021 128.3 5 PT sec 13.8 0.62 3 13.4 0.12 3 14.1 0.66 5 * 12.9 0.36 5 APTT sec 14.5 4.09 3 18.7 1.49 3 17.3 1.91 5 18.0 1.83 5 MIN 313/023622 TABLE 3 (Haematology - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 -- POSF-- 30 100 300 Group /sex Hct Hb RBC Retie L/L R/dL x10I2/L % 1M Mean SD n 0.410 0.0212 5 15.4 0.66 5 7.79 0.431 5 2.46 0.363 5 2M Mean SD n 0.395 0.0110 5 14.6 0.41 5 7.59 0.288 5 2.67 0.677 5 3M Mean SD n 0.413 0.0082 5 15.2 0.23 5 7.66 0.186 5 2.38 0.365 5 4M Mean SD n 0.414 0.0171 5 15.3 0.63 5 7.84 0.331 5 2.44 0.177 5 No differences of statistical significance (p> 0.05) MCH Pg 19.8 0.84 5 19.2 0.56 5 19.8 0.29 5 19.5 0.24 5 MCIIC g/dL 37.4 0.58 5 37.0 0.49 5 36.8 0.37 5 37.0 0.42 5 MCV fL 52.7 2.02 5 52.0 0.89 5 53.9 1.05 5 52.8 1.16 5 WBC x109/L 10.97 1.951 5 9.75 2.203 5 12.27 5.446 5 11.14 1.319 5 N x109/L 1.63 1.041 5 2.08 0.774 5 2.63 2.652 5 2.26 1.343 5 MIN 313/023622 TABLE 3 (Haematology - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 -- POSF-- 30 100 300 Group /sex L xlO'/L E x109/L B x109/L M x109/L 1M Mean SD n 8.70 1.204 5 0.15 0.041 5 0.03 0.018 5 0.25 0.068 5 2M Mean SD n 7.25 1.924 5 0.08 0.026 5 0.02 0.011 5 0.20 0.099 5 3M Mean SD n 9.11 3.060 5 0.15 0.087 5 0.04 0.026 5 0.22 0.148 5 4M Mean SD n 8.37 0.705 5 0.18 0.104 5 0.03 0.011 5 0.19 0.086 5 Level of significance - all comparisons made with control *p<, 0.05 (Williams' test) */?<0.05, ^1/75 0.01 (Dunnett's test) LUC x 109/L 0.21 0.074 5 0.11 0.032 5 0.12 0.105 5 0.12 0.044 5 Pit x109/L 1009 96.5 5 982 125.8 5 867 73.1 5 985 88.6 5 PT sec 12.9 0.22 5 ** 14.0 0.40 4 ** 14.6 0.49 5 * 13.7 0.50 5 AP1T sec 19.2 2.40 5 18.3 1.18 4 * 16.0 2.38 5 * 15.7 1.45 5 MIN 313/023622 TABLE 3 (Haematology - continued) Day 121 Group 1 Compound Control Exposure level (ppm) : 0 2 34 --POSF----- 30 100 300 Group /sex Met Hb RBC Retie L/L g/dL x10I2/L % IF Mean 0.391 14.8 7.18 2.23 SD 0.0102 0.10 0.093 0.405 n4 4 4 4 2F Mean 0.397 15.0 7.45 2.00 SD 0.0103 0.39 0.211 0.164 n4 44 5 3F Mean 0.373 14.1 6.96 2.24 SD 0.0370 1.55 0.603 0.169 n5 5 5 5 4F Mean 0.392 14.7 7.24 2.23 SD 0.0217 0.98 0.247 0.325 n5 555 Level of significance - all comparisons made with control *p<, 0.05 (Williams' test) MCH Pg 20.5 0.21 4 20.2 0.45 4 20.2 0.89 5 20.3 0.88 5 MCHC g/dL 37.5 0.36 4 37.8 0.35 4 37.8 0.49 5 37.5 0.67 5 MCV tL 54.5 0.90 4 53.3 1.13 4 53.5 2.44 5 54.2 1.56 5 WBC x109/L 8.03 1.085 4 6.04 0.972 4 5.46 1.892 5 5.27 1.748 5 N x 109/L 1.23 0.324 4 1.18 0.871 4 1.07 0.444 5 0.90 0.545 5 MIN 313/023622 TABLE 3 (Haematology - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 234 ---POSF-- 30 100 300 Group /sex L x109/L E x109/L B xio7 l M x 107L IF Mean 6.30 0.16 0.02 0.21 SD 1.301 0.085 0.008 0.019 n4 44 4 2F Mean 4.50 0.12 0.01 0.15 SD 0.896 0.050 0.005 0.074 n444 4 * ** 3F Mean 4.13 0.08 0.01 0.10 SD 1.378 0.028 0.007 0.068 n555 5 * ** 4F Mean 4.07 0.10 0.01 0.09 SD 1.101 0.056 0.004 0.042 n5555 Level of significance - all comparisons made with control * p<, 0.05 (Williams' test) LUC xlO'/L 0.12 0.034 4 0.10 0.047 4 0.07 0.048 5 0.10 0.055 5 Pit x109/L 950 110.4 4 840 133.2 4 * 683 164.1 5 * 750 157.6 5 PT sec 13.2 0.46 4 13.5 0.35 4 14.3 0.41 4 13.7 0.82 4 APIT sec 17.1 1.57 4 17.3 0.95 4 17.8 1.53 4 18.8 3.32 4 MIN 313/023622 TABLE 4 Biochemistry - group mean values Day 91 Group Compound Exposure level (ppm) i Control 0 2 34 --POSF...... 30 100 300 Group / sex ALP ALT AST LDH U/L U/L U/L Total U/L 1M Mean 202 SD 40.5 n5 2M Mean 257 SD 63.2 n5 * 3M Mean 279 SD 43.9 n5 ** 4M Mean 327 SD 67.6 n5 44 99 497 7.7 17.6 410.1 555 * 61 143 444 12.3 54.8 361.6 555 * 58 102 296 9.3 25.9 147.6 55 ** 5 67 103 244 12.5 13.1 59.7 555 Level of significance - all comparisons made with control * p < 0.05, **/?<0.01 (Williams' test) LDH1 U/L 27 24.7 5 28 7.3 5 22 18.0 5 16 6.0 5 LDH2 U/L 19 12.9 5 21 12.4 5 15 9.1 5 15 3.2 5 LDH3 U/L 12 9.4 5 13 13.9 5 7 2.3 5 7 1.3 5 LDH4 U/L LDH5 U/L 23 417 25.0 341.0 55 27 354 25.8 305.6 55 11 240 6.4 113.2 55 13 194 4.4 61.0 55 MIN 313/023622 TABLE 4 (Biochemistry - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 --POSE----- 30 100 300 Group / sex CPK SDH Bili Urea Total U/L pmol/L mmol/L U/L 1M Mean 242 SD 119.0 n5 15.3 2.67 5 1 0.5 5 6.05 1.124 5 2M Mean 1633 SD 3213.7 n5 3M Mean 202 SD 78.0 n5 4M Mean 168 SD 53.6 n5 13.1 2.40 5 * 11.5 2.67 5 * 12.0 1.36 5 1 0.5 5 1 0.5 5 1 0.0 5 6.56 0.848 5 5.80 0.254 5 6.81 0.492 5 Level of significance - all comparisons made with control * p < 0.05 (Williams' test) Creat pmo1/1. 53 3.8 5 52 2.2 5 * 48 3.6 5 * 48 2.7 5 Glue mmol/L Choi mmol/L Trig mmol/L 6.47 0.722 5 5.40 0.568 5 * 5.69 0.178 5 * 5.69 0.553 5 1.44 0.290 5 1.05 0.303 5 * 0.96 0.315 5 * 0.80 0.434 5 0.44 0.208 5 0.32 0.113 5 0.37 0.096 5 0.24 0.188 5 MIN 313/023622 TABLE 4 (Biochemistry - continued) MIN 313/023622 52 : Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 --POSF...... 30 100 300 Group /sex Na mmol/L K mmol/L Cl mmol/L Ca mmol/L Phos Total Prot mmol/L g/L 1M Mean 143 SD 1.9 n5 2M Mean 145 SD 0.8 n5 3M Mean 144 SD 1.8 n5 4M Mean 143 SD 1.6 n5 4.0 106 0.61 0.7 55 3.8 108 0.23 1.5 55 4.1 107 0.29 1.7 55 4.1 106 0.21 1.2 55 2.68 0.083 5 2.69 0.051 5 2.77 0.043 5 2.77 0.054 5 2.02 0.202 5 2.05 0.111 5 2.17 0.115 5 2.03 0.109 5 63 1.6 5 67 4.8 5 65 3.3 5 67 4.7 5 TABLE 4 (Biochemistry - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 --POSF----- 30 100 300 Group / sex Alb al a2 Beta g/L g/L g/L g/L 1M Mean 29 SD 0.8 n4 11 1.5 4 3 0.5 4 2M Mean 30 SD 1.1 n5 11 1.3 5 3 0.4 5 3M Mean 30 SD 1.8 n5 * 4M Mean 33 SD 3.6 n5 12 1.3 5 12 1.3 5 3 0.4 5 3 0.4 5 Level of significance - all comparisons made with control * p < 0.05 (Williams' test) 15 2.1 4 15 2.0 5 14 1.7 5 14 1.1 5 Gamma g/L 6 1.3 4 7 1.9 5 5 1.1 5 5 1.3 5 A/G Ratio 0.85 0.054 4 0.85 0.110 5 0.88 0.124 5 1.01 0.130 5 MIN 313/023622 TABLE 4 (Biochemistry - continued) MIN 313/023622 54 : Day 91 Group Compound Exposure level (ppm) 1 Control 0 2 34 --POSF--- 30 100 300 Group / sex ALP ALT AST LDH U/L U/L U/L Total U/L IF Mean 162 38 108 466 SD 36.3 6.0 40.8 242.8 n4 444 2F Mean 135 56 109 217 SD 26.6 26.6 28.8 46.6 n5 5 5 5 3F Mean 124 49 103 207 SD 26.3 37.6 52.0 71.3 n5 555 * 4F Mean 123 47 106 303 SD 30.4 20.1 28.3 111.9 n5 555 Level of significance - all comparisons made with control * p < 0.05 (Williams' test) LDH1 U/L 23 14.2 4 13 3.3 5 12 1.9 5 24 22.2 5 LDU2 U/L 27 15.9 4 14 5.4 5 11 3.5 5 22 17.9 5 LDI13 U/L 13 8.4 4 9 7.0 5 6 2.2 5 7 2.9 5 LDH4 U/L LDH5 U/L 25 378 17.7 195.4 44 * 10 171 2.8 53.5 55 * 9 169 3.2 62.5 55 * 13 237 6.4 72.2 55 TABLE 4 (Biochemistry - continued) MIN 313/023622 55 : Day 91 Group Compound Exposure level (ppm) i Control 0 2 34 --POSF--- 30 100 300 Group / sex CPK Total U/L SDH Bili Urea U/L pmol/L mmol/L IF Mean 564 SD 710.1 n4 * 2F Mean 130 SD 42.1 n5 * 3F Mean 132 SD 35.1 n5 * 4F Mean 143 SD 37.4 n5 13.6 3.42 4 12.4 4.03 5 10.7 2.60 5 15.5 1.54 5 2 0.0 4 3 0.5 5 2 0.0 5 2 0.4 5 7.80 1.077 4 7.15 0.535 5 6.21 0.604 5 8.32 2.625 5 Level of significance - all comparisons made with control */?<0.05, **p<0.01 (Williams' test) Creat pmol/L 66 6.6 4 * 58 2.7 5 * 55 2.7 5 * 60 8.8 5 Glue mmol/L Choi mmol/L Trig mmol/L 5.29 0.905 4 6.14 0.818 5 6.48 0.683 5 5.90 0.674 5 2.16 0.232 4 2.37 0.433 5 2.13 0.556 5 1.98 0.246 5 0.63 0.179 4 ** 0.40 0.067 5 ** 0.41 0.086 5 ** 0.39 0.082 5 TABLE 4 (Biochemistry - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 234 --POSF...... 30 100 300 Group / sex Na mmol/L K mmol/L Cl mmol/L Ca mmol/L Phos Total Prot mmol/L g/L IF Mean 142 SD 1.0 n4 2F Mean 143 SD 1.8 n5 3F Mean 143 SD 1.5 n5 4F Mean 143 SD 1.3 n5 3.5 106 0.39 0.6 44 3.2 107 0.21 0.8 55 3.0 105 0.18 1.8 55 3.4 105 0.24 0.9 55 2.81 0.064 4 2.84 0.112 5 2.75 0.079 5 2.79 0.028 5 1.60 0.292 4 1.50 0.118 5 1.58 0.094 5 1.61 0.148 5 70 1.5 4 75 3.0 5 70 4.0 5 * 76 3.6 5 MIN 313/023622 TABLE 4 (Biochemistry - continued) Day 91 Group Compound Exposure level (ppm) Group / sex 1 Control 0 Alb g/L 234 -- POSF-- 30 100 300 al a2 Beta g/L g/L g/L Gamma g/L A/G Ratio IF Mean 37 10 4 14 6 1.08 - - j SD 1.0 0.5 0.6 0.5 1.0 0.015 n4 4 4 4 4 4 2F Mean 40 11 4 14 7 1.15 SD 3.6 0.7 0.8 0.8 2.0 0.160 n55 5555 3F Mean 37 10 4 14 6 1.13 SD 2.3 0.5 0.5 0.5 1.5 0.057 n55 5555 ** ** 4F Mean 42 12 3 13 6 1.21 SD 2.0 0.5 0.5 1.8 1.6 0.099 n5 5 5 5 5 5 Level of significance - all comparisons made with control **p<0.0l (Williams' test) MIN 313/023622 TABLE 4 (Biochemistry - continued) Day 121 Group i Compound Control Exposure level (ppm) : 0 234 --POSF....... 30 100 300 Group / sex ALP ALT AST LDH U/L U/L U/L Total U/L 1M Mean 182 SD 25.4 n5 2M Mean 234 SD 34.4 n5 3M Mean 195 SD 48.3 n5 * 4M Mean 244 SD 44.0 n5 40 74 180 3.7 7.5 28.0 555 + 56 100 162 10.4 19.4 57.2 555 + 49 87 153 10.2 5.3 48.3 555 ++ 85 118 199 62.2 52.6 48.2 555 Level of significance - all comparisons made with control * p<, 0.05 (Williams' test) _p< 0.05 (Shirley's test) LDH1 U/L 11 1.5 5 14 3.8 5 15 3.0 5 17 7.3 5 LDH2 U/L 11 3.2 5 9 2.2 5 11 2.9 5 15 3.4 5 LDH3 U/L 14 8.6 5 5 1.6 5 5 2.9 5 8 1.4 5 LDH4 U/L 19 3.2 5 12 3.8 5 14 7.2 5 19 7.4 5 LDH5 U/L 125 19.8 5 122 51.7 5 108 37.5 5 141 32.8 5 MIN 313/023622 TABLE 4 (Biochemistry - continued) Day 121 Group 1 Compound Control Exposure level (ppm) : 0 234 --POSF......- 30 100 300 Group / sex CPK SDH Bili Urea Creat Total U/L (imol/L mmol/L pmol/L U/L 1M Mean 240 SD 146.1 n5 2M Mean 153 SD 45.5 n5 12.3 3.04 5 13.6 1.68 5 2 0.5 5 2 0.5 5 4.90 0.266 5 5.93 0.685 5 49 1.9 5 49 3.6 5 3M Mean 183 12.9 1 SD 116.5 4.77 0.8 n5 5 5 5.71 0.777 5 49 2.2 5 4M Mean 293 19.0 1 SD 119.0 9.61 0.4 n55 5 5.38 0.600 5 48 2.9 5 Level o f significance - all comparisons made with control * p<, 0.05 (Williams' test) _p< 0.05 (Shirley's test) Glue mmol/L Choi mmol/L Trig mmol/L 6.23 0.700 5 7.14 0.945 5 6.56 0.780 5 5.46 0.372 5 1.67 0.423 5 1.09 0.302 5 1.13 0.294 5 1.37 0.131 5 0.44 0.129 5 0.38 0.096 5 0.38 0.058 5 0.33 0.103 5 MIN 313/023622 TABLE 4 (Biochemistry - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 Group /sex Na mmoi/L 234 -- POSF-- 30 100 300 K mmoi/L Cl mmol/L Ca mmol/L Phos Total Prot mmol/L g/L 1M Mean 142 SD 0.5 n5 3.8 102 0.30 0.7 55 2M Mean 142 SD 1.1 n5 3M Mean 142 SD 0.9 n5 * 4M Mean 144 SD 0.7 n5 3.9 103 0.13 0.8 55 3.8 103 0.30 0.8 55 * 3.9 103 0.08 0.0 55 Level of significance - all comparisons made with control *p < 0.05 (Williams' test) * p < , 0.05 (Dunnett's test) 2.60 0.039 5 2.61 0.036 5 2.55 0.039 5 2.64 0.049 5 1.79 0.092 5 1.71 0.158 5 1.73 0.141 5 1.79 0.112 5 66 1.8 5 63 1.5 5 61 2.9 5 66 2.3 5 MIN 313/023622 TABLE 4 (Biochemistry - continued) Day 121 Group :1 Compound Control Exposure level (ppm) : 0 Group / sex Alb g/L 2 34 -- POSF-- 30 100 300 al a2 Beta g/L g/L g/L Gamma g/L A/G Ratio \ 1M Mean 31 12 5 14 4 0.92 SD 1.3 0.9 0.4 0.8 1.5 0.080 n55 5 5 55 * 2M Mean 29 10 5 15 5 0.84 SD 1.8 1.0 1.5 1.3 0.8 0.112 n5 5 5 5 55 ** 3M Mean 28 10 4 14 5 0.84 SD 0.7 1.1 0.4 1.1 1.6 0.059 n555 555 4M Mean 31 12 5 14 5 0.89 SD 1.8 0.9 0.4 0.8 0.8 0.070 n55 5555 Level of significance - all comparisons made with control *p< 0.05, ^ p 2 0.01 (Dunnett's test) MIN 313/023622 TABLE 4 (Biochemistry - continued) Day 121 Group 1 Compound Control Exposure level (ppm) : 0 234 --POSF----- 30 100 300 Group /sex ALP ALT AST LDH U/L U/L U/L Total U/L IF Mean 117 67 129 209 SD 26.8 60.5 113.0 163.1 n4 4 4 4 2F Mean 95 56 124 356 SD 28.9 37.7 94.1 216.3 n5 5 5 5 3F Mean 104 38 84 252 SD 13.4 12.7 19.9 147.5 n55 5 5 4F Mean 112 44 86 248 SD 22.3 11.7 17.7 121.3 n5 5 5 5 Level of significance - all comparisons made with control *p^0.05, ** p < 0.01 (Williams' test) LDH1 U/L 9 5.4 4 13 2.3 5 15 5.1 5 14 6.3 5 LDH2 U/L 8 4.2 4 10 5.8 5 9 5.1 5 11 4.8 5 LDH3 U/L 7 5.2 4 10 4.7 5 6 2.6 5 6 2.6 5 LDH4 U/L 10 7.7 4 21 17.8 5 17 13.5 5 16 6.6 5 LDH5 U/L 133 154.4 4 303 193.1 5 204 123.7 5 201 121.6 5 MIN 313/023622 TABLE 4 (Biochemistry - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 234 ---POSF-- 30 100 300 Group /sex CPK SDH Bili Urea Creat Total U/L pmol/L mniol/L pinol/L U/L IF Mean 104 19.1 3 SD 33.3 10.66 0.6 n444 * 2F Mean 170 16.9 2 SD 86.5 6.10 0.5 n555 * 3F Mean 285 13.1 2 SD 368.6 2.52 0.0 n5 55 ** 4F Mean 150 23.0 i SD 58.9 19.61 0.4 n5 5 5 6.85 0.882 4 6.23 0.770 5 6.13 0.375 5 6.87 1.226 5 59 3.2 4 57 3.3 5 57 4.9 5 59 6.0 5 Level of significance - all comparisons made with control */7^0.05, **/?<0.01 (Williams' test) Glue mmol/L Choi mmol/L Trig mmol/L 7.33 1.075 4 7.16 0.946 5 7.08 0.807 5 7.47 1.311 5 2.56 0.713 4 2.39 0.722 5 1.83 0.890 5 2.18 0.709 5 0.53 0.356 4 0.51 0.083 5 0.39 0.125 5 0.47 0.111 5 MIN 313/023622 TABLE 4 (Biochemistry - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Na mmol/L 234 -POSF----- 30 100 300 K mmol/L Cl mmol/L Ca mmol/L Phos Total Prot mmol/L g/L IF Mean 140 SD 1.3 n4 * 2F Mean 142 SD 0.9 n5 * 3F Mean 142 SD 1.1 n5 ** 4F Mean 143 SD 1.5 n5 3.3 101 0.14 1.0 44 3.4 102 0.32 1.3 55 * 3.3 103 0.22 0.9 55 ** 3.6 104 0.19 1.9 55 Level of significance - all comparisons made with control * p<, 005, ** p<, 0.01 (Williams' test) 2.78 0.093 4 2.74 0.086 5 2.64 0.089 5 2.71 0.080 5 1.58 0.217 4 1.32 0.157 5 1.42 0.227 5 1.63 0.369 5 74 1.3 4 71 4.0 5 70 4.9 5 71 2.5 5 MIN 313/023622 TABLE 4 (Biochemistry - continued) Day 121 Group :1 Compound Control Exposure level (ppm) 0 23 ..... POSF-- 30 100 4 300 Group / sex Alb al a2 Beta Gamma A/G g/L g/L g/L g/L g/L Ratio <OJl\ IF Mean 41 9 4 14 5 1.29 SD 3.1 0.8 0.0 0.5 0.8 0.171 n4 4 4 44 4 2F Mean 39 9 4 14 5 1.20 SD 2.5 1.2 0.5 0.8 1.3 0.102 n5 5 555 5 3F Mean 38 10 3 14 6 1.16 SD 2.3 1.1 0.5 0.5 2.2 0.096 n5 5 5555 4F Mean 39 9 4 14 5 1.21 SD 0.9 1.5 0.5 1.0 1.4 0.060 n5 5 5 55 5 No differences of statistical significance (p> 0.05) MIN 313/023622 TABLE 5 Urinalysis - group mean values Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Vol ml 2 34 -- POSF-- 30 100 300 pH SG Prot g/L g/L U-Na mmol/L U-K mmol/L U-Cl mmol/L 1M Mean SD n 7.2 7.3 1035 3.55 0.43 8.5 555 2M Mean SD n 5.3 7.3 1044 2.01 0.27 11.8 555 3M Mean SD n 4M Mean SD n 7.2 6.9 1036 1.49 0.33 5.8 555 ** ** 4.0 6.6 1058 1.59 0.15 13.8 555 Level of significance - all comparisons made with control * p<, 005, ** p < 0.01 (Williams' test) 0.94 0.362 5 1.00 0.315 5 0.85 0.096 5 1.15 0.366 5 75.2 5.71 5 85.7 46.88 5 68.3 25.17 5 101.9 62.90 5 200.9 49.15 5 247.7 44.38 5 195.3 40.74 5 * 268.7 53.63 5 88.2 16.03 5 87.1 30.99 5 77.2 19.32 5 94.0 27.71 5 MIN 313/023622 TABLE 5 (Urinalysis - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Vol ml 2 34 ---POSF-- 30 100 300 pH SG Prot g/L g/L U-Na mmol/L U-K mtnol/L U-Cl mmol/L Os IF Mean 4.3 6.6 1036 0.19 86.7 159.2 83.4 SD 2.56 0.18 8.4 0.061 28.95 40.42 28.60 n5 5 555 5 5 2F Mean SD n 3F Mean SD n 4F Mean SD n 4.2 6.7 1036 1.26 0.30 6.1 555 2.8 6.4 1054 0.91 0.16 9.8 555 ** 2.3 6.6 1068 0.61 0.38 22.3 555 0.21 0.053 5 0.25 0.041 5 ** 0.31 0.083 5 81.2 23.18 5 87.4 18.34 5 92.9 35.86 5 157.0 26.45 5 * 242.3 30.20 5 ** 310.0 96.56 5 79.7 12.41 5 93.3 21.99 5 112.2 31.87 5 Level of significance - all comparisons made with control * p < 005, ** p < 0.01 (Williams' test) MIN 313/023622 TABLE 5 (Urinalysis - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) 0 Group /sex Voi ml 2 34 ---POSE-- 30 100 300 pH SG Prot g/L g/L U-Na mmol/L U-K mmol/L U-Cl mmol/L 1M Mean SD n 7.1 7.1 1030 4.35 0.36 4.7 555 2M Mean SD n 3M Mean SD n 4M Mean SD n 6.2 7.0 1042 1.55 0.07 9.6 555 ** 5.8 6.7 1048 3.14 0.18 15.1 555 ** * 5.5 6.6 1048 2.03 0.12 12.7 555 Level of significance - all comparisons made with control *p < 005, ** p < 0.01 (Williams' test) 0.87 0.214 5 1.30 0.498 5 1.15 0.286 5 1.02 0.161 5 50.2 6.39 5 ** 92.2 19.26 5 ** 97.4 28.33 5 ** 69.0 12.55 5 155.9 38.18 5 209.4 61.83 5 218.5 63.78 5 231.4 61.31 5 52.0 11.04 5 79.2 22.80 5 71.3 21.21 5 67.4 11.61 5 MIN 313/023622 TABLE 5 (Urinalysis - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) 0 Group / sex Voi ml 2 34 --POSF-...... 30 100 300 PH SG Plot g/L g/L Na Total nnnol K Total mmol Cl Total mmol IF Mean SD n 4.0 6.1 1041 2.38 0.15 6.0 444 2F Mean SD n 4.7 6.2 1039 0.79 0.20 1.8 555 3F Mean 2.7 6.1 1049 SD 1.10 0.13 10.8 n555 4F Mean 3.4 6.2 1046 SD 1.03 0.11 9.6 n5 55 No differences of statistical significance (p> 0.05) 0.27 0.059 4 0.21 0.043 5 0.24 0.078 5 0.27 0.067 5 0.23 0.128 4 0.32 0.095 5 0.14 0.132 5 0.24 0.061 5 0.66 0.312 4 0.85 0.181 5 0.57 0.084 5 0.69 0.070 5 0.19 0.111 4 0.32 0.154 5 0.16 0.117 5 0.19 0.042 5 MIN 313/023622 TABLE 5 (Urinalysis - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 Group /sex U-Na mmol/L 2 34 -- POSF-- 30 100 300 U-K mmol/L U-Cl mmol/L IF Mean 58.8 177.6 49.3 SD 9.28 34.12 7.70 n4 44 2F Mean 67.9 181.2 66.8 SD 18.51 33.92 28.80 n5 55 3F Mean 47.3 239.0 54.1 SD 26.34 87.36 22.48 n5 55 4F Mean 72,1 218.5 57.3 SD 16.33 57.82 7.77 n5 55 MIN 313/023622 Main study animals GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 6 O rgan weights - group mean values (g) 2 34 -- POSF--- 30 100 300 Print No: 0021 Printed: 24-JUN-02 Xybion protocol number: MIN 313 ABSOLUTE VALUES SEX: GROUP : -- 2--- NUMBER: 5 5 5 5 N MEAN sd TERMINAL BODY WEIGHT (g) :5 : 452.6 30.2 5 449.7 43.0 5 440.5 25.3 5 371.4 26.9 BRAIN N MEAN sd : : : 5 2.02 0.08 5 2.03 0.09 5 1.98 0.09 5 2.00 0.07 ADRENALS N MEAN sd : : : 5 0.061 0.016 5 0.054 0.008 5 0.056 0.007 5 0.058 0.009 HEART N MEAN sd : : 5 1.460 0.213 5 1.377 0.184 5 1.380 0.161 5 1.327 0.217 KIDNEYS N MEAN sd : : : 5 2.59 0.23 5 2.61 0.26 5 2.78 0.34 5 2.45 0.25 Level of significance - all comparisons made with control * p<, 0.05, ** pS 0.01 (Williams' test) BODYWEIGHT ADJUSTED VALUES -- 2--- -- 4--- 5555 5 1.97 5 0.056 5 1.348 5 2.42 BRAIN 5 1.99 5 1.96 ADRENALS 5 0.050 5 0.054 HEART 5 1.279 5 1.325 KIDNEYS 5 2.45 5 2.70 5 2.10 5 0.069 5 1.592 * 5 2.86 MIN 313/023622 Main study animals GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 TABLE 6 (Organ weights - continued) 23 -- POSF-- 30 100 4 300 Print Mo: 0021 Printed: 24-JUN-02 Xybion protocol number: MIN 313 to ABSOLUTE VALUES SEX: GROUP : -- 2--- NUMBER: 5 5 5 5 LIVER N :5 MEAN : 14.82 sd : 2.02 5 14.57 1.94 5 16.27 1.61 5 18.41 1.78 LUNGS & BRONCHI N MEAN sd : : 5 1.520 0.115 5 1.468 0.149 5 1.592 0.264 5 1.979 0.152 Level of significance - all comparisons made with control * p<> 0.05, ** p<, 0.01 (Williams' test) BODWEIGHT ADJUSTED VALUES -- 2--555 5 5 13.70 LIVER * 55 13.59 15.71 ** 5 21.07 5 1.429 LUNGS & BRONCHI 5 1.388 5 1.547 ** 5 2.194 MIN 313/023622 Main study animals GROUP : COMPOUND : EXPOSURE LEVEL <PPM) : 1 CONTROL 0 TABLE 6 (Organ weights - continued) 234 -- POSF--- 30 100 300 Print No: 0022 Printed: 24-JUN-02 Xybion protocol number: MIN 313 ABSOLUTE VALUES sex: GROUP : -- 2-- -- 3-- - -- 4-- NUMBER: 4 5 5 5 N MEAN sd TERMINAL BODY WEIGHT (g) :4 : 265.3 : 7.7 5 242.8 14.5 5 256.7 31.6 5 241.0 15.3 BRAIN N MEAN sd : : : 4 1.92 0.10 5 1.86 0.03 5 1.88 0.07 5 1.92 0.09 ADRENALS N MEAN sd : : : 4 0.059 0.010 5 0.066 0.012 5 0.067 0.007 5 0.064 0.009 HEART N MEAN sd : : : 4 0.985 0.052 5 0.971 0.073 5 0.943 0.067 5 0.920 0.167 KIDNEYS N MEAN sd : : 4 1.62 0.18 5 1.69 0.11 5 1.76 0.12 5 1.64 0.23 No differences of statistical significance (p> 0.05) BODYWEIGHT ADJUSTED VALUES -- 2-- -- 3-- -- 4--- 4555 4 1.89 BRAIN 5 1.87 5 1.87 5 1.94 MIN 313/023622 TABLE 6 (Organ weights - continued) Main study animals GROUP : COMPOUND : EXPOSURE LEVEL(PPM) : 1 CONTROL 0 2 34 -- POSF-------- 30 100 300 -44^ ABSOLUTE VALUES SEX: GROUP: -- 2--- -- 4--- NUMBER: 4 5 5 5 LIVER N MEAN ad : : : 4 8.88 0.75 5 8.54 0.79 5 9.86 1.31 5 12.24 1.43 LUNGS & BRONCHI N MEAN sd : : 4 1.170 0.049 5 1.098 0.050 5 1.214 0.150 5 1.597 0.160 Level of significance - all comparisons made with control * pS 0.05, ** pS 0.01 (Williams' test) Printed: 24-JUN-02 Xybion protocol number: MIN 313 BODYWEIGHT ADJUSTED VALUES -- 3-4 55 5 4 8.29 LIVER * 5 8.86 5 9.62 ** 5 12.64 4 1.116 BUNGS S BRONCHI 5 1.127 5 1.193 ** 5 1.632 MIN 313/023622 Recovery phase animals GROUP : COMPOUND : EXPOSURE LEVEL(PPM): i CONTROL 0 TABLE 6 (Organ weights - continued) 234 -- POSF--- 30 100 300 Print No: 0014 Printed: 25-JUN-02 Xybion protocol number: MIN 313 SEX: GROUP : NUMBER: 5 ABSOLUTE VALUES -- 2--- -- 3--55 5 N MEAN sd TERMINAL BODY WEIGHT (9) :5 : 482.5 : 23.2 5 457.7 39.2 5 469.6 43.5 5 442.9 33.7 BRATN N MEAN sd : : : 5 2.11 0.09 5 2.13 0.09 5 2.08 0.08 5 2.05 0.12 ADRENALS N MEAN sd : : 5 0.057 0.010 5 0.052 0.008 5 0.052 0.004 5 0.050 0.007 HEART N MEAN sd : : : 5 1.471 0.150 5 1.449 0.156 5 1.513 0.143 5 1.546 0.226 KIDNEYS N MEAN sd : : : 5 2.80 0.06 5 2.97 0.25 5 2.89 0.41 5 2.86 0.26 Level of significance - all comparisons made with control * p<, 0.05 (Williams' test) BODYWEIGHT ADJUSTED VALUES -- 1--- -- 2 -55 -- 3--5 5 5 2.68 KIDNEYS 5 3.00 5 2.85 * 5 2.98 MIN 313/023622 Recovery phase animals GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 TABLE 6 (Organ weights - continued) 234 -- POSF-------- 30 100 300 Print No: 0014 Printed: 25-JUN-02 Xybion protocol number: MIN 313 SEX: GROUP : NUMBER: 5 ABSOLUTE VALUES 55 5 LIVER N MEAN sd : : : 5 12.14 0.67 5 12.48 1.43 5 13.94 1.72 5 15.53 1.18 LUNGS & BRONCHI N MEAN sd : : : 5 1.472 0.088 5 1.467 0.057 5 1.445 0.141 5 1.616 0.144 Level of significance - all comparisons made with control ** pS, 0.01 (Williams' test) BODYWEIGHT ADJUSTED VALUES -- 2 -- 55 5 5 5 11.62 LIVER ** 5 12.62 5 13.76 ** 5 16.07 5 1.426 LUNGS & BRONCHI 5 1.480 5 1.429 ** 5 1.665 MIN 313/023622 Recovery phase animals GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : i CONTROL 0 TABLE 6 (Organ weights - continued) 2 34 -- POSF--- 30 100 300 Print No: 0015 Printed: 25-JUN-02 Xybion protocol number: MIN 313 SEX: GROUP : NUMBER: 4 ABSOLUTE VALUES -- 3--55 -- 4-5 N MEAN sd TERMINAI, BODY WEIGHT (9) :4 : 291.4 : 18.1 5 291.5 22.0 5 258.0 11.2 5 260.9 23.9 BRAIN N MEAN sd : : : 4 1.97 0.05 5 2.05 0.09 5 1.92 0.07 5 1.93 0.06 ADRENALS N MEAN sd : : : 4 0.073 0.006 5 0.071 0.011 5 0.068 0.00 9 5 0.063 0.007 HEART N MEAN sd : : : 4 1.142 0.049 5 1.030 0.151 5 0.987 0.097 5 0.925 0.051 KIDNEYS N MEAN sd : : : 4 1.88 0.09 5 1.90 0.21 5 1.77 0.11 5 1.67 0.18 No differences of statistical significance (p> 0.05) BODYWEIGHT ADJUSTED VALUES -- 3-- 4555 4 1.94 4 0.069 4 1.105 4 1.79 BRAIN 5 2.02 5 1.95 ADRENALS 5 0.066 5 0.073 HEART 5 0.993 5 1.024 KIDNEYS 5 1.81 5 1.85 5 1.96 5 0.067 5 0.955 5 1.74 MIN 313/023622 Recovery phase animals GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 TABLE 6 (Organ weights - continued) 234 -- POSF-------- 30 100 300 Print No: 0015 Printed: 25-JUN-02 Xybion protocol number: MIN 313 SEX: GROUP : NUMBER: 4 ABSOLUTE VALUES 55 -- 4-5 LIVER N MEAN sd : : 4 8.99 0.94 5 8.80 0.67 5 8.44 0.46 5 9.78 1.11 LUNGS & BRONCHI N MEAN sd : : ! 4 1.290 0.067 4 1.200 0.072 5 1.089 0.072 5 1.187 0.074 Level of significance - all comparisons made with control ** 0.01 (Williams' test) BODYWEIGHT ADJUSTED VALUES 4 4 8.45 55 LIVER 5 8.23 5 9.00 5 ** 5 10.25 4 1.245 LUNGS & BRONCHI 5 1.151 5 1.131 5 1.221 MIN 313/023622 Satellite study animals GROUP : COMPOUND : EXPOSURE LEVEL(PPM): i CONTROL 0 TABLE 6 (Organ weights - continued) 234 -- POSP--- 30 100 300 SEX: GROUP: NUMBER : ABSOLUTE VALUES -------MALE------ -- 1 ---4 55 BODYWEIGHT ADJUSTED VALUES --------m a l e ------ 55 TERMINAL BODY WEIGHT (g) N MEAN sd :5 : 332.8 : 30.3 5 317.9 22.1 ADRENALS ADRENALS N MEAN sd : : : 5 0.053 0.015 5 0.056 0.009 5 0.050 5 0.059 BRAIN N MEAN sd : : : 5 1.94 0.12 5 1.97 0.06 HEART HEART N MEAN sd 5 1.284 0.223 5 1.304 0.154 5 1.235 5 1.353 KIDNEYS KIDNEYS N MEAN sd : : ; 5 2.32 0.23 5 2.33 0.19 5 2.27 5 2.38 No differences of statistical significance (p> 0.05) Printed: 24-JUN-02 Xybion protocol number: MIN 313A MIN 313/023622 Satellite study animals GROUP : COMPOUND : EXPOSURE LEVEL(PPM): i CONTROL 0 2 30 TABLE 6 (Organ weights - continued) 34 100 300 SEX: GROUP: NUMBER: ABSOLUTE VALUES ------ MALE-------- 1--- -- 4 55 BODYWEIGHT ADJUSTED VALUES --------MALE------ ---1 ---4 55 N MEAN sd LIVER 5 9.62 1.12 5 12.71 0.97 LIVER +++ 55 9.36 12.98 N MEAN sd LUNGS & BRONCHI 5 1.251 0.178 5 1.705 0.104 LUNGS & BRONCHI +++ 55 1.225 1.731 Level of significance - all comparisons made with control +++ p< 0.001 (Student's 't' test) Print No: 0005 Printed: 24-JUN-02 Xybion protocol number: MIN 313A Satellite study animals GROUP COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 6 (Organ weights - continued) 2 34 -- POSF--- 30 100 300 SEX : GROUP: NUMBER : ABSOLUTE VALUES ---- FEMALE------- 1--- -- 4--- 55 BODYWEIGHT ADJUSTED VALUES -------f e m a l e ------- 55 TERMINAL BODY WEIGHT (g) N MEAN sd :5 : 212.8 : 16.1 5 194.6 7.2 N MEAN sd : : : ADRENALS 5 0.066 0.012 5 0.069 0.010 ADRENALS + 55 0.060 0.076 BRAIN N MEAN sd 5 1.84 0.08 5 1.79 0.06 HEART HEART N MEAN sd 0.909 0.100 0.827 0.027 0.869 0.867 KIDNEYS N MEAN sd 1.54 0.11 1.48 0.08 Level of significance - all comparisons made with control + 0.05 (Student's 't' test) Print No: 0006 Printed: 24-JUN-02 Xybion protocol number : MIN 313A MIN 313/023622 Satellite study animals GROUP COMPOUND : EXPOSURE LEVEL(PPM): i CONTROL 0 TABLE 6 (Organ weights - continued) 234 -- POSF--- 30 100 300 oKo> ABSOLUTE VALUES SEX: ---- FEMALE------ GROUP: -- 1--- -- 4--- NUMBER : 5 5 BODYWEIGHT ADJUSTED VALUES ------ FEMALE------- 55 N MEAN sd : 5 7.20 0.62 LIVER + 5 7.93 0.27 N MEAN sd LUNGS & BRONCHI ++ 55 1.040 1.334 0.086 0.105 Level of significance - all comparisons made with control + p<, 0.05, ++ p<, 0.01 (Student's 't' test) Printed: 24-JUN-02 Xybion protocol number: MIN 313A MIN 313/023622 TABLE 7 Macroscopic pathology - group distribution Main study animals group : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 234 -- POSF--- 30 100 300 Print No: 0025 Printed: 25-JUN-02 Xybion protocol number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D --- ORGAN AND KEYWORD (S) OR PHRASE ** TOP OF LIST ** KIDNEYS ......................................... PELVIC DILATION LIVER ........................................... MEDIAN CLEFT PALE AREA(S) ENLARGED LUNGS & BRONCHI ................................. CONGESTED INCOMPLETE COLLAPSE DARK AREA(S) LN MANDIBULAR ................................... ENLARGED CONGESTED SEX: -- MALE-- -- FEMALE-- GROUP: -i- -2- -3- -4- -1- -2- -3- -4- NUMBER: 5 5 5 5 4 5 5 5 55554555 00000101 55554555 11110001 00030002 55554555 i ii0 0 00 0 000 00 00 3 00000001 555 5 4555 1 100 0 1 1 1 2 112 0 0 0 1 MISCELLANEOUS ................................... ANIMAL THIN FUR STAINING FUR BADLY GROOMED PREPUTIAL GLANDS ................................ ABSCESS (ES) PROSTATE ........................................ ABSCESS (ES) SKIN ............................................ HAIRLOSS SCAB (S) 5555 45 55 011 30 013 00001000 00 00 0 00 1 55 55 45 55 01000000 55 5 5 0 0 0 0 01000000 55554555 01 10 i 11 3 01000000 MIN 313/023622 TABLE 7 (Macroscopic pathology - continued) M ain study anim als GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 23 4 -- POSF--- 30 100 300 Print No: 0025 Printed: 25-JUN-02 Xybion protocol number: MIN 313 o4o^ ORGAN AND KEYWORD(S) OR PHRASE -- N U M B E R - O F - A N I M A L S - A F F E C T E D --- SEX: -- MALE -- FEMALE-- GROUP: -l- -2- -3- -4- -1- -2- -3- -4- NUMBER: 5 5 5 5 4 5 5 5 STOMACH ................... ANTRUM WHITE NODULE(S) CORPUS WHITE NODULE(S) FORESTOMACH DEPRESSION(S) T A I L ...................... SWELLING (S) THYMUS .................... DARK UTERUS .................... FLUID DISTENSION ** END OF LIST ** NUMBER EXAMINED: 5 5 5 5 4 5 5 5 10010110 00010000 000000 01 NUMBER EXAMINED: 5 5 5 5 4 5 5 5 000000 11 NUMBER EXAMINED: 5 5 5 5 4 5 5 5 00 0 00 10 0 NUMBER EXAMINED: 0 0 0 0 4 5 5 5 000003 12 MIN 313/023622 Recovery phase animals GROUP COMPOUND : EXPOSURE LEVEL (PPM) : i CONTROL 0 ORGAN AND KEYWORD (S) OR PHRASE ** TOP OF LIST ** KIDNEYS ...................... PELVIC DILATION LIVER ........................ MEDIAN CLEFT PALE AREA (S) LOBULAR PATTERN ACCENTUATED EPIDIDYMIDES ................. SWELLING (S) LN MANDIBULAR ................ ENLARGED CONGESTED MISCELLANEOUS ................ ANIMAL THIN SKIN ......................... HAIRLOSS SCAB (S) STOMACH ...................... ANTRUM WHITE NODULE (S) URETERS ...................... DISTENDED UTERUS ........................ FLUID DISTENSION ** END OF L IS T * * TABLE 7 (Macroscopic pathology - continued) 2 34 -- POSE--- 30 100 300 Printed: 25-JUN-02 Xybion protocol number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D --- SEX: -- -- MALE-- -- -- -- FEMALE-- GROUP: -i- -2- -3- -4- -1- -2- -3- -4- NUMBER: 5 5 5 5 4 5 5 5 5555 4555 00000100 555 545 55 00010 000 0 00 10 0 0 0 55 550000 0 010 0 0 00 5555 4555 00 10 1 100 00 0 0 0 0 0 2 55 5 5 4 5 5 5 00 0 10 00 0 5 5554555 22043210 01000000 5 5554555 02 100020 55 55 45 5 5 00 000 100 00 00 4 S 5 5 00000221 MIN 313/023622 TABLE 7 (Macroscopic pathology - continued) Satellite s t u d y animals GROUP : COMPOUND : EXPOSURE LEVEL(PPM): i CONTROL 0 234 -- POSF--- 30 100 300 Printed: 24-JUN-02 Xybion protocol number: MIN 313A -- N U M B E R - O F - A N I M A L S - A F F E C T E D --- ON ORGAN AND KEYWORD (S) OR PHRASE SEX: -- MALE-- -FEMALEGROUP: -1- -4- - 1 -- - 4 - NUMBER: 5 5 5 5 ** TOP OF LIST ** KIDNEYS ......... PELVIC DILATION NUMBER EXAMINED: 5 5 02 5 0 5 0 L I V E R ..................... MEDIAN CLEFT PALE AREA(S) ENLARGED NUMBER EXAMINED : 5 0 0 55 12 40 5 3 0 LN MANDIBULAR ENLARGED NUMBER EXAMINED: 5 5 00 55 30 LUNGS & BRONCHI PALE AREA(S) CONGESTED NUMBER EXAMINED : 5 5 5 5 0 0 10 00 0 1 T A I L ....... PUSTULE (S) UTERUS ........... FLUID DISTENTION ** END OF LIST ** NUMBER EXAMINED : 5 0 NUMBER EXAMINED : 0 0 55 02 05 00 5 0 5 1 MIN 313/023622 TABLE 8 Microscopie pathology - group distribution Main study animals GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 234 -- POSF--- 30 100 300 Print No: 0029 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D --- SEX: -- MALE-- -- -- -- FEMALE-- GROUP: -1- -2- -3- -4- -1- -2- -3- -4- ORGAN/TISSUE EXAMINED NUMBER: 5 5 s 5 4 5 5 5 ** TOP OF LIST ** HEART ................................................ . NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 1 0 0 5 4 0 0 5 --MYOCARDITIS, VENTRICULAR REGION Minimal> 4 0 0 0 0 0 0 0 Total> 4 0 0 0 0 0 0 0 KIDNEYS .............................................. . NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 2 0 0 3 1 0 0 4 -- CORTICAL TUBULAR BASOPHILIA Minimal> 2 0 0 1 1 0 0 1 Slight> 0 0 0 0 1 0 0 0 Total> 2 0 0 1 2 0 0 1 -- CORTICAL LYMPHOCYTIC INFILTRATION Minimal> 0 0 0 0 1 0 0 0 Total> 0 0 0 0 1 0 0 0 -- CORTICAL INTERSTITIAL FIBROBLAST PROLIFERATION/ FIBROSIS Slight> 0 0 0 0 1 0 0 0 Total> 0 0 0 0 1 0 0 0 -- UROTHELIAL HYPERPLASIA, UNILATERAL Minimal> 0 0 0 0 0 0 0 1 Total> 0 0 0 0 0 0 0 1 -- PELVIC DILATATION Moderate> 0 0 0 0 0 0 0 1 Total> 0 0 0 0 0 0 0 1 -- MINERALISATION, CORTICOMEDULLARY Minimal> 1 0 0 0 0 0 0 0 Total> 1 0 0 0 0 0 0 0 --MEDULLARY MINERALISATION Total> 0 0 0 0 0 0 0 0 MIN 313/023622 Main study animals GROUP COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 8 (Microscopic pathology 2 34 -- POSF--- 30 100 300 SEX: GROUP: ORGAN/TISSUE EXAMINED NUMBER: ** FROM PREVIOUS PAGE ** KIDNEYS ............................................... NUMBER EXAMINED: NOT REMARKABLE: 00 00 -- FOCAL CAPSULAR CONGESTION Minimal> Total> -- MEDULLARY TUBULAR BASOPHILIA Total> -- MEDULLARY TUBULAR FIBROSIS Total> L A R Y N X ................................................ NUMBER EXAMINED: NOT REMARKABLE: -- NECROSIS OF VENTRAL CARTILAGE Presents Total> -- ARYTENOID - EPITHELIAL HYPERPLASIA Minimal> Total> -- ARYTENOID - EPITHELIAL HYPERKERATOSIS PARAKERATOSIS Total> -- ARYTENOID - SUBEPITHELIAL INFLAMMATORY CELL INFILTRATION Minimal> Total> -- VENTROLATERAL EPITHELIAL HYPERPLASIA Total> -- VENTROLATERAL EPITHELIAL SQUAMOUS METAPLASIA Total> -- VENTRAL POUCH - EPITHELIAL HYPERPLASIA Minimal> Total> continued) Print No: 0029 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D -- ------ MALE------ ----- FEMALE-----1- -4- -1- -2- -3- -4554555 054005 031004 0 10000 010000 000000 000000 554555 4 13 4 40 0 40005 040005 021002 021002 000000 MIN 313/023622 in in 110000 1 10 0 0 0 000000 000000 000110 000110 Main study animals GROUP : COMPOUND : EXPOSURE LEVEL (PPM): 1 CONTROL 0 TABLE 8 (Microscopie pathology 2 34 --------POSF---30 100 300 SEX: GROUP: 0RGAN/TISSUE EXAMINED NUMBER: ** FROM PREVIOUS PAGE ** LARYNX ............................................... ,. NUMBER EXAMINED: OO NOT REMARKABLE: VO -- VENTRAL POUCH - INFLAMMATORY CELL INFILTRATION Total> -- VENTRAL POUCH - FORIGN BODY IN LUMEN, RELATED TO EPITHELIAL HYPERPLASIA/ INFLAMMATORY CELL INFILTRATION Total> --PROMINENT DILATED ACINI VENTRAL GLAND Total> LIVER ................................................ . NUMBER EXAMINED: NOT REMARKABLE: --CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA Slight Moderate Total> -- CENTRILOBULAR HEPATOCYTE HYPERTROPHY Slight Total> -- CENTRILOBULAR HEPATOCYTE NECROSIS/INFLAMMATORY CELL INFILTRATION/SINUSOIDAL CONGESTION Total> -- PROMINENT CENTRILOBULAR HEPATOCYTE PIGMENT Minimal Total -- FOCAL/MULTIFOCAL SINGLE CELL NECROSIS INFLAMMATORY CELL INFILTRATION Minimal Total continued) Print No: 0029 Printed: 18-3EP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D -- ----- HALE------ --- -- FEMALE-----1- -2- -3- -4- -1- -2- -3- -4- 555 5 4 5 55 in -t o o 45 55 3440 0 0 00 0 0 00 o in o 0 0 00 4555 24 40 0 4 0 00 0 0 5 50 0 4 5 50 0 10 0 10 00 00 0 0 0 00 0 0 0 0 0 0 03 02 05 00 00 00 0 00 0 0 0 0 00 0 0 0 0 00 1 0 0 000100 01 01 00 00 MIN 313/023622 Main study animals GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 8 (Microscopie pathology 23 ------- POSF--30 100 4 300 SEX: GROUP : ORGAN/TISSUE EXAMINED NUMBER: ** FROM PREVIOUS PAGE ** L I V E R .............................................. . VO NOT REMARKABLE : O -- PARENCHYMAL INFLAMMATORY CELL INFILTRATION Minimal Total -- PERIPORTAL LYMPHOCYTIC INFILTRATION Minimal Slight Total --CENTRILOBULAR INFLAMMATORY CELL INFILTRATION Total -- EXTRAMEDULLARY HAEMOPOE ISIS Minimal Total -- PERIPORTAL HEPATOCYTE VACUOLATION Minimal Total -- CENTRILOBULAR HEPATOCYTE VACUOLATION Minimal Slight Total -- HEPATOCYTE VACUOLATION, MEDIAN CLEFT Minimal Total LUNGS & BRONCHI ..................................... NOT REMARKABLE: -- FOCAL BRONCHOALVEOLAR HYPERPLASIA Total continued) Print No: 0029 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D -- ------ MALE------- --- -- FEMALE-----1- -2- -3- -4- -1- -2- -3- -4- 5555 4555 55 40 5 54 0 02 0 10 0 10 00 00 10 0 00 00 0 0 1 10 0 01 00 0 00 0 110 0 110 1 1 00 00 0 00 0 00 00 00 00 30 10 40 0 0 0 00 00 0 00 0 00 5 5 54 3 3 03 00 0 00 55 44 00 00 10 00 10 5 0 0 0 0 0 0 00 0 00 00 0 0 0 10 0 10 00 00 00 00 00 0 0 0 1 1 55 33 00 5 0 0 MIN 313/023622 in o Main study animals GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 8 (Microscopie pathology 2 34 --------POSF---30 100 300 SEX: GROUP: ORGAN/TISSUE EXAMINED NUMBER: ** FROM PREVIOUS PAGE ** LUNGS & BRONCHI ..................................... .. NUMBER EXAMINED: NOT REMARKABLE: -- FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES Minimal Slight> Moderate> Total> -- AGGREGATIONS OF ALVEOLAR MACROPHAGES Minimal Slight Total -- ALVEOLITIS Minimal Total -- ALVEOLAR HAEMORRHAGE Minimal Marked Total -- PERIVASCULAR INFLAMMATORY CELL INFILTRATION Minimal> Total> --PROMINENT AGGREGATIONS OF LYMPHOCYTES Mnimal> Total> -- VASCULAR CONGESTION Slight Total -- FOCAL OSSEOUS DEPOSITION Minimal> Total> continued) Print No: 0029 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D -- ------ MALE------- --- -- FEMALE-----1- -2- -3- -4- -1- -2- -3- -4- 555 5 4 555 MIN 313/023622 inO O O O O N O N <NCM N O N 55 5 4 33 0 3 00 00 00 00 30 10 10 50 00 00 00 10 10 00 01 01 10 10 10 2 0 10 112 1 0 1 0 00 0 0 0 00 0 0 0 0 10 0 01 0 0 0 0 0 10 0 0 01 0 0 0 00 0 0 0 00 0 0 55 33 5 0 0 12 00 2 00 0 0 14 110 00 0 110 00 00 0 0 00 00 00 2 0 2 10 10 1 1 00 00 0 0 00 00 0 0 10 10 3 3 Main study animals GROUP COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 8 (Microscopic pathology 234 -- POSF--- 30 100 300 SEX: GROUP: ORGAN/TISSUE EXAMINED NUMBER: ** FROM PREVIOUS PAGE ** LUNGS & BRONCHI ....................................... NUMBER EXAMINED: NOT REMARKABLE: VO N> -- EARLY AUTOLYSIS Total> T R A C H E A ............................................... NUMBER EXAMINED: NOT REMARKABLE: -- EPITHELIAL HYPERPLASIA Minimal> Total> -- EPITHELIAL INFLAMMATORY CELL INFILTRATION Minimal Total> -- SUBEPITHELIAL INFLAMMATORY AGGREGATIONS Minimal Total> -- SUBEPITHELIAL FIBROBLAST PROLIFERATION Minimal Total TRACHEAL BIFURC......................................... NUMBER EXAMINED: NOT REMARKABLE: URINARY BLADDER ....................................... NUMBER EXAMINED: NOT REMARKABLE: EPIDIDYMIDES .......................................... NUMBER EXAMINED: NOT REMARKABLE: LN MANDIBULAR ......................................... NUMBER EXAMINED: NOT REMARKABLE: continued) Print No: 0029 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D -- ------ MALE-- ---- - "" "-FEMALE"*"""" 1- -2- -3- -4- -1- -2- -3- -4- 555 5 4 55 5 5 55 5 4 555 0 33 0 3 3 30 0 00 0 0 0 00 5 00 5 4 0 05 5 0 0 3 4 0 05 0 00 2 0 0 0 0 0 00 2 0 0 00 0 00 10 0 00 0 00 1 0 000 0 0 0 1 0 000 0 00 1 0 000 0 00 1 0 0 00 0 00 1 0 0 00 5 00 5 4 0 05 5 00 5 4 0 05 5 00 5 4 0 05 5 00 5 4 0 05 0 0 0 0 0 0 00 0 0 0 0 0 0 00 0 00 0 0 0 00 0 00 0 0 0 00 MIN 313/023622 Main study animals GROUP COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 8 (Microscopic pathology 234 -- POSF--- 30 100 300 ORGAN/TISSUE EXAMINED SEX: GROUP: NUMBER: MISCELLANEOUS VUO) PREPUTIAL GLANDS NUMBER EXAMINED: NOT REMARKABLE: NUMBER EXAMINED: NOT REMARKABLE: PROSTATE .............................. NUMBER EXAMINED: NOT REMARKABLE: S K I N ................................................... NUMBER EXAMINED: NOT REMARKABLE: STOMACH ................................................ NUMBER EXAMINED: NOT REMARKABLE: T A I L ......................................... NUMBER EXAMINED: NOT REMARKABLE: THYMUS ................................................. NUMBER EXAMINED: NOT REMARKABLE: URETERS ............................................... NUMBER EXAMINED: NOT REMARKABLE: UTERUS ................................................ NUMBER EXAMINED: NOT REMARKABLE: --LUMINAL DILATATION ** END OF LIST ** Total> continued) Print No: 0029 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D -- ------- male--------- ---- -- FEMALE----- 1- 2- - 3- - 4- - 1- - -2- -3- -4- 5555455 5 0000000 0 0000000 0 0 00000 00 0 00 0 0 0 0 0 0 00 0 0 0 0 0 0 00 00 0 0 0 0 000 00 0 0 0000 00 0 0 0000 0 0 0 0 0000 00 0 0 000000 0 0 000000 0 0 000000 00 000000 00 00 000 0 0 0 000000 00 000000 0 0 00 00 0 0 0 0 00000000 MIN 313/023622 Recovery phase animals GROUP : COMPOUND EXPOSURE LEVEL(PPM): 1 CONTROL 0 TABLE 8 (Microscopie pathology 2 34 -- POSF--- -- 30 100 300 SEX: GROUP: ORGAN/TISSUE EXAMINED NUMBER: ** TOP OF LIST ** HEART ................................................. NUMBER EXAMINED: NOT REMARKABLE: V4O^ -- MYOCARDITIS, VENTRICULAR REGION Total> KIDNEYS ............................................... NUMBER EXAMINED: NOT REMARKABLE: -- CORTICAL TUBULAR BASOPHILIA Minimal Slight Total> -- CORTICAL LYMPHOCYTIC INFILTRATION Minimal Slight Total -- CORTICAL INTERSTITIAL FIBROBLASTPROLIFERATION/ FIBROSIS Total -- UROTHELIAL HYPERPLASIA, UNILATERAL Total -- PELVIC DILATATION Slight Total -- MINERALISATION, CORTICOMEDULLARY Minimal Total -- MEDULLARY MINERALISATION Minimal Total -- FOCAL CAPSULAR CONGESTION Total continued) Print No: 0031 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D ------- MALE------- --- -- FEMALE--- -- -1- -2- -3- -4- -1- -2- -3- -4- 5S554 555 540 540 000 540 220 2 00 100 3 00 00 00 00 000 05 05 00 05 04 00 00 00 00 00 00 00 00000 10000000 1 00 0 0 0 0 0 00002 0 0 1 00002 0 0 1 10 0 1 1 0 0 0 10 0 1 1 0 0 0 0000 0 000 MIN 313/023622 Recovery phase animals GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 8 (Microscopic pathology 2 34 -- POSF--- 30 100 300 SEX: GROUP: ORGAN/TISSUE EXAMINED NUMBER: ** FROM PREVIOUS PAGE ** KIDNEYS ............................................... NUMBER EXAMINED: VO NOT REMARKABLE: -- MEDULLARY TUBULAR BASOPHILIA Minimal> Total> -- MEDULLARY TUBULAR FIBROSIS Minimal> Total> LARYNX ................................................ NUMBER EXAMINED: NOT REMARKABLE: -- NECROSIS OF VENTRAL CARTILAGE Present> Total> -- ARYTENOID - EPITHELIAL HYPERPLASIA Minimal> Slight> Total> -- ARYTENOID - EPITHELIAL HYPERKERATOSIS PARAKERATOSIS Mlnimal> Total> -- ARYTENOID - SUBEPITHELIAL INFLAMMATORY CELL INFILTRATION Minimal> Total> -- VENTROLATERAL EPITHELIAL HYPERPLASIA Minimal> Total> --VENTROLATERAL EPITHELIAL SQUAMOUS METAPLASIA Minimal> Total> - continued) Print No: 0031 Printed: 18-SEP-02 P r o t o c o l Number: MIN 3 1 3 - N U M B E R - O F - A N I M A L S - A F F E C T E D ----- -2- -3- -4- -i- -2- -3- -4- 5 555 4 55 5 *" --S3 -- -- S3 -- --=s-- 500 5 4 00 5 0 00 2 2 00 4 0 00 1 0 00 0 0 00 1 0 00 0 000 1 0 000 000 1 0 000 555 5 4 555 5 44 1 4 4 42 000 40 003 000 4 0 00 3 011 00 0 0 0 0002 0 0 00 01 12 0 0 00 000 2 0 0 00 0002 0 0 00 0100 0 0 0 1 010 00 0 01 00 0 10 00 0 000 10 0 00 00 0 10 00 0 00 0 10 0 00 1 LO LO O OS to to Recovery phase animals GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 8 (Microscopie pathology 234 ------- POSF---30 100 300 SEX: GROUP: ORGAN/TISSUE EXAMINED NUMBER: ** FROM PREVIOUS PAGE ** LARYNX ............................................ ...,. NUMBER EXAMINED: NOT REMARKABLE: VO ON -- VENTRAL POUCH - EPITHELIAL HYPERPLASIA Minimal Total --VENTRAL POUCH - INFLAMMATORY CELL INFILTRATION Minimal Total -- VENTRAL POUCH - FORIGN BODY IN LUMEN, RELATED TO EPITHELIAL HYPERPLASIA/ INFLAMMATORY CELL INFILTRATION Slight Moderate Total -- PROMINENT DILATED ACINI VENTRAL GLAND Present Total LIVER ................................................ . NUMBER EXAMINED: NOT REMARKABLE: -- CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA Slight> Total> -- CENTRILOBULAR HEPATOCYTE HYPERTROPHY Minimal Slight Total -- CENTRILOBULAR HEPATOCYTE NECROSIS/INFLAMMATORY CELL INFILTRATION/SINUSOIDAL CONGESTION Moderate> Total> -- PROMINENT CENTRILOBULAR HEPATOCYTE PIGMENT Minimal Total - continued) Print No: 0031 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D --- ------ MALE------ ------FEMALE------1- -2- -3- -4- -1- -2- -3- -4- 55554555 555 5 45 5 5 544 1 4 44 2 0002 0 00 0 000 2 0 0 0 0 0002 0 0 0 0 000 2 0 0 0 0 0000 0 1 0 0 000 0 0 0 1 0 0000 0 1 1 0 00 0 0 0 0 0 1 00 0 0 0 0 0 1 55 5 5 4 5 5 5 5100 2 5 4 1 00 2 5 0 0 0 0 00 25 0 00 0 0130000 4 020 0 0 00 0 033 0 0 00 4 000 1 0 00 0 000 1 0 0 0 0 00 00 0 0 0 2 00 0 00 0 0 2 g u> u> o N> On to to Recovery phase animals GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 8 (Microscopie pathology 2 34 -- POSF--- 30 100 300 SEX: GROUP: ORGAN/TISSUE EXAMINED NUMBER: ** FROM PREVIOUS PAGE ** LIVER .................................................. NUMBER EXAMINED : NOT REMARKABLE: V*O-4 -- FOCAL/MULTIFOCAL SINGLE CELL NECROSIS INFLAMMATORY CELL INFILTRATION Total> -- PARENCHYMAL INFLAMMATORY CELL INFILTRATION Minimal> Total> -- PERIPORTAL LYMPHOCYTIC INFILTRATION Minimal Total -- CENTRILOBULAR INFLAMMATORY CELL INFILTRATION Minimal Total -- EXTRAMEDULLARY HAEMOPOEISIS Minimal Total -- PERIPORTAL HEPATOCYTE VACUOLATION Total -- CENTRILOBULAR HEPATOCYTE VACUOLATION Slight Total -- HEPATOCYTE VACUOLATION, MEDIAN CLEFT Slight Total LUNGS & BRONCHI ....................................... NUMBER EXAMINED: NOT REMARKABLE: -- FOCAL BRONCHOALVEOLAR HYPERPLASIA Minimal Total continued) Print No: 0031 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D --- ------ MALE------ ----- FEMALE------1- -2- -3- -4- -1- -2- -3- -4- 55 -- IH -- 5 -IH- 5 -H2- 4 -- S5-- 5 --= " 55 --= -- min o 555 4 10 0 2 000 0 5 55 5 41 0 00 o o o o o o o o o o o o o in cs 00 00 0 0 0 10 0 10 00 00 1 1 110 110 00 0 10 10 0 0 00 00 1 1 55 43 5 0 00 00 0 0 1 1 00 00 10 10 00 00 10 10 00 00 00 00 00 45 24 00 00 10 10 00 00 00 00 01 01 00 00 00 00 00 55 41 00 00 MIN 313/023622 Recovery phase animals GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 TABLE 8 (Microscopie pathology 234 --------POSF---30 100 300 SEX: GROUP: Or g a n /t i Ss u e EXAMINED NUMBER: ** FROM PREVIOUS PAGE ** ' LUNGS & BRONCHI ..................................... .. NUMBER EXAMINED : VO 00 NOT REMARKABLE: -- FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES Minimal> Slight> Total> -- AGGREGATIONS OF ALVEOLAR MACROPHAGES Mnmal> Total> -- ALVEOLITIS Minimal> Total> -- ALVEOLAR HAEMORRHAGE Minimal> Slight> Total> -- PERIVASCULAR INFLAMMATORY CELL INFILTRATION Minimal> Total> -- PROMINENT AGGREGATIONS OF LYMPHOCYTES Total> -- VASCULAR CONGESTION Total> --FOCAL OSSEOUS DEPOSITION Mnimal> Total> -- EARLY AUTOLYSIS Total> continued) Print No: 0031 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D ------- HALE------- --- -- FEMALE-----1- -2- -3- -4- -1- -2- -3- -4- 555 5 4 555 MIN 313/023622 cmco in 5 55 2 43 54 02 0 0 0 00 00 10 10 2 2 10 10 10 10 0 0 0 00 0 00 0 00 00 01 01 20 20 00 00 10 10 0 0 0 0 0 0 11 11 00 00 10 10 0 00 00 55 44 5 1 10 00 10 2 0 2 00 00 2 2 00 0 00 0 0 10 00 0 0 10 00 00 1 1 00 0 00 0 00 00 2 2 00 0 Recovery phase animals GROUP : COMPOUND : EXPOSURE LEVEL(PPM): i CONTROL 0 TABLE 8 (Microscopic pathology 234 -- POSF--- 30 100 300 QRGAN/TISSUE EXAMINED SEX: GROUP : NUMBER: T R A C H E A .................................................NUMBER EXAMINED: VO NOT REMARKABLE: VO -- EPITHELIAL HYPERPLASIA Total> -- EPITHELIAL INFLAMMATORY CELL INFILTRATION Total> -- SUBEPITHELIAL INFLAMMATORY AGGREGATIONS Slight> Total> --SUBEPITHELIAL FIBROBLAST PROLIFERATION Total> TRACHEAL BIFURC......................................... NUMBER EXAMINED : NOT REMARKABLE: URINARY BLADDER ........................................ NUMBER EXAMINED: NOT REMARKABLE: EPIDIDYMIDES ........................................... NUMBER EXAMINED : NOT REMARKABLE: LN MA N D I B U L A R .......................................... NUMBER EXAMINED: NOT REMARKABLE: MISCELLANEOUS .......................................... NUMBER EXAMINED: NOT REMARKABLE : PREPUTIAL GLANDS ...................................... NUMBER EXAMINED: NOT REMARKABLE: continued) Print No: 0031 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D -- ----- MALE------ ----- FEMALE-----1- -2- -3- -4- -1- -2- -3- -4- 55554555 5 005 4 005 5 004 4 005 00000000 0 00 0 0 00 0 00 0 1 0 0 0 0 00 0 1 0 0 0 0 00000 00 0 4 005 4 005 4 005 4 005 00 0 00000 00000000 00 0 00000 000 00000 0 00 00000 0 00 00000 00000000 00000000 MIN 313/023622 in in in in TABLE 8 (Microscopic pathology - continued) Recovery phase animals GROUP : COMPOUND : EXPOSURE UEVEL(PPM): i CONTROL 0 234 -- POSF--- 30 100 300 Printed: 18-SEP-02 Protocol Number: MIN 313 -- N U M B E R - O F - A N I M A L S - A F F E C T E D --- SEX: -- MALE-- -- -- -- FEMALE-- GROUP: -1- -2- -3- -4- -1- -2- -3- -4- ORGAN/TISSUE EXAMINED O O PROSTATE .................. NUMBER: 5 5 5 5 4 5 5 5 0 00 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 SKIN ...................... 0 000000 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 STOMACH ................... 00000000 NOT REMARKABLE: 0 0 0 0 0 0 0 0 TAIL ...................... 00000000 NOT REMARKABLE: 0 0 0 0 0 0 0 0 THYMUS .................... 0 0000000 NOT REMARKABLE: 0 0 0 0 0 0 0 0 URETERS ................... 0 0000 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 UTERUS .................... 0 0000 00 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 -- LUMINAL DILATATION ** END OF LIST ** Total> 0 0 0 0 0 0 0 0 MIN 313/023622 APPENDIX 1 Weekly physical examination - individual findings GROUP : EXPOSURE LEVEL(PPM): i CONTROL 0 2 30 3 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 1M 1 7 14 SKIN ENCRUSTATION (S) HEAD STAINING BROWN HEAD 3 7 14 COAT HAIRLOSS FORELIMBS 41 8 18 COAT HAIRLOSS HEAD 43 8 18 EYES PROMINENT RIGHT 45 8 18 SKIN ENCRUSTATION (S) HEAD 4 300 WEEKS 1-18 2-3 13-14 3-14 13-18 18 13-15 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 MIN 313/023622 MIN 313/023622 102 ; APPENDIX 1 (Weekly physical examination - continued) GROUP EXPOSURE LEVEL(PPM) i2 3 0 30 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 2M 8 7 14 SKIN ENCRUSTATION (S) TAIL STAINING BROWN DORSAL BODY SURFACE 9 7 14 COAT HAIRLOSS DORSAL BODY SURFACE 10 7 14 COAT HAIRLOSS HEAD 4 6 8 18 COAT HAIRLOSS HEAD SKIN ENCRUSTATION(S) HEAD 47 8 18 COAT HAIRLOSS DORSAL BODY SURFACE STAINING BROVIN HEAD 4 300 WEEKS 1-18 7-8 6-10 2-3 13-14 4-5, 8-18 8-9 2-4 10-18 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number : MIN 313 APPENDIX 1 (Weekly physical examination - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) i CONTROL 0 2 30 3 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 2M 48 8 49 8 50 8 18 SKIN ENCRUSTATION (S) NOSE DORSAL BODY SURFACE 18 STAINING BROWN HEAD 18 COAT HAIRLOSS DORSAL BODY SURFACE SKIN ENCRUSTATION(S) DORSAL BODY SURFACE STAINING BROWN HEAD 4 300 WEEKS 1-18 15-17 17-18 18 9-18 8 12-18 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 MIN 313/023622 MIN 313/023622 104 : APPENDIX 1 (Weekly physical examination - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) i CONTROL 0 2 30 3 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 3M 12 7 14 STAINING RED PENIS 13 7 14 COAT HAIRLOSS DORSAL BODY SURFACE 4 300 WEEKS 1-18 13-14 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 MIN 313/023622 105 : APPENDIX I (Weekly physical examination - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) i CONTROL 0 2 30 3 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 4M 16 7 14 STAINING BROWN HEAD DORSAL BODY SURFACE 17 7 14 STAINING BROWN HEAD 19 7 14 COAT HAIRLOSS HEAD DORSAL BODY SURFACE SKIN ENCRUSTATION (S) HEAD STAINING BROWN HEAD 20 7 14 STAINING BROWN HEAD 57 8 18 COAT HAIRLOSS HEAD 4 300 WEEKS 1-18 10-14 5-14 14 4 2-3 4 6-14 14 4-18 Print No: 0017 Printed: 24-JUN-02 X y b i o n protocol number: MIN 313 APPENDIX 1 (Weekly physical examination - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) i CONTROL 0 2 30 3 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 4M 58 8 59 8 60 8 18 COAT HAIRLOSS HEAD DORSAL BODY SURFACE STAINING BROWN DORSAL BODY SURFACE 18 COAT HAIRLOSS HEAD 18 COAT HAIRLOSS FORELIMBS 4 300 WEEKS 1-18 7-9, 17-18 2-18 6-18 5-18 5-18 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 MIN 313/023622 MIN 313/023622 107 : APPENDIX 1 (Weekly physical examination - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 234 --------POSF-------30 100 300 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: IF 21 7 14 STAINING BROWN HEAD 22 7 14 STAINING BROWN VENTRAL BODY SURFACE 23 F 13 COAT HAIRLOSS VENTRAL BODY SURFACE STAINING BROWN HEAD 24 7 14 STAINING BROWN HEAD 61 8 18 COAT HAIRLOSS DORSAL BODY SURFACE 62 8 18 COAT HAIRLOSS HEAD DORSAL BODY SURFACE WEEKS 1-18 8 12-14 12-13 12-13 3-8 16-18 6-11 16-18 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 APPENDIX 1 (Weekly physical examination - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) i CONTROL 0 2 30 34 100 300 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: IF 63 H 3 BUILD (DISPLACEMENT) LEFT HINDLIMB O RIGHT HINDLIMB oo BODY TEMPERATURE REDUCED COAT UNGROOMED EXCRETA FAECES LOOSE MUSCLE REACTION ABNORMAL GAIT SWAYING POSTURE TILTED HEAD LEANS TO THE RIGHT RIGHTING REFLEXES POOR STAINING RED NOSE 64 8 18 STAINING BROWN HEAD 65 8 18 COAT HAIRLOSS FORELIMBS WEEKS 1-18 3 3 3 3 3 3 3 3 3 3-13, 17-18 3-18 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 MIN 313/023622 MIN 313/023622 109 : APPENDIX 1 (Weekly physical examination - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) i CONTROL 0 2 30 3 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 2F 27 7 14 STAINING BROWN HEAD DORSAL BODY SURFACE 66 8 18 STAINING BROWN VENTRAL BODY SURFACE DORSAL BODY SURFACE 67 8 18 COAT HAIRLOSS DORSAL BODY SURFACE STAINING BROWN HEAD 68 8 18 COAT HAIRLOSS FORELIMBS HEAD STAINING BROWN HEAD 69 8 18 COAT HAIRLOSS HEAD SKIN ENCRUSTATION(S) HEAD 4 300 WEEKS 1-18 3- 6 11- 14 12- 17 18 4-10 12-18 15-18 10-18 12-15 8-18 8, 11-15, 17 Print No: 0017 Printed: 24-JUN-2 Xybion protocol number: MIN 313 MIN 313/023622 110 : APPENDIX 1 (Weekly physical examination - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): i CONTROL 0 234 -- POSF--- 30 100 300 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 2F 70 8 18 STAINING BROWN HEAD WEEKS 1-18 5-6 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 APPENDIX 1 (Weekly physical examination - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) i CONTROL 0 2 30 3 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 3F 31 7 14 STAINING BROWN HEAD 33 7 14 COAT HAIRLOSS HEAD STAINING BROWN HEAD 34 7 14 STAINING BROWN HEAD DEATH CODE TERMINAL KILL 35 7 14 STAINING BROWN HEAD 71 8 18 STAINING BROWN HEAD 72 8 18 STAINING BROWN HEAD 4 300 WEEKS 1-18 5-14 7-14 5-14 5-14 14 12-14 18 12-18 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 MIN 313/023622 MIN 313/023622 112 : APPENDIX 1 (Weekly physical examination - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) i CONTROL 0 2 30 3 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 3F 73 18 STAINING BROWN HEAD 75 18 COAT HAIRLOSS DORSAL BODY SURFACE 4 300 WEEKS 1-18 5-18 8-18 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 MIN 313/023622 113 : APPENDIX 1 (Weekly physical examination - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 234 -- POSF--- 30 100 300 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 4F 36 7 14 COAT HAIRLOSS RIGHT FORELIMB STAINING BROWN HEAD 37 7 14 BUILD (DEFORMITY) PROMINENT AREA TAIL STAINING BROWN VENTRAL BODY SURFACE DORSAL BODY SURFACE 38 7 14 COAT UNGROOMED HAIRLOSS LEFT HINDLIMB STAINING BROWN VENTRAL BODY SURFACE DORSAL BODY SURFACE 39 7 14 STAINING BROWN HEAD WEEKS 1-18 11-14 5- 14 9- 14 6- 14 5- 14 8-14 10- 14 4-14 4-14 6- 14 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 APPENDIX 1 (Weekly physical examination - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) i CONTROL 0 2 30 3 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 4F 40 7 14 STAINING BROWN HEAD VENTRAL BODY SURFACE DORSAL BODY SURFACE 77 8 18 STAINING BROWN VENTRAL BODY SURFACE DORSAL BODY SURFACE 78 8 18 STAINING BROWN HEAD VENTRAL BODY SURFACE 79 8 18 STAINING BROWN HEAD 80 8 18 STAINING BROWN HEAD 4 300 WEEKS 1-18 6-14 5-14 5-14 10-18 10-18 6-18 6-18 5-18 5-15 Print No: 0017 Printed: 24-JUN-02 Xybion protocol number: MIN 313 MIN 313/023622 APPENDIX 1 (Weekly physical examination - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) 1 CONTROL 0 2 30 3 100 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 1M 817 4 SKIN ENCRUSTATION (S) HEAD 4 300 WEEKS 1-4 2-3 Print No: 0008 Printed: 24-JUN-02 Xybion protocol number: MIN 313A MIN 313/023622 MIN 313/023622 116 : APPENDIX 1 (Weekly physical examination - continued) GROUP : COMPOUND ; EXPOSURE LEVEL(PPM): i CONTROL 0 23 -- POSF-- 30 100 4 300 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: IP 91 7 93 7 94 7 95 7 STAINING BROWN HEAD STAINING BROWN UPPER DORSAL THORAX DEATH CODE TERMINAL KILL 4 STAINING BROWN UPPER DORSAL THORAX WEEKS 1-4 4 3-4 4 4 Print No: 0008 Printed: 24-JUN-02 Xybion protocol number: MIN 313A 313/023622 117 : APPENDIX 1 (Weekly physical examination - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): i CONTROL 0 2 34 -- POSF--- 30 100 300 CATEGORY ANIMAL DEATH WK OF KEYWORD NUMBER CODE DEATH QUALIFIER GROUP: 4F 96 7 98 7 100 7 4 STAINING BROWN UPPER DORSAL THORAX 4 STAINING BROWN HEAD 4 STAINING BROWN HEAD WEEKS 1-4 4 4 4 Print No: 0008 Printed: 24-JUN-02 Xybion protocol number MIN 313A APPENDIX 2 Daily post - dose observation MIN 313/023622 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 --POSF-- 30 100 300 Main study animals Group Sign Animal Week No. (Number of days in week signs observed) 1 2 3 4 5 6 7 8 9 10 11 12 13 14* 1M Red/brown staining around eyes 1 1 1 11 i (Control) 21 2 32 2 11 1 4 3 2 2 4 12 1 13 2 1 51 Wet fur 1 354553434 3 1 11 2 2535535334 1 3 4535545333 1 3 4 2 5 3 5 5 4 5 3 3 4 12 2 1 5 25455344332 Brown staining on head 1 2 3 4 5 1 1 1 11 1 111 1 211 2 13 1 1 2 11 Brown staining muzzle 4 1 Brown staining body 1 4 2M Red/brown staining around eyes 6 3 1 1 242 1 1 (Low dose) 7 11 8 41 11 12 1 1 9 1 12 1 1 21 10 3 1 1 1 Wet fur 6 4434535453 2 7 3434435444 8 443443544 3 9 343443544 3 11 10 3 4 3 4 5 3 5 4 4 3 Brown staining body 6 3 Brown staining on head 6 211 7 11 81 4 11 91 1 1 10 1 1 Animals exposed for 5 days a week for 13 weeks * Animals sent for necropsy over first 3 days of Week 14 : 118 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 23 -- POSF 30 100 4 300 Main study animals Group Sign Animal No. 3M Red/brown staining around eyes (Inter, dose) 11 12 13 14 15 1 1 3 1 Week (Number of days in week signs observed) 2 3 4 5 6 7 8 9 10 11 12 13 14* 11 1 1 11 12 1 1 1 1 11 1 12 Wet fur 11 3 4 3 4 5 3 5 4 4 3 2 1 12 4 4 3 4 5 4 5 4 4 3 1 13 3 4 3 4 4 3 5 4 4 4 1 1 14 3 4 3 4 4 3 5 5 4 3 3 5 1 15 4 4 3 4 4 3 5 4 5 3 1 2 1 Brown staining on head 11 12 1 13 14 15 11 1 1 1 1 1 4M Red/brown staining around eyes 16 1 (High dose) 17 33 1542 23 1 2 22 21 18 3 12 1 19 4 1 2 2 2 2 1 5 3 4 2 2 20 1 1 Wet fur 16 4 5 3 4 5 3 4 5 5 4 5 4 4 1 17 3 5 3 4 5 3 4 4 5 3 11 18 3 5 3 4 5 3 4 4 5 4 3 2 2 19 5 5 3 4 5 4 4 5 5 5 5 4 3 1 20 3 5 3 4 5 3 4 4 5 3 1 1 1 Brown staining on head 16 1 2 1 2 3 2 17 1 1 18 1 19 1 1 1 1 20 1 12 52 1 1 1 12 111 2 Brown staining on muzzle 16 1 Brown staining on body 19 Animals exposed for 5 days a week for 13 weeks * Animals sent for necropsy over first 3 days of Week 14. 11 : 119 : APPENDIX 2 (Daily post - dose observation - continued) Group :1 Compound : Control Exposure level (ppm) : 0 2 34 -- POSF-- 30 100 300 MIN 313/023622 : 120 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 34 -- POSF-- 30 100 300 Main study animals Group Sign 3F Red/brown staining around eyes (Inter, dose) Animal No. 31 32 33 34 35 i 2 4 1 Week (Number of days in week signs observed) 2 3 4 5 6 7 8 9 10 11 12 13 14* 11 1 11 11 1 3452 2 12 1 14 2 4 3 2 3 2 1 14 1 11 422 11 1 Wet fur 31 5 5 3 4 5 3 5 4 5 5 5 5 4 2 32 5 4 3 4 5 3 5 4 5 3 3 5 5 1 33 3 5 3 4 5 4 5 4 5 5 5 5 5 2 34 5 5 4 4 5 4 5 4 5 5 5 5 5 2 35 5 5 4 4 5 4 5 4 5 5 4 5 5 2 Brown staining on head 31 1 3 1 1 2 1 1 2 32 1 2 1 33 1 2 1 15 21 34 1 2 1 2 3 1 2 1 3 2 2 1 3 2 35 11 222 1 4F Red/brown staining around eyes 36 (High dose) 37 2 1 1 38 1 2 39 40 3 21 1 111 12 12 12 2 1 14 11 2 3 1 Wet fur 36 4 5 3 4 4 3 5 4 5 4 4 5 3 2 37 5 5 3 5 5 5 5 5 5 4 4 5 5 2 38 5 5 4 5 5 5 4 5 5 5 4 4 4 2 39 5 5 3 4 5 4 5 5 5 5 4 5 3 2 40 5 5 3 4 4 3 5 5 5 5 4 5 2 2 Brown staining on head 36 37 1 38 39 1 40 1 1 12 3 1 31 1 21 1 11 1 1 12 1 1 Brown staining body 37 38 Animals exposed for 5 days a week for 13 weeks * Animals sent for necropsy over first 3 days of Week 14. 2 55 5 5555 52 : 121 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 -- POSF-- 30 100 300 Recovery phase animals Group Sign Animal Week No. (Number of days in week signs observed) 1 2 3 4 5 6 7 8 9 10 11 12 13 14* 1M Red/brown staining around eyes 41 1 332 222 111 (Control) 42 5 5 1 2 1 1 1 43 2 12 2 3 44 3 3 4 3 1 3 1 1 1 3 45 2 2 Wet fur 41 3 5 5 5 5 3 4 3 4 5 31 42 3 5 5 5 5 3 4 3 3 3 2 43 3 5 3 5 5 3 4 3 4 3 31 44 3 5 4 5 5 4 4 3 4 3 21 45 3 5 3 5 5 3 4 4 3 3 1 Brown staining on head 41 42 1 43 44 45 3 1 22 11 11 11 1 2 111 1 2M Red/brown staining around eyes 46 12 (Low dose) 47 3 1 1 12 1 1 48 2 112 49 1 1 1 3 4 3 2 3 1 1 11 50 2 1 2 1 1 331 232 Wet fur 46 3 4 3 4 4 3 5 4 4 3 1 47 3 4 3 4 4 3 5 4 4 3 1 48 3 4 5 4 5 3 5 4 4 3 2 1 49 4 4 3 4 5 3 5 4 5 4 1 1 1 50 5 4 3 4 5 4 5 5 5 4 4 4 3 Brown staining on head 46 47 48 49 50 Animals exposed for 5 days a week for 13 weeks 3 2 * Animals sent for necropsy over first 3 days of Week 14. 1 11 11 111 11 2 1 12 2 1 1 1 23 : 122 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 ----- POSF-- -- 30 100 300 Recovery phase animals Group Sign Animal Week No. (Number of days in week signs observed) 3M Red/brown staining around eyes 51 (Inter, dose) 52 1 2 3 4 5 6 7 8 9 10 11 12 13 14* 12 111 1 1 1 132 1 53 1 11 54 1 2 55 1 1 Wet fur 51 4 4 3 4 5 3 5 4 4 3 1 52 4 4 3 4 4 3 5 4 4 3 11 53 4 4 5 4 4 3 5 4 5 5 3 2 54 5 4 3 4 5 3 5 4 5 3 2 2 4 55 4 4 3 4 5 3 5 4 5 4 2 1 Brown staining on head 51 11 2 1 52 1 53 1 1 54 1 1 55 1 13 2 2 4M Red/brown staining around eyes 56 (High dose) 57 2 2 1 1 58 2 11 59 1 1 3 2 2 3 3 2 2 2 60 1 1 54 1 Wet fur 56 4 5 3 4 5 3 4 4 5 3 1 2 57 4 5 3 4 4 3 4 4 5 3 3 1 58 4 5 4 4 4 4 5 4 5 3 4 2 2 59 4 5 4 4 5 3 4 4 5 3 4 5 5 60 4 5 3 4 4 3 4 4 5 3 1 Brown staining on head 56 1 57 1 1 58 12 1 59 1 2 1 11 60 1 1 2 1 Animals exposed for 5 days a week for 13 weeks * Animals sent for necropsy over first 3 days of Week 14. : 123 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 ---- POSF----- 30 100 300 Recovery phase animals Group Sign Animal Week No. (Number of days in week signs observed) 1 2 3 4 5 6 7 8 9 10 11 12 13 14* IF Red/brown staining around eyes 61 4 1 1 2 3 1 1 (Control) 62 1 11 114 2 2 63 1 1 64 2 1 11 4 65 1 11 Wet fur 61 3 5 4 5 5 4 5 4 5 4 4 5 5 62 4 5 3 5 5 3 5 5 5 5 5 5 5 63a 5 5 1 64 2 5 3 5 5 3 5 5 5 4 4 5 5 65 3 5 4 5 5 2 5 4 5 4 5 5 5 Brown staining on head 61 12 13 3 4 2 42 13 62 1 2 2 2 1 3 3 1 5 3 63a 64 4 4 2 2 3 4 3 5 3 65 2 2 2 5 4 3 2 Loss of use of hind limbs 63a 1 Unsteady gait 63a 1 Brown staining muzzle 61 1 2F Red/brown staining around eyes 66 1 1 14 2 (Low dose) 67 2 3 1 2 2 21 68 12 13 69 2 70 2 1 1 1 12 13 12 Wet fur 66 5 4 3 4 5 4 5 5 5 4 5 5 5 67 4 5 3 4 5 3 5 4 5 4 5 5 5 68 4 5 3 5 5 5 5 5 5 5 5 5 4 69 4 5 3 4 5 3 5 4 5 4 4 5 5 70 4 5 3 5 5 5 5 5 5 5 5 5 5 Brown staining on head 66 2 1 1 1 1 67 1 1 2 2 1 2 2 2 2 68 2 13 1112 3 1 69 1 1 1 1 1 1 3 3 2 3 70 2 15422 12 Brown staining body 70 Animals exposed for 5 days a week for 13 weeks * Animals sent for necropsy over first 3 days of Week 14. a Animal 63 killed for humane reasons in Week 3. : 124 : 1 APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 ---POSF - 30 100 300 Recovery phase animals Group Sign Animal Week No. (Number of days in week signs observed) 1 2 3 4 5 6 7 8 9 10 11 12 13 14* 3F Red/brown staining around eyes 71 2 1 (Inter, dose) 72 12 1 73 1 1 2 74 1 75 1 Wet fur Brown staining on head 71 5 5 3 4 5 4 5 5 5 5 5 5 5 72 4 5 3 4 5 4 5 5 5 5 5 5 4 73 4 5 4 4 5 4 5 5 5 5 5 5 4 74 4 5 3 4 5 4 5 4 5 3 5 5 4 75 4 4 3 4 5 4 5 4 5 5 5 5 4 71 1 1 72 2 1 1 1 2 2 1 1 73 1 1 4 74 1 1 1 2 75 1 21 1 12 2 1 4F Red/brown staining around eyes 76 222 1 1 (High dose) 77 4 21 31 78 1 2 12 79 4 3 1 1 14 4 3 4 80 1 1 Wet fur 76 5 5 3 4 5 4 4 4 5 4 5 5 5 77 5 5 3 4 5 5 5 4 5 4 4 5 3 78 5 5 3 5 5 4 5 5 5 5 5 5 5 79 4 5 3 4 5 3 4 5 5 5 5 5 4 80 4 5 3 5 5 4 5 5 5 4 5 5 4 Brown staining on head 76 77 1 1 78 79 2 11 80 1 22 13 11 1 15 12 42 2 1 1 14 5 32 Brown staining body 76 78 Animals exposed for 5 days a week for 13 weeks * Animals sent for necropsy over first 3 days of Week 14. 