Document 70EMb7z98aenV1d8zaXgGBLkE

AR226-3573 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 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 ............................................................................................... 11 STUDY SCHEDULE AND STRUCTURE .............................................................................. 11 TEST SUBSTANCE AND FORMULATION.......................................................................... 12 ANIMAL MANAGEMENT..................................................................................................... 13 SERIAL OBSERVATIONS ..................................................................................................... 15 NECROPSY AND HISTOLOGY............................................................................................. 19 PATHOLOGY .......................................................................................................................... 22 DATA TREATMENT .............................................................................................................. 22 QUALITY ASSURANCE AND ARCHIVING PROCEDURES............................................. 26 RESULTS ..................................................................................................................................... 27 DISCUSSION ............................................................................................................................... 35 REFERENCES ............................................................................................................................. 36 FIGURES 1. Bodyweights - group mean values...................................................................................... 37 : 2 : MIN 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 : MIN313023622 COMPLIANCEWITH GOOD LABORATORY PRACTICE PERFLUOROOCTANESULFONYLFLUORIDE (POSF; T-7661.4) TOXICITYSTUDY BY INHALATION ADMINISTRATION TO CD RATS FOR 13 WEEKS FOLLOWEDBY A 4 WEEKRECOVERYPERIOD TPrhaectsitcuedsytdaensdacrrdisbeadndin|tchoisnsriedpeorrtthweadsatcaongdeunecrtaetdedintcobmepvlailaidn:ce with the following Good Laboratory "STthaetuUtoKryGIonosdraLmaebnotra2t0o0r4yNPor.ac9ti9c4e Regulations 1999(Statutory Instrument No 3106) as amended by OECD PrinciplesofGood Laboratory Practice (asrevised in 1997), ENVIMC/CHEM(GB)17, EECC CCoommmmiissssiioonn DDiirreeccttiivvee21090949//I101//EECC ooff11MFaerbcrhua1r9y9290(04Of(OfffaicJiaolurJnoaulrnNaol.NLo.7L7/8S)0,/a4s4a).mended by "STthaetseesopfriAncmieplreisocfanGdooJadpaLnaboonrtahteorbyaPsriacotficientaerergaocvceempmteendtbayl tahgerereemgeunltast.ory authorities ofthe United dAunraetxipoinroyftdhaetsftoudyt.he est substance was not given, The material was assumed o be stable for the `SGcLaPnnmionngitEolrecitnrgonprMoigcrraomsmce.opNy oandclXa-rmaoyfcaonmalpylsiisanwcaes wciotnhduGcLtePdibymadfeciIltryeswpheictchofithneosteppahraosfesthoe f the study. TerenceJ. Kenny, B.Sc. (Hons. Date MIN 313/023622 QUALITY ASSURANCE STATEMENT PERFLUOROOCTANESULFONYL FLUORIDE (POSF; T-7661.4) TOXICITY STUDY BY INHALATION ADMINISTRATION TO CD RATS FOR 13WEEKS FOLLOWED BY A 4 WEEK RECOVERY PERIOD "The followinginspectionsandaudits have been carried out in relation to this study: Study Phase DateofInspection Date ofReporting Protocol Audit 10 December2001 11 December2001 SSttuuddyypBraespaerdatIinosnpections TEexsptostuerme caonndtroglenaenrdatdiiospnosoiftitoenst aAttmmoosspphheerree sampling CClliinniiccaallssiiggnnss ((pwoesetk-ldyo)se) aFnraelsyhsisu.rine callcion for SEM Necropsy 2200DDeecceemmbbeerr 22000011 20 December 2001 2200DDeecceemmbbeerr 22000011 184JJaannuuaarryy22000022 21 March 2002 2200 DDeecceemmbbeerr22000011 20 December2001 2200 DDeecceemmbbeerr22000012 184JJaannuuaarryy22000022 26 March 2002 Report Audit 9 1J9unJeul2y0a0n4d-2252JuJluyne20200504 22 September 2005 2245 JJuunlye 22000054 22 September 2005 PDirroetcotcoorlaAnuddCito:mpAannyauMdiatnaogfetmheentproatsoicnodlicfaotredthaibsovset.udy was conducted and reported to the Study SSttuuddyyDbiarescetdorinasnpdecCtoiomnpsa:nyInMspaencatgioenmsenoftpahsaisnedsicoaftetdhiasbosvteu.dy were conducted and reported to the oPruotciensesabnadserdepientspieectipornosc:edAurtesoreambpoluotyetdheotnimtehisthitsypsetudoyf swuadsyinweprreogrceasmsieidnsopue.ctioTnhsesoef twheerre promptly reported toappropriate CompanyManagemen. cRoenpdourctteAdudaintd:rTehpiorsteredptoortthheasStbuedeynDiaruedcittoerdabnydtChoemQpuaalniytyMaAnsasugreamnecnetDaespairntdmiecantte.d aTbhovies.audit was 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 of 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 recovery 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 No. of animals Male Female 5 5 5 5 5 5 5 5 Main study (13 weeks) Animal numbers Male Female 1-5 21-25 6-10 11-15 16-20 26-30 31-35 36-40 Cage numbers Male Female 1 5 2 6 3 7 4 8 : 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 5 5 5 5 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 summary: 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 Crl:CD (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 recovery 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 recovery 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 H1E haematology analyser: Haematocrit (Hct) Haemoglobin (Hb) Red blood cell count (RBC) Reticulocytes (Retic) 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 (Plt) Reticulocyte count (Retic) - 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 (Gluc) Total cholesterol (Chol) Triglycerides (Trig) Sodium (Na) Potassium (K) Chloride (Cl) Calcium (Ca) Inorganic phosphorus (Phos) Total protein (Total Prot) Electrophoretic protein fractions; albumin (Alb), 1 globulin (a1), 2 globulin (a2), globulin (Beta), 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 (Gluc), ketones (Keto), bile pigments (Bili), haem pigments (Blood) by Multistix. Multistix are diagnostic reagents obtained from Bayer plc, 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 a Sections 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: [ad. fc] x w = (ad) [ad.