3 3 : 125 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 -- POSF----- 30 100 300 Satellite study animals Group Sign 1M Red/brown staining around eyes (Control) Animal No. 81 82 83 84 85 Week (Number of days in week signs observed) 12 3 4 4 12 13 12 42 12 334 1 Wet fur 81 4 5 4 5 82 3 5 3 5 83 3 5 5 5 84 4 5 5 5 85 5 5 5 5 Brown staining on head 81 1 82 83 1 84 85 1 21 1 1 31 42 IF Red/brown staining around eyes 91 2 2 3 1 (Control) 92 1 2 93 3 1 94 1 2 95 1 Wet fur 91 3 5 4 5 92 3 5 3 5 93 2 5 4 5 94 2 5 4 5 95 2 5 3 5 Brown staining on head Animals exposed for 5 days a week for 4 weeks 91 92 93 94 95 1 31 1 45 2 22 : 126 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group :1 Compound : Control Exposure level (ppm) : 0 234 ----- POSF----- 30 100 300 Satellite study animals Group Sign 4M (High dose) Red/brown staining around eyes Animal No. 86 87 88 89 90 Week (Number of days in week signs observed) 12 3 4 41 11 11 2 11 1 Wet fur 86 4 5 3 4 87 4 5 3 4 88 4 5 3 4 89 5 5 3 4 90 4 5 3 4 Brown staining on head 87 1 89 3 90 1 4F (High dose) Red/brown staining around eyes 97 98 99 100 2 1 1 1 Wet fur 96 4 5 3 4 97 4 5 4 4 98 5 5 3 4 99 3 5 3 4 100 3 5 3 4 Brown staining on head 96 1 98 1 99 100 Animals exposed for 5 days a week for 4 weeks 1 1 1 : 127 : APPENDIX 3 Bodyweights - individual values (g) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 234 --------POSF-------30 100 300 GROUP WEEK ANIMAL -1 WEEK 0 WEEK 1 WEEK 2 WEEK 3 1M i 248 292 328 356 386 2 224 252 276 307 338 3 227 263 284 311 337 4 226 248 266 291 314 5 232 253 275 296 313 41 229 252 281 296 312 42 247 275 296 324 355 43 229 246 273 302 323 44 236 257 279 298 316 45 228 268 300 316 337 2M 6 241 278 304 337 365 7 224 239 254 272 287 8 232 279 328 342 372 9 237 276 292 319 344 10 235 257 265 293 321 46 244 282 311 336 365 47 225 240 253 271 283 48 245 282 308 323 343 49 2 2 1 251 275 289 307 50 246 284 305 321 343 3M 1 1 235 264 290 308 333 12 238 272 295 318 332 13 237 270 293 302 341 14 239 263 269 288 317 15 234 255 277 291 310 51 245 274 291 317 339 52 236 271 294 313 329 53 232 265 289 320 342 54 245 275 297 315 332 55 214 227 242 255 266 WEEK 4 410 346 359 339 337 336 382 342 344 363 393 302 392 364 336 391 299 361 323 356 354 360 369 331 336 364 350 363 352 282 WEEK 5 428 363 378 355 365 352 408 365 368 379 409 319 416 374 352 409 309 375 336 371 376 380 395 349 353 380 360 374 363 294 WEEK 6 447 376 399 371 378 374 420 382 390 392 437 332 412 393 368 427 324 398 354 380 390 396 412 366 370 398 373 397 374 312 Print No: 0018 Printed: 24-JUN-02 Xybion protocol number: MIN 313 WEEK 7 WEEK 8 WEEK 9 460 474 492 396 404 412 408 423 435 390 402 422 398 402 417 387 402 412 434 451 461 404 416 436 409 425 432 404 409 416 452 469 490 352 366 380 430 451 466 408 416 426 391 404 419 442 459 472 335 344 351 422 434 442 366 373 376 392 408 409 405 416 421 413 424 441 430 451 459 379 392 400 380 399 415 406 421 4351 394 407 4101 401 410 413 390 402 418 325 336 347 MIN 313/023622 APPENDIX 3 (Bodyweights - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 34 -- POSF--- 30 100 300 GROUP WEEK ANIMAL -i WEEK 0 WEEK i WEEK 2 WEEK 3 WEEK 4 WEEK 5 WEEK 6 4M 16 234 262 289 309 330 351 364 17 220 241 254 267 285 298 301 18 227 258 274 303 324 343 356 19 238 267 283 304 329 351 357 20 234 260 276 295 310 330 339 56 269 261 266 292 319 338 353 57 227 263 288 321 346 368 382 58 240 261 277 291 311 328 339 59 217 243 255 280 293 306 314 60 234 261 275 294 314 334 358 IF 21 171 184 190 205 217 230 230 22 170 185 199 206 208 225 239 23 173 188 198 208 229 238 239 24 177 193 202 207 225 234 246 25 177 191 206 217 224 229 241 61 195 207 217 237 246 262 268 62 174 188 201 213 213 216 231 63 189 196 214 231 A 64 186 194 196 210 2 1 1 223 216 65 172 185 197 197 2 1 1 227 232 2F 26 177 195 206 213 215 229 240 27 178 176 189 199 202 195 207 28 180 190 198 202 208 206 219 29 165 172 178 189 202 208 209 30 164 170 184 187 194 196 206 66 188 208 227 238 252 262 271 67 163 191 210 221 237 252 263 68 177 184 204 208 217 210 227 69 183 199 215 220 226 236 249 70 174 183 203 208 233 246 250 Animal No. 63 sent for necropsy for humane reasons therefore no further bodyweights 375 316 373 373 347 372 395 348 320 381 250 237 251 240 252 276 241 227 243 247 206 232 225 216 274 269 233 256 250 Print Ho: 0018 Printed: 24 -JUN-02 Xybion protocol number: MIN 313 WEEK 7 WEEK 8 WEEK 9 379 387 384 322 32 6 32 9 378 38 6 387 387 389 402 348 359 361 386 399 404 409 419 423 362 388 377 331 339 345 393 402 410 259 263 258 238 245 255 254 257 259 257 261 265 260 250 268 282 287 294 251 246 263 239 253 243 252 256 266 243 256 257 jf 2 2 1 213 227239 240 247i. 230 231 225223 225 233 278 283 2881 280 293 287 c 232 234 241. 259 268 273 261 269 272 MIN 313/023622 MIN 313/023622 130 : APPENDIX 3 (Bodyweights - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 34 ---- -- POSF----30 100 300 GROUP WEEK ANIMAL -1 WEEK 0 WEEK 1 WEEK 2 WEEK 3 3F 31 171 182 193 197 200 32 186 201 209 215 226 33 168 172 200 191 201 34 194 209 227 240 251 35 169 189 187 207 219 71 169 187 203 201 217 72 187 187 194 196 200 73 181 201 2 1 1 216 221 74 182 185 199 205 200 75 166 176 184 195 202 4F 36 185 193 213 208 207 37 176 180 202 2 1 1 218 38 178 198 2 1 1 209 225 39 167 178 192 196 204 40 184 203 190 226 234 76 163 173 186 184 188 77 174 181 195 190 189 78 190 206 2 1 1 2 1 1 215 79 188 207 228 238 244 80 182 185 202 191 202 WEEK 4 219 243 202 252 227 226 208 236 214 210 219 219 231 205 244 192 195 215 249 204 WEEK 5 221 249 208 267 237 228 216 231 217 208 228 228 239 215 252 200 192 224 263 212 WEEK 6 227 249 214 280 243 238 223 239 223 221 234 240 239 218 256 199 205 226 268 224 Print Mo: 0018 Printed: 24-JUN-02 Xybion protocol number: MIN 313 WEEK 7 WEEK 8 WEEK 9 227 237 237 262 271 275 222 214 217 278 280 290 250 258 256 240 246 245 229 232 238 250 252 255 216 225 236 224 232 234 230 238 243 244 240 246 244 244 256 223 2 2 1 225 259 263 265 205 2 1 1 213 208 216 219 231 229 234 u) 271 271 277 238 235 2 3 5 ^ APPENDIX 3 (Bodyweights - continued) GROUP : COMPOUND : EXPOSURE LEVEL CPPM): 1 CONTROL 0 GROUP ANIM AXi WEEK 10 WEEK 11 1M 1 2 3 4 5 41 42 43 44 45 2M 6 7 8 9 10 46 47 48 49 50 3M 11 12 13 14 15 51 52 53 54 55 500 426 456 427 433 421 465 444 447 418 490 388 475 431 423 483 359 450 381 425 431 451 448 403 432 446 418 406 427 362 510 433 464 444 440 433 476 454 461 424 505 401 470 438 432 486 370 463 394 425 435 458 453 412 450 455 426 434 440 366 234 --------POSF-------30 100 300 WEEK 12 WEEK 13 WEEK 14 516 506 436 429 470 462 449 437 453 455 449 464 473 492 488 500 461 466 487 471 477 489 430 439 446 512 511 409 410 474 481 450 441 435 426 492 495 504 372 378 394 470 468 476 395 403 417 434 442 464 442 436 469 464 455 454 415 402 461 458 4 68 460 475 440 434 446 442 454 478 451 467 481 375 380 397 WEEK 15 485 516 500 502 455 509 409 488 431 471 501 459 492 493 409 WEEK 16 499 529 515 518 466 521 422 490 441 478 505 468 501 499 407 Print No: 0018 Printed: 24-JUN-02 Xybion protocol number: MIN 313 WEEK 17 506 542 531 533 472 533 435 500 444 491 514 487 509 504 413 MIN 313/023622 MIN 313/023622 132 : APPENDIX 3 (Bodyweights - continued) GROUP : EXPOSURE LEVEL(PPM): GROUP ANIMAL WEEK 10 4M 16 17 18 19 20 56 57 58 59 60 IF 21 22 23 24 25 61 62 64 65 2F 26 27 28 29 30 66 67 68 69 70 3F 31 32 33 34 389 336 395 408 372 413 433 378 358 422 271 254 266 274 272 297 267 260 268 262 234 251 232 239 286 301 242 273 266 238 265 224 300 i 0 WEEK 11 390 341 401 416 375 416 439 384 358 429 278 255 268 281 279 308 275 269 279 264 233 251 243 247 289 307 246 276 278 238 275 224 287 2 34 30 100 300 WEEK 12 WEEK 13 WEEK 14 394 378 349 337 397 392 422 416 379 357 421 429 444 442 444 461 393 395 406 363 366 386 429 431 431 284 270 261 258 270 282 270 274 274 307 301 313 269 275 287 269 265 277 281 274 288 269 261 230 226 2 45 243 239 232 245 243 2 94 293 300 310 301 317 240 234 278 282 287 297 283 281 282 244 233 284 271 2 2 1 216 300 300 WEEK 15 461 478 415 399 433 327 293 288 303 312 328 265 295 303 WEEK 16 466 501 426 424 452 326 294 289 312 320 330 274 303 303 Print No: 0018 Printed: 24-JUN-02 Xybion protocol number: MIN 313 WEEK 17 478 515 436 430 466 328 305 277 311 324 330 275 310 299 APPENDIX 3 (Bodyweights - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) i 1 CONTROL 0 GROUP ANIMAL WEEK 10 WEEK 11 3F 35 71 72 73 74 75 4F 36 37 38 39 40 76 77 78 79 80 256 249 243 255 239 240 244 256 248 227 269 215 222 237 279 241 264 255 247 258 236 240 243 258 257 228 269 217 226 238 280 241 2 34 -- POSF--- 30 100 300 WEEK 12 WEEK 13 WEEK 14 269 262 254 252 258 246 247 255 264 262 273 242 244 249 243 236 250 248 242 253 240 254 242 224 217 275 259 2 2 1 216 223 227 229 241 242 230 255 280 279 291 248 250 257 WEEK 15 268 262 281 249 258 229 251 267 302 268 WEEK 16 268 262 281 259 257 237 258 268 303 273 Print No: 0018 Printed: 24-JUN-02 Xybion protocol number : MIN 313 WEEK 17 274 266 288 259 256 242 262 278 318 272 MIN 313/023622 APPENDIX 3 (Bodyweights - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 GROUP ANIMAL WEEK -1 WEEK 0 1M 81 82 83 84 85 4M 86 87 88 89 90 IF 91 92 93 94 95 4F 96 97 98 99 100 245 277 244 263 224 242 235 271 237 264 232 270 232 272 235 262 249 277 238 278 172 187 188 194 182 194 174 181 186 197 183 190 182 192 177 190 169 180 167 183 2 34 -- POSF--- 30 100 300 WEEK 1 WEEK 2 WEEK 3 301 330 358 285 314 341 258 276 290 310 338 366 294 324 351 301 319 339 293 321 342 273 290 305 295 321 343 296 316 331 202 204 216 200 204 210 216 226 245 194 195 208 208 217 236 196 193 200 195 194 202 192 193 200 192 196 200 191 203 217 Print No: 0009 Printed: 24-JUNt02 Xybion protocol number : MIN 313A MIN 313/023622 MIN 313/023622 135 : GROUP COMPOUND EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 30 GROUP CAGE WEEK -1 WEEK 1 WEEK 2 1M 1 9 2M 2 10 3M 3 11 4M 4 12 IF 5 13 2F 6 14 3F 7 15 4F 8 16 181 183 192 195 192 190 186 186 136 137 131 150 138 135 144 140 180 163 169 173 175 168 157 150 132 133 126 147 134 130 137 128 196 176 184 184 185 172 173 171 136 142 126 149 153 133 137 122 APPENDIX 4 Food consumption - individual values (g/rat) 3 100 WEEK 3 205 186 197 190 191 192 179 179 143 145 132 157 147 136 140 125 4 300 WEEK 4 213 197 202 196 198 197 189 188 148 151 136 156 151 138 141 121 WEEK 5 213 198 204 194 199 194 181 185 149 151 138 157 153 137 144 126 WEEK 6 215 194 201 196 208 200 190 189 148 153 138 156 153 136 144 127 WEEK 7 214 192 208 198 209 198 180 191 147 151 137 154 151 136 143 125 Print No: 0019 Printed: 24-JUN-02 Xybion protocol number: MIN 313 WEEK e WEEK 9 WEEK 10 216 2 12 216 195 193 190 208 207 207 196 193 198 209 203 204 201 200 200 189 180 180 193 187 193 147 146 147 147 148 148 136 139 134 158 151 148 150 148 1 4 7 139 136 131 04 140 143 1 4 0 ^ 126 125 1 2 2 ( 0 APPENDIX 4 (Food consumption - continued) GROUP 1 234 COMPOUND EXPOSURE LEVEL(PPM): 0 30 100 300 GROUP CAGE WEEK 11 WEEK 12 WEEK 13 WEEK 14 WEEK 15 WEEK 16 WEEK 17 Co 1M 1 218 215 190 Os 9 195 200 199 198 197 201 203 2M 2 205 203 188 10 198 196 199 197 197 206 205 3M 3 202 204 181 1 1 202 199 199 205 205 206 205 4M 4 175 177 153 12 189 185 186 190 196 199 200 IF 5 149 145 132 13 146 146 147 152 158 190 147 2F 6 135 136 121 14 150 147 139 156 160 136 151 3F 7 145 150 127 15 135 134 132 138 142 140 138 4F 8 136 136 114 16 122 124 118 135 141 137 140 Print No: 0019 Printed: 24-JUN-02 Xybion protocol number: MIN 313 MIN 313/023622 MIN 313/023622 137 : APPENDIX 4 (Food consumption - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM): 1 CONTROL 0 GROUP CAGE WEEK -i WEEK i 1M 17 186 169 4M 16 196 161 IF 19 136 138 4F 20 137 122 23 -- POSP----- 30 100 WEEK 2 WEEK 3 185 197 177 188 145 149 131 136 Print No: 0010 Printed: 24-JUN-02 Xybion protocol number: MIN 313A APPENDIX 5 Haematology - individual values MIN 313/023622 138 : Day 91 Group 1 Compound Control Exposure level (ppm) : 0 Group / sex Animal number Hct L/L 1M 1 CTD 2 0.417 3 CTD 4 0.437 5 0.441 2M 6 0.451 7 0.438 8 0.455 9 0.437 10 0.423 3M 11 12 13 14 15 0.481 0.429 0.447 0.438 0.439 4M CTD 16 0.432 17 0.459 18 0.433 19 0.450 20 0.377 Clotted sample 2 34 --POSF....... 30 100 300 Hb RBC Retie E/dL x10I2/L % CTD CTD CTD 15.3 7.94 2.13 CTD CTD CTD 15.4 8.39 3.17 15.5 8.40 3.04 16.1 8.39 1.80 15.4 8.26 2.06 15.8 8.10 2.10 15.6 8.42 2.40 15.4 7.74 1.75 17.1 8.61 1.87 15.4 8.11 2.43 15.9 8.62 2.28 15.3 8.25 1.33 15.5 8.12 2.10 15.5 8.15 1.90 16.5 8.56 1.59 15.6 7.89 1.37 16.1 8.13 1.68 12.4 6.44 10.68 MCH Pg CTD 19.3 CTD 18.3 18.5 19.1 18.6 19.5 18.5 19.9 19.9 19.0 18.4 18.6 19.1 19.1 19.3 19.8 19.8 19.3 MCHC fi/dL CTD 36.6 CTD 35.1 35.2 35.6 35.1 34.8 35.8 36.4 35.7 36.0 35.5 35.0 35.3 36.0 36.0 36.1 35.9 32.9 MCV fL CTD 52.5 CTD 52.1 52.5 53.8 53.0 56.1 51.9 54.7 55.8 53.0 51.9 53.1 54.1 53.0 53.6 54.9 55.3 58.6 WBC x109/L CTD 16.92 CTD 9.87 9.86 12.24 11.36 21.76 10.76 11.21 10.61 9.88 11.61 11.75 11.07 11.32 10.12 12.33 12.44 12.36 N x109/L CTD 4.91 CTD 1.04 1.98 2.94 2.35 7.71 2.21 1.05 1.10 2.65 1.32 1.32 1.57 0.95 1.15 0.58 0.79 1.01 APPENDIX 5 (Haematology - continued) MIN 313/023622 139 : Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 234 -- POSF-- 30 100 300 Group Animal L E B M LUC Pit PT APTT /sex number x 109/L x109/L x109/L x109/l xlO'/L x 109/L sec sec 1M 1 CTD CTD CTD CTD CTD CTD CTD CTD 2 10.87 0.32 0.06 0.52 0.25 925 12.8 14.9 3 CTD CTD CTD CTD CTD CTD CTD CTD 4 8.24 0.11 0.03 0.27 0.19 957 13.2 23.1 5 7.39 0.10 0.03 0.28 0.08 537 CTD CTD 2M 6 8.81 0.19 0.03 0.21 0.06 888 13.5 15.9 7 8.43 0.18 0.03 0.31 0.06 961 14.0 16.9 8 13.10 0.23 0.10 0.47 0.15 223 CTD CTD 9 7.87 0.13 0.03 0.39 0.12 1108 CTD CTD 10 9.83 0.16 0.03 0.10 0.04 909 14.2 16.9 3M 11 12 13 14 15 9.07 0.19 0.03 0.18 0.05 580 6.89 0.11 0.02 0.18 0.04 869 9.76 0.28 0.04 0.17 0.04 1080 9.95 0.14 0.05 0.23 0.06 924 8.92 0.27 0.02 0.22 0.07 801 14.2 23.9 14.7 17.1 14.6 18.4 13.9 19.5 13.6 16.3 4M 16 10.15 0.09 0.02 0.06 0.04 803 17 8.72 0.09 0.03 0.10 0.03 952 18 11.40 0.07 0.05 0.17 0.06 1107 19 11.33 0.13 0.04 0.10 0.05 1058 20 10.71 0.15 0.04 0.34 0.11 1416 13.2 16.0 13.5 23.4 14.0 20.8 12.9 17.0 12.5 17.2 CTD Clotted sample APPENDIX 5 (Haematology - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 234 ---POSF-- 30 100 300 Group / sex Animal number Anisocytosis Microcytosis Macro- Hypo- Hyper- cytosis chromasia chromasia 1M 1 CTD CTD CTD CTD CTD 2 - +- - - 3 CTD CTD CTD CTD CTD 4 +- - - 5-+- - - 2M 6 _ +. . - 7-+ -" 8 9 _+ _ _ _ 10 - - - - " 3M 11 _ . . . - 12 - + - - - 13 - + - - - 14 - - - - - 15 - - - - 4M 16 _ . _ - - 17 - - - - - 18 - - - - - 19 - - - - - 20 - - - - - CTD Clotted sample MIN 313/023622 APPENDIX 5 (Haematology - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group /sex Animal number Hct L/L IF 21 CTD 22 0.435 23 NS 24 0.410 25 0.425 2F 26 27 28 29 30 0.440 0.411 0.432 0.423 0.420 3F 31 32 33 34 35 0.420 0.404 0.411 0.424 0.448 4F 36 37 38 39 40 0.437 0.437 0.443 0.417 0.423 CTD NS Clotted sample No sample obtained 2 34 -- POSF-- 30 100 300 Hb RBC Retie E/dL x10I2/L % CTD CTD CTD 15.6 7.90 2.04 NS NS NS 14.9 7.29 2.89 15.0 7.61 2.01 15.8 7.93 2.44 14.7 7.58 1.52 15.3 7.62 1.85 15.0 7.33 2.15 14.8 7.66 2.49 14.9 7.72 2.02 14.2 7.44 4.18 14.8 7.16 1.98 15.6 7.60 1.84 16.0 7.81 2.62 15.6 7.52 2.03 15.8 8.08 1.72 15.9 8.42 1.60 15.2 7.82 1.21 15.4 7.89 2.15 MCH PR CTD 19.7 NS 20.4 19.6 19.9 19.4 20.1 20.4 19.3 19.2 19.1 20.7 20.5 20.5 20.8 19.5 18.9 19.4 19.5 MCHC E/dL CTD 35.8 NS 36.3 35.2 35.8 35.8 35.5 35.4 35.3 35.4 35.1 36.0 36.7 35.8 35.7 36.2 35.8 36.4 36.3 MCV fL CTD 55.0 NS 56.2 55.8 55.5 54.2 56.6 57.7 54.8 54.4 54.3 57.4 55.8 57.4 58.2 54.0 52.7 53.3 53.6 WBC x109/L CTD 6.10 NS 7.11 11.72 7.26 10.82 8.15 8.91 10.22 8.08 9.74 6.97 14.35 8.28 11.38 7.75 8.44 9.66 10.39 N x109/L CTD 0.85 NS 0.90 2.12 0.96 2.89 1.26 1.80 0.90 1.45 1.15 0.68 3.44 1.45 1.92 0.67 1.50 0.90 1.27 MIN 313/023622 APPENDIX 5 (Haematology - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number L x109/L IF 21 CTD 22 4.84 23 NS 24 5.87 25 9.04 2F 26 27 28 29 30 5.77 7.17 6.53 6.70 8.40 3F 31 6.39 32 8.17 33 5.91 34 10.10 35 6.35 4F 36 37 38 39 40 8.74 6.79 6.68 8.39 8.72 CTD NS Clotted sample No sample obtained 2 34 ---POSE--- 30 100 300 E x109/L B x io 9/L M x 107l CTD 0.16 NS 0.12 0.17 CTD 0.01 NS 0.02 0.03 CTD 0.19 NS 0.15 0.25 0.15 0.02 0.26 0.23 0.02 0.40 0.09 0.01 0.21 0.15 0.02 0.19 0.36 0.02 0.35 0.10 0.02 0.08 0.14 0.03 0.19 0.07 0.02 0.23 0.30 0.06 0.36 0.19 0.01 0.22 0.24 0.03 0.35 0.12 0.01 0.12 0.05 0.02 0.14 0.21 0.03 0.09 0.16 0.02 0.16 LUC xlO'/L CTD 0.04 NS 0.05 0.10 0.10 0.12 0.05 0.05 0.19 0.04 0.06 0.05 0.08 0.06 0.10 0.04 0.05 0.05 0.06 Pit x109/L CTD 937 NS 1156 1080 871 948 955 906 997 888 948 839 954 831 1183 855 1005 956 1108 PT sec CTD 13.6 NS 14.5 13.3 13.3 CTD 13.5 13.5 CTD 14.6 13.1 14.4 14.6 13.7 13.2 12.9 12.4 13.3 12.8 APTT sec CTD 13.3 NS 19.1 11.2 17.6 CTD 20.4 18.1 CTD 17.4 14.9 16.4 20.1 17.7 18.3 20.2 19.3 15.8 16.6 MIN 313/023622 APPENDIX 5 (Haematology - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 ---POSF-- 30 100 300 Group / sex Animal number Anisocytosis Micro cytosis Macro- Hypo- Hyper- cytosis chromasia chromasia IF 21 CTD CTD CTD CTD CTD 22 - - - - - 23 NS NS NS NS NS 24 - - - - 25 - - - - - 2F 26 _ _ _ . 27 - - - 28 - - - 29 - - - 30 - - - - - - - 3F 31 _ _ _ _ - 32 - - - - - 33 - - - - - 34 - - - - - 35 - - 4F 36 _ _ _ _ _ 37 - - - - 38 - - - - 39 - - - - 40 - - - - - CTD Clotted sample NS No sample obtained MIN 313/023622 APPENDIX 5 (Haematology - continued) Day 121 Group 1 Compound Control Exposure level (ppm) : 0 Group / sex Animal number Hct L/L 1M 41 42 43 44 45 0.388 0.418 0.393 0.441 0.412 2M 46 47 48 49 50 0.390 0.379 0.406 0.394 0.404 3M 51 52 53 54 55 0.416 0.413 0.399 0.419 0.418 4M 56 57 58 59 60 0.413 0.391 0.439 0.415 0.410 2 34 --POSF....... 30 100 300 Hb RBC Retie R/dL x1012/L % 14.5 7.31 2.66 15.6 7.94 2.27 15.1 7.38 2.94 16.3 7.99 2.44 15.3 8.32 1.99 14.5 7.43 2.75 13.9 7.33 3.59 14.8 8.03 2.97 14.8 7.44 1.87 14.9 7.74 2.17 15.2 7.71 2.29 15.0 7.46 2.83 14.9 7.53 2.07 15.4 7.94 2.69 15.4 7.68 2.02 15.5 8.10 2.37 14.3 7.37 2.73 16.0 8.17 2.48 15.5 7.90 2.36 15.1 7.65 2.27 MCH PR 19.8 19.7 20.5 20.4 18.4 19.5 19.0 18.4 19.9 19.2 19.7 20.1 19.8 19.4 20.1 19.1 19.4 19.6 19.6 19.7 MCHC R/dL 37.4 37.4 38.4 36.9 37.1 37.2 36.8 36.4 37.7 36.8 36.6 36.3 37.3 36.8 36.9 37.5 36.7 36.5 37.3 36.8 MCV fL 53.0 52.6 53.3 55.2 49.6 52.5 51.7 50.6 52.9 52.2 53.9 55.3 52.9 52.8 54.4 50.9 53.0 53.8 52.5 53.6 WBC xlO'/L 9.30 9.12 10.82 11.77 13.86 7.40 9.98 13.26 8.53 9.59 14.42 20.80 7.38 8.83 9.92 13.34 10.97 10.28 11.13 9.98 N x109/L 0.78 0.84 2.01 1.24 3.27 1.57 2.78 2.58 2.51 0.98 1.53 7.34 0.95 1.49 1.85 4.02 3.35 1.03 1.65 1.24 MIN 313/023622 APPENDIX 5 (Haematology - continued) Day 121 Group Compound Exposure level (ppm) 1 Control 0 2 34 --POSF--- 30 100 300 Group Animal L E B M LUC Pit PT APTT /sex number x 109/L x109/L x109/L x 109/L x l 0 9/L x109/L sec sec 1M 41 7.88 0.10 0.01 0.30 0.23 1123 42 7.59 0.13 0.03 0.32 0.21 1045 43 8.02 0.20 0.04 0.27 0.28 1028 44 9.84 0.18 0.03 0.22 0.26 860 45 10.16 0.13 0.06 0.15 0.09 990 12.9 17.6 13.0 21.2 12.6 20.4 13.2 20.9 12.8 15.7 2M 46 5.51 0.13 0.02 0.10 0.06 1021 13.8 17.9 47 6.83 0.08 0.02 0.16 0.11 1147 CTD CTD 48 10.06 0.07 0.04 0.36 0.15 887 14.4 17.6 49 5.61 0.07 0.01 0.21 0.11 829 13.5 17.5 50 8.26 0.07 0.03 0.16 0.11 1028 14.2 20.0 3M 51 12.51 0.11 0.06 0.15 0.06 799 52 12.31 0.30 0.08 0.48 0.31 910 53 6.15 0.09 0.02 0.11 0.06 879 54 6.97 0.10 0.02 0.16 0.10 789 55 7,59 0,14 0.04 0.22 0.09 960 14.6 15.7 15.2 19.9 14.5 14.1 13.9 16.3 14.9 14.1 4M 56 57 58 59 60 8.52 0.28 0.05 0.29 0.19 1063 7.20 0.15 0.03 0.14 0.09 938 9.00 0.07 0.03 0.07 0.08 903 8.81 0.29 0.03 0.24 0.10 1098 8.31 0.09 0.02 0.19 0.13 925 13.8 14.7 13.6 14.3 14.4 18.0 13.0 15.5 13.8 16.1 CTD Clotted sample MIN 313/023622 APPENDIX 5 (Haematology - continued) MIN 313/023622 146 : Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 2 30 Group /sex 1M 2M Animal number 41 42 43 44 45 46 47 48 49 50 Anisocytosis - - _ + - - Microcytosis + + + - ++ + ++ + + 3M 51 52 53 54 55 4M 56 57 58 59 60 _ - - - ,, - . + - - + - - + - 34 POSF..... 100 300 Macro- Hypo- Hyper- cytosis chromasia chromasia -- - - -- - -- -- . -- - -- - -- -- - -" _ _- -- -- - --- --- _----- - APPENDIX 5 (Haematology - continued) Day 121 Group 1 Compound Control Exposure level (ppm) : 0 Group /sex Animal number Hct L/L IF 61 62 64 65 0.377 0.395 0.392 0.401 2F 66 0.403 67 0.387 68 0.408 69 CTD 70 0.389 3F 71 0.363 72 0.395 73 0.384 74 0.314 75 0.409 4F 76 0.413 77 0.365 78 0.375 79 0.397 80 0.412 CTD Clotted sample 2 34 --POSF....... 30 100 300 Hb RBC Retie g/dL x10I2/L % 14.7 7.04 1.91 14.9 7.19 2.74 14.7 7.25 2.37 14.8 7.22 1.90 15.3 7.34 14.8 7.36 15.4 7.76 CTD CTD 14.6 7.32 1.90 2.02 2.28 1.88 1.93 13.8 6.80 2.30 15.2 7.57 2.26 14.7 7.36 2.40 11.6 6.02 1.95 15.4 7.07 2.27 15.8 7.30 2.15 13.4 6.94 2.18 14.0 7.02 2.62 15.0 7.41 2.43 15.3 7.51 1.76 MCH Pg 20.3 20.7 20.3 20.6 20.8 20.1 19.8 CTD 19.9 20.3 20.0 19.9 19.3 21.7 21.7 19.3 19.9 20.3 20.4 MCHC g/dL 37.8 37.7 37.6 37.0 38.0 38.2 37.7 CTD 37.4 38.0 38.3 38.2 37.1 37.6 38.4 36.7 37.2 37.9 37.2 MCV fL 53.6 54.9 54.0 55.6 54.9 52.5 52.5 CTD 53.2 53.3 52.2 52.2 52.1 57.8 56.6 52.6 53.4 53.6 54.8 WBC xl09/L 8.85 6.62 7.72 8.91 4.90 6.85 5.57 CTD 6.85 7.11 5.15 4.18 3.21 7.65 4.54 3.45 7.32 4.11 6.95 N x109/L 0.92 1.41 1.59 0.99 0.71 0.80 0.71 CTD 2.48 1.39 1.04 1.10 0.34 1.46 0.54 0.39 1.52 0.58 1.46 MIN 313/023622 APPENDIX 5 (Haematology - continued) MIN 313/023622 148 : Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 2 34 -- POSF-- 30 100 300 Group Animal L E B M LUC Pit PT APTT /sex number xl09/L x109/L x109/L x 109/L xl09/L x109/L sec sec IF 61 62 64 65 7.44 0.12 0.02 0.23 0.12 947 4.84 0.07 0.01 0.19 0.11 869 5.56 0.27 0.02 0.19 0.09 1107 7.35 0.16 0.03 0.21 0.17 877 13.7 18.8 13.3 17.4 13.1 17.1 12.6 15.0 2F 66 3.94 0.09 0.01 0.10 0.06 685 13.7 16.7 67 5.73 0.11 0.01 0.11 0.10 877 CTD CTD 68 4.57 0.08 0.01 0.14 0.06 1001 13.9 18.7 69 CTD CTD CTD CTD CTD CTD 13.1 16.8 70 3.74 0.19 0.02 0.26 0.16 796 13.4 16.9 3F 71 72 73 74 75 5.41 0.10 0.01 0.12 0.08 672 3.90 0.07 0.01 0.09 0.04 491 2.95 0.05 0.01 0.05 0.03 564 2.69 0.07 0.00 0.04 0.06 890 5.69 0.12 0.02 0.21 0.15 800 14.6 17.7 13.8 15.7 CTD CTD 14.7 19.3 14.2 18.4 4F 76 77 78 79 80 3.62 0.19 0.01 0.08 0.10 865 2.93 0.05 0.00 0.04 0.04 745 5.39 0.09 0.01 0.13 0.18 831 3.33 0.06 0.01 0.07 0.06 479 5.10 0.12 0.01 0.14 0.12 828 14.8 23.4 12.9 15.6 13.3 18.6 CTD CTD 13.8 17.5 CTD Clotted sample APPENDIX 5 (Haematology - continued) Day 121 Group 1 Compound : Control Exposure level (ppm) : 0 2 34 -- POSF-- 30 100 300 Group / sex Animal number Anisocytosis Micro cytosis Macro- Hypo- Hyper- cytosis chromasia chromasia IF 61 62 - - - - 64 - - - - - 65 - - - - - 2F 66 __ 67 - - - - - 68 - - - - - 69 CTD CTD CTD CTD CTD 70 - - - - - 3F 71 __,, _ 72 - - - - - 73 +- - - - 74 - - - - - 75 - - - - - 4F 76 _ _ _ _ 77 - - - - 78 - - - - 79 - - - - 80 - - - - - CTD Clotted sample MIN 313/023622 APPENDIX 6 Biochemistry - individual values Day 91 Group Compound Exposure level (ppm) Group / sex Animal number 1M 1 2 3 4 5 2M 6 7 8 9 10 3M 11 12 13 14 15 1 Control 0 ALP U/L 150 217 186 198 260 290 212 213 218 354 228 302 259 264 342 2 34 --POSF...... 30 100 300 ALT AST LDH U/L U/L Total U/L 32 81 693 49 128 1131 43 92 232 52 101 176 46 94 254 61 105 187 67 114 266 56 235 1078 45 109 304 78 153 384 66 143 547 43 76 172 64 97 297 62 108 249 56 86 215 LDH1 U/L 30 68 11 7 17 21 28 34 21 37 54 10 16 14 17 LDH2 U/L 29 37 12 7 12 13 19 43 14 17 31 9 12 11 11 LDH3 U/L 21 23 6 4 5 5 7 38 8 8 11 5 7 6 7 LDH4 U/L 21 66 7 5 15 12 15 73 21 15 21 5 12 12 6 LDH5 U/L 592 938 196 153 206 136 197 889 240 308 430 143 249 206 173 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number CPK Total U/L 1M 1 298 2 424 3 129 4 187 5 174 2M 6 220 7 152 8 7382 9 227 10 186 3M 11 238 12 149 13 155 14 323 15 144 234 --POSF-...... 30 100 300 SDH Bili Urea U/L pmol/L mmol/L 17.2 2 18.8 2 14.3 1 12.0 1 14.4 1 14.3 1 12.9 2 10.7 1 16.6 2 11.2 1 15.5 1 12.4 2 11.5 1 9.7 1 8.6 2 5.90 8.02 5.61 5.47 5.26 5.86 5.87 7.59 6.12 7.37 6.07 5.55 6.02 5.53 5.82 Creat pmol/L 47 55 51 55 56 54 54 53 51 49 53 51 46 46 45 Glue mmol/L 6.62 6.75 5.78 7.45 5.73 5.07 5.23 4.82 5.61 6.28 5.59 5.58 6.00 5.68 5.60 Choi mmol/L 1.70 1.35 1.27 1.10 1.78 0.99 1.27 1.39 0.97 0.61 0.65 0.71 0.92 1.43 1.09 Trig mmol/L 0.55 0.23 0.42 0.26 0.73 0.20 0.40 0.21 0.45 0.35 0.33 0.25 0.35 0.46 0.48 Na mmol/L 145 140 142 142 144 145 144 146 145 144 146 142 145 146 143 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) MIN 313/023622 152 : Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 234 -- POSF-- 30 100 300 Group Animal K Cl Ca Phos Total Prot Alb al a2 Beta / sex number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L g/L g/L 1M 2M 3M INS 1 4.0 2 5.1 3 3.7 4 3.6 5 3.8 6 3.5 7 4.0 8 4.0 9 3.8 10 3.6 11 4.5 12 3.9 13 4.4 14 4.0 15 3.9 Insufficient sample 107 106 105 106 106 107 108 110 106 108 110 106 107 108 106 2.79 1.90 61 2.57 2.23 62 2.72 1.73 64 2.68 2.14 65 2.64 2.10 64 2.70 2.00 68 2.74 2.07 65 2.62 1.97 68 2.67 1.99 73 2.74 2.24 60 2.77 2.20 61 2.84 2.08 70 2.77 2.28 63 2.72 2.03 65 2.77 2.28 65 29 12 4 12 INS INS INS INS 29 12 3 14 30 12 3 15 28 9 3 17 32 12 30 10 29 13 31 12 30 10 3 15 4 15 3 17 3 17 3 12 32 10 32 13 28 12 29 13 30 13 3 12 3 16 4 12 3 14 3 14 APPENDIX 6 (Biochemistry - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) 0 Group / sex Animal number Gamma g/L 2 34 ---POSE-- 30 100 300 A/G Ratio Alb % al % a2 Beta Gamma %%% 1M 1 4 2 INS 36 45 57 2M 6 7 76 86 9 10 10 5 3M 11 4 12 6 13 6 14 6 15 4 INS Insufficient samples 0.91 INS 0.83 0.86 0.78 0.89 0.86 0.74 0.74 1.00 1.10 0.84 0.80 0.81 0.86 47.7 INS 45.6 45.7 44.4 46.7 45.8 42.6 42.9 49.6 52.1 45.2 45.1 44.7 46.7 20.1 INS 19.1 18.5 14.5 17.2 16.0 18.9 15.9 17.1 16.7 18.8 18.7 19.5 19.6 5.8 19.5 INS INS 4.4 21.6 4.7 23.1 4.5 26.0 4.4 21.5 5.6 23.0 4.7 24.4 3.8 23.1 4.7 20.7 4.5 20.2 4.8 22.3 6.5 19.6 5.1 21.8 4.7 22.3 6.8 INS 9.3 7.9 10.6 10.2 9.5 9.5 14.3 8.0 6.5 9.0 10.0 8.8 6.6 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) MIN 313/023622 154 : Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number ALP U/L 4M 16 366 17 346 18 302 19 399 20 224 234 -- POSF-- 30 100 300 ALT AST LDH U/L UL Total U/L 53 100 270 68 125 286 66 91 268 62 96 258 87 101 139 LDH1 U/L 11 26 12 15 15 LDH2 U/L 14 18 10 16 17 LDH3 U/L 9 6 7 6 6 LDH4 U/L 12 11 7 19 14 LDH5 U/L 224 226 232 201 87 APPENDIX 6 (Biochemistry - continued) Day 91 Group Compound Exposure level (ppm) Group / sex Animal number 4M 16 17 18 19 20 1 Control 0 CPK Total U/L 258 175 137 124 147 2 34 ---POSF----- 30 100 300 SDH Bili Urea U/L pmol/L mmol/L 13.7 1 12.2 1 12.7 1 11.4 1 10.1 1 6.59 6.17 6.68 7.34 7.27 Creat pmol/L 45 52 46 48 48 Glue mmol/L 6.47 5.26 5.07 5.88 5.79 Choi mmol/L 0.44 0.60 0.54 0.91 1.51 Trig mmol/L 0.05 0.17 0.29 0.15 0.54 Na mmol/L 141 145 143 144 142 APPENDIX 6 (Biochemistry - continued) Day 91 Group 123 4 Compound Control --POSF----- Exposure level (ppm) 0 30 100 300 L /1 ON Group Animal K Cl Ca Phos Total Prot Alb al a2 Beta / sex number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L g/L g/L 4M 16 17 18 19 20 4.4 106 4.3 108 4.1 105 3.9 105 4.0 106 2.83 2.02 65 35 12 2.72 2.18 67 34 11 2.81 1.94 70 36 13 2.79 2.10 72 35 14 2.71 1.92 60 27 11 2 12 3 14 3 13 3 14 3 15 MIN 313/023622 MIN 313/023622 157 : APPENDIX 6 (Biochemistry - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number Gamma g/L 234 -- POSE-- 30 100 300 MG Ratio Alb % al % 4M 16 17 18 19 20 4 6 6 7 4 1.17 53.1 1.03 50.0 1.06 50.8 0.95 48.2 0.82 45.6 18.6 16.9 18.0 18.8 18.5 a2 Beta Gamma %%% 3.5 18.5 6.4 4.1 20.5 8.5 3.7 18.9 8.6 3.9 19.1 10.0 5.1 24.9 5.9 APPENDIX 6 (Biochemistry - continued) Day 91 Group Compound Exposure level (ppm) 1 Control 0 2 34 --POSF...... 30 100 300 Group Animal ALP ALT AST LDH / sex number UL U/L U/L Total U/L IF 21 197 43 121 780 22 189 36 160 516 23 NS NS NS NS 24 128 30 76 355 25 133 42 75 214 2F 26 167 39 85 269 27 148 42 104 181 28 136 71 132 214 29 95 33 78 163 30 129 96 144 259 3F 31 97 28 70 167 32 167 29 79 163 33 127 38 80 143 34 117 116 195 309 35 113 35 91 255 NS No sample Animal 23 died at bleed LDH1 U/L 40 29 NS 14 9 13 19 11 11 12 10 12 13 15 11 LDH2 U/L 41 39 NS 17 9 13 23 12 10 10 7 12 9 16 13 LDI13 U/L 15 24 NS 8 5 3 21 10 5 8 7 4 3 7 8 LDH4 U/L 42 39 NS 11 9 8 6 12 12 12 6 6 7 11 13 LDH5 U/L 643 385 NS 304 181 232 112 169 126 218 138 129 111 259 209 APPENDIX 6 (Biochemistry - continued) MIN 313/023622 159 : Day 91 Group 1 Compound Control Exposure level (ppm) : 0 2 34 --POSF----- 30 100 300 Group / sex Animal number CPK Total U/L SDH Bili Urea U/L pmol/L mmol/L IF 21 298 9.4 2 9.20 22 1625 14.5 2 8.07 23 NS NS NS NS 24 181 12.8 2 7.09 25 152 17.6 2 6.83 2F 26 200 11.4 3 27 100 9.1 2 28 111 11.0 3 29 101 10.9 3 30 140 19.4 2 6.86 6.88 8.08 6.80 7.15 3F 31 107 7.9 2 32 116 11.1 2 33 100 9.2 2 34 155 14.8 2 35 182 10.7 2 7.01 5.39 5.90 6.36 6.38 NS No sample Animal 23 died at bleed Crt pmol/L 74 69 NS 63 59 57 56 62 55 58 55 54 53 55 60 Glue mmol/L 4.59 4.42 NS 6.00 6.13 6.38 4.83 5.91 6.65 6.91 6.47 6.87 6.27 7.30 5.48 Choi mmol/L 2.08 2.35 NS 2.33 1.86 2.50 1.64 2.52 2.42 2.79 1.91 L.96 2.28 2.99 L.50 Trig mmol/L 0.56 0.82 NS 0.41 0.71 0.36 0.33 0.50 0.43 0.38 0.30 0.35 0.45 0.52 0.43 Na mmol/L 142 143 NS 143 141 141 142 144 145 145 144 141 144 141 143 APPENDIX 6 (Biochemistry - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number K mmol/L 2 34 -- POSF-- 30 100 300 Cl mmol/L Ca mmol/L Phos mmol/L Total Prot g/L Alb g/L al a2 Beta g/L g/L IF 21 4.1 105 2.75 1.49 71 37 ll 4 13 22 3.4 105 2.90 1.88 71 37 10 3 14 23 NS NS NS NS NS NS NS NS NS 24 3.2 106 2.80 1.79 68 35 10 4 14 25 3.4 106 2.79 1.24 71 37 10 3 14 2F 26 27 28 29 30 3.4 106 3.5 107 3.1 108 3.0 107 3.2 106 2.85 1.43 75 43 10 2.69 1.50 71 34 11 3.00 1.51 76 42 12 2.80 1.38 79 41 11 2.86 1.69 73 39 11 4 14 3 15 3 14 5 13 4 13 3F 31 2.8 107 32 2.9 104 33 3.2 107 34 3.2 103 35 3.0 105 2.83 1.72 74 38 10 2.68 1.47 65 34 10 2.73 1.58 73 40 10 2.84 1.53 72 38 11 2.68 1.61 67 36 11 4 14 3 13 4 13 4 14 3 14 NS No sample Animal 23 died at bleed MIN 313/023622 APPENDIX 6 (Biochemistry - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number Gamma g/L 234 -- POSF-- 30 100 300 A/G Ratio Alb % al % a2 Beta Gamma %%% IF 21 7 1.09 51.8 15.4 5.6 18.0 9.2 o\ 22 6 1.09 52.1 14.2 4.8 20.0 8.9 23 NS NS NS NS NS NS NS 24 5 1.06 51.8 14.8 5.2 20.9 7.2 25 7 1.09 52.2 13.8 4.1 19.9 10.0 2F 26 5 27 9 28 5 29 9 30 6 1.34 57.2 0.92 47.5 1.24 54.8 1.08 52.5 1.15 54.0 13.5 14.8 15.6 13.4 14.6 4.7 18.1 6.5 4.5 20.8 12.4 4.1 19.0 6.5 5.7 16.5 12.0 5.0 17.9 8.5 3F 31 8 32 5 33 6 34 6 35 4 1.06 50.9 1.10 53.0 1.21 54.8 1.12 52.5 1.16 53.3 13.9 14.7 14.0 15.5 15.9 5.3 18.7 11.2 4.9 19.5 7.9 5.4 18.2 7.6 5.0 19.3 7.7 4.1 20.2 6.6 NS No sample Animal 23 died at bleed 313/023622 APPENDIX 6 (Biochemistry - continued) 162 : Day 91 Group Compound Exposure level (ppm) Group / sex Animal number 4F 36 37 38 39 40 1 Control 0 ALP UL 92 142 124 94 162 2 34 --POSF--- 30 100 300 ALT AST LDH U/L U/L Total U/L 28 73 325 34 95 197 45 98 287 48 115 482 80 149 225 LDH1 U/L 16 12 14 64 16 LDH2 U/L 14 17 11 54 15 LDH3 U/L 8 6 5 12 5 LDH4 U/L 14 10 6 23 11 LDH5 U/L 273 152 251 330 178 APPENDIX 6 (Biochemistry - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number CPK Total U/L 4F 36 137 37 96 38 158 39 197 40 129 23 -- POSF-- 30 100 4 300 SDH Bili Urea U/L pmol/L mmol/L 12.9 2 16.5 2 16.8 2 15.6 1 15.6 2 5.75 8.15 7.63 12.73 7.32 Creat pmol/L 51 61 55 74 58 Glue mmol/L 5.20 5.25 6.67 5.91 6.46 Choi mmol/L 1.65 1.94 1.87 2.17 2.27 Trig mmol/L 0.49 0.33 0.28 0.41 0.42 Na mmol/L 144 144 144 141 143 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) Day 91 Group 12 34 Compound : Control --POSF....... Exposure level (ppm) : 0 30 100 300 o\ Group Animal K Cl Ca Plios Total Prot Alb al a2 Beta /se x number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L g/L g/L 4F 36 37 38 39 40 3.2 104 3.2 104 3.4 106 3.8 104 3.4 105 2.83 1.41 