(fc - xw) 2 ] sd = (ad) - 1 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: A/G = Albumin concentration 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 F1 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 F1 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 H1 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 H1 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 of 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: Mean chamber concentration (ppm) Group 2 3 4 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 of 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 when adjusted for bodyweight. Following a 4 week recovery period the recovery 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 recovery 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) 0 Necrosis of ventral cartilage Total 0 Males 30 100 300 0 0 0 4a 0 Females 30 100 300 0 0 5b Number of larynx examined 5 5 5 5 5 5* 5 5 a - p<0.05, b - p<0.01 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 - p<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) 0 Foamy alveolar macrophages/ septal thickening/scattered alveolar macrophages Minimal 0 Slight 0 Moderate 0 Total 0 Males 30 100 300 0 0 0 3 0 0 0 1 0 0 0 1 0 0 0 5b 0 Females 30 100 300 0 1 2 0 0 2 0 0 0 0 1 4a Number of lungs examined 5 5 5 5 5 5* 5 5 a - p<0.05, b - p<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) 0 Foamy alveolar macrophages/ septal thickening/scattered alveolar macrophages Minimal 1 Slight 0 Total 1 Males 30 100 300 0 1 1 2 0 0 0 3 0 1 1 5a 0 Females 30 100 300 1 0 2 0 0 0 1 0 2 Number of lungs examined 5 5 5 5 5* 5 5 5 a - p<0.05 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 of 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 Dosage (ppm) 0 Centrilobular hepatocyte hypertrophy extending to midzonal area Slight 0 Moderate 0 Total 0 Centrilobular hepatocyte hypertrophy Slight 0 Total 0 Prominent centrilobular hepatocyte pigment Minimal 0 Total 0 Males 30 100 300 0 4 0 0 0 0 5 5 0 4a 5b 5b 0 1 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 Females 30 100 300 0 0 3 0 0 2 0 0 5b 0 0 0 0 0 0 0 0 1 0 0 1 Number of livers examined 5 5 5 5 5 5* 5 5 a - p<0.05, b - p<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) 0 Centrilobular hepatocyte hypertrophy extending into midzonal area Slight 0 Total 0 Centrilobular hepatocyte hypertrophy Minimal 0 Slight 0 Total 0 Prominent centrilobular hepatocyte pigment Minimal 0 Total 0 Number of livers examined 5 Males 30 100 300 0 Females 30 100 300 0 2 5 0 0 0 0 0 2 5b 0 0 0 0 1 3 0 0 0 0 4 2 0 0 0 0 0 0 3 3 0 0 0 0 4a 0 0 0 0 0 0 2 0 0 0 0 0 0 2 5 5 5 5* 5 5 5 a - p<0.05, b - <p<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 larynx 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 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. 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% 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 of 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 Methods for Research Workers. Oliver and Boyd, Edinburgh. HEALEY, G.F. (1999) The F1 approximate parameter test for monotonicity. Statistical Specification ST9725, Department of Statistics, Huntingdon Life Sciences. HEALEY, G.F. (1999) The H1 approximate non-parameter test for monotonicity. Specification ST9726, Department of Statistics, Huntingdon Life Sciences. Statistical 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) Bodyweight (g) MIN 313/023622 : 37 : 520 500 480 460 440 420 400 Males 380 360 340 320 300 280 260 240 Females 220 200 180 160 140 120 100 0 1 2 3 4 5 6 7 8 9 10 11 12 13 R1 R2 R3 R4 Week Group 1 : Control Group 2 : 30 ppm Group 3 : 100 ppm Group 4 : 300 ppm TABLE 1 MIN 313/023622 : 38 : BODYWEIGHT - GROUP MEAN VALUES (G) Print No: 0001 Main study and recovery phase animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ SEX: ---------------------MALE--------------------- --------------------FEMALE-------------------- GROUP: 1 2 3 4 1 2 3 4 WEEK ------------------------------------------------------------------------------------------------------------------------------------ -1 233 235 236 234 178 175 177 179 0 261 267 264 258 191 187 189 190 1 286 290 284 274 202 201 201 203 2 310 310 303 296 213 208 206 206 3 333 333 324 316 220 219 214 213 4 356 352 346 335 232 224 224 217 5 376 367 362 346 238 234 228 225 6 393 383 379 360 246 241 236 231 7 409 399 392 369 255 246 240 235 8 421 412 406 379 258 251 245 237 9 433 423 416 382 264 255 248 241 10 444 430 422 390 270 259 251 244 11 454 438 433 395 277 263 252 246 12 463 444 442 399 277 264 257 247 13 462 445 441 395 274 260 252 240 14a 479 451 455 425 291 295 257 253 15a 492 462 470 437 303 301 264 263 16a 506 470 476 454 305 306 265 268 17a 517 481 485 465 305 308 269 274 ------------------------------------------------------------------------------------------------------------------------------------ Gain * * ** * ** Week 0-13 202 179 177 137 83 73 63 50 % of Control - 89 88 69 - 88 76 60 Weeks 13-17 50 44 47 52 27 28 20 33 % of Control - 88 94 104 - 104 74 122 Level of significance - all comparisons made with control * p 0.05, ** p 0.01 (Williams' test) a Recovery animals only TABLE 1 MIN 313/023622 : 39 : (Bodyweight - continued) Print No: 0002 Satellite study animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ SEX: ----------MALE--------- --------FEMALE-------- GROUP: 1 4 1 4 WEEK ------------------------------------------------------------------------------------------------------------------------------------ -1 237 237 180 176 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) TABLE 2 MIN 313/023622 : 40 : FOOD CONSUMPTION - GROUP MEAN VALUES (G/RAT/WEEK) Print No: 0002 Main study and recovery phase animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ SEX: ---------------------MALE--------------------- --------------------FEMALE-------------------- GROUP: 1 2 3 4 1 2 3 4 WEEK ------------------------------------------------------------------------------------------------------------------------------------ -1 182 194 191 186 137 141 137 142 1 171 171 172 154 133 137 132 132 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 14a 198 197 205 190 152 156 138 135 15a 197 197 205 196 158 160 142 141 16a 201 206 206 199 190 136 140 137 17a 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) a Recovery animals only TABLE 2 MIN 313/023622 : 41 : (Food consumption - continued) Print No: 0003 Satellite study phase GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ SEX: ---------------------MALE--------------------- --------------------FEMALE-------------------- GROUP: 1 2 3 4 1 2 3 4 WEEK ------------------------------------------------------------------------------------------------------------------------------------ 1 169 161 138 122 2 185 177 145 131 3 197 188 149 136 MIN 313/023622 : 42 : TABLE 3 Haematology - group mean values Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex Hct Hb RBC Retic L/L g/dL x1012/L % 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 4M Mean SD n 0.447 0.0202 5 0.430 0.0319 5 15.8 0.74 5 15.2 1.63 5 8.34 0.255 5 7.83 0.816 5 2.00 0.430 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 : 43 : TABLE 3 (Haematology - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex L x109/L E x109/L B x109/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 8.92 0.20 0.03 0.20 SD 1.216 0.076 0.013 0.027 n 5 5 5 5 * 4M Mean 10.46 0.11 0.04 0.15 SD 1.098 0.033 0.011 0.111 n 5 5 5 5 Level of significance - all comparisons made with control * p 0.05, ** p 0.01 (Williams' test) * p 0.05 (Dunnett's test) LUC x109/L 0.17 0.086 3 * 0.09 0.047 5 ** 0.05 0.013 5 ** 0.06 0.031 5 Plt x109/L 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 : 44 : TABLE 3 (Haematology - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex Hct Hb RBC Retic L/L g/dL x1012/L % 1F Mean 0.423 15.2 7.60 2.31 SD 0.0126 0.38 0.305 0.500 n 3 3 3 3 2F Mean 0.425 15.1 7.62 2.09 SD 0.0112 0.44 0.214 0.409 n 5 5 5 5 3F Mean 0.421 15.1 7.55 2.53 SD 0.0168 0.71 0.256 0.971 n 5 5 5 5 4F Mean 0.431 15.6 7.95 1.74 SD 0.0109 0.29 0.333 0.372 n 5 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 : 45 : TABLE 3 (Haematology - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex L x109/L E x109/L B x109/L M x109/L 1F Mean 6.58 0.15 0.02 0.20 SD 2.189 0.026 0.010 0.050 n 3 3 3 3 2F Mean 6.91 0.20 0.02 0.28 SD 0.972 0.104 0.004 0.090 n 5 5 5 5 3F Mean 7.38 0.16 0.03 0.22 SD 1.748 0.090 0.019 0.100 n 5 5 5 5 4F Mean 7.86 0.16 0.02 0.17 SD 1.041 0.075 0.008 0.103 n 5 5 5 5 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 Plt 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 : 46 : TABLE 3 (Haematology - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex Hct Hb RBC Retic L/L g/dL x1012/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 MCHC 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 : 47 : TABLE 3 (Haematology - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex L x109/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) * p 0.05, ** p 0.01 (Dunnett's test) LUC x109/L 0.21 0.074 5 0.11 0.032 5 0.12 0.105 5 0.12 0.044 5 Plt 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 APTT sec 19.2 2.40 5 18.3 1.18 4 * 16.0 2.38 5 * 15.7 1.45 5 MIN 313/023622 : 48 : TABLE 3 (Haematology - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex Hct Hb RBC Retic L/L g/dL x1012/L % 1F Mean 0.391 14.8 7.18 2.23 SD 0.0102 0.10 0.093 0.405 n 4 4 4 4 2F Mean 0.397 15.0 7.45 2.00 SD 0.0103 0.39 0.211 0.164 n 4 4 4 5 3F Mean 0.373 14.1 6.96 2.24 SD 0.0370 1.55 0.603 0.169 n 5 5 5 5 4F Mean 0.392 14.7 7.24 2.23 SD 0.0217 0.98 0.247 0.325 n 5 5 5 5 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 fL 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 x109/L 1.23 0.324 4 1.18 0.871 4 1.07 0.444 5 0.90 0.545 5 MIN 313/023622 : 49 : TABLE 3 (Haematology - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex L x109/L E x109/L B x109/L M x109/L 1F Mean 6.30 0.16 0.02 0.21 SD 1.301 0.085 0.008 0.019 n 4 4 4 4 2F Mean 4.50 0.12 0.01 0.15 SD 0.896 0.050 0.005 0.074 n 4 4 4 4 * * * 3F Mean 4.13 0.08 0.01 0.10 SD 1.378 0.028 0.007 0.068 n 5 5 5 5 * * * 4F Mean 4.07 0.10 0.01 0.09 SD 1.101 0.056 0.004 0.042 n 5 5 5 5 Level of significance - all comparisons made with control * p 0.05 (Williams' test) LUC x109/L 0.12 0.034 4 0.10 0.047 4 0.07 0.048 5 0.10 0.055 5 Plt 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 APTT sec 17.1 1.57 4 17.3 0.95 4 17.8 1.53 4 18.8 3.32 4 MIN 313/023622 : 50 : TABLE 4 Biochemistry - group mean values Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex ALP ALT AST LDH U/L U/L U/L Total U/L 1M Mean 202 44 99 497 SD 40.5 7.7 17.6 410.1 n 5 5 5 5 * 2M Mean 257 61 143 444 SD 63.2 12.3 54.8 361.6 n 5 5 5 5 * * 3M Mean 279 58 102 296 SD 43.9 9.3 25.9 147.6 n 5 5 5 5 ** ** 4M Mean 327 67 103 244 SD 67.6 12.5 13.1 59.7 n 5 5 5 5 Level of significance - all comparisons made with control * p 0.05, ** p 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 23 25.0 5 27 25.8 5 11 6.4 5 13 4.4 5 LDH5 U/L 417 341.0 5 354 305.6 5 240 113.2 5 194 61.0 5 TABLE 4 (Biochemistry - continued) MIN 313/023622 : 51 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex CPK Total U/L SDH Bili Urea U/L mol/L mmol/L 1M Mean 242 15.3 1 SD 119.0 2.67 0.5 n 5 5 5 6.05 1.124 5 2M Mean 1633 13.1 1 SD 3213.7 2.40 0.5 n 5 5 5 * 3M Mean 202 11.5 1 SD 78.0 2.67 0.5 n 5 5 5 * 4M Mean 168 12.0 1 SD 53.6 1.36 0.0 n 5 5 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 mol/L 53 3.8 5 52 2.2 5 * 48 3.6 5 * 48 2.7 5 Gluc mmol/L Chol 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 TABLE 4 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex Na mmol/L K mmol/L Cl mmol/L Ca mmol/L Phos mmol/L Total Prot g/L MIN 313/023622 : 52 : 1M Mean 143 SD 1.9 n 5 2M Mean 145 SD 0.8 n 5 3M Mean 144 SD 1.8 n 5 4M Mean 143 SD 1.6 n 5 4.0 106 0.61 0.7 5 5 3.8 108 0.23 1.5 5 5 4.1 107 0.29 1.7 5 5 4.1 106 0.21 1.2 5 5 2.68 0.083 5 2.69 0.051 5 * 2.77 0.043 5 * 2.77 0.054 5 2.02 63 0.202 1.6 5 5 2.05 67 0.111 4.8 5 5 2.17 65 0.115 3.3 5 5 2.03 67 0.109 4.7 5 5 MIN 313/023622 : 53 : TABLE 4 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group Alb / sex g/L 2 3 4 ------POSF------ 30 100 300 a1 a2 Beta g/L g/L g/L 1M Mean 29 11 SD 0.8 1.5 n 4 4 3 15 0.5 2.1 4 4 2M Mean 30 11 SD 1.1 1.3 n 5 5 3 15 0.4 2.0 5 5 3M Mean 30 12 SD 1.8 1.3 n 5 5 * 4M Mean 33 12 SD 3.6 1.3 n 5 5 3 14 0.4 1.7 5 5 3 14 0.4 1.1 5 5 Level of significance - all comparisons made with control * p 0.05 (Williams' test) 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 : 54 : TABLE 4 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex ALP ALT AST LDH U/L U/L U/L Total U/L 1F Mean 162 38 108 466 SD 36.3 6.0 40.8 242.8 n 4 4 4 4 * 2F Mean 135 56 109 217 SD 26.6 26.6 28.8 46.6 n 5 5 5 5 * 3F Mean 124 49 103 207 SD 26.3 37.6 52.0 71.3 n 5 5 5 5 * 4F Mean 123 47 106 303 SD 30.4 20.1 28.3 111.9 n 5 5 5 5 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 LDH2 U/L 27 15.9 4 14 5.4 5 11 3.5 5 22 17.9 5 LDH3 U/L 13 8.4 4 9 7.0 5 6 2.2 5 7 2.9 5 LDH4 U/L 25 17.7 4 10 2.8 5 9 3.2 5 13 6.4 5 LDH5 U/L 378 195.4 4 * 171 53.5 5 * 169 62.5 5 * 237 72.2 5 TABLE 4 (Biochemistry - continued) MIN 313/023622 : 55 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex CPK Total U/L SDH Bili Urea U/L mol/L mmol/L 1F Mean 564 13.6 2 SD 710.1 3.42 0.0 n 4 4 4 * 2F Mean 130 12.4 3 SD 42.1 4.03 0.5 n 5 5 5 * 3F Mean 132 10.7 2 SD 35.1 2.60 0.0 n 5 5 5 * 4F Mean 143 15.5 2 SD 37.4 1.54 0.4 n 5 5 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 * p 0.05, ** p 0.01 (Williams' test) Creat mol/L 66 6.6 4 * 58 2.7 5 * 55 2.7 5 * 60 8.8 5 Gluc mmol/L Chol 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 : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex Na mmol/L K mmol/L Cl mmol/L Ca mmol/L Phos mmol/L Total Prot g/L MIN 313/023622 : 56 : 1F Mean 142 SD 1.0 n 4 2F Mean 143 SD 1.8 n 5 3F Mean 143 SD 1.5 n 5 4F Mean 