82 2.77 1.67 73 2.76 1.81 75 2.78 1.61 74 2.80 1.55 78 45 40 42 42 40 12 11 11 12 12 4 13 3 14 4 13 3 11 3 16 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number Gamma g/L 2 34 -- POSF-- 30 100 300 A/G Ratio Alb % al % 4F 36 37 38 39 40 9 6 5 5 6 1.22 54.5 1.21 54.7 1.27 55.8 1.31 56.7 1.05 51.8 14.6 15.0 14.4 16.3 15.9 a2 Beta Gamma %%% 4.7 15.6 3.9 18.7 5.8 16.9 4.4 15.5 4.0 20.8 10.7 7.7 7.1 7.0 7.5 MIN 313/023622 Day 121 Group Compound Exposure level (ppm) Group / sex Animal number 1M 41 42 43 44 45 2M 46 47 48 49 50 3M 51 52 53 54 55 1 Control 0 ALP U/L 172 208 143 198 187 178 240 268 231 254 171 239 200 126 240 APPENDIX 6 (Biochemistry - continued) 234 --POSF--- 30 100 300 ALT AST LDH U/L U/L Total U/L 43 84 205 40 66 151 39 77 149 43 76 189 34 67 205 56 99 200 51 85 241 74 133 140 50 95 129 49 87 100 49 84 127 52 86 157 43 88 123 37 81 124 64 95 236 LDH1 U/L 13 12 11 10 10 21 11 13 12 14 13 16 19 12 14 LDH2 U/L 16 10 10 8 9 13 9 8 10 7 9 8 12 8 15 LDH3 U/L 25 7 4 21 11 6 6 4 6 3 3 4 3 8 9 LDII4 U/L 22 17 15 22 21 17 15 11 12 7 10 14 9 10 26 LDH5 U/L 129 104 109 130 154 144 200 105 89 70 92 114 80 85 171 MIN 313/023622 Day 121 Group :1 Compound : Control Exposure level (ppm) 0 Group /sex Animal number CPK Total U/L 1M 41 434 42 102 43 157 44 358 45 151 2M 46 188 47 144 48 136 49 205 50 90 3M 51 271 52 102 53 89 54 111 55 344 APPENDIX 6 (Biochemistry - continued) 2 34 ---POSF-- 30 100 300 SDH Bili Urea U/L pmol/L mmol/L Creat pmol/L 17.1 2 9.6 1 10.8 2 10.6 1 13.5 2 14.9 2 15.0 1 14.4 2 11.1 1 12.7 2 7.6 1 13.0 2 12.2 1 11.1 0 20.6 2 5.10 48 5.07 49 5.10 50 4.52 46 4.73 51 6.87 54 5.33 45 5.20 46 6.23 50 6.03 49 5.49 49 4.74 49 6.20 46 5.39 50 6.75 52 Glue mmol/L 5.80 6.76 5.86 7.17 5.54 5.98 6.93 8.59 6.91 7.30 7.74 6.39 6.29 6.75 5.61 Choi mmol/L 1.54 1.55 2.40 1.29 1.58 1.24 1.05 1.24 1.35 0.59 1.24 0.93 0.84 1.58 1.04 Trig mmol/L 0.33 0.55 0.54 0.27 0.50 0.55 0.33 0.32 0.36 0,34 0.37 0.40 0.31 0.47 0.37 Na mmol/L 142 143 143 142 142 144 142 141 142 143 142 142 142 144 142 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 234 -- POSF-- 30 100 300 Group Animal K Cl Ca Phos Total Prot Alb al a2 Beta /sex number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L g/L g/L 1M 41 4.1 102 2.62 1.80 66 32 ii 5 15 Os 00 42 3.6 101 2.58 1.74 66 32 13 4 14 43 3.5 102 2.61 1.90 65 29 12 5 15 44 4.2 103 2.64 1.84 63 32 11 5 13 45 3.8 102 2.54 1.66 68 32 11 5 14 2M 46 47 48 49 50 3.8 104 3.9 103 4.1 102 3.8 103 3.8 102 2.60 1.70 65 29 11 2.58 1.86 63 31 9 2.65 1.62 65 26 10 2.65 1.50 62 29 11 2.58 1.87 62 29 9 5 16 4 15 8 16 5 13 5 14 3M 51 52 53 54 55 4.1 102 3.5 103 3.5 102 4.1 104 3.9 103 2.51 1.81 62 29 10 2.55 1.49 65 28 10 2.52 1.77 58 27 9 2.61 1.85 63 28 12 2.56 1.73 59 28 10 4 14 3 16 4 14 4 14 4 13 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 2 30 3 POSF 100 Group / sex Animal number Gamma g/L A/G Ratio Alb % al % a2 Beta Gamma %%% 1M 41 42 43 44 45 2M 46 47 48 49 50 3M 51 52 53 54 55 3 4 4 2 6 5 4 6 4 5 5 8 5 4 4 0.94 48.1 0.94 47.8 0.81 44.6 1.03 50.2 0.89 47.5 16.3 19.4 18.4 17.5 16.1 7.7 22.7 5.9 21.4 8.0 22.7 7.6 21.0 7.2 20.8 5.2 5.5 6.3 3.7 8.4 0.81 44.0 16.5 7.3 24.8 0.97 48.8 14.3 6.7 23.3 0.67 39.4 14.8 12.3 24.5 0.88 46.9 17.3 8.4 21.5 0.88 46.5 14.4 8.5 21.9 7.3 6.9 9.1 6.0 8.7 0.88 46.9 0.76 43.6 0.87 46.2 0.80 45.2 0.90 48.2 16.5 14.9 15.8 19.7 17.0 6.9 21.8 7.9 5.3 24.2 11.9 6.9 23.3 7.9 5.9 23.0 6.2 6.6 21.2 7.0 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) Day 121 Group Compound Exposure level (ppm) Group / sex Animal number 1 Control 0 ALP U/L 4M 56 248 57 226 58 219 59 318 60 208 234 --POSF----- 30 100 300 ALT AST LDH U/L U/L Total U/L 192 206 228 52 83 146 39 82 191 56 92 166 85 129 266 LDH1 U/L LDI12 U/L LDH3 U/L LDH4 U/L LDH5 U/L 15 13 8 19 174 10 11 8 14 103 17 15 7 15 136 13 14 8 15 116 29 20 11 32 174 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) Day 121 Group Compound Exposure level (ppm) Group / sex Animal number 4M 56 57 58 59 60 1 Control 0 CPK Total U/L 163 309 214 304 475 234 --POSF----- 30 100 300 SDH Bili Urea U/L pmol/L mmol/L 31.6 1 26.1 I 8.4 2 12.6 1 16.4 1 4.66 5.18 5.25 6.30 5.52 Crt pmol/L 48 51 44 50 46 Glue mmol/L 5.33 6.04 5.60 5.19 5.13 Choi mmol/L 1.42 1.58 1.30 1.25 1.32 Trig mmol/L 0.25 0.35 0.41 0.19 0.43 Na mmol/L 144 143 144 144 145 APPENDIX 6 (Biochemistry - continued) Day 121 Group 12 34 Compound Control --POSF----- Exposure level (ppm) 0 30 100 300 -toj Group Animal K Cl Ca Phos Total Prot Alb al a2 Beta / sex number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L g/L 4M 56 57 58 59 60 3.9 103 3.8 103 3.9 103 3.8 103 4.0 103 2.69 1.63 66 29 11 2.57 1.75 66 32 11 2.68 1.88 63 29 12 2.62 1.91 69 33 13 2.65 1.76 64 31 11 5 15 5 14 5 13 4 14 5 13 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) Day 121 Group :1 2 3 4 Compound : Control -- POSF-- Exposure level (ppm) : 0 30 100 300 u> Group Animal Gamma A/G Alb al a2 Beta Gamma /sex number g/L Ratio % % % % % 4M 56 57 58 59 60 6 5 4 5 4 0.78 44.4 0.94 48.0 0.85 46.5 0.92 47.8 0.94 48.7 17.1 16.2 18.7 19.4 17.2 7.0 22.4 6.9 21.6 7.3 21.3 5.2 20.9 7.6 20.6 9.0 7.3 6.2 6.8 5.9 MIN 313/023622 Day 121 Group Compound Exposure level (ppm) Group / sex Animal number IF 61 62 64 65 2F 66 67 68 69 70 3F 71 72 73 74 75 4F 76 77 78 79 80 APPENDIX 6 (Biochemistry - continued) 1 Control 0 ALP U/L 106 157 105 100 112 65 129 105 65 90 107 101 96 125 81 104 123 113 141 234 --POSF--- 30 100 300 ALT AST LDH U/L U/L Total U/L 48 84 140 26 67 86 157 298 449 38 67 161 46 89 437 123 291 392 34 82 88 43 90 651 35 66 213 31 77 226 38 119 488 34 80 248 60 76 81 28 69 215 63 108 284 34 64 111 38 98 239 48 88 431 38 74 176 LDH1 U/L 11 13 NR 10 13 15 10 13 15 11 15 23 13 9 14 11 7 24 12 16 L.DH2 U/L 8 10 NR 10 10 20 6 6 11 8 7 19 8 7 7 9 7 18 9 14 LDH3 U/L 7 3 NR 10 13 15 10 3 12 8 6 11 4 5 6 7 3 6 4 10 LDH4 U/L 11 7 NR 21 14 31 10 4 47 11 10 41 11 11 14 6 12 22 20 20 LDH5 U/L 104 54 NR 398 112 356 356 62 566 176 188 394 212 50 174 251 82 169 386 116 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number CPK Total U/L IF 61 78 62 77 64 147 65 112 2F 66 285 67 129 68 77 69 236 70 125 3F 71 129 72 943 73 123 74 82 75 148 4F 76 120 77 79 78 163 79 150 80 238 2 34 -- POSF-- 30 100 300 SDH Bili Urea U/L pmol/L mmol/L 15.1 2 10.7 2 34.7 3 15.8 3 13.2 2 26.7 1 10.8 2 17.9 2 16.0 1 14.6 2 15.3 2 12.8 2 8.9 2 13.9 2 18.9 2 11.5 1 57.6 1 16.2 1 10.9 1 7.86 5.94 6.31 7.29 6.68 5.77 7.35 5.47 5.88 6.45 5.62 5.83 6.38 6.35 8.86 5.68 6.66 7.05 6.12 Creat pmol/L 64 58 58 57 56 54 55 62 59 59 64 55 51 55 58 55 69 59 54 Glue mmol/L 7.02 7.35 6.18 8.76 6.03 6.74 7.12 8.61 7.32 7.65 5.95 7.98 7.16 6.65 6.62 7.79 9.17 7.99 5.77 Choi mmol/L 2.35 1.84 3.54 2.52 2.57 2.78 1.55 1.77 3.29 1.27 0.77 1.61 2.62 2.86 1.99 1.83 1.92 3.43 1.71 Trig mmol/L 0.45 0.19 0.44 1.03 0.65 0.53 0.49 0.44 0.46 0.28 0.32 0.30 0.55 0.50 0.45 0.40 0.35 0.64 0.49 Na mmol/L 141 140 140 138 141 141 142 143 141 142 140 143 142 142 143 142 143 145 141 MIN 313/023622 APPENDIX 6 (Biochemistry - continued) MIN 313/023622 176 : Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 234 ---POSF--- 30 100 300 Group Animal K Cl Ca Phos Total Prot Alb al a2 Beta /sex number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L g/L g/L IF 61 62 64 65 2F 66 67 68 69 70 3F 71 72 73 74 75 4F 76 77 78 79 80 3.2 100 3.5 102 3.3 100 3.2 100 3.6 103 3.3 101 2.9 101 3.7 104 3.5 102 3.3 103 3.7 101 3.2 103 3.2 103 3.2 103 3.5 104 3.4 104 3.5 102 3.9 107 3.6 103 2.90 1.84 75 43 10 2.70 1.35 73 41 9 2.71 1.46 72 37 9 2.80 1.66 74 44 8 2.82 1.56 68 38 8 2.68 1.34 69 35 8 2.66 1.31 70 39 9 2.68 1.14 70 39 9 2.84 1.23 78 42 11 2.63 1.43 66 36 8 2.56 1.52 67 37 9 2.55 1.64 68 35 10 2.75 1.04 72 40 11 2.71 1.47 78 40 10 2.62 1.20 70 38 9 2.73 1.39 67 38 7 2.70 2.05 70 38 8 2.66 1.53 73 40 9 2.83 1.97 73 39 11 4 14 4 14 4 15 4 14 5 13 5 14 4 14 4 13 4 15 4 13 3 13 3 14 3 14 4 14 4 13 4 15 5 15 5 14 4 13 APPENDIX 6 (Biochemistry - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number Gamma g/L IF 61 62 64 65 4 5 6 5 2F 66 67 68 69 70 4 7 4 5 6 3F 71 5 72 5 73 5 74 5 75 10 4F 76 77 78 79 80 5 3 5 5 7 234 -- POSF-- 30 100 300 A/G Ratio Alb % al % 1.34 57.7 1.28 56.6 1.06 51.3 1.47 59.2 1.27 55.5 1.03 50.6 1.26 55.1 1.26 55.8 1.17 54.2 1.20 55.2 1.23 54.5 1.06 51.3 1.25 55.4 1.05 51.2 1.19 54.9 1.31 56.0 1.19 54.3 1.21 54.3 1.15 53.4 12.8 12.0 13.1 10.4 12.0 11.9 12.9 13.4 14.7 12.6 13.4 15.3 15.2 13.2 13.1 11.1 11.2 12.8 14.4 a2 Beta Gamma %%% 5.6 18.9 5.0 19.4 5.9 20.8 5.2 18.7 5.0 7.0 9.0 6.5 6.9 19.3 7.0 20.6 5.8 20.3 5.8 17.9 4.9 19.2 6.2 9.9 5.9 7.2 7.1 5.5 19.7 7.0 4.6 19.9 7.6 5.0 20.9 7.5 4.0 19.1 6.4 4.7 18.4 12.5 5.6 18.8 6.7 21.7 7.1 20.9 6.6 19.4 4.8 18.0 7.6 4.6 6.5 6.8 9.4 MIN 313/023622 APPENDIX 7 Urinalysis - individuai values MIN 313/023622 178 : Day 29 Group 1 Compound : Control Exposure level (ppm) : 0 2 30 Group / sex Animal number App. Voi ml 3 -- POSF-- 100 PH 4 300 SG g/L 1M 81 MY 4.0 82 PY 4.2 83 MY 3.8 84 PY 5.6 85 PY 4.0 4M 86 MY 2.6 87 PY 4.0 88 MY 2.0 89 MY 3.4 90 PY 4.6 IF 91 MY 2.6 92 PY 3.4 93 PY 5.8 94 MY 2.4 95 PY 4.0 4F 96 MY 2.0 97 MY 1.8 98 MY 2.2 99 PY 4.8 100 MY 2.2 6.5 1092 6.8 1042 6.6 1041 6.7 1034 6.6 1047 6.2 1074 6.5 1046 6.2 1074 6.6 1062 6.6 1039 6.3 1045 6.1 1041 6.5 1036 6.6 1050 6.5 1039 6.2 1020 6.2 1074 5.7 1040 6.1 1033 6.0 1086 Prot g/L 1.01 1.29 1.41 1.19 0.78 1.05 0.93 0.97 1.44 1.20 0.56 0.42 0.32 0.40 0.45 0.46 0.46 0.69 0.30 0.44 Glue 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Keto 0 TR TR TR 0 TR TR TR TR TR 0 0 0 0 0 0 0 0 0 0 Bili Blood 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 APPENDIX 7 (Urinalysis - continued) Day 29 Group :1 Compound : Control Exposure level (ppm) : 0 2 30 Group / sex Animal number Epi Leuc 3 ---POSF-- 100 RBC 4 300 Cryst Sperm 1M 81 0 0 0 0 0 82 0 0 0 0 0 83 0 1 0 0 1 84 1 0 0 0 1 85 0 0 0 0 1 4M 86 0 0 0 0 0 87 0 0 0 0 0 88 0 0 0 0 2 89 0 0 0 0 0 90 0 0 0 0 1 IF 91 0 I 0 0 0 92 0 0 0 0 0 93 0 1 0 0 0 94 0 0 0 0 0 95 1 0 0 0 0 4F 96 1 0 0 0 0 97 0 0 0 0 0 98 1 0 0 0 0 99 0 0 0 0 0 100 0 0 0 0 0 Casts 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Abn. Na Total K Total mmol mmol 0 0.26 1.32 0 0.40 1.40 0 0.37 0.80 0 0.75 1.31 0 0.50 1.19 0 0.23 0.95 0 0.29 1.16 0 0.16 0.35 0 0.34 0.73 0 0.64 1.10 0 0.34 0.54 0 0.31 0.54 0 0.48 0.97 0 0.39 0.46 0 0.44 0.88 0 0.20 0.57 0 0.13 0.60 0 0.17 0.55 0 0.39 0.78 0 0.21 0.72 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) Day 29 Group :1 Compound : Control Exposure level (ppm) : 0 234 ......POSF-- 30 100 300 Group / sex Animal number Cl Total mmol U-Na mmol/L U-K mmol/L U-Cl mmol/L 1M 81 0.29 66.1 331.0 71.4 82 0.41 94.5 332.5 97.6 83 0.39 96.7 211.0 101.7 84 0.58 133.1 234.0 103.1 85 0.32 124.5 297.5 80.3 4M 86 0.30 87.4 366 114.2 87 0.33 73.5 290 83.2 88 0.15 79.9 176.5 75.3 89 0.26 98.9 215.0 76.5 90 0.49 139.5 239.5 105.9 IF 91 0.29 132.4 208.0 112.2 92 0.29 91.6 159.5 84.1 93 0.34 82.0 167.0 59.0 94 0.25 162.7 190.5 103.1 95 0.36 109.1 221 90.2 4F 96 0.21 98.4 282.5 105.3 97 0.13 73.4 334.5 71.6 98 0.19 76.4 252.0 84.8 99 0.37 80.7 161.5 77.7 100 0.24 96.3 327.5 108.3 APPENDIX 7 (Urinalysis - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) 0 Group / sex Animal number App. 2 34 ---POSF-- 30 100 300 Voi PH SG ml g/L 1M 1 CPY 9.0 7.0 1031 2 CMY 4.2 7.7 1037 3 CMY 6.2 6.8 1043 4 CMY 4.0 7.4 1041 5 PY 12.4 7.8 1022 2M 6 CPY 6.8 7.2 1036 7 CMY 2.8 7.1 1062 8 CPY 7.8 7.5 1032 9 CPY 4.8 7.1 1042 10 CMY 4.2 7.7 1049 3M 11 CPY 8.0 7.2 1031 12 CPY 9.0 7.3 1030 13 CPY 7.4 6.9 1039 14 CPY 6.2 6.5 1036 15 CPY 5.2 6.7 1044 Prot g/L 0.53 0.98 1.17 1.40 0.64 0.62 1.12 0.90 1.47 0.91 0.82 0.70 0.91 0.89 0.94 Glue 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Keto 0 TR TR TR 0 TR TR TR TR TR TR TR TR + TR Bili Blood 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 MIN 313/023622 MIN 313/023622 182 : APPENDIX 7 (Urinalysis - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number Epi 234 -- POSF-- 30 100 300 Leuc RBC Cryst 1M 1 0 0 0 0 200 0 0 30000 40000 50 10 0 2M 6 1 0 0 0 7 10 0 0 800 00 90 0 0 0 10 0 0 0 0 3M 11 0 0 0 0 12 0 0 0 0 13 0 0 0 0 14 0 0 0 0 15 0 0 0 0 Sperm 1 0 0 0 0 0 0 0 0 0 0 0 1 2 1 Casts 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Abn. Na Total K Total mmol mmol 0 0.66 1.63 0 0.34 0.99 0 0.42 1.43 0 0.29 0.94 0 0.99 1.53 0 0.50 1.45 0 0.32 0.81 0 0.44 1.49 0 0.16 1.23 0 0.64 1.21 0 0.68 1.28 0 0.58 1.57 0 0.75 1.54 0 0.23 1.07 0 0.28 1.36 APPENDIX 7 (Urinalysis - continued) Day 91 Group 1 Compound : Control Exposure level (ppm) : 0 Group /sex Animal number Cl Total mmol 234 --POSF....... 30 100 300 U-Na mmol/L U-K mmol/L U-Cl mmol/L 1M 1 2 3 4 5 2M 6 7 8 9 10 3M 11 12 13 14 15 0.68 73.8 0.45 82.0 0.51 67.9 0.41 72.6 0.89 79.9 0.46 74.1 0.28 112.5 0.48 56.6 0.33 33.4 0.57 151.7 0.61 84.8 0.59 64.7 0.81 100.7 0.38 36.5 0.38 54.6 181.0 235.5 231.0 234.0 123.0 213.0 290.0 191.0 257.0 287.5 160.5 174.0 208.0 173.0 261.0 76.1 107.9 82.5 102.3 72.0 67.5 101.6 62.0 69.4 135.1 76.6 65.9 110.1 61.1 72.3 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) Day 91 Group 12 3 4 Compound : Control ---POSF-- Exposure level (ppm) : 0 30 100 300 o4o^ Group Animal App. Voi pH SG Prot Glue Keto Bili Blood / sex number ml g/L g/L 4M 16 CMY 2.8 6.8 1066 17 CMY 2.8 6.7 1068 18 CPY 5.2 6.7 1043 19 CPY 6.2 6.6 1043 20 CMY 3.0 6.4 1070 0.90 0 1.60 0 0.72 0 1.11 0 1.44 0 TR 0 TR 0 TR 0 TR 0 TR 0 0 0 0 0 0 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) 0 Group / sex Animal number Epi 2 34 --POSF--- 30 100 300 Leuc RBC Cryst 4M 16 0 0 0 0 17 0 0 0 0 18 0 0 0 0 19 0 0 0 0 20 0 0 0 0 Sperm 0 0 1 1 2 Casts 0 0 0 0 0 Abn. Na Total K Total mmol mmol 0 0.58 0.77 0 0.30 0.79 0 0.36 1.13 0 0.55 1.36 0 0.12 1.04 APPENDIX 7 (Urinalysis - continued) MIN 313/023622 186 : Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 234 -- POSF-- 30 100 300 Group / sex Animal number Cl Total mmol U-Na mmol/L U-K mmol/L U-Cl mmol/L 4M 16 17 18 19 20 0.38 205.6 0.23 105.8 0.49 69.8 0.60 88.4 0.18 39.9 276.5 282.5 217.0 220.0 347.5 136.0 83.0 94.3 97.2 59.7 MIN 313/023622 187 : APPENDIX 7 (Urinalysis - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number App. 2 34 ---POSE--- 30 100 300 Voi pH SG ml g/L IF 21 PY 3.2 6.6 1033 22 MY 2.6 6.4 1050 23 PY 1.8 6.9 1029 24 PY 8.0 6.6 1031 25 PY 5.8 6.6 1037 2F 26 PY 4.0 6.6 1040 27 PY 2.6 6.4 1044 28 PY 6.0 6.8 1032 29 PY 3.8 7.2 1029 30 CPY 4.8 6.6 1034 3F 31 MY 2.0 6.2 1049 32 PY 2.6 6.5 1050 33 MY 2.0 6.6 1064 34 MY 3.0 6.4 1064 35 PY 4.2 6.3 1042 Prot g/L 0.14 0.28 0.16 0.14 0.22 0.23 0.29 0.18 0.15 0.20 0.26 0.31 0.25 0.23 0.20 Glue 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Keto 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Bili Blood 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 APPENDIX 7 (Urinalysis - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group /sex Animal number Epi 2 34 -- POSF-- 30 100 300 Leuc RBC Cryst IF 21 0 10 0 22 0 0 0 0 23 0 0 0 0 24 0 0 0 0 25 0 0 0 0 2F 26 0 0 0 0 27 0 0 0 0 28 0 0 0 0 29 0 0 0 0 30 0 0 0 0 3F 31 0 0 0 0 32 0 0 0 0 33 0 0 0 0 34 0 0 0 0 35 0 0 0 0 Sperm 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Casts 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Abn. Na Total K Total mmol mmol 0 0.20 0.55 0 0.35 0.58 0 0.12 0.23 0 0.76 0.95 0 0.43 0.90 0 0.44 0.77 0 0.22 0.43 0 0.47 0.73 0 0.33 0.55 0 0.22 0.78 0 0.20 0.41 0 0.25 0.64 0 0.19 0.55 0 0.28 0.79 0 0.23 0.93 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 234 --POSF....... 30 100 300 Group / sex Animal number Cl Total mmol U-Na mmol/L U-K mmol/L U-Cl mmol/L IF 21 0.27 63.9 171.0 83.6 22 0.34 134.0 221.5 132.5 23 0.11 67.5 129.5 63.4 24 0.58 94.4 119.0 73.0 25 0.37 73.8 155.0 64.6 2F 26 0.30 109.5 192.5 74.4 27 0.24 85.9 164.0 93.9 28 0.37 78.2 121.0 61.8 29 0.33 87.2 144.5 87.8 30 0.39 45.4 163.0 80.4 3F 31 0.21 97.5 203.0 103.2 32 0.21 96.6 247.0 82.0 33 0.21 95.9 276.0 106.5 34 0.34 92.3 264.5 114.4 35 0.25 54.8 221.0 60.3 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) Day 91 Group Compound Exposure level (ppm) Group / sex Animal number 1 Control 0 App. 2 34 ---POSF...... 30 100 300 Voi pH SG ml g/L 4F 36 CMY 3.2 6.4 1043 37 MY 2.6 7.2 1045 38 MY 1.8 6.5 1084 39 MY 1.8 6.6 1078 40 MY 2.0 6.2 1090 Prot g/L 0.39 0.23 0.30 0.23 0.40 Glue 0 0 0 0 0 Keto 0 0 0 0 0 Bili Blood 00 00 00 00 00 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) Day 91 Group :1 Compound : Control Exposure level (ppm) : 0 Group /sex Animal number Epi 2 34 --POSF---- 30 100 300 Leuc RBC Cryst Sperm 4F 36 0 10 0 0 37 0 0 0 0 0 38 0 0 0 0 0 39 0 0 0 0 0 40 0 0 0 0 0 Casts 0 0 0 0 0 Abn. Na Total K Total mmol mmol 0 0.28 0.55 0 0.15 0.66 0 0.10 0.73 0 0.25 0.68 0 0.24 0.68 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) MIN 313/023622 192 : Day 91 Group :1 Compound : Control Exposure level (ppm) 0 234 ---POSF...... 30 100 300 Group / sex Animal number Ci Total mmol U-Na mmol/L U-K mmol/L U-Cl mmol/L 4F 36 37 38 39 40 0.33 88.2 0.17 59.6 0.21 57.8 0.24 137.0 0.29 121.8 170.5 254.5 406.5 376.5 342.0 102.1 63.5 115.9 134.7 144.9 APPENDIX 7 (Urinalysis - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number App. 234 ---POSF-- 30 100 300 Voi PH SG ml g/L Prot Glue Keto g/L Bili Blood IM 41 CPY 3.2 6.8 1033 1.07 0 TR 0 0 U) 42 PY 8.0 6.7 1036 0.94 0 TR 0 0 43 CPY 12.0 7.6 1025 0.55 0 TR 0 0 44 CPY 10.2 7.2 1026 0.77 0 + 0 0 45 CPY 2.0 7.2 1032 1.03 0 +0 + 2M 46 CPY 8.8 7.0 1033 47 CPY 5.4 6.9 1043 48 CMY 4.8 7.0 1058 49 CPY 5.8 7.0 1040 50 CPY 6.4 7.1 1037 2.18 0 +0 1.08 0 TR 0 1.01 0 TR 0 1.22 0 TR 0 1.02 0 TR 0 0 0 0 0 0 3M 51 CPY 9.2 6.9 1032 52 CPY 5.0 6.7 1044 53 MY 2.6 6.5 1058 54 PY 9.0 6.9 1036 55 MY 3.2 6.6 1068 0.82 0 0.92 0 1.41 0 1.46 0 1.12 0 TR 0 TR 0 TR 0 TR 0 TR 0 0 0 0 0 0 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number Epi 2 34 ---POSF-- 30 100 300 Leuc RBC Cryst 1M 41 0 0 0 0 42 1 1 0 0 43 0 0 0 0 44 0 0 0 0 45 0 0 1 0 2M 46 0 0 0 0 47 0 1 0 0 48 1 0 0 0 49 0 0 0 0 50 0 0 0 0 3M 51 0 0 0 0 52 0 0 0 0 53 0 0 0 0 54 0 0 0 0 55 0 0 0 0 Sperm 1 2 0 2 2 1 0 0 1 1 1 1 2 0 1 Casts 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Abn. Na Total K Total mmol mmol 0 0.18 0.44 0 0.34 1.77 0 0.66 1.72 0 0.56 1.24 0 0.09 0.31 0 0.52 1.21 0 0.51 1.25 0 0.45 1.45 0 0.61 1.06 0 0.69 1.23 0 0.86 1.25 0 0.48 1.20 0 0.25 0.74 0 0.56 1.52 0 0.45 0.84 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) Day 121 Group :1 Compound Control Exposure level (ppm) 0 234 --POSF----- 30 100 300 Group / sex Animal number Cl Total mmol U-Na mtnol/L U-K mmol/L U-CI mmol/L 1M 41 42 43 44 45 2M 46 47 48 49 50 3M 51 52 53 54 55 0.19 54.7 0.49 42.5 0.45 54.9 0.44 55.0 0.12 44.0 0.37 59.6 0.45 94.9 0.50 93.5 0.46 104.7 0.55 108.2 0.56 93.4 0.43 95.6 0.18 94.5 0.39 62.1 0.31 141.4 139.0 221.0 143.5 122.0 154.0 138.0 232.0 302.5 183.0 191.5 135.5 239.0 285.5 169.0 263.5 60.5 61.4 37.4 43.0 57.7 41.9 83.2 104.0 80.0 86.7 61.1 86.9 67.5 43.7 97.3 MIN 313/023622 MIN 313/023622 196 : APPENDIX 7 (Urinalysis - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 Group /sex Animal number App. 4M 56 PY 57 PY 58 MY 59 PY 60 MY 234 ---POSF---- 30 100 300 Voi pH SG ml g/L 5.8 6.6 1041 8.4 6.8 1036 4.0 6.5 1056 6.2 6.6 1040 3.2 6.5 1066 Prot g/L 0.98 0.76 1.18 1.10 1.07 Glue 0 0 0 0 0 Keto TR TR TR TR TR Bili Blood 00 00 00 00 00 APPENDIX 7 (Urinalysis - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 Group /sex Animal number Epi 234 --POSF--- 30 100 300 Leuc RBC Cryst 4M 56 57 58 59 60 0 0 0 0 0 10 00 00 00 00 0 0 0 0 0 Sperm 1 1 1 2 0 Casts 0 0 0 0 0 Abn. Na Total K Total mmol mmol 0 0.35 1.22 0 0.48 1.61 0 0.27 1.14 0 0.55 1.01 0 0.23 0.98 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) 198 : Day 121 Group 1 Compound : Control Exposure level (ppm) : 0 234 --POSF----- 30 100 300 Group /sex Animal number Cl Total mmol U-Na mmol/L U-K mmol/L U-Cl mmol/L 4M 56 57 58 59 60 0.38 59.5 210.5 0.55 57.3 192.0 0.31 68.6 285.0 0.31 89.0 163.5 0.25 70.5 306.0 66.0 64.9 76.9 50.1 79.3 APPENDIX 7 (Urinalysis - continued) MIN 313/023622 199 : Day 121 Group 1 Compound : Control Exposure level (ppm) : 0 Group / sex Animal number App. 2 34 -- POSF-- 30 100 300 Voi pH SG ml g/L IF 61 PY 5.4 6.2 1039 62 PY 1.6 6.0 1047 64 PY 6.6 6.2 1033 65 MY 2.4 5.9 1043 2F 66 MY 4.2 6.0 1039 67 PY 5.0 6.2 1037 68 PY 4.2 6.4 1041 69 PY 4.2 6.4 1040 70 PY 6.0 6.0 1037 3F 71 PY 2.4 6.0 1060 72 PY 3.2 6.0 1042 73 MY 1.2 6.2 1060 74 MY 2.6 6.1 1047 75 PY 4.2 6.3 1036 4F 76 MY 3.0 6.1 1045 77 MY 3.8 6.2 1047 78 MY 2.0 6.2 1060 79 PY 4.8 6.2 1033 80 PY 3.2 6.4 1047 Prot g/L 0.27 0.30 0.18 0.31 0.17 0.28 0.20 0.19 0.23 0.22 0.15 0.34 0.18 0.29 0.19 0.28 0.26 0.23 0.37 Glue 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Keto 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Bili Blood 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 00 APPENDIX 7 (Urinalysis - continued) Day 121 Group :1 Compound : Control Exposure level (ppm) : 0 Group /sex Animal number Epi 2 34 -- POSF-- 30 100 300 Leuc RBC Cryst IF 61 0 62 0 64 0 65 0 00 00 00 10 0 0 0 0 2F 66 0 0 0 0 67 0 0 0 0 68 0 0 0 0 69 1 0 0 0 70 0 0 0 0 3F 71 0 0 0 0 72 0 0 0 0 73 0 0 0 0 74 0 0 0 0 75 0 0 0 0 4F 76 0 0 0 0 77 0 0 0 0 78 0 0 0 0 79 0 0 0 0 80 0 0 0 0 Sperm 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Casts 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Abn. Na Total K Total mmol mmol 0 0.31 0.88 0 0.08 0.36 0 0.36 0.97 0 0.17 0.42 0 0.18 0.59 0 0.36 1.09 0 0.40 0.80 0 0.26 0.87 0 0.39 0.91 0 0.08 0.63 0 0.06 0.63 0 0.07 0.44 0 0.10 0.63 0 0.37 0.54 0 0.19 0.63 0 0.29 0.75 0 0.16 0.63 0 0.24 0.78 0 0.30 0.66 MIN 313/023622 APPENDIX 7 (Urinalysis - continued) MIN 313/023622 : 201 Day 121 Group 1 Compound : Control Exposure level (ppm) 0 234 --POSF----- 30 100 300 Group /sex Animal number Cl Total mmol U-Na mmol/L U-K mmol/L U-Cl mmol/L IF 61 62 64 65 2F 66 67 68 69 70 3F 71 72 73 74 75 4F 76 77 78 79 80 0.31 57.5 0.07 51.5 0.26 53.9 0.13 72.2 0.11 43.9 0.52 72.7 0.35 95.0 0.24 62.3 0.38 65.6 0.10 34.3 0.09 19.1 0.05 56.9 0.20 38.4 0.34 87.8 0.20 62.7 0.21 75.7 0.12 80.1 0.23 49.9 0.18 92.3 162.5 225.5 146.5 176.0 140.5 218.0 189.5 206.5 151.5 260.5 197.5 365.5 243.5 128.0 211.0 196.5 316.0 162.0 207.0 56.5 45.9 40.0 54.6 27.3 103.5 83.8 56.3 63.3 40.8 28.9 45.4 75.6 80.0 68.1 55.1 60.8 47.1 55.4 APPENDIX 8 Absolute organ weights - individual values (g) Print No: 0026 GROUP :1 2 34 Printed: 23-JUL- COMPOUND : CONTROL -- POSF---- EXPOSURE LEVEL (PPM) : 0 30 100 300 KoN>> TERMINAL Xybion protocol number : MIN 3 GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR 1M 1 2 3 4 5 500.7 421.8 457.3 433.8 449.6 2.14 2.03 1.98 1.99 1.94 0.086 0.050 0.069 0.045 0.054 1.803 1.227 1.383 1.406 1.481 3.00 2.46 2.55 2.44 2.52 17.89 13.39 15.49 14.55 12.76 1.673 1.384 1.467 1.601 1.474 2M 6 7 8 9 10 509.2 403.3 478.1 433.0 424.9 2.06 1.90 2.16 2.01 2.01 0.063 0.052 0.047 0.060 0.046 1.554 1.282 1.428 1.514 1.105 2.92 2.23 2.74 2.66 2.48 15.57 12.30 17.33 13.52 14.12 1.668 1.264 1.510 1.498 1.397 MIN 313/023622 APPENDIX 8 (Absolute organ weights - continued) Print No: 0026 GROUP :1 2 34 Printed: 23-JUL- COMPOUND : CONTROL -- POSF---- EXPOSURE LEVEL (PPM) : 0 30 100 300 Oto U J TERMINAL Xybion protocol number: MIN 3 GROUP ANIMAL BODY WT <g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR 3M 11 12 13 14 15 430.6 465.0 448.0 401.6 457.4 1.83 2.03 2.01 1.96 2.07 0.055 0.050 0.065 0.047 0.060 1.557 1.492 1.270 1.166 1.415 2.40 3.18 2.82 2.48 3.04 14.24 17.99 17.26 14.91 16.94 1.450 1.540 1.561 1.366 2.043 4M 16 17 18 19 20 377.6 332.9 385.0 403.3 358.4 2.03 1.88 2.06 2.04 1.97 0.069 0.055 0.045 0.065 0.058 1.395 1.025 1.318 1.625 1.270 2.72 2.15 2.63 2.51 2.23 16.91 16.77 19.38 20.97 18.03 1.878 1.765 2.044 2.131 2.077 MIN 313/023622 APPENDIX 8 (Absolute organ weights - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM) : 1 CONTROL 0 2 34 -- POSF---- 30 100 300 GROUP IF ANIMAL 21 22 24 25 TERMINAL BODY WT (g) 269.8 253.9 269.9 267.8 BRAIN 1.84 2.04 1.94 1.85 ADRENALS 0.049 0.060 0.053 0.073 HEART 1.028 0.913 1.018 0.979 2F 26 27 28 29 30 265.1 226.0 244.5 235.2 243.0 1.88 1.81 1.86 1.89 1.86 0.066 0.066 0.063 0.051 0.085 1.067 0.908 1.023 0.899 0.960 KIDNEYS 1.53 1.41 1.70 1.82 1.70 1.57 1.86 1.70 1.61 LIVER 8.71 8.30 8.53 9.98 LUNGS & BR 1.125 1.142 1.236 1.177 9.09 7.23 8.81 8.41 9.15 1.171 1.037 1.118 1.074 1.090 Print No: 0026 Printed: 23-JUL-02 Xybion protocol number: MIN 313 MIN 313/023622 APPENDIX 8 (Absolute organ weights - continued) U> 1 fO O1 1 1 1 GROUP : COMPOUND : EXPOSURE LEVEL(PPM) : 1 CONTROL 0 34 -- POSF---- 100 300 GROUP 3F ANIMAT 31 32 33 34 35 TERMINAL BODY WT <g) 235.3 270.1 218.2 299.8 260.1 BRAIN 1.79 1.93 1.86 1.98 1.84 ADRENALS 0.069 0.076 0.057 0.061 0.071 HEART 1.032 0.946 0.843 0.952 0.943 4F 36 37 38 39 40 243.4 242.6 241.7 217.3 260.2 1.88 1.93 1.97 1.79 2.03 0.063 0.054 0.070 0.057 0.076 1.203 0.893 0.880 0.757 0.865 KIDNEYS 1.61 1.91 1.78 1.84 1.67 LIVER 8.90 10.00 8.61 11.96 9.82 LUNGS & BR 1.063 1.220 1.070 1.410 1.309 1.98 1.41 1.65 1.46 1.71 11.69 11.08 14.47 11.14 12.85 1.703 1.322 1.598 1. 647 1.714 Print No: 0026 Printed: 23-JUL-02 Xybion protocol number: MIN 313 MIN 313/023622 APPENDIX 8 (Absolute organ weights - continued) Print No: 002 GROUP 1 234 Printed: 23-JUL- COMPOUND : CONTROL -- POSF---- EXPOSURE LEVEL (PPM) : 0 30 100 300 to OoN TERMINAL Xybion protocol number: MIN 3 GROUP ANIMAL BODY v n (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR 1M 41 42 43 44 45 466.6 506.1 496.4 493.2 450.3 2.17 2.09 1.97 2.13 2.21 0.049 0.075 0.053 0.057 0.051 1.320 1.417 1.413 1.718 1.484 2.76 2.75 2.78 2.91 2.80 13.02 11.85 12.36 12.24 11 .2 2 1.398 1.599 1.519 1.389 1.455 2M 46 47 48 49 50 498.8 409.2 485.8 423.7 470.8 2.23 1.99 2.13 2.17 2.14 0.045 0.047 0.050 0.054 0.065 1.405 1.685 1.481 1.254 1.417 2.98 2.75 3.34 2.73 3.06 14.24 11.50 13.44 10.70 12.50 1.549 1.422 1.487 1.406 1.470 MIN 313/023622 APPENDIX 8 (Absolute organ weights - continued) Print No: 0028 GROUP COMPOUND :1 : CONTROL 2 34 -- POSF---- Printed: 23-JUL-02 EXPOSURE LEVEL (PPM) : 0 30 100 300 oto TERMINAL Xybion protocol number: MIN 313 GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR 3M 51 52 53 54 55 502.1 465.1 497.0 487.9 396.1 2.09 2.09 2 .1 1 2.18 1.96 0.049 0.050 0.053 0.058 0.048 1.568 1.596 1.378 1.677 1.348 2.83 2.99 3.07 3.33 2.24 13.72 14.91 14.63 15.37 11.05 1.555 1.512 1.457 1.499 1.2 0 1 4M 56 57 58 59 60 451.5 491.2 414.0 407;7 449.9 1.94 2 .1 1 1.97 2.00 2.22 0.058 0.054 0.054 0.042 0.044 1.735 1.771 1.555 1.218 1.449 2.92 3.26 2.76 2.55 2.79 16.08 16.25 13.44 16.07 15.79 1.514 1.831 1.457 1.637 1.642 MIN 313/023622 APPENDIX 8 (Absolute organ weights - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM) : 1 CONTROL 0 2 34 -- POSF---- 30 100 300 GROUP IF ANIMAL 61 62 64 65 TERMINAL BODY WT (g) 313.0 285.2 270.2 297.2 BRAIN 2.05 1.94 1.94 1.95 ADRENALS 0.078 0.078 0.073 0.064 HEART 1.196 1.083 1.164 1.125 2F 66 67 68 69 70 304.1 316.9 258.3 286.4 291.7 2.17 2.01 1.95 2.12 2.01 0.065 0.078 0.055 0.072 0.083 0.864 1.223 0.908 1.136 1.020 KIDNEYS 1.86 1.92 1.75 1.97 1.83 2.27 1.77 1.78 1.84 LIVER 9.45 9.62 7.58 9.29 LUNGS & BR 1.365 1.276 1.206 1.314 8.35 9.64 7.93 8.97 9.12 1.242 1.279 1.128 UNSUITABLE 1.150 Print No: 0028 Printed: 23-JUL-02 Xybion protocol number: MIN 313 MIN 313/023622 APPENDIX 8 (Absolute organ weights - continued) Print No: 0028 GROUP :1 2 3 4 Printed: 23-JUL-02 COMPOUND : CONTROL -- POSF---- EXPOSURE LEVEL (PPM) : 0 30 100 300 oVroO TERMINAL Xybion protocol number: MIN 313 GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR 3F 71 72 73 74 75 260.4 251.6 276.7 251.5 249.9 1.89 1.88 2.00 1.85 2.00 0.065 0.075 0.080 0.062 0.060 0.889 0.940 1.137 1.023 0.944 1.66 1.73 1.96 1.73 1.76 8.15 8.59 8.95 7.81 8.71 1.187 1.132 1.079 1.046 1.003 4F 76 77 78 79 80 232.2 250.8 259.0 297.6 265.0 1.90 1.87 2.03 1.94 1.94 0.055 0.057 0.063 0.071 0.071 0.849 0.988 0.918 0.950 0.918 1.43 1.75 1.53 1.74 1.88 8.63 9.02 9.49 11.37 10.40 1.082 1.142 1.259 1.242 1.212 MIN 313/023622 APPENDIX 8 (Absolute organ weights - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 234 -- POSF--- 30 100 300 GROUP 1M ANIMAL 81 82 83 84 85 TERMINAL BODY WT (g) 353.0 334.2 280.2 350.5 346.2 ADRENALS 0.076 0.045 0.037 0.054 0.051 4M 86 87 88 89 90 333.4 325.3 281.2 335.2 314.4 0.063 0.05 9 0.040 0.061 0.057 BRAIN 1.88 2.10 1.79 2.01 1.91 1.98 1.89 2.02 2.03 1.92 HEART 1.510 1.214 0.933 1.408 1.353 1.481 1.391 1.141 1.361 1.144 KIDNEYS 2.42 2.41 1.91 2.47 2.41 LIVER 10.74 9.47 7.83 10.31 9.78 LUNGS & BR 1.424 1.126 1.005 1.379 1.319 2.37 2.28 2.03 2.53 2.43 12.42 13.17 11.18 13.72 13.05 1.852 1.596 1.699 1.620 1.757 Print No: 0013 Printed: 23-JUL- Xybion protocol number: MIN 31 MIN 313/023622 MIN 313/023622 : 211 APPENDIX 8 (Absolute organ weights - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 234 -- POSF--- 30 100 300 GROUP IF ANIMAL 91 92 93 94 95 TERMINAL BODY WT (g) 206.8 204.5 234.1 194.3 224.5 ADRENALS 0.061 0.053 0.084 0.061 0.072 BRAIN 1.84 1.80 1.78 1.81 1.98 HEART 0.795 0.923 1.061 0.850 0.914 4P 96 97 98 99 100 191.9 193.2 193.2 188.0 206.9 0.064 0.07 9 0.070 0.056 0.078 1.69 1.82 1.80 1.78 1.87 0.835 0.828 0.842 0.781 0.849 KIDNEYS 1.62 1.41 1.49 1.52 1.68 1.42 1.55 1.39 1.46 1.56 LIVER 6.72 6.76 7.19 7.10 8.24 LUNGS & BR 1.021 1.088 0.979 0.948 1.161 7.96 8.33 7.59 7.90 7.87 1.310 1.467 1.286 1.201 1.407 Print No: 0013 Printed: 23-JUL-02 Xybion protocol number: MIN 313A APPENDIX 9 Individual pathological findings MIN 313/023622 The microscopic pathology was carried out by two pathologists. The initial examination was undertaken by the study pathologist, the results of which were then subjected to a routine peer review by a second pathologist. The diagnoses reported here represent the consensus opinions of both pathologists. Study pathologist: Peer review: Morven I. Petersen-Jones, B.V.M.S., M.R.C.V.S., Pathologist, Department of Pathology. Samuel. G. McCormick, M.V.B., M.R.C.V.S., PhD., F.R.C.Path, Director of Pathology, Department of Pathology. 