143 SD 1.3 n 5 3.5 106 0.39 0.6 4 4 3.2 107 0.21 0.8 5 5 3.0 105 0.18 1.8 5 5 3.4 105 0.24 0.9 5 5 2.81 0.064 4 2.84 0.112 5 2.75 0.079 5 2.79 0.028 5 1.60 70 0.292 1.5 4 4 1.50 75 0.118 3.0 5 5 1.58 70 0.094 4.0 5 5 * 1.61 76 0.148 3.6 5 5 MIN 313/023622 : 57 : TABLE 4 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group Alb / sex g/L 2 3 4 ------POSF------ 30 100 300 a1 a2 Beta g/L g/L g/L 1F Mean 37 10 SD 1.0 0.5 n 4 4 4 14 0.6 0.5 4 4 2F Mean 40 11 SD 3.6 0.7 n 5 5 4 14 0.8 0.8 5 5 3F Mean 37 10 SD 2.3 0.5 n 5 5 ** ** 4F Mean 42 12 SD 2.0 0.5 n 5 5 4 14 0.5 0.5 5 5 3 13 0.5 1.8 5 5 Level of significance - all comparisons made with control ** p 0.01 (Williams' test) Gamma g/L 6 1.0 4 7 2.0 5 6 1.5 5 6 1.6 5 A/G Ratio 1.08 0.015 4 1.15 0.160 5 1.13 0.057 5 1.21 0.099 5 MIN 313/023622 : 58 : TABLE 4 (Biochemistry - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex ALP ALT AST LDH U/L U/L U/L Total U/L 1M Mean 182 40 74 180 SD 25.4 3.7 7.5 28.0 n 5 5 5 5 + 2M Mean 234 56 100 162 SD 34.4 10.4 19.4 57.2 n 5 5 5 5 + 3M Mean 195 49 87 153 SD 48.3 10.2 5.3 48.3 n 5 5 5 5 * + + 4M Mean 244 85 118 199 SD 44.0 62.2 52.6 48.2 n 5 5 5 5 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 : 59 : TABLE 4 (Biochemistry - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex CPK Total U/L SDH Bili Urea Creat U/L mol/L mmol/L mol/L 1M Mean 240 12.3 2 SD 146.1 3.04 0.5 n 5 5 5 4.90 49 0.266 1.9 5 5 2M Mean 153 13.6 2 SD 45.5 1.68 0.5 n 5 5 5 5.93 49 0.685 3.6 5 5 3M Mean 183 12.9 1 SD 116.5 4.77 0.8 n 5 5 5 5.71 49 0.777 2.2 5 5 4M Mean 293 19.0 1 SD 119.0 9.61 0.4 n 5 5 5 5.38 48 0.600 2.9 5 5 Level of significance - all comparisons made with control * p 0.05 (Williams' test) + p 0.05 (Shirley's test) Gluc mmol/L Chol 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 TABLE 4 (Biochemistry - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex 1 Control 0 Na mmol/L 2 3 4 ------POSF------ 30 100 300 K mmol/L Cl mmol/L Ca mmol/L Phos mmol/L Total Prot g/L MIN 313/023622 : 60 : 1M Mean 142 SD 0.5 n 5 3.8 102 0.30 0.7 5 5 2M Mean 142 SD 1.1 n 5 3.9 103 0.13 0.8 5 5 3M Mean 142 SD 0.9 n 5 * 4M Mean 144 SD 0.7 n 5 3.8 103 0.30 0.8 5 5 * 3.9 103 0.08 0.0 5 5 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 66 0.092 1.8 5 5 1.71 63 0.158 1.5 5 5 * 1.73 61 0.141 2.9 5 5 1.79 66 0.112 2.3 5 5 MIN 313/023622 : 61 : TABLE 4 (Biochemistry - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group Alb / sex g/L 2 3 4 ------POSF------ 30 100 300 a1 a2 Beta g/L g/L g/L 1M Mean 31 12 SD 1.3 0.9 n 5 5 * 2M Mean 29 10 SD 1.8 1.0 n 5 5 ** 3M Mean 28 10 SD 0.7 1.1 n 5 5 5 14 0.4 0.8 5 5 5 15 1.5 1.3 5 5 4 14 0.4 1.1 5 5 4M Mean 31 12 SD 1.8 0.9 n 5 5 5 14 0.4 0.8 5 5 Level of significance - all comparisons made with control * p 0.05, ** p 0.01 (Dunnett's test) Gamma g/L 4 1.5 5 5 0.8 5 5 1.6 5 5 0.8 5 A/G Ratio 0.92 0.080 5 0.84 0.112 5 0.84 0.059 5 0.89 0.070 5 MIN 313/023622 : 62 : TABLE 4 (Biochemistry - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex ALP ALT AST LDH U/L U/L U/L Total U/L 1F Mean 117 67 129 209 SD 26.8 60.5 113.0 163.1 n 4 4 4 4 2F Mean 95 56 124 356 SD 28.9 37.7 94.1 216.3 n 5 5 5 5 3F Mean 104 38 84 252 SD 13.4 12.7 19.9 147.5 n 5 5 5 5 4F Mean 112 44 86 248 SD 22.3 11.7 17.7 121.3 n 5 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 : 63 : TABLE 4 (Biochemistry - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex CPK Total U/L SDH Bili Urea Creat U/L mol/L mmol/L mol/L 1F Mean 104 19.1 3 SD 33.3 10.66 0.6 n 4 4 4 * 2F Mean 170 16.9 2 SD 86.5 6.10 0.5 n 5 5 5 * 3F Mean 285 13.1 2 SD 368.6 2.52 0.0 n 5 5 5 ** 4F Mean 150 23.0 1 SD 58.9 19.61 0.4 n 5 5 5 6.85 59 0.882 3.2 4 4 6.23 57 0.770 3.3 5 5 6.13 57 0.375 4.9 5 5 6.87 59 1.226 6.0 5 5 Level of significance - all comparisons made with control * p 0.05, ** p 0.01 (Williams' test) Gluc mmol/L Chol 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 TABLE 4 (Biochemistry - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex 1 Control 0 Na mmol/L 2 3 4 ------POSF------ 30 100 300 K mmol/L Cl mmol/L Ca mmol/L Phos mmol/L Total Prot g/L MIN 313/023622 : 64 : 1F Mean 140 SD 1.3 n 4 * 2F Mean 142 SD 0.9 n 5 * 3F Mean 142 SD 1.1 n 5 ** 4F Mean 143 SD 1.5 n 5 3.3 101 0.14 1.0 4 4 3.4 102 0.32 1.3 5 5 * 3.3 103 0.22 0.9 5 5 ** 3.6 104 0.19 1.9 5 5 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 74 0.217 1.3 4 4 1.32 71 0.157 4.0 5 5 1.42 70 0.227 4.9 5 5 1.63 71 0.369 2.5 5 5 MIN 313/023622 : 65 : TABLE 4 (Biochemistry - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group Alb / sex g/L 2 3 4 ------POSF------ 30 100 300 a1 a2 Beta g/L g/L g/L 1F Mean 41 9 4 14 SD 3.1 0.8 0.0 0.5 n 4 4 4 4 2F Mean 39 9 4 14 SD 2.5 1.2 0.5 0.8 n 5 5 5 5 3F Mean 38 10 SD 2.3 1.1 n 5 5 3 14 0.5 0.5 5 5 4F Mean 39 9 4 14 SD 0.9 1.5 0.5 1.0 n 5 5 5 5 No differences of statistical significance (p> 0.05) Gamma g/L 5 0.8 4 5 1.3 5 6 2.2 5 5 1.4 5 A/G Ratio 1.29 0.171 4 1.20 0.102 5 1.16 0.096 5 1.21 0.060 5 TABLE 5 Urinalysis - group mean values Day 91 Group : Compound : Exposure level (ppm) : Group / sex 1 Control 0 Vol ml 2 3 4 ------POSF------ 30 100 300 pH SG Prot g/L g/L U-Na mmol/L U-K mmol/L U-Cl mmol/L MIN 313/023622 : 66 : 1M Mean SD n 7.2 7.3 1035 3.55 0.43 8.5 5 5 5 2M Mean SD n 5.3 7.3 1044 2.01 0.27 11.8 5 5 5 3M Mean SD n 4M Mean SD n 7.2 6.9 1036 1.49 0.33 5.8 5 5 5 ** ** 4.0 6.6 1058 1.59 0.15 13.8 5 5 5 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 TABLE 5 (Urinalysis - continued) Day 91 Group : Compound : Exposure level (ppm) : Group / sex 1 Control 0 Vol ml 2 3 4 ------POSF------ 30 100 300 pH SG Prot g/L g/L U-Na mmol/L U-K mmol/L U-Cl mmol/L MIN 313/023622 : 67 : 1F Mean 4.3 6.6 1036 SD 2.56 0.18 8.4 n 5 5 5 2F Mean 4.2 6.7 1036 SD 1.26 0.30 6.1 n 5 5 5 3F Mean 2.8 6.4 1054 SD 0.91 0.16 9.8 n 5 5 5 ** 4F Mean 2.3 6.6 1068 SD 0.61 0.38 22.3 n 5 5 5 Level of significance - all comparisons made with control * p 005, ** p 0.01 (Williams' test) 0.19 0.061 5 0.21 0.053 5 0.25 0.041 5 ** 0.31 0.083 5 86.7 28.95 5 81.2 23.18 5 87.4 18.34 5 92.9 35.86 5 159.2 40.42 5 157.0 26.45 5 * 242.3 30.20 5 ** 310.0 96.56 5 83.4 28.60 5 79.7 12.41 5 93.3 21.99 5 112.2 31.87 5 TABLE 5 (Urinalysis - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex 1 Control 0 Vol ml 2 3 4 ------POSF------ 30 100 300 pH SG Prot g/L g/L U-Na mmol/L U-K mmol/L U-Cl mmol/L MIN 313/023622 : 68 : 1M Mean SD n 7.1 7.1 1030 4.35 0.36 4.7 5 5 5 2M Mean SD n 3M Mean SD n 4M Mean SD n 6.2 7.0 1042 1.55 0.07 9.6 5 5 5 * * 5.8 6.7 1048 3.14 0.18 15.1 5 5 5 ** * 5.5 6.6 1048 2.03 0.12 12.7 5 5 5 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 TABLE 5 (Urinalysis - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex 1 Control 0 