212 APPENDIX 9 213 : Individual pathological findings Print No: 0034 GROUP : COMPOUND : EXPOSURE LEVEL(PPM) : 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0001 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 500.7 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LIVER : -MEDIAN CLEFT PALE AREA<S) ; ONE, SUBCAPSOIAR, 1MM. LIVER : >NO S IG N IF IC A N T LE SIO N LUNGS & BRONCHI : -A L V E O L IT IS , -MINIMAL -ALVEOLAR HAEMORRHAGE, -M IN IM A L STOMACH : -ANTRUM WHITE NODULE (S); MUCOSA, ONE, NEAR TO LIMITING RIDGE, 1MM. APPENDIX 9 MIN 313/023622 214 (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0002 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT : 421.8 GRAMS NECROPSY PA T H O LOGY O B SERVAT.IONS HISTOPATHOLOGY HEART : -MYOCARDITIS, VENTRICULAR REGION,-MINIMAL, MULTI-FOCAL LN MANDIBULAR : -ENLARGED; 1OMM. -CONGESTED, MINIMAL LUNGS & BRONCHI : -ALVEOLAR HAEMORRHAGE, -M IN IM A L APPENDIX 9 MIN 313/023622 : 215 (Individual pathological findings - continued) Print No: 0034 GROUP COMPOUND EXPOSURE LEVEL (PPM): 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0003 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH : 14 TERMINAL BODY WEIGHT : 457.3 GRAMS NECROPSY PA T HO LOGY OB SERVATION S HISTOPATHOLOGY LN MANDIBULAR : -CONGESTED HEART : -M Y O CA RD ITIS, VENTRICULAR R EG IO N ,-M IN IM A L , FOCAL KIDNEYS : -M IN ER A LISA TIO N , CORTICOMEDULLARY, -M INIM AL LARYNX : -ARYTENOID - E P IT H E L IA L H Y PERPLA SIA ,-M IN IM A L LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES, -M INIM AL APPENDIX 9 (Individual pathological Findings - continued) Print No: 0034 GROUP : COMPOUND : EXPOSURE LEVEL(PPM): i CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0004 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 433.8 GRAMS NECROPSY PA T HO LOGY O B S E R V AT I O N S HISTOPATHOLOGY HEART : -MYOCARDITIS, VENTRICULAR REGION,-MINIMAL, MULTI-FOCAL KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL LUNGS & BRONCHI : -CONGESTED; A FEW AREAS, DIFFUSE. LUNGS & BRONCHI : -ALVEOLITIS,-MINIMAL MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) GROUP COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0005 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 449.6 GRAMS to PATHOLOGY O B S RVAT IONS NECROPSY HI STOPATHOLOGY HEART : -M YOCARDITIS, VENTRICULAR R EG IO N , -M IN IM A L, FOCAL KIDNEYS : -CO RTICA L TUBULAR B A S O P H IL IA ,-M IN IM A L , FOCAL LIVER : -PER IPO R TA L LYMPHOCYTIC IN F IL T R A T IO N ,-M IN IM A L LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES , -M INIM AL MIN 313/023622 * * * ANIMAL HAS NO GROSS OBSERVATIONS RECORDED * * * APPENDIX 9 218 (Individual pathological findings - continued) Print Ko: 0034 GROUP COMPOUND EXPOSURE LEVEL (PPM) 1 CONTROL 0 234 ---------------- PO SE----------------30 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0041 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 466.6 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY KIDNEYS : -C O R TIC A L TUBULAR B A S O P H IL IA ,-M IN IM A L , FOCAL -MEDULLARY M IN E R A L IS A T IO N ,-M IN IM A L LUNGS & BRONCHI : -PER IV A SC U LA R INFLAMMATORY C ELL IN F IL T R A T IO N , -M IN IM A L SKIN : -HAIRLOSS, MINIMAL ; LEFT SIDE OF FACE, DIFFUSE. MIN 313/023622 : 219 APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0042 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 506.1 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY KIDNEYS : -CO RTICA L TUBULAR B A S O P H IL IA ,-S L IG H T , FOCAL LUNGS & BRONCHI : -PER IV A SC U LA R INFLAMMATORY C ELL IN F IL T R A T IO N , -M IN IM A L *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 34 -- POSF--- 30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number : MIN 313 ANIMAL NUMBER: 0043 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULEDr FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 496.4 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY KIDNEYS : -CORTICAL TUBULAR BASOPHILIA, -MINIMAL, MULTI-FOCAL -CORTICAL LYMPHOCYTIC INFILTRATION,-SLIGHT, MULTI-FOCAL LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL -ALVEOLITIS ,-MINIMAL -FOCAL OSSEOUS DEPOSITION,-MINIMAL SKIN : -HAIRLOSS, M INIM AL; RIGHT SID E OF FACE D IFFU SE APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP :1 2 34 Printed: 09-SEP-03 COMPOUND CONTROL EXPOSURE LEVEL(PPM): 0 30 100 300 ttoo ANIMAL NUMBER: 0044 SEX: MALE DOSE GROUP: 1 Protocol Number: MIN 313 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC DATE OF DEATH: 19-APR-02 STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 493.2 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY KIDNEYS : -C O R TIC A L TUBULAR B A S O P H IL IA ,-M IN IM A L , FOCAL -CO RTICA L LYMPHOCYTIC IN F IL T R A T IO N , -M IN IM A L, M ULTI-FOCAL *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0045 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED r FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 450.3 GRAMS NECROPSY PA T HO LOGY OB S E R V A T I O N S HISTOPATHOLOGY KIDNEYS : -PELVIC DILATATION ,-SLIGHT 222 *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) Pxint No: 0034 GROUP Kto> COMPOUND EXPOSURE LEVEL <PPM) : 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0006 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT 509.2 GRAMS NECROPSY PA T HO LOGY OB SERVATION S HISTOPATHOLOGY LIVER : -MEDIAN CLEFT PALE AREA(S); ONE, SUBCAPSULAR, 1MM. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-SLIGHT -PARENCHYMAL INFLAMMATORY CELL INFILTRATION,-MINIMAL, FOCAL 224 GROUP : EXPOSURE LEVEL(PPM): ANIMAL NUMBER: 0007 DATE OF DEATH: 21-MAR-02 LN MANDIBULAR : -CONGESTED, MINIMAL APPENDIX 9 (Individual pathological findings - continued) Print Mo: 0034 1 234 Printed: 09-SEP-03 0 30 100 300 Protocol Number: MIN 313 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 403.3 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LIVER : -CEN TRILO BU LA R HEPATOCYTE HYPERTROPHY EXTENDING IN TO MIDZONAL AREA, -SLIGHT APPENDIX 9 MIN 313/023622 : 225 (Individual pathological findings - continued) Print No: 0034 GROUP COMPOUND EXPOSURE LEVEL (PPM): 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0008 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT : 478.1 GRAMS NECROPSY PA T HO LOGY O B SERVATION S HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL A R E A ,-SLIGHT LUNGS & BRONCHI : -A L V E O L IT IS , -MINIMAL MISCELLANEOUS : -ANIMAL THIN, MINIMAL PREPUTIAL GLANDS : -ABSCESS(ES); RIGHT, ONE, 3MM. PROSTATE : -ABSCESS<ES); RIGHT SIDE, ONE, 13X11X11MM. SKIN : -SCAB (S) ; GENITAL REGION, ONE, 3X2MM. APPENDIX 9 MIN 313/023622 226 (Individual pathological findings - continued) Print No: 0034 GROUP COMPOUND EXPOSURE LEVEL (PPM): 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0009 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT : 433.0 GRAMS NECROPSY P A T H O LOGY 0 B S E R V A I ION S HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT LN MANDIBULAR : -ENLARGED; UP TO 13MM. LUNGS & BRONCHI : -ALVEOLAR HAEMORRHAGE,-MINIMAL APPENDIX 9 227 (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND ! EXPOSURE LEVEL (PPM) : 1 CONTROL 0 234 ---------------- P O S F ----------------30 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0010 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DELATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 424.9 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT -EXTRAMEDULLARY HAEMOPOEISIS,-MINIMAL LUNGS & BRONCHI : -CONGESTED, MINIMAL; PATCHY. LUNGS & BRONCHI : >NO SIGNIFICANT LESION SKIN : -HAIRLOSS, MINIMAL; SIDES OF FACE, DIFFUSE. MIN 313/023622 ii O 11 X : 228 APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP COMPOUND EXPOSURE LEVEL (PPM): 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0046 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 498.8 GRAMS NECROPSY PA 1 LOGY 0 B SERVATIONS HISTOPATHOLOGY SKIN : -HAIRLOSS, MINIMAL; SIDES OF FACE, DIFFUSE. *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** APPENDIX 9 229 : (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0047 DATE OP DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 409.2 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE VACUOLATION, -S L IG H T LUNGS & BRONCHI : -FOAMY ALVEOLAR M ACROPHAGES/SEPTAL T H IC K E N IN G / SCATTERED ALVEOLAR MACROPHAGES, -M INIM AL STOMACH : -ANTRUM WHITE NODULE (S) ; MUCOSA, ONE, NEAR TO LIMITING RIDGE, 1MM. APPENDIX 9 MIN 313/023622 230 (Individual pathological findings - continued) GROUP ; COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0048 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 485.8 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-SLIGHT SKIN : -SCAB(S); RIGHT DORSAL LUMBAR REGION, ONE, 3X1MM. STOMACH : -ANTRUM WHITE NODULE (S); MUCOSA, ONE, NEAR TO LIMITING RIDGE, 1MM. APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0049 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 423-7 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-MINIMAL *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 9 MIN 313/023622 232 (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND : EXPOSURE LEVEL (PPM): 1 CONTROL 0 234 --------PQSF-------30 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0050 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 470.8 GRAMS NECROPSY PA T HO LOGY O B SERVAT IONS HISTOPATHOLOGY LARYNX : -ARYTENOID - EPITHELIAL HYPERPLASIA, -MINIMAL -ARYTENOID - SUBEPITHELIAL INFLAMMATORY CELL INFILTRATION, MINIMAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-SLIGHT -EXTRAMEDULLARY HAEMOPOEISIS,-MINIMAL SKIN : -HAIRLOSS, MODERATE; LEFT AND RIGHT SCAPULAR REGIONS. APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP :1 2 34 Printed: 09-SEP-03 COMPOUND : CONTROL EXPOSURE LEVEL(PPM): uuto>> ANIMAL NUMBER: 0011 0 30 SEX: MALE 100 300 DOSE GROUP: 3 Protocol Number: MIN 313 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH : 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 430.6 GRAMS NECROPSY PA T HO LOGY 0 B SERVATIONS HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA, -MODERATE -CENTRILOBULAR HEPATOCYTE VACUOLATION,-MINIMAL MISCELLANEOUS : -ANIMAL T H IN , MINIMAL : 234 GROUP COMPOUND EXPOSURE LEVEL (PPM): ANIMAL NUMBER: 0012 DATE OF DEATH: 21-MAR-02 LN MANDIBULAR : -CONGESTED, MINIMAL APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 1 2 34 Printed: 09-SEP-03 0 30 100 300 Protocol Number: MIN 313 SEX: MALE DOSE GROUP : 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 465.0 GRAMS NECROPSY PA T H O LOGY 0 B SERVATIONS HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL A R EA , -MODERATE -CENTRILOBULAR HEPATOCYTE VACUOLATION, -MINIM AL APPENDIX 9 MIN 313/023622 : 235 (Individual pathological findings - continued) Print No: 0034 GROUP ; COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0013 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 448.0 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA, -MODERATE -CENTRILOBULAR HEPATOCYTE VACUOLATION, -M INIM AL SKIN : -H A IR LO SS, MINIMAL; REGION, D IFFU SE. S ID E S OF FACE AND R IG H T DORSAL THORACIC APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP :1 2 34 Printed: 09-SEP-03 CONTROL EXPOSURE LEVEL (PPM) : 0 30 100 300 K) UJ Os ANIMAL NUMBER: 0014 SEX: MALE DOSE GROUP: 3 Protocol Number: MIN 313 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH : 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 401.6 GRAMS NECROPSY PA T HO LOGY 0 B SERVAT IONS HISTOPATHOLOGY LIVER -MEDIAN CLEFT PALE AREA(S) / ONE, SUBCAPSULAR, 1MM. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL A R E A ,-MODERATE -CENTRILOBULAR HEPATOCYTE VACUOLATION,-SLIGHT LUNGS & BRONCHI : -PROMINENT AGGREGATIONS OF LYMPHOCYTES , -M IN IM A L -VASCULAR C O N G E ST IO N ,-S L IG H T MIN 313/023622 APPENDIX 9 MIN 313/023622 237 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0015 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 457.4 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY LARYNX : -ARYTENOID - SU B E P IT H E L IA L INFLAMMATORY CELL IN F IL T R A T IO N , M IN IM A L LIVER -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA j -MODERATE -EXTRAMEDULLARY H A EM O PO EISIS, -M INIM AL LUNGS & BRONCHI : -CONGESTED, MINIMAL LUNGS & BRONCHI : -ALVEOLAR HAEMORRHAGE,-MARKED MIN 313/023622 : 238 APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 34 -- POSF--- 30 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0051 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH:: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 502.1 GRAMS NECROPSY PA T HO LOGY 0B SERVATION S HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY, -MINIM AL LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES, -M INIM AL * * * ANIMAL HAS NO GROSS OBSERVATIONS RECORDED * * * APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 34 -- POSF--- 30 xoo 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0052 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS:: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH : 16 TERMINAL BODY WEIGHT: 465.1 GRAMS NECROPSY PA T H O LOGY OB SERVATION S HISTOPATHOLOGY LN MANDIBULAR : -ENLARGED; RIGHT, ONE, 12MM. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY, -MINIM AL -EXTRAMEDULLARY H A EM O PO EISIS, -M INIM AL LUNGS & BRONCHI : -FOAMY ALVEOLAR M ACROPHAGES/SEPTAL ALVEOLAR MACROPHAGES, -M INIM AL -A L V E O LIT IS , -MINIMAL THICKENING/ SCATTERED STOMACH ; -ANTRUM W HITE NODULE <S) ; RID G E, PUNCTATE. MUCOSA, ONE, NEAR TO L IM IT IN G MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP COMPOUND :1 2 3 4 : CONTROL Printed: 09-SEP-03 E X P O S U R E L E V E L (PPM) : 0 30 100 300 K> O ANIMAL NUMBER: 0053 DATE OF DEATH: 19-APR-02 Protocol Number: MIN 313 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 497.0 GRAMS NECROPSY PA 1 HO LOGY 0B SERVATIONS HISTOPATHOLOGY LIV E R : -CENTRILO BULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA, -SLIG H T -PER IPO R TA L LYMPHOCYTIC IN F IL T R A T IO N , -M IN IM A L * * * A N IM AL HAS NO GROSS O BSERVATIO NS RECORDED * * * MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND : E X P O S U R E L E V E L (PPM) : 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0054 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 487.9 GRAMS NECROPSY PA T H O LOGY O B SERVAT IONS HISTOPATHOLOGY LARYNX : -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 9 242 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0055 DATE OF DEATH: I9-APR-02 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 396.1 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY EPIDIDYMIDES : -SWELLING(S) ; R I G H T CAUDA, ONE, 3MM. LIV E R : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY, -M IN IM A L APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND : E X P O S U R E L E V E L (PPM) : 1 CONTROL 0 234 --- P O S F --- 30 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0016 DATE O F DEATH: 2 1 -MAR-02 SEX: MALE DOSE GROUP: 4 rS ACRIFICE STATUS: S C H E D U L E D T E R M I N A L SACR I F I C E STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 377.6 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY UKt>*> LARYNX : - NECROSIS O F V E N T R A L C A R T I L A G E ,-PRESENT LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL A R E A ,-MODERATE LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPKAGES/SEPTAL THICKENING/ SCATTERED A L V E O L A R M A C R O P H A G E S ,-MODERATE TRACHEA : -EPITHELIAL HYPERPLASIA,-MINIMAL, FOCAL -E P I T H E L I A L I N F L A M M A T O R Y C E L L I N F I L T R A T I O N ,- M I N I M A L , FOCAL -SUBEPITHELIAL INFLAMMATORY AGGREGATIONS ,-MI N I M A L , FOCAL -SUBEPITHELIAL FIBROBLAST PROLIFERATION,-MINIMAL, FOCAL MIN 313/023622 *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 9 MIN 313/023622 244 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): ANIMAL NUMBER: 0017 DATE OF DEATH: 21-MAR-02 Print No: 0034 1 2 34 Printed: 09-SEP-03 0 30 100 300 Protocol Number: MIN 313 SEX: MALE DOSE GROUP : 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY HEIGHT: 332.9 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL A R E A ,-MODERATE LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-SLIGHT -ALVEOLITIS ,-MINIMAL MISCELLANEOUS : -ANIMAL THIN STOMACH : -CORPUS WHITE NODULE (S); MUCOSA, ONE NEAR TO LIMITING RIDGE, 1MM. APPENDIX 9 MIN 313/023622 245 (Individual pathological findings - continued) Print No: 0034 GROUP : COMPOUND EXPOSURE LEVEL <PPM) : 1 CONTROL 0 2 34 -- POSF--- 30 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0018 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS:: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH:: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 385.0 GRAMS NECROPSY PA T HO LOGY OB SERVATION s HISTOPATHOLOGY KIDNEYS : -FOCAL CAPSULAR CONGESTION,-MINIMAL liver : -ENLARGED; 19.38G. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL A R E A ,-MODERATE LN MANDIBULAR : -CONGESTED, MODERATE LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL MISCELLANEOUS : -ANIMAL THIN APPENDIX 9 MIN 313/023622 246 : (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0019 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OP DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 403.3 GRAMS NECROPSY PA T HO LOGY 0 B SERVATIONS HISTOPATHOLOGY KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LIVER : -MEDIAN CLEFT PALE AREA(S) ; ONE, SUBCAPSULAR, 2MM. -ENLARGED; 20.97G. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-MODERATE LN MANDIBULAR : -CONGESTED, MINIMAL LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL -ALVEOLAR HAEMORRHAGE,-MINIMAL MISCELLANEOUS : -ANIMAL THIN, MINIMAL APPENDIX 9 (Individual pathological findings - continued) Print Mo: 0034 GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0020 DATE OF DEATH: 21-MAR-02 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 358.4 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY to LARYNX : ^0 -NECROSIS OF VENTRAL CARTILAGE,-PRESENT -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL -ARYTENOID - SUBEPITHELIAL INFLAMMATORY CELL INFILTRATION, - MINIMAL LIVER : -ENLARGED; 18.03G. LIVER I -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL A R E A ,-MODERATE -FOCAL/MULTIFOCAL SINGLE CELL NECROSIS INFLAMMATORY CELL INFILTRATION,-MINIMAL LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL -ALVEOLAR HAEMORRHAGE,-MINIMAL STOMACH : -ANTRUM WHITE NODULE (S); MUCOSA, ONE, NEAR TO LIMITING RIDGE, 1MM. TRACHEA : -EPITHELIAL HYPERPLASIA,-MINIMAL, FOCAL MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) Print No: 0034 GROUP COMPOUND EXPOSURE LEVEL(PPM) J CONTROL -POSF- 0 30 100 300 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0056 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS:: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 451.5 GRAMS NECROPSY PA T H O LOGY OB SERVATION S HISTOPATHOLOGY KIDNEYS : -CORTICAL TUBULAR BASOPHILIA, -MINIMAL, FOCAL N> OO LARYNX ! -NECROSIS OF VENTRAL CARTILAGE,-PRESENT -ARYTENOID - EPITHELIAL HYPERPLASIA,-SLIGHT -ARYTENOID - EPITHELIAL HYPERKERATOSIS PARAKERATOSIS,-MINIMAL -VENTROLATERAL EPITHELIAL SQUAMOUS METAPLASIA,-MINIMAL -VENTRAL POUCH - EPITHELIAL HYPERPLASIA,-MINIMAL -VENTRAL POUCH - INFLAMMATORY CELL INFILTRATION,-MINIMAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT -CENTRILOBULAR HEPATOCYTE NECROSIS/-INFLAMMATORY CELL INFILTRATION/SINUSOIDAL CONGESTION,-MODERATE, MULTI-FOCAL LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL -FOCAL OSSEOUS DEPOSITION,-MINIMAL MIN 313/023622 *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) GROUP :1 2 34 COMPOUND EXPOSURE LEVEL(PPM): 0 30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0057 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY HEIGHT: 491.2 GRAMS N> P A T H O L O G Y O B S E R V A T I O N S 'O NECROPSY HISTOPATHOLOGY KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, MULTI-FOCAL -MEDULLARY TUBULAR BASOPHILIA,-MINIMAL, FOCAL -MEDULLARY TUBULAR FIBROSIS,-MINIMAL, FOCAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-SLIGHT SKIN : -HAIRLOSS, MODERATE; RIGHT CRANIAL REGION. MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL (PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0058 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 414.0 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-SLIGHT, MULTI-FOCAL ot o Ul -MEDULLARY MINERALISATION,-MINIMAL LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT -ARYTENOID - EPITHELIAL HYPERPLASIA,-SLIGHT -ARYTENOID - EPITHELIAL HYPERKERATOSIS PARAKERATOSIS,-MINIMAL -VENTROLATERAL EPITHELIAL HYPERPLASIA,-MINIMAL -VENTRAL POUCH - EPITHELIAL HYPERPLASIA, --MINIMAL -VENTRAL POUCH - INFLAMMATORY CELL INFILTRATION,-MINIMAL LIVER : -MEDIAN CLEFT PALE AREA(S) ; ONE, SUBCAPSULAR, 2MM. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT -HEPATOCYTE VACUOLATION , MEDIAN CLEFT,-SLIGHT LUNGS & BRONCHI ! -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-SLIGHT -PERIVASCULAR INFLAMMATORY CELL INFILTRATION,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; RIGHT SCAPULAR REGION AND CRANIAL REGION. MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0059 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 407.7 GRAMS P A T H O L O G Y OB S E R V A T I O N S ot o* NECROPSY HISTOPATHOLOGY LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LIVER : -LOBULAR PATTERN ACCENTUATED, MINIMAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT -CENTRILOBULAR INFLAMMATORY CELL INFILTRATION,-MINIMAL, FOCAL LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-SLIGHT SKIN : -HAIRLOSS, MODERATE; CRANIAL REGION, RIGHT SIDE OF FACE AND RIGHT SCAPULAR REGION, DIFFUSE. TRACHEA : -SUBEPITHELIAL INFLAMMATORY AGGREGATIONS,-SLIGHT MIN 313/023622 APPENDIX 9 MIN 313/023622 O X 252 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL (PPM) CONTROL -POSF- 0 30 100 300 Print No: 0034 Protocol Number: MIN 313 ANIMAL NUMBER: 0060 DATE OF DEATH: 19-APR-02 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH:: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 449.9 GRAMS NECROPSY PA LOGY O B SERVA T I O N S HISTOPATHOLOGY LARYNX r -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT LUNGS & BRONCHI : -FOCAL BRONCHOALVEOLAR HYPERPLASIA,-MINIMAL -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL MISCELLANEOUS : -ANIMAL THIN, MINIMAL SKIN : -HAIRLOSS, MINIMAL; FORELIMBS. APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0021 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 to L /l U) STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: PATHOLOGY OBSERVATIONS 269.8 GRAMS NECROPSY HISTOPATHOLOGY KIDNEYS : -CORTICAL LYMPHOCYTIC INFILTRATION,-MINIMAL, FOCAL LARYNX : -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL LIVER : -PERIPORTAL LYMPHOCYTIC INFILTRATION,-SLIGHT *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM): 1 CONTROL 0 234 --------POSF-------30 100 300 Print Ho; 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0022 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS:: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH:: 92 STUDY WEEK OF DEATH : 14 TERMINAL BODY WEIGHT 253.9 GRAMS NECROPSY MISCELLANEOUS : -FUR STAINING1; DORSUM, BROWN. PA T HO LOGY OB SERVATION S HISTOPATHOLOGY 254 *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) GROUP ; COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0023 DATE OP DEATH: 20-MAR-02 SEX: [TEMALE DOSE GROUP : 1 SACRIFICE STATUS: UNSCHEDULED (F) STUDY DAY OF DEATH: 91 STUDY WEEK OF DEATH: 13 TERMINAL BODY WEIGHT: 242.4 GRAMS PA T HO LOGY 0 B S E R V A T I O N S t o NECROPSY HISTOPATHOLOGY o KIDNEYS : -MEDULLARY MINERALISATION,-MINIMAL LIVER -MEDIAN CLEFT PALE AREA (S) ; ONE, SUBCAPSULAR, 2MM. LIVER : -PARENCHYMAL INFLAMMATORY CELL INFILTRATION,-MINIMAL , FOCAL -HEPATOCYTE VACUOLATION, MEDIAN CLEFT,-MINIMAL LUNGS & BRONCHI : -CONGESTED, SEVERE -INCOMPLETE COLLAPSE LUNGS & BRONCHI : -ALVEOLAR HAEMORRHAGE,-MARKED -VASCULAR CONGESTION,-MODERATE -EARLY AUTOLYSIS,-PRESENT SKIN : -HAIRLOSS, MINIMAL; VENTRAL SURFACE, PATCHY, DIFFUSE. UTERUS : -FLUID DISTENSION, MINIMAL UTERUS : -LUMINAL DILATATION,-MINIMAL MIN 313/023622 GROUP EXPOSURE LEVEL (PPM) : ANIMAL NUMBER: 0024 DATE OF DEATH: 21-MAR-02 APPENDIX 9 (Individual pathological findings - continued) 1 2 34 Print No: 0034 Printed: 09-SEP-03 0 30 100 300 Protocol Number: MIN 313 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT : 269.9 GRAMS NECROPSY PA T HO LOGY O B S E R V A T ION S HISTOPATHOLOGY KIDNEYS : -CO RTICAL TUBULAR B A S O P H IL IA ,-S L IG H T , FOCAL -CO RTICAL IN T E R S T IT IA L FIBRO BLAST PR O LIFER A TIO N / F IB R O S IS ,SLIG H T, FOCAL LIV E R : -EXTRAMEDULLARY HAEM O PO EISIS, -M IN IM A L LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES , -S L IG H T -ALVEOLAR HAEMORRHAGE, -M IN IM A L * * * ANIM AL HAS NO GROSS OBSERVATIONS RECORDED * * * MIN 313/023622 APPENDIX 9 MIN 313/023622 257 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM): 1 CONTROL 0 234 --------POSE-------30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0025 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS:: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH:: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 267.8 GRAMS NECROPSY PA T HO LOGY 0 B S E R V A T ION S HISTOPATHOLOGY KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL SKIN : -HAIRLOSS, MINIMAL; VENTRAL SURFACE AND RIGHT HINDLIMB, DIFFUSE. APPENDIX 9 MIN 313/023622 258 : (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 34 -------- POSF-------30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0061 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 313.0 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES ,-MINIMAL -PERIVASCULAR INFLAMMATORY CELL INFILTRATION,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; RIGHT SCAPULAR AND RIGHT DORSAL THORACIC REGION, DIFFUSE. APPENDIX 9 : 259 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM ) j. j CONTROL -POSF- 0 30 100 300 Print No: 0034 Printed: U9-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0062 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS:: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 285.2 GRAMS NECROPSY PA T H O LOGY O B S E R V A T ION S HISTOPATHOLOGY SKIN : -H A IR LO S S , M IN IM A L; DORSAL THORACIC REG IO N, D IF F U S E . KIDNEYS : -MINERALISATION, CORTICOMEDULLARY,-MINIMAL APPENDIX 9 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL (PPM) CONTROL -POSF- 0 30 100 300 Print No: 0034 Printed: uy-SEP-uj Protocol Number: MIN 313 ANIMAL NUMBER: 0063 DATE OF DEATH: 03-JAN-02 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: UNSCHEDULED (H) STUDY DAY OF DEATH:: 15 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: 213.3 GRAMS NECROPSY PA T H O LOGY O B SERVATIONS HISTOPATHOLOGY KIDNEYS : -CO RTICAL TUBULAR B A S O P H ILIA ,-M IN IM A L, FOCAL -C O R TIC A L IN T E R S T IT IA L FIBROBLAST P R O LIFE R A TIO N / F IB R O S IS ,M IN IM A L, FOCAL -MEDULLARY M IN E R A LIS A TIO N , -M IN IM A L * * * A N IM A L HAS NO GROSS O BS E R V A TIO N S RECORDED * * * MIN 313/023622 APPENDIX 9 : 261 (Individual pathological findings - continued) GROUP COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 34 -- POSF--- 30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0064 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 270.2 GRAMS NECROPSY PA T HO LOGY 0 B SERVATIONS HISTOPATHOLOGY LIVER : -PERIPORTAL LYMPHOCYTIC INFILTRATION,-MINIMAL LN MANDIBULAR : -ENLARGED; 9MM. LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES ,-MINIMAL -ALVEOLAR HAEMORRHAGE,-SLIGHT APPENDIX 9 (Individual pathological Undings - continued) GROUP COMPOUND EXPOSURE LEVEL (PPM) 1 CONTROL 0 234 --------POSF-------30 100 300 Print Mo: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0065 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 297.2 GRAMS NECROPSY PA T HO LOGY 0B SERVATION S HISTOPATHOLOGY KIDNEYS : -MINERALISATION, CORTICOMEDULLARY,-MINIMAL -MEDULLARY MINERALISATION,-MINIMAL LIVER -EXTRAMEDULLARY HAEMOPOEISIS ,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; FORELIMBS, DIFFUSE. MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): to ON 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0026 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULEDr TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 265.1 GRAMS NECROPSY PA T HO LOGY 0 B SERVATIONS HISTOPATHOLOGY KIDNEYS : -P E L V IC D ILA TIO N , M IN IM A L; R IG H T. LUNGS & BRONCHI : -FOCAL OSSEOUS D EPO SITIO N ,-M IN IM A L MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND ; EXPOSURE LEVEL (PPM) : 1 CONTROL 0 23 -- POSF-30 100 4 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0027 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP : 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 226.0 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY SKIN : -HAIRLOSS, MINIMAL; DORSAL THORACIC REGION, DIFFUSE. UTERUS -FLUID DISTENSION, MODERATE 264 * * * ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** APPENDIX 9 (Individual pathological Undings - continued) GROUP COMPOUND EXPOSURE LEVEL (PPM) : ANIMAL NUMBER: 0028 DATE OF DEATH : 21-MAR-02 1 2 34 Print No: 0034 Printed: 09-SEP-03 0 30 100 300 Protocol Number: MIN 313 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 244.5 GRAMS NECROPSY UTERUS : -F L U ID D IS T E N S IO N , MODERATE PATHOLOGY OBSERVATIONS HISTOPATHOLOGY *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** APPENDIX 9 266 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 234 --------POSE-------30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0029 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS:: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH : 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 235.2 GRAMS LN MANDIBULAR : -ENLARGED; ONE, 12MM. NECROPSY PA T HO LOGY O B S E R V A T ION s HISTOPATHOLOGY LUNGS & BRONCHI : -AGGREGATIONS OP ALVEOLAR MACROPHAGES,-MINIMAL -PERIVASCULAR INFLAMMATORY CELL INFILTRATION,-MINIMAL APPENDIX 9 (Individual pathological findings - continued) GROUP :i2 34 COMPOUND EXPOSURE LEVEL(PPM): 0 30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0030 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 243.0 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY LARYNX : -VENTRAL POUCH - E P IT H E L IA L H Y PER PL A SIA ,-M IN IM A L LIVER : -PER IPO R TA L LYMPHOCYTIC IN F IL T R A T IO N , -M INIM AL STOMACH : -ANTRUM WHITE NODULE (S); MUCOSA, ONE, NEAR TO LIMITING RIDGE, 1MM. THYMUS : -DARK UTERUS ! -FLUID DISTENSION, MINIMAL MIN 313/023622 APPENDIX 9 (Individual pathological Findings - continued) GROUP : (O COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 34 --------POSF-------30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0066 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 304.1 GRAMS * * * ANIMAL HAS NO GROSS OBSERVATIONS RECORDED * * * *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): ANIMAL NUMBER: 0067 DATE OF DEATH: 19-APR-02 1 2 34 Print No: 0034 Printed: 09-SEP-03 0 30 100 300 Protocol Number: MIN 313 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 316.9 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY KIDNEYS : -P E L V IC D IL A T IO N , MODERATE; R IG H T. LARYNX : -VENTRAL POUCH - FO RIG N BODY IN LUMEN, RELATED TO E P IT H E L IA L H Y P E R P L A S IA / INFLAMMATORY C ELL IN F IL T R A T IO N ,-S L IG H T URETERS : -DISTENDED; RIGHT, 1MM. APPENDIX 9 MIN 313/023622 270 : (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0068 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 258.3 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; RIGHT, DIFFUSE. UTERUS : -FLUID DISTENSION, MODERATE APPENDIX 9 (Individual pathological Findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 34 -- POSF--- 30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number : MIN 313 ANIMAL NUMBER: 0069 DATE OF DEATH: 19-APR.