Vol ml 2 3 4 ------POSF------ 30 100 300 pH SG Prot g/L g/L Na Total mmol K Total mmol Cl Total mmol MIN 313/023622 : 69 : 1F Mean 4.0 6.1 1041 SD 2.38 0.15 6.0 n 4 4 4 2F Mean 4.7 6.2 1039 SD 0.79 0.20 1.8 n 5 5 5 3F Mean 2.7 6.1 1049 SD 1.10 0.13 10.8 n 5 5 5 4F Mean 3.4 6.2 1046 SD 1.03 0.11 9.6 n 5 5 5 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 TABLE 5 (Urinalysis - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex 1 Control 0 U-Na mmol/L 2 3 4 ------POSF------ 30 100 300 U-K mmol/L U-Cl mmol/L MIN 313/023622 : 70 : 1F Mean 58.8 177.6 49.3 SD 9.28 34.12 7.70 n 4 4 4 2F Mean 67.9 181.2 66.8 SD 18.51 33.92 28.80 n 5 5 5 3F Mean 47.3 239.0 54.1 SD 26.34 87.36 22.48 n 5 5 5 4F Mean 72.1 218.5 57.3 SD 16.33 57.82 7.77 n 5 5 5 TABLE 6 Organ weights - group mean values (g) Print No: 0021 Main study animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 71 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: -----------------MALE----------------- -----------------MALE----------------- GROUP: ---1--- ---2--- ---3--- ---4--- ---1--- ---2--- ---3--- ---4--- NUMBER: 5 5 5 5 5 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL BODY WEIGHT (g) N : 5 5 5 5 MEAN : 452.6 449.7 440.5 371.4 sd : 30.2 43.0 25.3 26.9 ------------------------------------------------------------------------------------------------------------------------------------ BRAIN BRAIN * N : 5 5 5 5 5 5 5 5 MEAN : 2.02 2.03 1.98 2.00 1.97 1.99 1.96 2.10 sd : 0.08 0.09 0.09 0.07 ------------------------------------------------------------------------------------------------------------------------------------ ADRENALS ADRENALS N : 5 5 5 5 5 5 5 5 MEAN : 0.061 0.054 0.056 0.058 0.056 0.050 0.054 0.069 sd : 0.016 0.008 0.007 0.009 ------------------------------------------------------------------------------------------------------------------------------------ HEART HEART N : 5 5 5 5 5 5 5 5 MEAN : 1.460 1.377 1.380 1.327 1.348 1.279 1.325 1.592 sd : 0.213 0.184 0.161 0.217 ------------------------------------------------------------------------------------------------------------------------------------ KIDNEYS KIDNEYS * ** N : 5 5 5 5 5 5 5 5 MEAN : 2.59 2.61 2.78 2.45 2.42 2.45 2.70 2.86 sd : 0.23 0.26 0.34 0.25 ------------------------------------------------------------------------------------------------------------------------------------ Level of significance - all comparisons made with control *p 0.05, ** p 0.01 (Williams' test) TABLE 6 (Organ weights - continued) Print No: 0021 Main study animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 72 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: -----------------MALE----------------- -----------------MALE----------------- GROUP: ---1--- ---2--- ---3--- ---4--- ---1--- ---2--- ---3--- ---4--- NUMBER: 5 5 5 5 5 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ LIVER LIVER * ** N : 5 5 5 5 5 5 5 5 MEAN : 14.82 14.57 16.27 18.41 13.70 13.59 15.71 21.07 sd : 2.02 1.94 1.61 1.78 ------------------------------------------------------------------------------------------------------------------------------------ LUNGS & BRONCHI LUNGS & BRONCHI ** N : 5 5 5 5 5 5 5 5 MEAN : 1.520 1.468 1.592 1.979 1.429 1.388 1.547 2.194 sd : 0.115 0.149 0.264 0.152 ------------------------------------------------------------------------------------------------------------------------------------ Level of significance - all comparisons made with control * p 0.05, ** p 0.01 (Williams' test) TABLE 6 (Organ weights - continued) Print No: 0022 Main study animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 73 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: ----------------FEMALE---------------- -----------------FEMALE--------------- GROUP: ---1--- ---2--- ---3--- ---4--- ---1--- ---2--- ---3--- ---4--- NUMBER: 4 5 5 5 4 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL BODY WEIGHT (g) N : 4 5 5 5 MEAN : 265.3 242.8 256.7 241.0 sd : 7.7 14.5 31.6 15.3 ------------------------------------------------------------------------------------------------------------------------------------ BRAIN BRAIN N : 4 5 5 5 4 5 5 5 MEAN : 1.92 1.86 1.88 1.92 1.89 1.87 1.87 1.94 sd : 0.10 0.03 0.07 0.09 ------------------------------------------------------------------------------------------------------------------------------------ ADRENALS N : 4 5 5 5 MEAN : 0.059 0.066 0.067 0.064 sd : 0.010 0.012 0.007 0.009 ------------------------------------------------------------------------------------------------------------------------------------ HEART N : 4 5 5 5 MEAN : 0.985 0.971 0.943 0.920 sd : 0.052 0.073 0.067 0.167 ------------------------------------------------------------------------------------------------------------------------------------ KIDNEYS N : 4 5 5 5 MEAN : 1.62 1.69 1.76 1.64 sd : 0.18 0.11 0.12 0.23 ------------------------------------------------------------------------------------------------------------------------------------ No differences of statistical significance (p> 0.05) TABLE 6 (Organ weights - continued) Print No: 0022 Main study animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 74 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: ----------------FEMALE---------------- -----------------FEMALE-------------- GROUP: ---1--- ---2--- ---3--- ---4--- ---1--- ---2--- ---3--- ---4--- NUMBER: 4 5 5 5 4 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ LIVER LIVER * ** N : 4 5 5 5 4 5 5 5 MEAN : 8.88 8.54 9.86 12.24 8.29 8.86 9.62 12.64 sd : 0.75 0.79 1.31 1.43 ------------------------------------------------------------------------------------------------------------------------------------ LUNGS & BRONCHI LUNGS & BRONCHI ** N : 4 5 5 5 4 5 5 5 MEAN : 1.170 1.098 1.214 1.597 1.116 1.127 1.193 1.632 sd : 0.049 0.050 0.150 0.160 ------------------------------------------------------------------------------------------------------------------------------------ Level of significance - all comparisons made with control * p 0.05, ** p 0.01 (Williams' test) TABLE 6 (Organ weights - continued) Print No: 0014 Recovery phase animals GROUP : 1 2 3 4 Printed: 25-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 75 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: -----------------MALE----------------- -----------------MALE----------------- GROUP: ---1--- ---2--- ---3--- ---4--- ---1--- ---2--- ---3--- ---4--- NUMBER: 5 5 5 5 5 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL BODY WEIGHT (g) N : 5 5 5 5 MEAN : 482.5 457.7 469.6 442.9 sd : 23.2 39.2 43.5 33.7 ------------------------------------------------------------------------------------------------------------------------------------ BRAIN N : 5 5 5 5 MEAN : 2.11 