-02 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 286.4 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY SKIN : -HAIRLOSS, MINIMAL; RIGHT SIDE IF FACE, DIFFUSE. UTERUS : -FLUID DISTENSION, MODERATE *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL (PPM) : i CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0070 DATE OF DEATH: 19-APR -02 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, FIRST POST -RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 291.7 GRAMS NECROPSY PA T HO LOGY O B SERVAT I O N S HISTOPATHOLOGY LN MANDIBULAR -ENLARGED; 9MM. MIN 313/023622 272 *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** MIN 313/023622 : 273 APPENDIX 9 (Individual pathological findings - continued) GROUP : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0031 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 235.3 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LN MANDIBULAR : -ENLARGED; UP TO 10MM. * * * ANIMAL HAS NO M ICROSCOPIC FIN D IN G S RECORDED * * * APPENDIX 9 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL <PPM) CONTROL -POSF- 0 30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0032 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS:: SCHEDULED r TERMINAL SACRIFICE STUDY DAY OF DEATH:: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT : 270.1 GRAMS NECROPSY PA T H O LOGY 0B SERVATION S HISTOPATHOLOGY LIVER : -PERIPORTAL HEPATOCYTE VACUOLATION,-MINIMAL MIN 313/023622 274 *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) GROUP EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0033 SEX: [TEMALE DOSE GROUP: 3 SACRIFICE STATUS SCHEDULED, TERMINAL SACRIFICE t o DATE OF DEATH: 21-MAR-02 -o Oh STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT : 218.2 GRAMS PA I HO LOGY O B SERVATION S NECROPSY HISTOPATHOLOGY LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; INTER-ORBITAL REGION, DIFFUSE. TAIL : -SWELLING (S) ; ONE, 3X2X2MM, (TAKEN WITH TATTOO) . UTERUS -FLUID DISTENSION, MODERATE MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0034 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 299.8 GRAMS NECROPSY PA T HO LOGY O B S E R V A T ION S HISTOPATHOLOGY STOMACH : -ANTRUM WHITE NODULE (S) ; MUCOSA, TWO, NEAR TO LIMITING RIDGE, PUNCTATE. *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 34 -- POSF--- 30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0035 DATE OF DEATH: 21-MAR-02 SEX: FEMAJLE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED,. TERMINAL SACRIFICE STUDY DAY OF DEATH:: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 260.1 GRAMS NECROPSY PA T HO LOGY OB SERVATION S HISTOPATHOLOGY LARYNX : -VENTRAL POUCH - EPITHELIAL HYPERPLASIA,-MINIMAL LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES ,-MINIMAL MISCELLANEOUS : -ANIMAL THIN, MINIMAL APPENDIX 9 278 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL (PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0071 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS SCHEDULED, FIRST POST -RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 260.4 GRAMS NECROPSY PAT H O L O G Y OB SERVATION S HISTOPATHOLOGY LARYNX : -VENTRAL POUCH - FORIGN BODY IN LUMEN, RELATED TO EPITHELIAL HYPERPLASIA/ INFLAMMATORY CELL INFILTRATION,-MODERATE LIVER : -MEDIAN CLEFT PALE AREA (S) ; ONE, SUB CAPSULAR. LIVER : >NO SIGNIFICANT LESION STOMACH : -ANTRUM WHITE NODULE (S) ; MUCOSA, TWO, NEAR TO LIMTING RIDGE, PUNCTATE. APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 234 ---------------- P O S F ----------------30 100 300 Print Ho: 0034 Printed: 09-SEP-03 to o VO ANIMAL NUMBER: 0072 DATE OF DEATH: 19-APR-02 Protocol Number: MIN 313 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 251.6 GRAMS 3 X O NECROPSY PA LOGY OBS E R V A T I O N S HISTOPATHOLOGY LIVER : -PARENCHYMAL INFLAMMATORY CELL INFILTRATION,-MINIMAL, FOCAL *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED ** MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL <EPM) : 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0073 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 276.7 GRAMS NECROPSY PATHOLOGY STOMACH : -ANTRUM WHITE NODULE (S) ; MUCOSA, THREE, NEAR TO LIMITING RIDGE, 1MM. OBSERVATIONS HISTOPATHOLOGY UTERUS : -FLUID DISTENSION, MINIMAL 280 : *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0074 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 251.5 GRAMS NECROPSY PA T H O LOGY O B SERVAT I O N S HISTOPATHOLOGY LUNGS & BRONCHI : -ALVEOLAR HAEMORRHAGE,-MINIMAL UTERUS : -FLUID DISTENSION, MODERATE MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : i CONTROL 0 2 30 34 100 300 Print Mo: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0075 DATE OF DEATH: 19-APR--02 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 249.9 GRAMS NECROPSY PA T HO LOGY O B SERVATIONS HISTOPATHOLOGY SKIN : -HAIRLOSS, MINIMAL; LEFT DORSUM, DIFFUSE MIN 313/023622 282 *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** APPENDIX 9 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL (PPM) : 1 CONTROL 0 2 34 -------- POSF-------30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0036 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 243.4 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY to KIDNEYS : uo>o -PELVIC DILATION, MINIMAL; RIGHT. KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL -UROTHELIAL HYPERPLASIA, UNILATERAL,-MINIMAL, FOCAL -PELVIC DILATATION,-MODERATE LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL, FOCAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT LN MANDIBULAR : -ENLARGED; 9M M . -CONGESTED; RIGHT, ONE. LUNGS & BRONCHI : -INCOMPLETE COLLAPSE, MINIMAL LUNGS & BRONCHI : -FOCAL OSSEOUS DEPOSITION,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; RIGHT FORELIMB, DIFFUSE. MIN 313/023622 APPENDIX 9 MIN 313/023622 284 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL(PPM) l CONTROL -POSF- 0 30 100 300 Print No: 0034 printed: u ^-s e p -u j Protocol Number: MIN 313 ANIMAL NUMBER: 0037 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP : 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 242.6 GRAMS NECROPSY PA T HO LOGY OBSERVATIONS HISTOPATHOLOGY LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL MISCELLANEOUS : -ANIMAL THIN, MINIMAL -FUR BADLY GROOMED, MINIMAL TAIL : -SWELLING(S) ; ONE, FIRM, 3X1MM. UTERUS : -FLUID DISTENSION, MODERATE APPENDIX 9 MIN 313/023622 : 285 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL (PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0036 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT : 241.7 GRAMS NECROPSY PA T HO LOGY O B SERVAT IONS HISTOPATHOLOGY LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL, FOCAL LIVER -ENLARGED; 14.47G. LUNGS & BRONCHI : -INCOMPLETE COLLAPSE, MINIMAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA j -MODERATE -PROMINENT CENTRILOBULAR HEPATOCYTE PIGMENT,-MINIMAL LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-SLIGHT -ALVEOLAR HAEMORRHAGE,-MINIMAL -FOCAL OSSEOUS DEPOSITION,-MINIMAL MISCELLANEOUS : -ANIMAL THIN SKIN : -HAIRLOSS, MINIMAL; HINDLIMBS AND LEFT CERVICAL REGION, DIFFUSE. APPENDIX 9 MIN 313/023622 286 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE liEVEL ( PPM) jL CONTROL -POSF- 0 30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0039 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 217.3 GRAMS NECROPSY P A T H O L O G O B SE R V A T ION S HISTOPATHOLOGY LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LIVER : -MEDIAN CLEFT PALE AREA(S) / ONE, SUB CAPSULAR, 3MM. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL A R E A ,-MODERATE -HEPATOCYTE VACUOLATION , MEDIAN CLEFT,-MINIMAL MISCELLANEOUS : -ANIMAL THIN LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-SLIGHT SKIN I -HAIRLOSS, MODERATE; LEFT HINDLIMB. STOMACH : -FORESTOMACH DEPRESSION(S>; EPITHELIAL ASPECT, ONE, CRATERLIKE, 1MM, IN OESOPHAGEAL GROOVE. UTERUS : -FLUID DISTENSION, MODERATE APPENDIX 9 MIN 313/023622 287 (Individual pathological findings - continued) GROUP 1 2 34 Print No: 0034 Printed: 09-SEP-03 EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ANIMAL NUMBER: 0040 DATE OF DEATH: 21-MAR-02 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS SCHEDULED, TERMINAL SACRIFICE STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT 260.2 GRAMS 1 X1 O 1i t* O O X NECROPSY PAT O B S E R V A T ION S HISTOPATHOLOGY i 1 1i LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LIVER : -ENLARGED; 12.85G. LIVER -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT LUNGS & BRONCHI : -INCOMPLETE COLLAPSE, MINIMAL; RIGHT. -DARK AREA(S); RIGHT, MIDDLE LOBE, ONE, SUBPLEURAL, 3MM. LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL -ALVEOLAR HAEMORRHAGE,-MINIMAL -PERIVASCULAR INFLAMMATORY CELL INFILTRATION,-MINIMAL -FOCAL OSSEOUS DEPOSITION,-MINIMAL APPENDIX 9 288 (Individual pathological findings - continued) GROUP : COMPOUND ; EXPOSURE LEVEL(PPM) : 1 CONTROL 0 2 34 -- POSE1--- 30 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0076 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 232.2 GRAMS NECROPSY PA T HO LOGY 0B SERVATION S HISTOPATHOLOGY KIDNEYS : -MINERALISATION, CORTICOMEDULLARY ,-MINIMAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-MINIMAL -PROMINENT CENTRILOBULAR HEPATOCYTE PIGMENT,-MINIMAL -EXTRAMEDULLARY HAEMOPOEISIS,-MINIMAL LN MANDIBULAR -CONGESTED APPENDIX 9 MIN 313/023622 289 (Individual pathological findings - continued) GROUP EXPOSURE LEVEL(PPM): i CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0077 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 250.8 GRAMS NECROPSY P A T H O LOGY O B SERVAT IONS HISTOPATHOLOGY LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT -ARYTENOID - SUBEPITHELIAL INFLAMMATORY CELL INFILTRATION, MINIMAL, FOCAL -PROMINENT DILATED ACINI VENTRAL GLAND,-PRESENT LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-MINIMAL -PROMINENT CENTRILOBULAR HEPATOCYTE PIGMENT,-MINIMAL LN MANDIBULAR : -CONGESTED LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-MINIMAL APPENDIX 9 MIN 313/023622 290 (Individual pathological findings - continued) GROUP : COMPOUND : EXPOSURE LEVEL(PPM): 1 CONTROL 0 2 30 34 100 300 Print No: 0034 Printed: 09-SEP-03 Protocol Number: MIN 313 ANIMAL NUMBER: 0078 DATE OF DEATH: 19-APR-02 SEX: f e m a l e DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 259.0 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-MINIMAL -FOCAL OSSEOUS DEPOSITION, -MINIMAL UTERUS : -FLUID DISTENSION, MODERATE APPENDIX 9 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL (PPM) CONTROL -POSF- 0 30 100 300 Print No: 0034 P r m te d : U 9-SEP-U 3 Protocol Number: MIN 313 ANIMAL NUMBER: 0079 DATE OF DEATH: 19-APR-02 VKO> SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH: 121 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 297.6 GRAMS NECROPSY PATHOLOGY OBSERVATIONS HISTOPATHOLOGY LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-MINIMAL LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** MIN 313/023622 APPENDIX 9 (Individual pathological findings - continued) GROUP COMPOUND EXPOSURE LEVEL (PPM) CONTROL -PO SF- 0 30 100 300 Print No: 0034 P r in te d : uy-S E P-U J Protocol Number: MIN 313 ANIMAL NUMBER: 0080 DATE OF DEATH: 19-APR-02 SEX: FEMALE DOSE GROUP : 4 SACRIFICE STATUS: SCHEDULED, FIRST POST-RECOV SAC STUDY DAY OF DEATH:: 1 2 1 STUDY WEEK OF DEATH: 18 TERMINAL BODY WEIGHT: 265.0 GRAMS NECROPSY PA T HO LOGY O B SERVATION s HISTOPATHOLOGY LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY, -M INIM AL LUNGS & BRONCHI -PER IV A SC U LA R INFLAMMATORY C ELL IN F IL T R A T IO N ,-M IN IM A L -FOCAL OSSEOUS D EPO SITIO N ,-M IN IM A L MIN 313/023622 292 * * * ANIMAL HAS NO GROSS O BSERVATIONS RECORDED * * * APPENDIX 10 pH values for terminal urine - individual values MIN 313/023622 Group :1 Compound : Control Exposure level (ppm) : 0 234 ----- POSF----- 30 100 300 Main study animals Group Males Animal pH number Females Animal pH number 1 1 8.0 21 7.3 2 - 22 8.2 3 - 23 - 4 7.5 24 7.7 5 7.8 25 - 2 6 8.4 26 7.8 7 8.0 27 7.9 8 7.6 28 8.0 9 8.4 29 - 10 7.4 30 7.7 3 11 7.7 31 - 12 - 32 - 13 8.0 33 6.7 14 - 34 7.2" 15 8.4 35 7.3 4 16 8.1 36 8.1 17 - 37 - 18 19 - 38 8.4 39 8.0 20 8.1 40 8.2 Sample taken from bladder at necropsy Sample volume < 8 pi required to obtain accurate pH reading/No sample : 293 : APPENDIX 10 (pH values for terminal urine - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 234 ----- POSF----- 30 100 300 Recovery study animals Group Males Animal number pH Females Animal pH number 1 41 61 7.5 42 6.6 62 43 7.8 63 Dead 44 64 45 6.2 65 2 46 7.8 66 47 67 7.6 48 68 49 69 50 8.8 70 3 51 7.3 71 52 7.9 72 53 7.5 73 54 74 6.3 55 7.2 75 7.8 4 56 76 57 77 58 78 59 6.6 79 60 80 Sample volume < 8 pi required to obtain accurate pH reading/No sample Animal 63 - unscheduled sacrifice : 294 : APPENDIX 10 (pH values for terminal urine - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 234 ----- POSF----- 30 100 300 Satellite animals Group Males Animal number pH Females Animal number pH 1 81" 7.1 91 . 82 - 92 7.6 83" 7.5 93 - 84 - 94 6.0 85 - 95 6.5 4 86 6.4 96 6.1 87 7.7 97 5.9 88 6.0 98 6.3 89 7.1 99 6.0 90 - 100 - Sample volume < 8 pi required to obtain accurate pH reading Sample pH'd approximately 1.5 hours following sample collection due to failure of pH hardware 295 : APPENDIX 11 Cell proliferation findings Group :1 Compound : Control Exposure level (ppm) : 0 234 ---POSE--- 30 100 300 Satellite animals Group Animal Level 1 No. Positive Negative Total C.P.I.* 1M 81 60 940 1000 6.0 82 34 966 1000 3.4 83 31 969 1000 3.1 84 15 985 1000 1.5 85 46 954 1000 4.6 4M 86 70 930 1000 7.0 87 46 954 1000 4.6 88 27 973 1000 2.7 89 31 969 1000 3.1 90 25 975 1000 2.5 IF 91 49 951 1000 4.9 92 41 959 1000 4.1 93 51 949 1000 5.1 94 35 965 1000 3.5 95 64 936 1000 6.4 4F 96 63 937 1000 6.3 97 35 965 1000 3.5 98 42 958 1000 4.2 99 40 960 1000 4.0 100 41 959 1000 4.1 Cell proliferation index ("/positive cells) Level 2 Level 3 Positive Negative Total C.P.I.* Positive Negative Total 57 943 1000 5.7 59 941 1000 40 960 1000 4.0 33 967 1000 26 974 1000 2.6 34 966 1000 24 976 1000 2.4 23 977 1000 47 953 1000 4.7 73 927 1000 64 936 1000 6.4 81 919 1000 60 940 1000 6.0 49 951 1000 36 964 1000 3.6 38 962 1000 34 966 1000 3.4 27 973 1000 31 969 1000 3.1 39 961 1000 54 946 1000 5.4 46 954 1000 60 940 1000 6.0 32 968 1000 46 954 1000 4.6 53 947 1000 37 963 1000 3.7 36 964 1000 59 941 1000 5.9 56 944 1000 55 945 1000 5.5 61 939 1000 50 950 1000 5.0 46 954 1000 51 949 1000 5.1 53 947 1000 45 955 1000 4.5 38 962 1000 55 945 1000 5.5 45 955 1000 C.P.I.* 5.9 3.3 3.4 2.3 7.3 8.1 4.9 3.8 2.7 3.9 4.6 3.2 5.3 3.6 5.6 6.1 4.6 5.3 3.8 4.5 Group Mean cell proliferation index ("/positive cells) Male Female 1 4.0 4 4.4 4.8 4.8 MIN 313/023622 APPENDIX 11 (Cell proliferation findings - continued) Group :1 Compound : Control Exposure level (ppm) : 0 234 -- POSF-- 30 100 300 Main group animals Group Animal Level 1 No. Positive Negative Total C.P.I.* 1M 1 NR NR NR NR 2 NR NR NR NR 3 NR NR NR NR 4 NR NR NR NR 5 NR NR NR NR 4M 16 NR NR NR NR 17 NR NR NR NR 18 NR NR NR NR 19 NR NR NR NR 20 NR NR NR NR IF 21 NR NR NR NR 22 NR NR NR NR 23 NR NR NR NR 24 NR NR NR NR 25 NR NR NR NR 4F 36 NR NR NR NR 37 NR NR NR NR 38 NR NR NR NR 39 NR NR NR NR 40 NR NR NR NR Cell proliferation index (%positive cells) Positive NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR Level 2 Negative Total NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR C.P.I.* NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR Positive NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR Level 3 Negative Total NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR C.P.I.* NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR MIN 313/023622 Group Mean cell proliferation index ( % positive cells) Male Female 1 NR 4 NR NR NR NR Not recorded MIN 313/023622 ADMINISTRATION OF POSF BY INHALATION TO RATS : 298 Author Simon Moore CONTENTS MIN 313/023622 Page TEST SUBSTANCE AND ADMINISTRATION Test substance...................................................................................................................................300 Administration...................................................................................................................................300 Test Atmosphere Generation............................................................................................................ 301 Exposure chambers.......................................................................................................................... 301 Procedure...........................................................................................................................................302 Test Atmosphere analysis................................................................................................................. 303 Chamber Monitoring system............................................................................................................ 304 Target concentrations....................................................................................................................... 305 Exposure chamber conditions.......................................................................................................... 305 RESULTS Vapour concentration....................................................................................................................... 306 Nominal concentration..................................................................................................................... 306 Chamber Temperature.......................................................................................................................307 Discussion.............................. 307 Calculations.......................................................................................................................................307 FIGURES A. Schematic of a rodent inhalation dosing system...................................................................308 B. Schematic of a Fourier Transform Infrared Spectrophotometer and thesampling system 309 TABLES A. Operating conditions of the inhalation exposure system.................................................... 310 B. Chamber concentrations of POSF (ppm) - daily mean values........................................... 311 C. Nominal concentrations of POSF(ppm) - individual and mean exposure values...............313 D. Chamber temperature - exposure mean values....................................................................320 APPENDICES A. Methods of sample collection and analysis for POSF........................................................ 322 B. Individual POSF concentration measurements....................................................................332 : 299 : TEST SUBSTANCE AND ADMINISTRATION MIN 313/023622 TEST SUBSTANCE The test substance, POSF, is a liquid with boiling point of 154C. A consignment of the Test Article (4 x 20 kg, Lot number 040227), was received from the Sponsor on 14 June 2001. The test substance was stored securely in the original containers at room temperature until it was transferred to the atmosphere generation system. Information provided by the Sponsor indicated that the test substance was stable for the intended duration of use on the study. The stated purity was >95.5%. Information regarding the purity and stability of the test substance is the responsibility of the Sponsor. ADMINISTRATION The test material was administered to the rats using snout-only exposure chambers, as described below: The chamber atmosphere was produced by metering the liquid test substance into a glass vapour generator through which dried air was passed at a flow rate of 29 1/minute for administration to the rats, by inhalation from snout only exposure chambers. The target chamber concentrations were achieved by metering the test substance supply using polypropylene syringes mounted on syringe drivers into the chamber air. Different concentrations of POSF were produced by the selection of different syringe feed rates. The initial settings required to obtain the target chamber concentrations were determined during the preliminary generation trials without animals present (based on the Fourier Transform Infrared (FT-ER) analysis of chamber atmosphere samples). Minor adjustments were made to the test material delivery rates in order to maintain the chamber concentration close to target. Animals assigned to Group 1 (Air control) received an exposure to air only, from the same compressed air source as used for the generation of the test atmospheres. The usage of POSF was determined, for each day of treatment, for each of the three dose group from the weight of the test material used during each exposure. The duration of administration was a single 6-hour exposure, daily, for 13 weeks for 5 days each week and two further exposures for termination purposes. : 300 : TEST ATMOSPHERE GENERATION MIN 313/023622 The vapour delivery system for the dose groups comprised individual polypropylene syringes located on syringe drivers (Precidor, Model 5003), which delivered the liquid test material via PolyTetraFluoroEthylene (PTFE) tubing to a glass frit contained in a glass vessel. The syringe sizes and syringe drivers settings required to achieve the target concentrations were established during the preliminary phase of the study. Air was passed through the vapouriser at a rate of 29 1/minute. The vapour/air mixture passed out of the vapouriser into the chamber inlet ducting through a 22 mm diameter flexible pipe. All equipment was housed in individual enclosed extracted cabinets. Each test group inhalation chamber was connected to an atmosphere generation system supplying a flow of the appropriate dilution in air of POSF vapour mixture. A calibrated flow of 30 1/minute was used as extract for each Group. For the dose groups this comprised 29 1/minute from the base of the chamber and 1 1/minute drawn for analytical sampling. A 1 1/minute differential was maintained between the inlet and outlet air flows to provide a small negative pressure within the exposure system. The in-line flowmeters were calibrated daily against high quality tapered tube rotameters measuring the free flow of air at points of attachment of the supply and extract lines to each chamber. The air control group received clean air only at a rate of 29 1/minute and a calibrated flow of 30 1/minute from the base of the chamber. Four exposure systems were used, one for each of the three dose groups and the other used solely for the control group. For all dose groups, the syringes were filled with sufficient POSF to complete the six-hour exposure period. Schematics of the inhalation exposure chamber and exposure system are presented in Figures A and B respectively and the exposure system operating conditions are presented in Table A. EXPOSURE CHAMBERS The inhalation exposure system comprised snout-only inhalation exposure chambers and rat restraining tubes. Accessories included air supply and extract lines, which attached to the top and bottom of the chamber respectively. A filtration system was incorporated into the extract line. A schematic diagram of an exposure chamber is shown in Figure A. The component parts of the system are described in further detail below: Inhalation chamber ADG snout-only inhalation chamber (ADG Developments Ltd, Hitchin, Hertfordshire, England). This is a modular apparatus of aluminium alloy construction comprising of a base unit, a variable number of animal exposure sections, each having 20 exposure ports, and a top section incorporating a central vapour inlet surrounded by a tangential air inlet. The chambers used on this study were assembled using 3 rodent exposure sections, identified as levels 1 (top) to 3 (bottom). All exposure sections had 20 ports and formed a 28 cm diameter cylinder with a volume of approximately 47 litres. During dosing, each chamber was housed in an enclosed ventilated cabinet. : 301 : Rats restraining tubes MIN 313/023622 Moulded polycarbonate tubes, tapered at one end to allow the snout only to project from the tapered end. The other end is normally closed by insertion of an expanded plastic bung. A push rod passes through the centre of the bung and is adjustable to maintain the position of the rats during restraint. Tubes were attached to a chamber by means of push-fit "O" ring seals located in the exposure ports of the animal exposure sections. Chamber levels two and three were used to expose the animals. All exposure ports not in use were sealed with an expanded plastic bung. Air supply and extract Air supplies were provided by a compressor. The air was filtered to remove any residual particulate and was dried (dew point ~2C). The calibrated exhaust airflow from the base of the exposure system was passed through a trapping system comprising a Fluosorber carbon vapour filter and a silica gel column before it was passed to exhaust. The airflow extract required for the vapour concentration analysis was solely passed to exhaust after being diluted with the room air extract. The inlet and exhaust flows used for each group are detailed in Table A. The chamber air extract was provided by vacuum pumps located outside the building that vented to atmosphere via an exhaust stack. PROCEDURE Four exposure systems were used, one for each group. The procedure followed for each group was similar except that, for the control group, the rats received air only. Therefore in the following description the comments relating to the sampling and the Fourier Transform Infrared spectrophotometer apply to the dose groups only. The FT-IR data capture programme was initialised and the path difference was checked to ensure the value was zero, subsequently, a background scan was recorded and the cell path length was calibrated. The syringes were selected, filled with the test substance, the weights of liquids were recorded and the syringes were mounted on the syringe drivers. The syringes were connected to the vapourisers by PTFE tubing. The pumps were turned on briefly in order to engage the gearing and to move the liquid along the PTFE tubing almost to the surface of the glass frit. The injection rate to be used was then set. The rats were removed from their cages and placed into restraining tubes, which were colour coded for the treatment group and numbered for each animal. The tubes were then attached to levels 2 and 3 of the chamber. Unused exposure ports were sealed with blanking plugs. The inlet and outlet airflows of the exposure system were calibrated using precision made tapered glass tube flowmeters. The connections of the generation and extract systems to the chamber were checked to ensure they were correct and working. The barometric pressure was recorded manually. : 302 : MIN 313/023622 Generation commenced as the syringe drivers were turned on, the exposure start times were then recorded. The time-controlled chamber switching system was turned on simultaneously. A sample pump continually delivered the chamber atmosphere from each test chamber to the time-controlled chamber switching system at a flow rate of 1 1/minute. At 7 minutes into the exposure, the data capture system was activated. At intervals of 10 minutes from the start of exposure, the time-controlled chamber switching system redirected the test mixture from each exposure chamber in the order Group 4, 3 and 2 to the FT-IR analyser. Analysis for the determination of POSF concentration occurred at exactly 30 minute intervals throughout each exposure. All chamber atmospheres were pumped to waste from both the FT-IR analyser and timecontrolled chamber switching system. At intervals of 30 minutes, any reactions by the rats to exposure, together with checks of generation and chamber operational parameters (including temperature and inlet and outlet air flowrate) were manually recorded. The airflows were measured throughout the exposures using in-line flowmeters. The temperature was recorded through a modified blanking plug situated at the same level in the chambers as the animals. At the end of six hours generation, the syringe drivers were turned off and the weight of the remaining syringe contents were recorded. The time-controlled chamber switching system was switched off and reset. Generation airflow was turned off and the chamber extract allowed to clear the vapour for 5 - 10 minutes (equilibration time Xgg is 7 minutes). The data capture programme halted automatically after collecting 12 samples from each dose group. At the end of this time, the rats were unloaded from the chambers and returned to their respective holding cages. The chambers were washed with hot water. The IR cell path length was retested to ensure that the initial value prior to the exposure was consistent throughout the exposure. TEST ATMOSPHERE ANALYSIS (FIGURE B) Air samples were removed from the inhalation exposure system in order to estimate the vapour concentration of the test atmosphere. The concentrations of POSF in air within the inhalation chambers of the dose groups were measured using a FT-IR Spectrophotometer. Definitive details of the FT-IR, its standardisation and validation are given in Appendix A. The Spectrophotometer was located adjacent to the exposure chambers in a separately housed ventilated cabinet. The instrument was connected to each selected sampling port by programmed switching of valves constructed as part of the time-controlled chamber switching system. All gas sampling lines were 0.6 cm diameter Teflon tubing. Each sample was removed from a spare animal exposure port through a modified blanking plug at chamber level 2, the same as the treated animals. : 303 : MIN 313/023622 CHAMBER MONITORING SYSTEM A PC running AutoQuant 3.01 software was used to monitor and record the system performance during each exposure. The data collection sequence and display were controlled by a personal computer (PC) and all information collected was displayed on a monitor. Simultaneously, the data was stored electronically. This programme was composed of three basic stages of operation: an initial setting up (pre-exposure) phase, an exposure monitoring phase and the post exposure data collation and presentation phase. The programme and FTIR hardware were loaned by the Sponsor for the duration of the study. Raw data printed as a hard copy. Setting-up phase In the initial phase, prompted by the programme screen display, the instrument response was checked, followed by a background scan. An ultra pure nitrogen cylinder was used for this purpose. The pathlength was then calibrated using a certified cylinder of ethylene in nitrogen, A regulated flowrate of 2 1/minute was used for the nitrogen and ethylene in nitrogen cylinders, the analyser drew 1 1/minute of the gas stream and the remainder was vented to waste. The flow meters used in conjunction with the cylinders were monitored using a calibrated in-line tapered tube gas flowmeter. The study details (including the daily pathlength, number of scans and samples and frequency of sampling) were entered and stored into the data capture programme prior to the exposure. Exposure monitoring phase This phase was started 7 minutes later than the start of atmosphere generation. Group 4 was analysed and subsequently after exactly 10 minutes Group 3 was analysed followed by Group 2. Each test chamber's environment was monitored during a 30-minute cycle when the analysed concentration was recorded. The data were displayed on screen, printed and stored on the local hard drive of the computer and an external zip disc. A total of twelve sample points per dose group were recorded, each of which represented 32 coadded scans. The chamber selection was controlled by an time-controlled chamber switching system. Post exposure phase At the end of the exposure, the data collected during exposure was collated and printed. The mean values, together with standard deviation were calculated for each parameter recorded. Midac Grams 32 version 4.11 software was solely used to generate a hard copy o f the infrared data. The IR cell path length calibration of the FT-IR was reassessed to ensure that the initial value prior to the exposure was consistent throughout the exposure. This was to provide a second or backup calibration if any problems with the analysis had occurred during the exposure. Details of the analytical methodologies used are given in Appendix A. : 304 : TARGET CONCENTRATIONS MIN 313/023622 The target concentrations of POSF were: Group 2 3 4 Designation Low dose Intermediate dose High dose Concentration (ppm) 30 100 300 The target concentrations were selected in consultation with the Sponsor, following the review of available data and Huntingdon Life Sciences report MIN312/014272. EXPOSURE CHAMBER CONDITIONS Chamber analysed concentration of POSF The test atmosphere was sampled from level 2 in each test atmosphere chamber. Each atmosphere was continually drawn through a transfer line at a flowrate of 1 1/minute, which was therefore in equilibrium with the mean concentration from the test chamber. Every ten minutes, air from each transfer line was sampled by the software for automated analysis in die order of descending dose group for data logging activated by the FT-IR AutoQuant software. Each dose chamber was analysed every 30 minutes. When not being sampled, these transfer lines were pumped to waste (Figure B). The achieved chamber concentrations o f POSF are presented in terms of parts per million of air and were derived from Beer Lambert's Law. The values obtained from the Sponsor's POSF calibration were standardised against validated gasbags by applying a conversion factor to give the vapour concentrations. The methodology and further detail are presented in Appendix A. Chamber spatial distribution All the animals were situated on levels two and three of the three level exposure chamber. The analytical sampling point was situated on level two of the exposure chamber. The spatial distribution of vapour in the chamber was checked between chamber levels 2 and 3 during preliminary trials for this study. A difference in concentration of less than 2% for all dose groups with varying chamber level was observed, which was well within the 10% tolerance limits normally allowed for the analysis of such exposure systems. Nominal concentration of chamber atmospheres Each chamber nominal concentration was calculated from the mass of liquid used over the six-hour exposure period and the exposure mean airflow. The ideal gas equation was used with the molecular weight of the liquid and the measured chamber conditions to compute the volume of vapour produced from the mass of liquid used. The calculation is detailed in Table C. The nominal concentration is presented in parts per million of air. Airflow and temperature These parameters were recorded manually, as described previously under the Test Atmosphere generation and Procedure sections. : 305 : RESULTS MIN 313/023622 VAPOUR CONCENTRATION The data are presented as follows: Daily mean values Individual values Table B Appendix B The study mean concentrations (the mean of daily mean values) for each group exposed to POSF are presented below: Group 2 3 4 Chamber concentration (ppm) Target Analysed 30 26 100 104 300 299 The analysed concentrations were in reasonably good agreement with target concentrations. Group 2 (Low dose) was only marginally below 10% from the target concentration, partly due to unusually low analysis results during exposures 8 and 10. The coefficients of variation of the daily mean were 15.5, 6.4 and 5.3% for Groups 2, 3 and 4 (Low, Intermediate and High doses) respectively and reflect the increasing difficulty of controlling the test material feed rate with decreasing vapour concentration. NOMINAL CONCENTRATION The data and equation are presented in Table C and summarised below: Group 2 3 4 Nominal concentration (ppm) 31 103 305 A/N ratio (%) 86 101 99 A/N _ { Analysed concentration'jxlOO Nominal concentration The nominal concentration for each exposure period was calculated for the test group from the mass of POSF delivered into the generator. The expected mean ratios of analysed to nominal concentration (A/N), expressed as a percentage for the study, should be approximately 100% for all dose groups and Groups 3 and 4 (Intermediate and High doses) showed excellent agreement with this value. The coefficients of variations for the mean nominal concentrations for Groups 3 and 4 (Intermediate and High doses) were less than 7.5% for both groups. This indicates consistency vapourisation of the test material and very little likelihood of test substance residues being retained within the generation system. The A/N ratio for Group 2 (Low dose) was lower than expected but can be accounted for by the loss of test material during the initial priming of the inlet lines to the vapouriser prior to the start of the : 306 : MIN 313/023622 exposure. This effect is more marked for Group 2, where less test material is used overall during an exposure. CHAMBER TEMPERATURE The daily mean chamber temperatures axe presented in Table D. The chamber temperatures were similar for all groups on each day of the study. DISCUSSION Control of the POSF vapour deliveiy to the exposure chambers was good especially for Groups 3 and 4 (Intermediate and High doses). The low vapour concentration and the 15.5% coefficient of variation for Group 2 (Low dose) reflect the difficulty in controlling the test material input at a very low feed rate. On occasion problems with the analysis occurred. Most of these problems relate directly to automated data capture. Where no analytical data was available, it is reasonable to assume (using the consistent A/N ratio) that the animals were exposed to the correct concentration, as a good agreement between the analysed and nominal concentrations was observed. CALCULATIONS In order to minimise the cumulative errors, which result from repeated rounding of numbers, much of the data in this report has been calculated continuously using unrounded numbers and only rounded for printing. Consequently, these rounded numbers may include rounding errors in the last significant figure, possibly leading to small apparent discrepancies with other data in the report. : 307 : FIGURE A Schematic of a rodent inhalation dosing system MIN 313/023622 Key a Polypropylene syringe b Syringe driver c Test compound feed line d Glass Vapouriser e Air supply f Sinter diameter g Exposure chamber h Observation port i Blanking plugs j Rodent restraining tubes (standard sections with 20 exposure ports on levels 1 to 3) k Exhaust plenum 1 Filtration m Air extract : 308 FIGURE B MIN 313/023622 Schematic of a Fourier Transform Infrared spectrophotometer and the sampling system Group4 d Group2 d h Key a Nitrogen cylinder b Ethylene in nitrogen cylinder c Flowmeter regulated to 1 1/minute d Test compound sample line from chamber e 10 cm IR cell f FT-IR spectrophotometer g Flowmeter regulated to 2 1/minute h Diaphragm pump i Sample line return to waste j Toggle Valves k Time-controlled chamber switching system : 309 TABLEA Operating conditions for the inhalation exposure system MIN 313/023622 Parameter Target concentration of POSF (ppm) Chamber airflows (1/minute) Elutriator output (chamber inlet) Chamber extract Vapour generator settings Sinter diameter (mm) Test material feed Syringe size (ml) N/A Not applicable 1 (Air control) 0 Group 23 (Low dose) (Intermediate dose) 30 100 4 (High dose) 300 29 29 29 29 30 29 29 29 N/A 114 114 114 N/A Precidor 5003 syringe pump N/A 5 20 50 310 : TABLE B MIN 313/023622 Chamber concentration of POSF (ppm) - daily mean values Exposure no. i 2 3 4A 1 5 2,3 Analysei Concentration (ppm) Gp2 Gp3 Gp4 35 114 327 33 117 350 36 132 302 Actual Concentration (ppm) Gp2 Gp3 Gp4 22 87 261 21 89 279 23 101 240 6' 7 42 147 393 8 22 127 363 Q1 28 114 315 12 97 290 10 21 130 359 11 41 147 380 12 41 132 370 13 43 132 369 14 42 124 365 15 35 140 378 16 46 146 390 17 45 123 376 18 42 125 371 19 43 125 381 20 44 126 376 21 4 40 117 354 22 40 120 385 23 42 129 386 24 58 143 393 25 38 123 364 26 41 135 365 27 41 136 363 28 42 133 373 29 42 134 378 30 44 137 371 31 41 140 382 32 39 135 368 33 40 139 371 34 1 11 100 287 27 114 304 27 101 296 29 102 295 28 95 292 22 108 303 31 112 313 30 94 301 28 96 297 29 95 305 29 96 301 26 89 283 26 91 308 28 99 310 41 111 315 25 94 291 27 104 292 27 105 290 28 102 299 27 103 303 29 105 297 27 108 306 25 104 295 26 107 297 35 41 135 387 27 104 310 36 39 134 369 26 103 295 37 40 134 382 26 103 306 38 42 137 389 28 105 311 39 41 146 396 27 113 317 40 42 137 386 28 106 309 No data collected due to computer problem with data capture programme Problem with ethylene in nitrogen calibration cylinder, no data analytical data collected Ethylene in nitrogen calibration cylinder ran out New Ethylene in nitrogen calibration cylinder : 311 : TABLEB MIN 313/023622 (Chamber concentration of POSF (ppm) - daily mean values - continued) Exposure Analyser Concentration (ppm) Actual Concentration (ppm) no. Gp2 Gp3 Gp4 Gp2 Gp3 Gp4 41 37 127 352 24 97 281 42 38 134 342 25 103 273 43 41 139 365 27 107 292 44 36 131 359 23 101 287 45 40 138 358 26 106 286 46 40 138 360 26 107 288 47 40 141 362 26 109 290 48 3 39 129 364 25 99 291 49 37 133 357 24 102 285 50 37 136 367 23 104 293 51 40 141 374 26 109 299 52 42 142 373 27 109 299 53 40 146 405 26 113 324 54 40 144 380 26 111 304 55 39 140 374 25 108 299 56 39 142 378 26 110 303 57 42 145 371 28 112 297 58 41 139 375 27 107 300 59 36 140 383 23 108 306 60 39 145 393 25 112 315 61 48 147 422 32 114 339 62 44 138 378 29 106 302 63 44 141 413 29 109 331 64 46 138 411 31 106 329 65 43 137 406 28 106 326 66 42 139 415 28 107 333 67 40 146 403 26 113 323 Overall Mean 40 135 375 26 104 300 sd CV (%) 4.9 12.1 8.1 20.2 4.0 6.0 5.4 15.2 6.6 16.5 6.4 5.5 Mean (1-65) 40 135 374 26 104 299 sd CV (%) 5.0 12.3 8.1 19.4 4.1 6.0 5.2 15.5 6.6 15.9 6.4 5.3 Ethylene in nitrogen calibration cylinder ran out sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 312 TABLE C MIN 313/023622 Nominal concentrations of POSF (ppm) - individual and mean exposure values The following footnotes relate to Table C and are as follows: No data collected due to computer problem with data capture programme Problem with ethylene in nitrogen calibration cylinder, no data analytical data collected Nominal concentrations were calculated from the following equations: Concentration (ppm) = ----V---- x 10 6 Va + V V= W xRxT 760 mm Hg x -------------- -- M Atm where V W M R T Atm = V. the exposure gaseous volume of POSF mass of POSF molecular weight POSF (502.14 g/mole) gas constant (0.08205 1atm mol'1K.'1) temperature (K), = temperature (C, see Table D) + 273 atmospheric pressure (mmHg) volume of air (litres) passing through the chamber during A/N sd CV (%) Analysed/nominal concentration ratio expressed as a percentage Standard deviation Coefficient of variation (sd x 100/mean) 313 : TABLEC MIN 313/023622 (Nominal concentrations of POSF (ppm) - individual exposure values - continued) Group 2 (Low dose) - Target concentration 30 ppm Exposure Time Period Barometric Test Compound No. (mins) Pressure (mmHg) usage (g) 1 360 773 5.42 2 360 759 5.58 3 360 759 5.59 4 360 765 6.19 5 360 757 6.13 6 360 751 6.05 7 360 756 6.04 8 360 756 6.45 9 360 756 5.91 10 360 773 6.00 11 360 771 6.06 12 360 770 5.91 13 360 774 6.66 14 360 773 5.67 15 360 775 5.25 16 360 775 6.84 17 360 770 6.62 18 360 771 6.53 19 360 766 6.71 20 360 776 6.43 21 360 761 6.82 22 360 762 7.01 23 360 755 7.00 24 360 754 6.33 25 360 755 6.57 26 360 744 6.74 27 360 767 6.27 28 360 765 6.90 29 360 767 7.00 30 360 761 6.89 31 360 759 7.07 32 360 760 7.05 33 360 760 7.12 34 360 760 6.86 35 360 765 6.75 Analysed (ppm) 22 21 23 l 2 1 28 12 1 11 27 27 29 28 22 31 30 28 29 29 26 26 28 41 25 27 27 28 27 29 27 25 26 1 27 Nominal (ppm)5 24 25 25 27 27 27 27 29 26 26 27 26 29 25 23 30 29 29 30 28 30 31 31 28 29 31 28 30 31 31 31 31 32 30 30 A/N ratio % 92 83 93 103 40 41 101 104 98 112 97 104 103 97 96 104 85 83 88 143 84 87 96 91 89 96 86 81 83 90 : 314 : TABLEC MIN 313/023622 (Nominal concentrations of POSF (ppm) - individual exposure values - continued) Group 2 (Low dose) - Target concentration 30 ppm Exposure Time Period Barometric Test Compound No. (mins) Pressure (mmHg) usage (g) 36 360 762 6.92 37 360 761 7.11 38 360 751 6.96 39 360 757 13.79 40 360 748 6.77 41 360 773 7.21 42 360 775 7.02 43 360 779 6.98 44 360 768 7.03 45 360 765 6.92 46 360 768 6.85 47 360 768 6.85 48 360 768 6.98 49 360 768 6.83 50 360 767 6.67 51 360 745 7.30 52 360 757 7.16 53 360 762 7.31 54 360 764 7.29 55 360 765 7.25 56 360 760 7.19 57 360 768 7.28 58 360 757 7.37 59 360 758 7.63 60 360 756 6.67 61 360 761 8.01 62 360 755 7.94 63 360 757 7.99 64 360 756 7.98 65 360 748 7.98 66 360 750 7.86 67 360 758 7.32 Overall Mean 360 762 6.91 sd 0.0 8.0 1.059 CV (%) 0.0 1.1 15.33 Analysed (ppm) 26 26 28 27 28 24 25 27 23 26 26 26 25 24 23 26 27 26 26 25 26 28 27 23 25 32 29 29 31 28 28 26 26 4.0 15.2 Nominal (ppm)5 31 32 31 61 31 31 31 30 31 31 30 30 31 30 29 33 32 32 32 32 32 32 33 34 30 36 35 36 36 36 35 32 31 4.8 15.7 A/N ratio % 83 82 88 44 91 76 81 90 73 84 87 87 82 78 80 77 86 80 80 78 80 87 83 67 84 91 82 81 88 78 79 81 86 15.1 17.5 : 315 : TABLE C MIN 313/023622 (Nominal concentrations of POSF (ppm) - individual exposure values - continued) Group 3 (Intermediate dose) - Target concentration 100 ppm Exposure Time Period Barometric Test Compound No. (mins) Pressure (mmHg) usage (g) 1 360 773 19.77 2 360 759 19.88 3 360 759 22.27 4 360 765 19.59 5 360 757 25.55 6 360 751 25.57 7 360 756 23.57 8 360 756 23.23 9 360 756 23.01 10 360 773 23.12 11 360 771 22.81 12 360 770 21.67 13 360 774 20.53 14 360 773 19.68 15 360 775 21.91 16 360 775 22.51 17 360 770 19.18 18 360 771 19.34 19 360 766 20.10 20 360 776 19.89 21 360 761 20.96 22 360 762 20.81 23 360 755 21.34 24 360 754 20.80 25 360 755 22.48 26 360 744 23.19 27 360 767 23.65 28 360 765 22.63 29 360 767 23.10 30 360 761 23.06 31 360 759 24.23 32 360 760 23.73 33 360 760 24.96 34 360 760 23.72 35 360 765 24.28 Analysed (ppm) 87 89 101 l 2 1 114 97 1 100 114 101 102 95 108 112 94 96 95 96 89 91 99 111 94 104 105 102 103 105 108 104 107 1 104 Nominal (ppm)5 86 88 99 86 114 115 105 104 103 101 100 95 90 86 95 98 84 85 89 86 93 92 95 93 100 105 104 100 102 102 108 105 111 105 107 A/N ratio % 100 101 103 108 94 99 114 107 113 110 113 115 112 113 107 111 96 99 103 119 93 99 100 102 101 103 100 99 96 97 : 316 : TABLEC MIN 313/023622 (Nominal concentrations of POSF (ppm) - individual exposure values - continued) Group 3 (Intermediate dose) - Target concentration 100 ppm Exposure Time Period Barometric Test Compound No. (mins) Pressure (mmHg) usage (g) 36 360 762 23.91 37 360 761 23.52 38 360 751 23.08 39 360 757 24.10 40 360 748 22.49 41 360 773 23.13 42 360 775 24.36 43 360 779 24.50 44 360 768 23.87 45 360 765 24.32 46 360 768 24.68 47 360 768 25.88 48 360 768 22.68 49 360 768 23.43 50 360 767 24.83 51 360 745 25.30 52 360 757 25.01 53 360 762 26.15 54 360 764 24.59 55 360 765 24.67 56 360 760 25.31 57 360 768 25.66 58 360 757 24.37 59 360 758 25.91 60 360 756 25.48 61 360 761 26.38 62 360 755 25.10 63 360 757 25.69 64 360 756 24.21 65 360 748 24.77 66 360 750 25.28 67 360 758 26.72 Overall Mean 360 762 23.37 sd 0.0 8.0 1.958 CV (%) 0.0 1.1 8.4 Analysed (ppm) 103 103 105 113 106 97 103 107 101 106 107 109 99 102 104 109 109 113 111 108 110 112 107 108 112 114 106 109 106 106 107 113 104 6.6 6.4 Nominal (ppm)5 106 104 104 107 101 101 106 106 105 107 108 114 100 103 109 115 111 115 109 109 112 113 109 115 114 117 112 114 108 112 113 119 103 9.0 8.7 A/N ratio % 97 99 101 105 104 96 97 101 96 99 98 96 99 99 96 95 98 98 102 99 98 99 98 94 98 97 95 95 99 95 94 95 101 6.2 6.1 : 317 : TABLE C MIN 313/023622 (Nominal concentrations of POSF (ppm) - individual exposure values - continued) Group 4 (High dose) - Target concentration 300 ppm Exposure Time Period Barometric Test Compound No. (mins) Pressure (mmHg) usage (g) 1 360 773 65.85 2 360 759 60.94 3 360 759 50.67 4 360 765 68.06 5 360 757 69.55 6 360 751 70.10 7 360 756 64.39 8 360 756 63.86 9 360 756 62.01 10 360 773 65.30 11 360 771 64.25 12 360 770 60.48 13 360 774 64.10 14 360 773 61.10 15 360 775 68.94 16 360 775 64.94 17 360 770 65.16 18 360 771 67.11 19 360 766 66.48 20 360 776 64.43 21 360 761 66.92 22 360 762 71.42 23 360 755 68.98 24 360 754 68.06 25 360 755 68.86 26 360 744 67.64 27 360 767 67.39 28 360 765 68.07 29 360 767 68.92 30 360 761 66.93 31 360 759 71.21 32 360 760 68.58 33 360 760 69.77 34 360 760 71.58 35 360 765 72.73 Analysed (ppm) 261 279 240 i 2 1 315 290 l 287 304 296 295 292 303 313 301 297 305 301 283 308 310 315 291 292 290 299 303 297 306 295 297 l 310 Nominal (ppm)5 288 272 226 301 311 316 288 286 278 286 282 265 280 267 301 284 286 294 294 281 297 317 309 305 308 307 297 301 304 297 317 305 310 317 321 A/N ratio % 90 103 107 109 101 100 108 111 105 109 101 110 105 101 104 107 95 97 100 103 94 95 98 99 100 100 96 97 96 96 : 318 : TABLEC MIN 313/023622 (Nominal concentrations of POSF (ppm) - individual exposure values - continued) Group 4 (High dose) - Target concentration 300 ppm Exposure Time Period Barometric Test Compound No. (mins) Pressure (mmHg) usage (g) 36 360 762 69.30 37 360 761 71.16 38 360 751 68.29 39 360 757 69.06 40 360 748 68.35 41 360 773 68.52 42 360 775 67.52 43 360 779 69.51 44 360 768 69.42 45 360 765 68.12 46 360 768 68.72 47 360 768 67.79 48 360 768 66.76 49 360 768 67.16 50 360 767 69.87 51 360 745 72.90 52 360 757 70.84 53 360 762 53.96 54 360 764 72.94 55 360 765 70.93 56 360 760 72.86 57 360 768 71.65 58 360 757 74.47 59 360 758 79.85 60 360 756 73.32 61 360 761 78.25 62 360 755 76.65 63 360 757 77.41 64 360 756 73.64 65 360 748 77.28 66 360 750 78.12 67 360 758 77.61 Overall Mean 360 762 68.76 sd 0.0 8.0 5.141 CV (%) 0.0 1.1 7.5 Analysed (ppm) 295 306 311 317 309 281 273 292 287 286 288 290 291 285 293 299 299 324 304 299 303 297 300 306 315 339 302 331 329 326 333 323 300 16.5 5.5 Nominal (ppm)5 307 316 307 308 308 299 294 302 305 301 302 298 293 295 308 330 316 238 322 313 324 315 332 355 327 346 342 345 328 348 350 344 305 23.8 7.8 A/N ratio % 96 97 101 103 100 94 93 97 94 95 95 97 99 97 95 91 94 136 94 96 93 94 90 86 96 98 88 96 100 94 95 94 99 7.2 7.3 : 319 : TABLE D Chamber temperature - exposure mean values MIN 313/023622 Temperature (C) Gpl Gp2 Gp3 Gp4 (Control) (Low dose) (Inter dose) (High dose) 1 20.8 20.5 20.4 21.3 2 20.3 20.9 20.4 21.3 3 20.5 20.6 20.2 20.9 4 20.7 20.4 20.3 21.1 5 20.4 20.5 20.3 21.2 6 20.6 20.3 20.0 20.9 7 22.0 20.3 20.3 21.3 8 21.7 20.4 20.4 21.2 9 21.3 20.2 20.3 21.5 10 21.8 20.2 20.4 21.0 11 22.3 20.5 20.3 21.2 12 22.0 20.8 20.2 20.7 13 21.3 21.0 20.4 21.0 14 21.3 21.0 20.3 21.0 15 21.3 20.8 20.0 20.9 16 21.3 20.0 20.3 21.2 17 21.3 20.0 20.4 20.9 18 21.3 20.0 20.0 21.0 19 21.4 20.9 20.4 21.2 20 21.3 20.0 19.9 20.9 21 20.9 20.0 20.0 20.8 22 20.8 20.0 20.0 20.8 23 21.2 20.8 20.3 20.8 24 21.0 20.0 20.0 20.9 25 20.9 20.6 20.0 20.8 26 21.0 20.5 20.0 20.9 27 21.0 20.8 20.3 20.8 28 21.3 20.7 20.0 20.8 29 21.7 20.7 20.2 20.9 30 20.8 20.8 20.2 20.9 31 20.8 20.7 20.1 20.6 32 20.8 20.7 19.9 20.5 33 20.8 20.5 20.0 20.4 34 20.8 20.3 19.9 20.0 35 20.8 20.0 20.0 20.5 36 20.9 20.4 19.9 20.4 37 20.0 20.2 20.0 20.4 38 20.2 20.4 20.4 20.8 39 20.0 20.3 20.0 20.5 40 20.0 20.2 20.0 20.4 : 320 : TABLED MIN 313/023622 (Chamber temperature - exposure mean values - continued) Exposure Temperature (C) no. Gpl Gp2 Gp3 (Control) (Low dose) (Inter dose) 41 20.0 20.2 20.1 42 20.0 20.5 20.1 43 20.3 20.4 20.4 44 20.0 20.5 20.3 45 20.3 20.4 20.2 46 20.3 20.4 20.1 47 19.9 20.3 19.9 48 20.0 20.7 20.2 49 20.3 20.4 20.3 50 20.2 20.3 20.0 51 19.9 20.4 20.2 52 20.0 20.3 20.0 53 19.3 19.4 19.2 54 20.0 20.4 20.2 55 20.0 20.0 19.9 56 19.9 19.9 20.0 57 20.0 20.2 20.0 58 19.9 20.3 20.2 59 20.1 20.6 20.3 60 20.0 20.1 20.2 61 19.9 20.4 19.9 62 19.9 20.3 19.9 63 19.9 20.3 20.0 64 19.9 20.0 20.0 65 19.9 20.2 19.9 66 19.3 19.8 19.5 67 19.0 19.5 19.6 Overall Mean 20.6 20.4 20.1 sd 0.71 0.33 0.23 CV (%) 3.4 1.6 1.1 Mean (1-65) 20.6 20.4 20.1 sd 0.67 0.31 0.21 CV (%) 3.2 1.5 1.0 sd standard ceviation CV (%) Coefficient of variation (sd x 100/mean) Gp4 (High dose) 20.3 20.4 20.9 20.5 20.4 20.8 20.7 20.3 20.4 20.5 20.4 20.7 19.6 20.6 20.5 20.8 20.8 20.1 20.3 20.0 19.9 19.8 19.9 20.0 19.9 19.0 19.0 20.6 0.50 2.4 20.7 0.42 2.0 : 321 : APPENDIX A Methods of sample collection and analysis for POSF MIN 313/023622 SAMPLE COLLECTION Chamber concentration Each chamber atmosphere was continually drawn from each test chamber to the time-controlled chamber switching system at a regulated flow rate of 1 1/minute using a laboratory pump. The chamber atmosphere was drawn through the ERcell at a regulated flow rate positioned downstream of the IR cell and subsequently pumped to waste. The concentration of POSF was determined at exactly 30 minute intervals throughout each exposure. The sampled atmosphere was pumped to waste. The airflow to the spectrophotometer was monitored throughout each of the exposures using a calibrated in-line tapered tube gas flowmeter. Gas sampling lines were PTFE tubing (0.6 cm diameter). The sample line from each test chamber was located on level 2 of the chamber. METHOD OF ANALYSIS Chamber atmosphere samples were analysed by extractive FT-IR. The method of sample analysis is detailed, together with a summary of the method validation, in the Inhalation Analytical Procedure at the end of this Appendix. : 322 : APPENDIX A MIN 313/023622 (Methods of sample collection and analysis for POSF - continued) CALCULATIONS FT-ER analysis The samples of chamber atmosphere were passed through the IR cell, which was calculated using the Sponsors electronic standard from Beer-Lambert's Law. A conversion factor was applied to the AutoQuant concentrations obtained with the Sponsors electronic standard to convert these values to the actual vapour concentrations. This arose from a discrepancy of between 23% to 47%, increasing with vapour concentration between validated gasbags, nominal chamber concentrations and the AutoQuant concentration. The equation used for the conversion calculated from validated gasbags and given below as equation 1, had a correlation coefficient of 0.9995 over the POSF concentration range. ,Vrapour C_oncentrati.on (ppm) = -A--u-t-o-Q--u--a-n--t-C--o-n^centr-a--ti-o--n---8--.-2-4--1-4- (.1..) This difference in values was also apparent during Huntingdon Life Sciences report no MIN312/014272 but to a lesser degree, therefore, the introduction of the time-controlled chamber sampling system may have partly resulted in the increased discrepancy. Other difference in these values is attributed to a combination of uncertainties as follows: - POSF reference spectra and subsequent AutoQuant method calculations. These have errors of 5% associated with them (responsibility of the Sponsor); - The cylinder of ethylene in nitrogen standard has certified errors of 2%; - Pressure drop in the IR cell; - The effect of using ultra pure nitrogen for the background scans (rather than the control chamber atmosphere) is unknown. The uncertainties relating to the analysed and room air concentrations were estimated by the data capture programme based on the least squares statistics. The method for calculating the concentration of POSF from the mass used to prepare each vapour standard using the nominal feed rate is given below in equations 2 and 3. where Concentration = -------- x 1,0 0 0 ,0 0 0 ppm V. + V V - w x R x T 760 mm HgX ---------------------------------- M Atm V= gaseous volume of POSF (ml) w= mass of POSF (g) M= molecular weight of POSF (502.14 g/mole) R = Gas constant (0.08205 ml atm mmol' 1 K'1) T = temperature (K) Atm = atmospheric pressure (mmHg) va = volume o f air (ml) (2 ) : 323 : APPENDIX A MIN 313/023622 (Methods of sample collection and analysis for POSF - continued) COMPOUND SPECIFIC INHALATION ANALYTICAL PROCEDURE FOR POSF The analysis of POSF (Perfluorooctane sulfonyl fluoride) in air sample substrate The method outlined in this document has been validated and is considered fit for the purpose of monitoring conditions in an Inhalation Toxicology study. This document details the basic procedures for the analysis of POSF sampled by extractive FT-IR from test atmospheres. The resulting samples, of approximate concentration 240 to 360 ppm, are analysed using the MEDAC I-Series spectrophotometer and AutoQuant software. Study specific amendments and additions will be detailed within a supplementary document. NOTE Throughout this document, the symbol indicates that the relevant information is not available at present, but will be included in a Study specific supplement. EFFECTIVE FROM: 29 August 2001 Test substance POSF, perfluorooctane sulfonyl fluoride has the following formula: C8F17SO2F. Appearance Clear liquid Storage Ambient temperature Reagents Air In House Compressed Ethylene in nitrogen Sponsor Calibration gas (certified 2%) Nitrogen Sponsor Calibration gas (certified >99.9995%) Liquid nitrogen BOC Cooling Medium : 324 : APPENDIX A MIN 313/023622 (Methods of sample collection and analysis for POSF - continued) Equipment Balance and Data printer Sartorius R160P with YDP-01 Flow meter Gilmont Instruments 0-4 Lpm FT-IR Midac Corporation 12001 series (serial no. 151) Computer Compaq LTE5400 Sotware Midac Corporation AutoQuant Version 3.01 Midac Grams 32 Version 4.11 Chamber ADG 3 level snout only Syringe Driver Precidor Type 5003 UV Quartz Cells Midac Corporation ZnSe Gas Cell General laboratory glassware Accessories also employed are: a vacuum pump; cylinder regulators, power connectors; regulated exhaust pumps. Consumables Syringes Aldrich 50 ml polypropylene Method of Sample Measurement A volume of POSF is dispensed from the test drum into a 50ml syringe. The syringe driver is set to give a concentration of 30, 100 or 300ppm in an airstream of 30 1/minute airflow, mixed thoroughly in the vapouriser and fed directly into the chamber. The test atmosphere is drawn from the exposure chamber along the sample line at a flow rate of 1 1/minute directly through the IR cell. Production of a Background Scan A flowrate of 11/minute ultra-pure nitrogen (cylinder No. C0C7700219 (nitrogen >99.9995 2%) is drawn though the IR cell. After a period of 3-5 minutes, an FT-IR single beam spectrum is generated approximately every 2 seconds and continuously collected for 32 scans to produce a co-added spectrum. : 325 : APPENDIX A MIN 313/023622 (Methods of sample collection and analysis for POSF - continued) Storage of standards and samples Stability experiments were not required for on-line analysis. Calibration and Quantification The AutoQuant system calculated a concentration for each absorbance measurement as follows. For each sample measurement, the amount present is determined from the absorbance sample using the equation below (Beer's Law): Where A a b Amount(ppw) = ------ axb absorbance at a given frequency of POSF in the sample spectrum absorption coefficient (absorptivity) of POSF in the sample spectrum, which is derived from the reference calibration transfer standard spectrum path length of the cell derived from calibration data 326 : APPENDIX A MIN 313/023622 (Methods of sample collection and analysis for POSF - continued) Calibration of Cell Path Length Calibration of the cell path length is performed daily before each exposure. A certified concentration of ethylene in nitrogen (cylinder No. C0C9700874 (ethylene = 2020ppm 2 %) is passed through the sampling system prior to sample collection to check instrument response and determine the calibrated cell path-length. Cell path-length is a critical input to the analytical method used to perform each spectral analysis. The ethylene value is read from the test certificate on the calibration cylinder. A test atmosphere of ethylene in nitrogen at a flowrate of 1 1/minute is drawn through the DR. cell. After a period of 3-5 minutes, a FT-ER single beam spectrum is generated approximately every 2 seconds and continuously collected for 32 scans to produce a less noisy spectrum. This produces a calibrated path length. This value is then entered into the POSF method file and manually recorded. QC Path length Limits: -The calibration will be repeated by the user if no peak is present in the 1400-650 cm' 1 region or if the path length value is outside the range 0.100 to 0.115 m. If the calibration still does not conform, all of the calibration set up parameters will be checked. QA Instrument Calibration Check: The calibration is reverified and subsequently recorded at the end of the exposure. Instrument calibration checks are within 5%. Repetition of two four hornperiods showed that the final calibration gave the path length values differed by less than 2 % from the initial calibration value o f the ethylene in nitrogen cylinder (cylinder No. C0C9700874 (ethylene = 2020ppm 2%). The 2% value is inside the 5% tolerance limits allowed as quality check standards. The temperature of the test gas stream and the barometric pressure will be monitored manually. FT-IR analyser specification The Midac 12000 series FT-ER spectrophotometer has a wavelength accuracy of 0.5cm'1. A MCT detector monitors the absorbance of various chemical bonds. AutoQuant version 3.01 (MIDAC, Irvine, CA) software is used for all infrared data acquisition and quantification. The length of the DR. cell or path length (approximately 10 cm) dictates the concentration range that will give a linear relationship between absorbance and concentration. The optimum flow rate for the ER cell is 1 1/minute, which is set by a flow meter and all the analysis is conducted at that flow rate to avoid differences of cell pressure. Further details can be obtained from the operating procedure stored proximal to the FT-ER system. The analyser draws 1 1/minute from the gas stream and the remainder is returned to the exposure system exhaust. All exhaust lines are monitored by precision stainless steel and glass in-line rotameters. : 327 : APPENDIX A MIN 313/023622 (Methods of sample collection and analysis for POSF - continued) Raw Data Raw data is defined as the first hard copy output obtained immediately following data acquisition by the FT-ER system. The raw data is printed using the MIDAC Grams 32 software and will be signed and dated immediately after printing. An electronic backup copy of this is saved to the local zip drive. If a run time hard copy was not obtained through a printer fault, the electronic copy from the zip drive directory may be used to generate a hard copy, which will be signed and dated. Exposure data is stored in the file format YYMMDD.txt. Computer hardware: Computers and VDUs are maintained by the Sponsor. Printer is maintained by the IT department. FT-IR System hardware: The Sponsor has supplied the hardware. Arrangements for servicing and maintenance would be made in consultation with the Sponsor. Software: Documentation is the responsibility of the Sponsor. Safety The COSHH assessment provides details regarding the toxicity of the test material POSF. It is considered that the standard handling procedures used within the department are suitable to prevent exposure to the test material. In case of a total power failure, switch off the FT-IR and test compound generation equipment immediately. : 328 : APPENDIX A MIN 313/023622 (Methods of sample collection and analysis for POSF - continued) Summary of method validation The raw data for the method validation is located in study MIN/312. Uncertainty of POSF analysis Uncertainties were said (by the Sponsor) to be less than 5% for the AutoQuant method calculations. The POSF reference spectrum is from the 3M quantitative library and was generated at 3M using EPA guidelines. Other principal uncertainties were: ethylene path length Calibration (2%); interference from alkyl sufonyl fluorides (<4.5% v/v, but interference unknown). 329 : APPENDIX A MIN 313/023622 (Methods of sample collection and analysis for POSF - continued) MIN/313 - STUDY SPECIFIC SUPPLEMENT TO THE INHALATION ANALYTICAL PROCEDURE FOR POSF (PERFLUORO-OCTANE SULFONYL FLUORIDE) This supplement details additions and amendments to the procedure to be used for the FT-IR assay of POSF obtained from air samples collected on the above study. The assay, incorporating the additions and amendments, is suitable for the analysis of POSF, in air, at concentrations within the range of 20 to 400ppm. The IAP was prepared as part of MIN/312. Details given in this supplement supersede those in the compound specific IAP. EFFECTIVE DATE : 18 December 2001 Analytical standard Name Batch number Purity Expiry date Supplier FX-8B, POSF, T-7661.1, Perfluoro-octane sulfonyl fluoride 040227 >95.5% (Perfluoroalkyl sulfonylfluorides <4.5%) Not Supplied Sponsor Fourier Transform - Infrared Spectrophotometer The analysis is performed using a FT-ER analyser supplied by the sponsor. Preparation of standard solutions Prepare gasbags in the range 20 to 400 ppm. Calibration and Quantification Calibration of the pathlength by an ethylene in nitrogen standard (cylinder No. C0C9700874 (ethylene = 2020ppm 2%). Quantification using the 3M quantitative library generated at 3M using EPA guidelines for generating reference spectra (v). Calibration Conversion Factor A conversion factor is to be used between nominal and analyser concentrations. Gasbag cone (ppm) = (AutoQuant Cone (ppm) - 8.2414)/ 1.2219 R2= 0.9995 : 330 : APPENDIX A MIN 313/023622 (Methods of sample collection and analysis for POSF - continued) EFFECTIVE DATE : 17 January 2002 Calibration and Quantification Calibration of the pathlength by an ethylene in nitrogen standard (cylinder No. 9701599 (ethylene = 2QQ0ppm 2%). Quantification using the 3M quantitative library generated at 3M using EPA guidelines for generating reference spectra (v). : 331 : APPENDIX B Individual POSF concentration measurements MIN 313/023622 Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 1 1 35.90 92.55 69.28 2 38.33 113.27 315.96 3 36.15 115.69 331.68 4 33.10 113.96 335.12 5 34.54 113.89 332.97 6 33.79 113.48 327.27 7 34.49 113.72 318.74 8 33.98 113.53 321.30 9 34.64 113.90 324.21 10 34.86 115.03 330.44 11 34.16 115.18 331.42 12 34.81 115.07 329.33 Mean6 34.80 114.25 327.13 sd 1.396 0.833 6.249 CV (%) 4.0 0.7 1.9 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 332 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 2 13 7 31.68 90.80 54.20 14 7 33.01 100.55 8.95 Mean 32.34 95.68 31.57 ~TP~ 35.15 117.75 299.11 16 7 35.74 119.89 321.14 Mean 35.44 118.82 310.13 177 36.03 122.00 360.78 18 7 36.54 121.99 364.65 Mean 36.28 121.99 362.71 19 33.57 121.54 365.83 20 31.83 116.18 352.78 21 32.40 116.61 347.51 22 33.44 116.50 352.83 23 32.46 115.05 356.40 24 32.46 115.36 348.13 25 32.77 116.04 345.75 26 33.17 116.84 351.15 27 32.77 115.55 353.97 28 33.17 29 33.48 Mean * 33.33 117.32 349.74 sd 1.573 2.416 14.502 CV (%) 4.7 2.1 4.1 Sampling time selected at 5 rather than every 10 minutes intervals The initial 2 concentration measurement of each exposure were excluded from all calculations sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 333 APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 30 34.90 130.74 16.23 31 38.33 142.20 48.93 32 35.32 145.53 52.30 33 36.18 138.76 341.38 34 34.32 135.83 374.05 35 36.67 131.15 378.02 36 36.51 127.34 371.22 37 35.28 126.71 352.88 38 36.56 127.23 352.70 39 37.92 126.06 350.11 40 35.80 125.75 348.27 41 37.45 128.14 352.90 Mean6 36.39 132.25 302.07 sd 1.201 7.136 124.866 CV (%) 3.3 5.4 41.3 7 42 -0.33 -0.30 * -0.32 * 43 40.82 143.55 349.91 44 40.67 146.12 388.24 45 42.73 146.27 397.31 46 41.54 146.87 402.32 47 42.35 146.42 402.25 48 41.53 150.84 400.80 49 43.12 148.34 396.29 50 40.08 147.35 394.59 51 43.16 147.42 395.53 52 43.37 146.53 395.09 53 44.45 148.59 396.89 Mean6 42.16 147.12 392.66 sd 1.347 1.817 14.728 CV (%) 3.2 1.2 3.8 The initial concentration measurement of each exposure was excluded from all calculations Chamber sample lines not connect to chamber sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 334 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 8 54 55 56 57 58 59 60 25.27 105.90 122.93 TO 140.34 10 154.99 10 170.37 10 183.67 10 195.64 10 207.48 10 61 24.07 126.02 357.72 62 23.24 129.13 372.61 63 21.57 127.61 372.90 64 21.78 124.02 361.56 65 22.61 126.55 359.06 66 20.66 126.59 361.39 67 20.32 124.14 359.34 68 22.93 126.77 360.38 69 23.31 128.38 360.28 70 23.59s 126.39 362.33 71 22.09 s 128.83 362.78 Mean 22.33 a 126.776 362.76 sd 1.213 1.682 5.156 CV (%) 5.4 1.3 1.4 The initial concentration measurement of each exposure was excluded from all calculations Sampling time selected at 1 rather than every 10 minutes intervals, values excluded from mean and standard deviation sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) Overall mean is time weighted for samples 70 and 71. The mean of these two samples is used in the overall caluclations. : 335 APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 10 72 73 19.86 li 95.85 11 69.43 330.27 74 20.67 131.48 360.69 75 20.38 130.55 360.93 76 21.51 129.94 360.96 77 21.19 128.79 359.95 78 20.36 129.67 358.44 79 21.52 129.41 356.23 80 21.89 129.40 358.68 81 22.36 130.69 361.78 82 21.04 132.40 372.17 83 23.19 129.88 365.15 Mean6 21.41 130.22 358.66 sd 0.896 1.083 10.307 CV (%) 4.2 0.8 2.9 The initial concentration measurement of each exposure was excluded from all calculations Computer malfunction with data capture programme during exposure as it was observed that the backup drive was full. Rectified by changing to fresh zip disk, sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 336 APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 11 84 39.60 137.85 256.25 85 40.91 154.56 336.79 86 40.87 159.78 377.66 87 41.67 163.38 385.94 88 40.74 163.10 387.87 89 42.12 165.72 388.93 90 40.52 164.94 392.01 91 42.29 152.18 389.73 92 40.79 124.65 396.41 93 40.23 125.31 405.70 94 41.44 124.03 363.92 95 41.11 123.08 356.98 Mean 6 41.15 147.34 380.17 sd 0.654 18.751 20.016 CV (%) 1.6 12 96 39.05 12.7 84.53 5.3 -------- 13--------- 97 41.78 128.93 325.45 98 40.74 133.38 362.16 99 41.44 133.81 378.47 100 40.44 133.96 380.42 101 41.09 132.09 376.41 102 41.05 132.16 376.09 103 40.69 131.55 375.12 104 41.00 129.85 370.99 105 41.69 130.53 372.55 106 41.56 133.59 373.38 107 40.82 131.64 373.78 Mean6 41.12 131.95 369.53 sd 0.443 1.676 15.366 CV (%) 1.1 1.3 4.2 The initial concentration measurement of each exposure was excluded from all calculations Sample missed as analysis started late in error sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 337 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 13 108 39.94 97.15 136.85 109 43.16 116.79 328.28 110 42.73 120.36 354.56 111 41.84 120.12 356.49 112 42.95 126.85 352.76 113 43.39 141.33 380.05 114 43.15 141.21 379.63 115 43.68 137.95 379.08 116 43.10 136.71 380.39 117 43.23 136.76 380.04 118 43.62 137.16 383.01 119 44.16 140.35 381.80 Mean 6 43.18 132.33 368.73 sd 0.593 9.391 17.985 CV (%) 1.4 7.1 4.9 14 120 3.44 2.84 4.53 121 20.56 19.96 327.08 122 45.85 131.04 373.76 123 44.88 134.56 365.24 124 44.18 131.30 366.61 125 44.52 134.09 364.23 126 43.48 135.35 364.13 127 44.03 136.22 368.66 128 43.89 135.93 369.21 129 45.40 134.68 370.31 130 44.82 134.65 371.43 131 44.12 136.03 373.60 Mean6 42.34 123.98 364.93 sd 7.257 34.546 13.011 CV (%) 17.1 27.9 3.6 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 338 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 15 132 6.20 u 113.67 311.48 133 12.34 13 137.61 367.35 134 6.8613 143.16 382.52 135 35.91 143.10 388.48 136 41.80 139.85 383.03 137 40.23 141.18 383.36 138 41.76 140.74 378.17 139 42.35 137.69 375.66 140 41.35 139.23 375.23 141 41.35 139.32 374.48 142 43.37 141.19 376.17 143 42.17 140.46 375.38 Mean6 35.41 140.32 378.16 sd 12.961 1.846 5.773 CV (%) 36.6 1.3 1.5 16 144 44.73 119.42 109.02 145 46.44 143.35 367.09 146 45.40 145.90 400.96 147 45.09 146.04 392.96 148 46.21 145.41 390.15 149 45.12 144.29 385.77 150 45.94 144.94 387.23 151 45.66 146.25 388.61 152 46.20 146.21 389.78 153 46.57 146.55 394.96 154 46.87 146.89 398.52 155 47.96 146.32 399.36 Mean6 46.13 145.65 390.49 sd 0.843 1.066 9.293 CV (%) 1.8 0.7 2.4 The initial concentration measurement of each exposure was excluded from all calculations Syringe driver incorrectly set at start of exposure sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 339 : MIN 313/023622 APPENDIX B (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 17 156 42.98 115.04 133.52 157 43.48 122.76 349.23 158 44.37 123.47 368.05 159 44.15 119.40 365.70 160 44.63 120.33 365.13 161 44.42 121.94 371.88 162 44.01 122.91 383.05 163 45.88 121.80 384.45 164 44.88 119.55 384.35 165 45.25 124.05 386.08 166 45.26 128.18 388.02 167 46.34 128.27 389.64 Mean6 44.79 122.97 375.96 sd 0.843 3.010 12.822 CV (%) 1.9 2.4 3.4 18 168 40.04 104.86 218.39 169 40.50 120.73 347.40 170 40.74 122.04 366.15 171 41.78 122.67 365.99 172 42.08 124.61 369.35 173 42.57 125.99 365.23 174 42.12 126.21 363.72 175 42.47 126.93 361.94 176 42.87 125.19 368.71 177 42.77 126.88 385.43 178 42.90 127.85 393.29 179 42.80 129.60 398.75 Mean6 42.15 125.34 371.45 sd 0.838 2.653 15.016 CV (%) 2.0 2.1 4.0 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 340 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 19 180 14 181 14 182 43.31 125.29 399.81 183 42.40 125.64 411.94 184 43.09 124.45 380.88 185 43.47 120.64 373.43 186 44.93 123.18 366.86 187 42.31 124.02 363.96 188 41.39 123.33 364.05 189 43.58 125.47 378.18 190 43.62 126.77 386.73 191 43.55 126.59 388.45 Mean6 43.165 124.54 381.43 sd 0.9569 1.84 15.78 CV (%) 2.2 1.5 4.1 20 192 2.24 100.04 84.58 193 73.48 123.30 352.36 194 39.57 126.59 387.60 195 39.90 125.30 391.28 196 38.44 125.42 389.29 197 41.29 124.10 382.83 198 40.64 123.95 371.92 199 41.51 123.55 370.23 200 41.18 123.75 369.34 201 41.38 125.30 373.65 202 42.59 131.93 375.02 203 41.47 130.40 374.77 Mean6 43.77 125.78 376.21 sd 9.918 2.861 11.188 CV (%) 22.7 2.3 3.0 The initial concentration measurement of each exposure was excluded from all calculations Time-controlled chamber switching device out of synchrony with measurement, sample collected over 2 groups, values excluded from mean and standard deviation sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 341 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 21 204 36.69 94.76 83.95 205 38.54 113.61 312.36 206 39.89 117.11 352.13 207 39.70 115.33 355.36 208 38.95 113.83 366.07 209 39.28 113.43 335.20 210 38.37 115.61 318.35 211 40.59 119.13 362.39 212 40.56 119.31 373.52 213 40.59 119.82 372.14 214 40.20 120.53 373.54 215 40.06 121.05 372.45 Mean6 39.70 117.16 353.96 sd 0.811 2.924 22.389 CV (%) 2.0 2.5 6.3 22 216 39.30 100.58 114.06 217 39.86 117.88 354.92 218 39.74 120.56 376.31 219 40.10 117.48 385.68 220 39.06 117.83 386.24 221 38.69 120.75 382.81 222 39.69 121.86 391.60 223 39.89 119.89 390.29 224 38.67 119.04 392.70 225 40.16 119.18 391.00 226 39.98 120.02 390.68 227 41.64 124.38 391.23 Meanb 39.77 119.90 384.86 sd 0.820 2.012 11.067 CV (%) 2.1 1.7 2.9 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 342 : MIN 313/023622 APPENDIX B (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 23 228 41.70 102.53 257.45 229 42.14 126.01 367.43 230 40.26 129.21 400.37 231 40.89 129.67 409.41 232 41.22 128.41 396.45 233 41.75 127.74 380.48 234 40.95 126.51 382.36 235 43.11 126.68 380.13 236 41.91 129.06 384.69 237 42.68 131.20 383.94 238 43.15 131.09 382.87 239 42.81 130.97 382.97 Mean 6 41.90 128.78 386.46 sd 0.980 1.885 11.444 CV (%) 2.3 1.5 3.0 24 240 59.85 129.61 148.70 241 63.11 147.22 375.61 242 60.23 149.54 407.34 243 61.31 141.12 400.47 244 57.70 142.37 393.96 245 56.57 143.23 391.62 246 56.28 144.13 391.74 247 57.35 144.86 392.28 248 54.99 139.36 391.70 249 56.08 140.61 392.38 250 56.06 143.21 394.66 251 56.62 141.56 393.35 Mean6 57.84 143.38 393.19 sd 2.564 2.979 7.571 CV (%) 4.4 2.1 1.9 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 343 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 25 252 30.84 111.70 93.22 253 33.23 119.18 334.79 254 38.65 119.84 363.66 255 37.64 120.48 366.52 256 36.95 120.98 366.55 257 38.55 118.42 365.26 258 40.24 117.84 366.08 259 39.18 126.60 366.96 260 39.10 127.90 367.12 261 38.92 125.76 366.40 262 39.85 126.45 366.91 263 40.73 126.59 369.85 Mean6 38.46 122.73 363.65 sd 2.042 3.889 9.682 CV (%) 5.3 3.2 2.7 26 264 38.73 105.65 124.48 265 39.50 132.68 345.22 266 40.62 136.33 368.37 267 40.64 131.73 367.59 268 40.73 131.68 367.15 269 40.55 136.58 366.16 270 39.82 137.20 366.56 271 40.78 134.76 365.63 272 40.56 135.74 366.14 273 42.63 135.96 365.94 274 41.33 135.94 369.44 275 41.16 137.71 369.57 Mean6 40.76 135.12 365.25 sd 0.811 2.141 6.783 CV (%) 2.0 1.6 1.9 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 344 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 27 276 40.18 108.26 97.60 277 40.31 132.81 333.61 278 41.30 138.52 365.51 279 39.56 136.87 374.62 280 41.24 136.02 368.45 281 39.65 134.74 364.71 282 40.44 134.80 361.69 283 40.80 135.44 360.84 284 41.21 135.77 362.29 285 40.63 136.42 365.36 286 42.95 137.09 365.96 287 40.85 137.40 370.96 Mean6 40.81 135.99 363.09 sd 0.921 1.549 10.595 CV (%) 2.3 1.1 2.9 28 288 40.18 115.18 101.56 289 41.78 129.92 366.93 290 42.11 131.01 375.46 291 41.62 134.33 375.24 292 40.15 133.65 373.83 293 42.80 132.06 373.03 294 41.20 131.52 371.78 295 42.68 132.97 368.54 296 42.44 133.71 375.67 297 42.83 133.86 372.59 298 41.49 133.42 376.12 299 42.72 132.04 375.06 Mean6 41.98 132.59 373.11 sd 0.838 1.384 3.019 CV (%) 2.0 1.0 0.8 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 345 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 29 300 39.00 115.96 281.48 301 40.30 142.70 357.92 302 42.39 141.69 383.39 303 41.40 131.70 381.43 304 41.20 130.98 378.05 305 41.57 131.07 376.59 306 41.58 132.06 378.17 307 42.10 132.38 379.36 308 41.65 131.68 383.07 309 41.24 131.67 378.66 310 42.51 132.17 379.34 311 43.69 136.46 384.56 Mean6 41.79 134.05 378.23 sd 0.878 4.294 7.200 CV (%) 2.1 3.2 1.9 30 312 43.08 110.18 104.51 313 44.88 127.62 342.01 314 44.00 141.97 370.71 315 43.32 140.98 385.94 316 43.83 138.47 373.85 317 44.23 137.21 368.95 318 44.47 136.18 369.93 319 44.62 135.88 370.89 320 45.41 136.86 373.25 321 43.72 137.36 376.54 322 44.19 135.71 374.70 323 43.94 138.59 377.14 Mean 6 44.24 136.98 371.26 sd 0.584 3.702 10.786 CV (%) 1.3 2.7 2.9 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 346 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 31 324 41.23 122.56 92.77 325 41.38 138.49 348.71 326 41.93 136.16 389.29 327 41.36 141.71 391.07 328 41.99 140.66 376.82 329 40.85 140.85 377.81 330 40.98 140.13 379.77 331 41.25 134.52 384.00 332 39.97 140.82 385.50 333 41.59 142.92 386.36 334 42.10 141.63 386.60 335 42.54 141.35 390.94 Mean6 41.45 139.93 381.53 sd 0.705 2.545 11.945 CV (%) 1.7 1.8 3.1 32 336 36.89 108.07 68.97 337 38.14 128.15 328.86 338 38.61 135.52 366.71 339 37.62 134.33 369.47 340 38.01 134.73 370.31 341 38.61 133.52 368.87 342 39.18 135.24 372.46 343 40.90 137.13 375.19 344 39.06 137.00 377.62 345 40.52 136.98 373.86 346 40.64 136.77 373.68 347 41.64 139.90 375.60 Mean6 39.36 135.39 368.42 sd 1.346 2.964 13.520 CV (%) 3.4 2.2 3.7 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 347 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 33 348 37.69 115.51 77.87 349 39.83 137.08 345.41 350 40.32 139.17 372.03 351 40.59 137.72 374.35 352 40.12 136.18 372.51 353 40.27 136.20 372.40 354 40.74 136.92 370.83 355 40.80 137.86 372.19 356 39.09 139.94 373.02 357 41.54 142.87 373.66 358 40.55 140.10 375.38 359 41.44 139.56 376.69 Mean6 40.48 138.51 370.77 sd 0.692 2.037 8.573 CV (%) 1.7 1.5 2.3 35 360 39.81 109.35 61.78 361 40.82 134.10 346.78 362 40.84 137.26 388.17 363 40.35 136.13 389.64 364 40.97 134.37 387.76 365 39.86 134.03 388.65 366 40.84 134.33 390.82 367 42.11 134.26 391.30 368 40.22 134.44 392.30 369 41.71 135.68 392.48 370 41.34 135.37 391.04 371 39.74 137.57 393.68 Mean 40.80 135.23 386.60 sd 0.736 1.283 13.340 CV (%) 1.8 0.9 3.45 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 348 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 36 372 38.42 121.10 202.70 373 39.23 131.97 367.10 374 39.40 134.49 389.06 375 38.78 133.54 375.05 376 38.41 131.27 366.55 377 39.38 132.75 363.17 378 39.31 135.10 364.81 379 39.99 134.92 361.86 380 38.81 134.00 364.23 381 39.83 135.03 368.88 382 40.83 134.50 369.28 383 40.13 136.51 370.27 Mean6 39.46 134.01 369.11 sd 0.693 1.524 7.585 CV (%) 1.8 1.1 2.1 37 384 37.17 115.55 119.43 385 39.19 140.86 354.04 386 38.78 142.03 379.80 387 40.23 139.28 382.51 388 39.56 133.42 386.25 389 40.25 132.33 388.03 390 39.41 131.81 386.03 391 39.50 131.66 384.07 392 40.19 132.59 384.71 393 40.06 132.90 380.54 394 39.51 131.69 389.82 395 40.24 128.53 390.12 Mean6 39.72 134.28 382.36 sd 0.503 4.365 9.985 CV (%) 1.3 3.3 2.6 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 10Q/mean) : 349 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 38 396 44.26 103.94 134.80 397 41.71 133.83 370.04 398 40.52 134.46 392.60 399 42.41 136.21 392.35 400 41.58 138.35 391.19 401 41.22 137.96 390.95 402 41.68 136.82 386.97 403 42.30 134.46 386.02 404 41.45 135.47 387.15 405 43.00 135.72 388.95 406 42.56 140.81 394.32 407 43.64 138.35 394.35 Mean6 42.01 136.59 388.63 sd 0.880 2.117 6.812 CV (%) 2.1 1.6 1.8 39 408 347.10 129.38 277.10 409 20.65 146.13 388.04 410 46.79 145.34 396.02 411 43.03 147.11 394.47 412 43.13 146.81 396.60 413 42.12 144.46 397.08 414 42.59 143.14 396.75 415 42.69 145.55 398.55 416 43.79 145.91 400.86 417 43.48 146.06 396.23 418 43.62 147.08 395.93 419 44.12 145.97 394.04 Mean 6 41.45 145.78 395.87 sd 7.009 1.172 3.188 CV (%) 16.9 0.8 0.8 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 350 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 40 420 41.39 111.51 97.23 421 43.16 135.88 363.21 422 42.89 137.89 383.43 423 42.40 138.52 386.36 424 41.55 138.76 381.98 425 41.44 137.60 382.03 426 42.75 136.32 385.98 427 42.51 137.54 391.57 428 41.52 138.38 391.55 429 41.76 136.37 391.40 430 42.76 135.97 390.97 431 42.64 136.10 393.04 Mean6 42.31 137.21 385.59 sd 0.623 1.107 8.494 CV (%) 1.5 0.8 2.2 41 432 36.22 117.90 126.18 433 36.20 134.74 344.49 434 36.78 131.04 352.89 435 37.02 125.50 353.18 436 36.96 124.69 349.02 437 37.41 125.96 348.81 438 37.55 126.35 350.20 439 37.37 125.59 353.12 440 37.67 127.02 352.55 441 37.42 123.69 355.82 442 38.67 123.88 357.96 443 38.26 124.90 353.65 Mean6 37.39 126.67 351.97 sd 0.678 3.335 3.677 CV (%) 1.8 2.6 1.0 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 351 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 42 444 37.02 107.71 125.72 445 38.49 135.89 282.30 446 37.64 136.10 322.67 447 38.62 135.68 343.65 448 37.21 133.49 348.74 449 37.87 133.24 350.07 450 38.95 133.32 351.09 451 37.89 133.96 351.22 452 39.01 133.02 352.05 453 38.78 132.47 352.13 454 37.62 135.38 352.32 455 40.63 135.89 350.37 Mean6 38.43 134.40 341.51 sd 0.947 1.382 21.440 CV (%) 2.5 1.0 6.3 43 456 ,s 457 41.81 138.78 336.91 458 41.33 141.36 360.84 459 40.76 139.00 366.44 460 41.56 138.38 367.70 461 41.87 136.97 366.83 462 40.36 137.62 368.55 463 41.50 139.47 369.00 464 41.00 138.79 365.29 465 42.38 137.28 368.64 466 41.84 137.45 369.23 467 41.40 141.33 374.68 Mean6 41.44 138.766 364.92 sd 0.5670 1.4997 9.862 CV (%) 1.4 1.1 2.7 The initial concentration measurement of each exposure was excluded from all calculations Computer malfunction with data capture programme during exposure sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 352 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 44 468 36.25 107.61 1V 145.81 u 469 36.70 144.64 355.78 470 35.19 135.31 366.72 471 34.72 132.79 358.87 472 35.57 131.69 357.86 473 36.14 109.64 355.45 474 35.39 134.55 353.91 475 35.32 130.04 355.31 476 35.55 130.18 355.81 477 37.15 132.07 359.99 478 35.29 129.76 362.08 479 36.42 131.71 363.46 Mean6 35.77 131.12 358.66 sd 0.736 8.257 4.031 CV (%) 2.1 45 480 36.88 6.3 98.70 1.1 -------- 15--------- 481 40.03 137.29 324.31 482 39.54 139.74 348.06 483 39.37 140.54 360.02 484 39.65 137.62 363.46 485 38.64 136.34 363.59 486 40.13 135.74 364.67 487 39.69 134.98 362.17 488 40.15 136.73 360.70 489 39.26 137.75 360.69 490 40.05 138.59 362.70 491 138.63 364.80 Mean 6 39.65 137.63 357.74 sd 0.477 1.674 12.013 CV (%) 1.2 1.2 3.4 The initial concentration measurement of each exposure was excluded from all calculations Computer data capture programme froze and had to be reset during exposure Ethylene in nitrogen calibration cylinder left on in error sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 353 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 46 492 37.72 113.77 108.27 493 39.69 137.02 335.70 494 40.60 138.54 358.72 495 39.79 139.06 366.89 496 38.88 138.06 363.84 497 39.64 137.87 358.95 498 40.23 137.10 364.08 499 40.44 136.43 357.23 500 41.44 141.63 361.55 501 41.22 140.92 364.44 502 40.23 137.49 365.50 503 42.03 139.07 365.07 Mean 6 40.38 138.47 360.18 sd 0.909 1.623 8.702 CV (%) 2.3 1.2 2.4 47 504 12 505 12 506 12 507 40.09 12 12 508 38.95 148.33 362.91 509 38.96 145.82 360.01 510 39.78 145.92 360.67 511 39.88 146.64 360.14 512 41.46 143.04 361.02 513 39.69 132.86 362.21 514 40.19 132.01 365.13 515 41.83 132.85 367.19 Mean0 40.09 140.93 362.41 sd 0.987 7.080 2.574 CV (%) 2.5 5.0 0.7 The initial concentration measurement of each exposure was excluded from all calculations Sample missed as analysis started late in error sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 354 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 48 516 36.02 113.08 255.13 517 38.58 133.51 343.18 518 39.46 127.63 363.12 519 38.90 127.44 368.26 520 38.48 126.95 364.47 521 38.91 127.01 363.93 522 38.79 124.13 364.28 523 38.81 123.50 363.67 524 39.70 122.26 365.28 525 39.06 135.84 366.73 526 39.70 134.99 367.04 527 40.26 134.75 369.94 Mean6 39.15 128.91 363.63 sd 0.554 4.978 7.104 CV (%) 1.4 3.9 3.0 49 528 35.70 109.84 73.51 529 37.44 137.12 341.36 530 37.02 139.57 364.71 531 37.60 145.98 368.11 532 36.94 132.51 359.55 533 37.14 129.64 357.14 534 36.85 129.81 355.17 535 36.53 129.62 353.37 536 36.40 129.72 353.83 537 36.22 126.62 353.95 538 37.24 128.04 356.18 539 37.33 129.82 359.01 Mean6 36.97 132.59 356.58 sd 0.442 5.883 6.876 CV (%) 1.2 4.4 1.9 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 355 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 50 540 25.70 103.22 76.33 541 34.77 130.73 311.87 542 35.63 132.81 349.47 543 35.45 133.29 356.10 544 35.94 133.02 355.08 545 37.44 135.13 366.75 546 37.18 137.08 382.49 547 37.71 137.89 381.77 548 37.58 140.14 381.62 549 36.88 140.41 380.89 550 38.17 137.22 382.62 551 38.22 137.31 384.60 Mean6 36.82 135.91 366.66 sd 1.182 3.153 22.416 CV (%) 3.2 2.3 6.1 51 552 36.73 116.01 89.34 553 38.71 139.96 341.25 554 39.35 142.41 372.60 555 40.23 141.92 373.48 556 40.70 142.66 373.62 557 38.50 141.13 376.43 558 38.09 138.42 377.82 559 39.69 138.61 377.28 560 38.78 138.92 376.47 561 39.86 140.44 375.90 562 40.12 142.80 381.29 563 40.53 142.62 383.57 Mean6 39.50 140.90 373.61 sd 0.880 1.715 11.222 CV (%) 2.2 1.2 3.0 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 356 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 52 564 38.83 105.46 93.48 565 40.88 142.00 346.87 566 40.56 146.12 373.05 567 40.98 144.98 375.68 568 41.17 141.01 373.27 569 41.09 140.11 372.69 570 42.30 141.28 377.88 571 42.40 143.43 378.45 572 40.86 143.93 375.85 573 42.72 138.33 378.21 574 41.38 136.85 376.73 575 43.14 138.80 376.45 Mean6 41.59 141.53 373.19 sd 0.882 2.907 8.969 CV (%) 2.1 2.1 2.4 53 576 35.26 113.40 92.75 577 37.08 142.41 377.63 578 38.99 144.11 408.97 579 38.23 143.50 410.05 580 39.95 143.36 407.35 581 40.81 147.78 417.63 582 39.91 155.81 430.02 583 38.98 151.08 408.38 584 39.76 147.26 398.82 585 39.06 147.63 396.67 586 40.11 142.01 395.94 587 45.38 145.88 399.94 Mean6 39.84 146.44 404.67 sd 2.098 4.160 13.442 CV (%) 5.3 2.8 3.3 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 357 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 54 588 38.13 108.39 103.62 589 39.75 142.78 358.22 590 38.79 144.35 415.38 591 40.57 145.69 385.58 592 38.72 145.68 369.73 593 40.02 145.58 376.46 594 39.97 145.09 380.00 595 39.40 144.00 378.43 596 39.93 142.50 378.03 597 38.82 140.85 377.80 598 39.12 141.39 379.55 599 43.01 141.63 381.31 Mean 6 39.83 143.59 380.04 sd 1.214 1.837 13.741 CV (%) 3.0 1.3 3.6 55 600 37.27 102.05 69.33 601 37.33 136.20 330.48 602 37.05 138.89 364.23 603 37.99 139.47 371.75 604 38.31 140.36 377.64 605 39.54 139.40 375.53 606 38.23 140.17 375.46 607 38.13 141.19 377.60 608 38.59 142.20 386.92 609 38.65 141.83 387.09 610 40.42 141.61 382.42 611 40.19 142.25 381.92 Mean6 38.59 140.32 373.73 sd 1.075 1.804 15.800 CV (%) 2.8 1.3 4.2 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 358 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 56 612 38.10 -0.45 90.59 613 37.74 137.57 341.84 614 40.23 140.48 369.75 615 40.20 140.21 378.04 616 40.23 141.45 379.22 617 40.15 144.40 382.88 618 40.33 142.69 379.52 619 39.91 142.89 381.79 620 37.99 144.07 389.34 621 37.83 146.34 389.20 622 39.07 143.92 388.07 623 40.50 142.01 382.02 Mean6 39.47 142.37 378.33 sd 1.104 2.406 13.373 CV (%) 2.8 1.7 3.5 57 624 39.14 ---------- n ------------ ---------- 12------------ 625 41.83 138.91 251.89 626 41.51 145.30 371.68 627 41.29 145.89 383.20 628 41.21 145.38 384.95 629 41.44 145.61 379.96 630 42.60 144.44 381.84 631 42.71 144.64 377.96 632 43.03 145.48 384.94 633 42.95 145.08 384.45 634 43.30 147.40 383.69 635 43.75 148.50 391.78 Mean 0 42.33 145.15 370.58 sd 0.899 2.387 39.677 CV (%) 2.1 1.6 10.7 The initial concentration measurement of each exposure was excluded from all calculations Sample missed as analysis started late in error sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 359 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 58 636 39.83 118.41 122.15 637 41.89 150.19 299.12 638 42.16 149.20 344.73 639 40.62 133.72 378.10 640 40.14 133.32 383.34 641 41.40 133.80 388.67 642 42.32 138.04 388.85 643 41.26 146.13 388.29 644 41.27 150.60 391.02 645 41.92 134.84 390.22 646 41.05 131.03 385.96 647 42.22 127.41 384.83 Mean6 41.48 138.93 374.83 sd 0.699 8.473 28.307 CV (%) 1.7 6.1 7.6 59 648 0.20 18 116.68 204.63 649 0.23 18 138.55 304.27 650 35.04 139.96 367.38 651 38.84 142.17 384.71 652 38.92 138.00 388.95 653 40.03 138.72 393.51 654 39.44 139.79 394.78 655 39.56 140.69 393.48 656 40.41 140.26 393.38 657 40.82 139.95 394.27 658 41.06 140.91 395.97 659 44.51 142.54 397.84 Mean 6 36.26 140.14 382.59 sd 12.155 1.418 27.317 CV (%) 33.5 1.0 7.1 The initial concentration measurement of each exposure was excluded from all calculations Low concentration due to air bubble in test material feed line sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 360 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 60 660 -0.33 110.62 160.67 661 37.53 139.46 363.69 662 38.13 143.48 381.80 663 37.31 144.36 389.25 664 37.06 145.10 397.01 665 38.85 145.11 393.56 666 39.29 145.18 394.04 667 39.66 145.25 394.12 668 39.28 145.54 398.33 669 39.04 147.20 402.95 670 41.29 146.10 402.39 671 39.87 144.33 402.71 Mean 6 38.85 144.65 392.71 sd 1.262 1.973 11.516 CV (%) 3.2 1.4 2.9 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 361 APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 61 672 1U 96.65 673 10 115.11 674 10 675 132.16 14 14 676 10 80.00 677 10 85.09 678 10 89.69 679 10 94.05 680 10 97.80 681 10 101.53 682 10 105.34 683 10 109.05 684 10 112.31 685 10 686 14 115.67 14 687 10 45.88 688 10 45.57 689 10 45.52 690 10 45.88 691 10 46.14 692 10 46.31 693 50.15 147.87 424.28 694 51.61 153.81 427.93 695 50.72 145.76 432.32 696 47.25 145.89 428.16 697 45.23 145.52 424.75 698 44.55 145.23 411.27 699 45.70 145.42 419.83 700 45.38 146.25 414.58 701 45.76 148.07 417.72 702 46.93 148.20 420.21 703 49.61 148.50 Mean 47.53 147.32 422.10 sd 2.524 2.489 6.572 CV (%) 5.3 1.7 1.6 M sd CV (%) Sampling time selected at 1 rather than every 10 minutes intervals. Values excluded from mean and standard deviation Time-controlled chamber switching device out of synchrony with measurement, sample collected over 2 groups, values excluded from mean and standard deviation Standard deviation Coefficient of variation (sd x 100/mean) : 362 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 62 704 -0.23 -0.20 -0.16 705 43.64 134.40 126.87 706 43.49 139.92 394.16 707 43.36 139.84 403.15 708 43.18 137.71 400.62 709 44.21 137.89 403.42 710 43.90 137.15 403.90 711 43.74 137.05 403.34 712 44.65 138.31 402.69 713 45.58 138.15 403.90 714 46.69 138.65 406.52 715 40.82 138.14 405.98 Mean 6 43.93 137.93 377.69 sd 1.476 1.492 83.252 CV (%) 3.4 1.1 22.0 63 716 43.60 109.48 109.10 717 44.85 137.25 380.19 718 44.89 139.46 410.27 719 46.28 139.66 413.29 720 44.72 142.53 413.02 721 45.82 142.64 417.57 722 45.53 141.35 416.17 723 46.09 141.43 415.58 724 47.00 142.30 418.35 725 47.66 141.87 417.52 726 48.91 141.99 417.82 727 17.15 142.20 420.97 Mean6 43.54 141.15 412.80 sd 8.844 1.679 11.212 CV (%) 20.3 1.2 2.7 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 363 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 64 728 45.20 101.07 20.79 729 46.15 137.11 347.32 730 47.13 139.70 418.78 731 47.31 139.65 426.18 732 46.47 139.91 423.25 733 46.52 138.04 414.69 734 47.01 138.84 414.02 735 46.80 136.37 414.42 736 45.43 144.27 414.72 737 44.30 136,13 413.70 738 45.88 136.52 414.75 739 47.13 135.42 416.88 Mean6 46.37 138.36 410.79 sd 0.900 2.524 21.447 CV (%) 1.9 1.8 5.2 65 740 40.07 117.10 75.70 741 40.60 136.15 366.40 742 42.38 137.77 400.42 743 42.33 137.77 412.38 744 42.55 137.85 411.73 745 42.95 137.36 409.56 746 43.11 137.60 406.32 747 42.40 139.11 408.85 748 44.71 139.46 408.09 749 44.21 135.51 413.05 750 46.10 135.55 414.21 751 37.61 137.77 415.27 Mean6 42.63 137.44 406.02 sd 2.208 1.277 13.789 CV (%) 5.2 0.9 3.4 The initial concentration measurement of each exposure was excluded from all calculations sd Standard deviation. CV (%) Coefficient of variation (sd x 100/mean) : 364 : APPENDIX B MIN 313/023622 (Individual POSF concentration measurements - continued) Exposure Sample Chamber Concentration (ppm) No. No. Group 2 Group 3 Group 4 (Low dose) (Inter, dose) (High dose) 66 752 re re 12 753 14 754 38.76 136.70 403.38 755 41.05 136.23 406.44 756 41.64 136.44 409.08 757 41.35 138.19 411.40 758 42.10 138.76 416.98 759 41.48 138.53 415.86 760 43.32 140.80 420.91 761 42.45 139.96 421.06 762 45.29 140.23 419.66 763 46.05 140.25 425.00 Mean 42.348 138.609 414.98 sd 2.1126 1.7052 7.106 CV (%) 5.0 1.2 1.7 67 764 -0.34 103.26 52.41 765 37.28 143.26 358.26 766 38.88 145.92 403.83 767 38.47 145.82 407.26 768 39.33 145.95 404.32 769 40.29 146.00 406.00 770 40.78 144.80 407.33 771 40.35 145.49 406.13 772 41.62 146.29 406.36 773 42.18 146.92 410.03 774 41.70 146.40 411.34 756 43.57 149.24 413.76 Mean 40.40 146.01 403.15 sd 1.829 1.443 15.184 CV (%) 4.5 1.0 3.8 Sample missed as analysis started late in error Time-controlled chamber switching device out of synchrony with measurement, sample collected over 2 groups, values excluded from mean and standard deviation sd Standard deviation CV (%) Coefficient of variation (sd x 100/mean) : 365 : United Kingdom Huntingdon Life Sciences Ltd W oolley Road A lc o n b u ry H u n tin g d o n Cambridgeshire PE28 4HS England Huntingdon Life Sciences Ltd Eye S u ffolk IP23 7PX England USA H u ntingdon Life Sciences Inc PO Box 2360 Mettlers Road East M illstone NJ 08875-2360 USA Tel: +1 (732) 873-2550 Fax: +1 (732) 873-3992 e-mail: sales@ princeton.huntingdon.com Japan Huntingdon Life Sciences Co Ltd Bancho Kaikan Korea Shin Won Scientific Company. Ltd. # 603, Kum bong Technovalley II 93-5, Dangjeong-dong, Gunpo-si. Gyeonggi-do 435-831. Korea. Tel: +82 31 477 9771 Fax: +82-31 477 9772 Denmark Huntingdon Life Sciences