2.13 2.08 2.05 sd : 0.09 0.09 0.08 0.12 ------------------------------------------------------------------------------------------------------------------------------------ ADRENALS N : 5 5 5 5 MEAN : 0.057 0.052 0.052 0.050 sd : 0.010 0.008 0.004 0.007 ------------------------------------------------------------------------------------------------------------------------------------ HEART N : 5 5 5 5 MEAN : 1.471 1.449 1.513 1.546 sd : 0.150 0.156 0.143 0.226 ------------------------------------------------------------------------------------------------------------------------------------ KIDNEYS KIDNEYS * N : 5 5 5 5 5 5 5 5 MEAN : 2.80 2.97 2.89 2.86 2.68 3.00 2.85 2.98 sd : 0.06 0.25 0.41 0.26 ------------------------------------------------------------------------------------------------------------------------------------ Level of significance - all comparisons made with control * p 0.05 (Williams' test) TABLE 6 (Organ weights - continued) Print No: 0014 Recovery phase animals GROUP : 1 2 3 4 Printed: 25-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 76 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: -----------------MALE----------------- -----------------MALE----------------- GROUP: ---1--- ---2--- ---3--- ---4--- ---1--- ---2--- ---3--- ---4--- NUMBER: 5 5 5 5 5 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ LIVER LIVER ** ** N : 5 5 5 5 5 5 5 5 MEAN : 12.14 12.48 13.94 15.53 11.62 12.62 13.76 16.07 sd : 0.67 1.43 1.72 1.18 ------------------------------------------------------------------------------------------------------------------------------------ LUNGS & BRONCHI LUNGS & BRONCHI ** N : 5 5 5 5 5 5 5 5 MEAN : 1.472 1.467 1.445 1.616 1.426 1.480 1.429 1.665 sd : 0.088 0.057 0.141 0.144 ------------------------------------------------------------------------------------------------------------------------------------ Level of significance - all comparisons made with control ** p 0.01 (Williams' test) TABLE 6 (Organ weights - continued) Print No: 0015 Recovery phase animals GROUP : 1 2 3 4 Printed: 25-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 77 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: ----------------FEMALE---------------- -----------------FEMALE-------------- GROUP: ---1--- ---2--- ---3--- ---4--- ---1--- ---2--- ---3--- ---4--- NUMBER: 4 5 5 5 4 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL BODY WEIGHT (g) N : 4 5 5 5 MEAN : 291.4 291.5 258.0 260.9 sd : 18.1 22.0 11.2 23.9 ------------------------------------------------------------------------------------------------------------------------------------ BRAIN BRAIN N : 4 5 5 5 4 5 5 5 MEAN : 1.97 2.05 1.92 1.93 1.94 2.02 1.95 1.96 sd : 0.05 0.09 0.07 0.06 ------------------------------------------------------------------------------------------------------------------------------------ ADRENALS ADRENALS N : 4 5 5 5 4 5 5 5 MEAN : 0.073 0.071 0.068 0.063 0.069 0.066 0.073 0.067 sd : 0.006 0.011 0.009 0.007 ------------------------------------------------------------------------------------------------------------------------------------ HEART HEART N : 4 5 5 5 4 5 5 5 MEAN : 1.142 1.030 0.987 0.925 1.105 0.993 1.024 0.955 sd : 0.049 0.151 0.097 0.051 ------------------------------------------------------------------------------------------------------------------------------------ KIDNEYS KIDNEYS N : 4 5 5 5 4 5 5 5 MEAN : 1.88 1.90 1.77 1.67 1.79 1.81 1.85 1.74 sd : 0.09 0.21 0.11 0.18 ------------------------------------------------------------------------------------------------------------------------------------ No differences of statistical significance (p> 0.05) TABLE 6 (Organ weights - continued) Print No: 0015 Recovery phase animals GROUP : 1 2 3 4 Printed: 25-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 78 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: ----------------FEMALE---------------- -----------------FEMALE-------------- GROUP: ---1--- ---2--- ---3--- ---4--- ---1--- ---2--- ---3--- ---4--- NUMBER: 4 5 5 5 4 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ LIVER LIVER ** N : 4 5 5 5 4 5 5 5 MEAN : 8.99 8.80 8.44 9.78 8.45 8.23 9.00 10.25 sd : 0.94 0.67 0.46 1.11 ------------------------------------------------------------------------------------------------------------------------------------ LUNGS & BRONCHI LUNGS & BRONCHI N : 4 4 5 5 4 5 5 5 MEAN : 1.290 1.200 1.089 1.187 1.245 1.151 1.131 1.221 sd : 0.067 0.072 0.072 0.074 ------------------------------------------------------------------------------------------------------------------------------------ Level of significance - all comparisons made with control ** p 0.01 (Williams' test) TABLE 6 (Organ weights - continued) Print No: 0005 Satellite study animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 79 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: -------MALE------- --------MALE------- GROUP: ---1--- ---4--- ---1--- ---4--- NUMBER: 5 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL BODY WEIGHT (g) N : 5 5 MEAN : 332.8 317.9 sd : 30.3 22.1 ------------------------------------------------------------------------------------------------------------------------------------ ADRENALS ADRENALS N : 5 5 5 5 MEAN : 0.053 0.056 0.050 0.059 sd : 0.015 0.009 ------------------------------------------------------------------------------------------------------------------------------------ BRAIN N : 5 5 MEAN : 1.94 1.97 sd : 0.12 0.06 ------------------------------------------------------------------------------------------------------------------------------------ HEART HEART N : 5 5 5 5 MEAN : 1.284 1.304 1.235 1.353 sd : 0.223 0.154 ------------------------------------------------------------------------------------------------------------------------------------ KIDNEYS KIDNEYS N : 5 5 5 5 MEAN : 2.32 2.33 2.27 2.38 sd : 0.23 0.19 ------------------------------------------------------------------------------------------------------------------------------------ No differences of statistical significance (p> 0.05) TABLE 6 (Organ weights - continued) Print No: 0005 Satellite study animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 80 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: -------MALE------- --------MALE------- GROUP: ---1--- ---4--- ---1--- ---4--- NUMBER: 5 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ LIVER LIVER +++ N : 5 5 5 5 MEAN : 9.62 12.71 9.36 12.98 sd : 1.12 0.97 ------------------------------------------------------------------------------------------------------------------------------------ LUNGS & BRONCHI LUNGS & BRONCHI +++ N : 5 5 5 5 MEAN : 1.251 1.705 1.225 1.731 sd : 0.178 0.104 ------------------------------------------------------------------------------------------------------------------------------------ Level of significance - all comparisons made with control +++ p 0.001 (Student's `t' test) TABLE 6 (Organ weights - continued) Print No: 0006 Satellite study animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 81 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: -----FEMALE------- -------FEMALE------- GROUP: ---1--- ---4--- ---1--- ---4--- NUMBER: 5 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL BODY WEIGHT (g) N : 5 5 MEAN : 212.8 194.6 sd : 16.1 7.2 ------------------------------------------------------------------------------------------------------------------------------------ ADRENALS ADRENALS + N : 5 5 5 5 MEAN : 0.066 0.069 0.060 0.076 sd : 0.012 0.010 ------------------------------------------------------------------------------------------------------------------------------------ BRAIN N : 5 5 MEAN : 1.84 1.79 sd : 0.08 0.06 ------------------------------------------------------------------------------------------------------------------------------------ HEART HEART N : 5 5 5 5 MEAN : 0.909 0.827 0.869 0.867 sd : 0.100 0.027 ------------------------------------------------------------------------------------------------------------------------------------ KIDNEYS N : 5 5 MEAN : 1.54 1.48 sd : 0.11 0.08 ------------------------------------------------------------------------------------------------------------------------------------ Level of significance - all comparisons made with control + p 0.05 (Student's `t' test) TABLE 6 (Organ weights - continued) Print No: 0006 Satellite study animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ MIN 313/023622 : 82 : ABSOLUTE VALUES BODYWEIGHT ADJUSTED VALUES SEX: -----FEMALE------- -------FEMALE------- GROUP: ---1--- ---4--- ---1--- ---4--- NUMBER: 5 5 5 5 ------------------------------------------------------------------------------------------------------------------------------------ LIVER + N : 5 5 MEAN : 7.20 7.93 sd : 0.62 0.27 ------------------------------------------------------------------------------------------------------------------------------------ LUNGS & BRONCHI ++ N : 5 5 MEAN : 1.040 1.334 sd : 0.086 0.105 ------------------------------------------------------------------------------------------------------------------------------------ Level of significance - all comparisons made with control + p 0.05, ++ p 0.01 (Student's `t' test) TABLE 7 MIN 313/023622 : 83 : Macroscopic pathology - group distribution Print No: 0025 Main study animals GROUP : 1 2 3 4 Printed: 25-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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: -1- -2- -3- -4- -1- -2- -3- -4- ORGAN AND KEYWORD(S) OR PHRASE NUMBER: 5 5 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** TOP OF LIST ** KIDNEYS ............................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 PELVIC DILATION 0 0 0 0 0 1 0 1 LIVER ................................................. NUMBER EXAMINED: 5 5 5 5 4 5 5 5 MEDIAN CLEFT PALE AREA(S) 1 1 1 1 0 0 0 1 ENLARGED 0 0 0 3 0 0 0 2 LUNGS & BRONCHI ....................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 CONGESTED 1 1 1 0 0 0 0 0 INCOMPLETE COLLAPSE 0 0 0 0 0 0 0 3 DARK AREA(S) 0 0 0 0 0 0 0 1 LN MANDIBULAR ......................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 ENLARGED 1 1 0 0 0 1 1 1 CONGESTED 2 1 1 2 0 0 0 1 MISCELLANEOUS ......................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 ANIMAL THIN 0 1 1 3 0 0 1 3 FUR STAINING 0 0 0 0 1 0 0 0 FUR BADLY GROOMED 0 0 0 0 0 0 0 1 PREPUTIAL GLANDS ...................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 ABSCESS(ES) 0 1 0 0 0 0 0 0 PROSTATE .............................................. NUMBER EXAMINED: 5 5 5 5 0 0 0 0 ABSCESS(ES) 0 1 0 0 0 0 0 0 SKIN .................................................. NUMBER EXAMINED: 5 5 5 5 4 5 5 5 HAIRLOSS 0 1 1 0 1 1 1 3 SCAB(S) 0 1 0 0 0 0 0 0 TABLE 7 MIN 313/023622 : 84 : (Macroscopic pathology - continued) Print No: 0025 Main study animals GROUP : 1 2 3 4 Printed: 25-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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: -1- -2- -3- -4- -1- -2- -3- -4- ORGAN AND KEYWORD(S) OR PHRASE NUMBER: 5 5 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- STOMACH ............................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 ANTRUM WHITE NODULE(S) 1 0 0 1 0 1 1 0 CORPUS WHITE NODULE(S) 0 0 0 1 0 0 0 0 FORESTOMACH DEPRESSION(S) 0 0 0 0 0 0 0 1 TAIL .................................................. NUMBER EXAMINED: 5 5 5 5 4 5 5 5 SWELLING(S) 0 0 0 0 0 0 1 1 THYMUS ................................................ NUMBER EXAMINED: 5 5 5 5 4 5 5 5 DARK 0 0 0 0 0 1 0 0 UTERUS ................................................ NUMBER EXAMINED: 0 0 0 0 4 5 5 5 FLUID DISTENSION 0 0 0 0 0 3 1 2 ** END OF LIST ** TABLE 7 MIN 313/023622 : 85 : (Macroscopic pathology - continued) Print No: 0016 Recovery phase animals GROUP : 1 2 3 4 Printed: 25-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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: -1- -2- -3- -4- -1- -2- -3- -4- ORGAN AND KEYWORD(S) OR PHRASE NUMBER: 5 5 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** TOP OF LIST ** KIDNEYS ............................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 PELVIC DILATION 0 0 0 0 0 1 0 0 LIVER ................................................. NUMBER EXAMINED: 5 5 5 5 4 5 5 5 MEDIAN CLEFT PALE AREA(S) 0 0 0 1 0 0 0 0 LOBULAR PATTERN ACCENTUATED 0 0 0 1 0 0 0 0 EPIDIDYMIDES .......................................... NUMBER EXAMINED: 5 5 5 5 0 0 0 0 SWELLING(S) 0 0 1 0 0 0 0 0 LN MANDIBULAR ......................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 ENLARGED 0 0 1 0 1 1 0 0 CONGESTED 0 0 0 0 0 0 0 2 MISCELLANEOUS ......................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 ANIMAL THIN 0 0 0 1 0 0 0 0 SKIN .................................................. NUMBER EXAMINED: 5 5 5 5 4 5 5 5 HAIRLOSS 2 2 0 4 3 2 1 0 SCAB(S) 0 1 0 0 0 0 0 0 STOMACH ............................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 ANTRUM WHITE NODULE(S) 0 2 1 0 0 0 2 0 URETERS ............................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 DISTENDED 0 0 0 0 0 1 0 0 UTERUS ................................................ NUMBER EXAMINED: 0 0 0 0 4 5 5 5 FLUID DISTENSION 0 0 0 0 0 2 2 1 ** END OF LIST ** TABLE 7 MIN 313/023622 : 86 : (Macroscopic pathology - continued) Print No: 0007 Satellite study animals GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 --- SEX: --MALE-- -FEMALEGROUP: -1- -4- -1- -4- ORGAN AND KEYWORD(S) OR PHRASE NUMBER: 5 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- ** TOP OF LIST ** KIDNEYS ............................................... NUMBER EXAMINED: 5 5 5 5 PELVIC DILATION 0 2 0 0 LIVER ................................................. NUMBER EXAMINED: 5 5 5 5 MEDIAN CLEFT PALE AREA(S) 0 1 2 3 ENLARGED 0 4 0 0 LN MANDIBULAR ......................................... NUMBER EXAMINED: 5 5 5 5 ENLARGED 0 0 3 0 LUNGS & BRONCHI ....................................... NUMBER EXAMINED: 5 5 5 5 PALE AREA(S) 0 0 1 0 CONGESTED 0 0 0 1 TAIL .................................................. NUMBER EXAMINED: 5 5 5 5 PUSTULE(S) 0 0 2 0 UTERUS ................................................ NUMBER EXAMINED: 0 0 5 5 FLUID DISTENTION 0 0 0 1 ** END OF LIST ** TABLE 8 MIN 313/023622 : 87 : Microscopic pathology - group distribution Print No: 0029 Main study animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 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 TABLE 8 MIN 313/023622 : 88 : (Microscopic pathology - continued) Print No: 0029 Main study animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** FROM PREVIOUS PAGE ** KIDNEYS ............................................... NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 2 0 0 3 1 0 0 4 --FOCAL CAPSULAR CONGESTION Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 --MEDULLARY TUBULAR BASOPHILIA Total> 0 0 0 0 0 0 0 0 --MEDULLARY TUBULAR FIBROSIS Total> 0 0 0 0 0 0 0 0 LARYNX ................................................ NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 4 5 4 1 3 4 4 0 --NECROSIS OF VENTRAL CARTILAGE Present> 0 0 0 4 0 0 0 5 Total> 0 0 0 4 0 0 0 5 --ARYTENOID - EPITHELIAL HYPERPLASIA Minimal> 1 0 0 2 1 0 0 2 Total> 1 0 0 2 1 0 0 2 --ARYTENOID - EPITHELIAL HYPERKERATOSIS PARAKERATOSIS Total> 0 0 0 0 0 0 0 0 --ARYTENOID - SUBEPITHELIAL INFLAMMATORY CELL INFILTRATION Minimal> 0 0 1 1 0 0 0 0 Total> 0 0 1 1 0 0 0 0 --VENTROLATERAL EPITHELIAL HYPERPLASIA Total> 0 0 0 0 0 0 0 0 --VENTROLATERAL EPITHELIAL SQUAMOUS METAPLASIA Total> 0 0 0 0 0 0 0 0 --VENTRAL POUCH - EPITHELIAL HYPERPLASIA Minimal> 0 0 0 0 0 1 1 0 Total> 0 0 0 0 0 1 1 0 TABLE 8 MIN 313/023622 : 89 : (Microscopic pathology - continued) Print No: 0029 Main study animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** FROM PREVIOUS PAGE ** LARYNX ................................................ NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 4 5 4 1 3 4 4 0 --VENTRAL POUCH - INFLAMMATORY CELL INFILTRATION Total> 0 0 0 0 0 0 0 0 --VENTRAL POUCH - FORIGN BODY IN LUMEN, RELATED TO EPITHELIAL HYPERPLASIA/ INFLAMMATORY CELL INFILTRATION Total> 0 0 0 0 0 0 0 0 --PROMINENT DILATED ACINI VENTRAL GLAND Total> 0 0 0 0 0 0 0 0 LIVER ................................................. NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 4 0 0 0 2 4 4 0 --CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA Slight> 0 4 0 0 0 0 0 3 Moderate> 0 0 5 5 0 0 0 2 Total> 0 4 5 5 0 0 0 5 --CENTRILOBULAR HEPATOCYTE HYPERTROPHY Slight> 0 1 0 0 0 0 0 0 Total> 0 1 0 0 0 0 0 0 --CENTRILOBULAR HEPATOCYTE NECROSIS/INFLAMMATORY CELL INFILTRATION/SINUSOIDAL CONGESTION Total> 0 0 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 --FOCAL/MULTIFOCAL SINGLE CELL NECROSIS INFLAMMATORY CELL INFILTRATION Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 TABLE 8 MIN 313/023622 : 90 : (Microscopic pathology - continued) Print No: 0029 Main study animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** FROM PREVIOUS PAGE ** LIVER ................................................. NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 4 0 0 0 2 4 4 0 --PARENCHYMAL INFLAMMATORY CELL INFILTRATION Minimal> 0 1 0 0 0 0 0 0 Total> 0 1 0 0 0 0 0 0 --PERIPORTAL LYMPHOCYTIC INFILTRATION Minimal> 1 0 0 0 0 1 0 0 Slight> 0 0 0 0 1 0 0 0 Total> 1 0 0 0 1 1 0 0 --CENTRILOBULAR INFLAMMATORY CELL INFILTRATION Total> 0 0 0 0 0 0 0 0 --EXTRAMEDULLARY HAEMOPOEISIS Minimal> 0 1 1 0 1 0 0 0 Total> 0 1 1 0 1 0 0 0 --PERIPORTAL HEPATOCYTE VACUOLATION Minimal> 0 0 0 0 0 0 1 0 Total> 0 0 0 0 0 0 1 0 --CENTRILOBULAR HEPATOCYTE VACUOLATION Minimal> 0 0 3 0 0 0 0 0 Slight> 0 0 1 0 0 0 0 0 Total> 0 0 4 0 0 0 0 0 --HEPATOCYTE VACUOLATION, MEDIAN CLEFT Minimal> 0 0 0 0 0 0 0 1 Total> 0 0 0 0 0 0 0 1 LUNGS & BRONCHI ....................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 0 3 3 0 3 3 3 0 --FOCAL BRONCHOALVEOLAR HYPERPLASIA Total> 0 0 0 0 0 0 0 0 TABLE 8 MIN 313/023622 : 91 : (Microscopic pathology - continued) Print No: 0029 Main study animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** FROM PREVIOUS PAGE ** LUNGS & BRONCHI ....................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 0 3 3 0 3 3 3 0 --FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES Minimal> 0 0 0 3 0 0 1 2 Slight> 0 0 0 1 0 0 0 2 Moderate> 0 0 0 1 0 0 0 0 Total> 0 0 0 5 0 0 1 4 --AGGREGATIONS OF ALVEOLAR MACROPHAGES Minimal> 2 0 0 0 0 1 1 0 Slight> 0 0 0 0 1 0 0 0 Total> 2 0 0 0 1 1 1 0 --ALVEOLITIS Minimal> 2 1 0 1 0 0 0 0 Total> 2 1 0 1 0 0 0 0 --ALVEOLAR HAEMORRHAGE Minimal> 2 1 0 2 1 0 0 2 Marked> 0 0 1 0 0 0 0 0 Total> 2 1 1 2 1 0 0 2 --PERIVASCULAR INFLAMMATORY CELL INFILTRATION Minimal> 0 0 0 0 0 1 0 1 Total> 0 0 0 0 0 1 0 1 --PROMINENT AGGREGATIONS OF LYMPHOCYTES Minimal> 0 0 1 0 0 0 0 0 Total> 0 0 1 0 0 0 0 0 --VASCULAR CONGESTION Slight> 0 0 1 0 0 0 0 0 Total> 0 0 1 0 0 0 0 0 --FOCAL OSSEOUS DEPOSITION Minimal> 0 0 0 0 0 1 0 3 Total> 0 0 0 0 0 1 0 3 TABLE 8 MIN 313/023622 : 92 : (Microscopic pathology - continued) Print No: 0029 Main study animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** FROM PREVIOUS PAGE ** LUNGS & BRONCHI ....................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 0 3 3 0 3 3 3 0 --EARLY AUTOLYSIS Total> 0 0 0 0 0 0 0 0 TRACHEA ............................................... NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 5 0 0 3 4 0 0 5 --EPITHELIAL HYPERPLASIA Minimal> 0 0 0 2 0 0 0 0 Total> 0 0 0 2 0 0 0 0 --EPITHELIAL INFLAMMATORY CELL INFILTRATION Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 --SUBEPITHELIAL INFLAMMATORY AGGREGATIONS Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 --SUBEPITHELIAL FIBROBLAST PROLIFERATION Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 TRACHEAL BIFURC. ...................................... NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 5 0 0 5 4 0 0 5 URINARY BLADDER ....................................... NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 5 0 0 5 4 0 0 5 EPIDIDYMIDES .......................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 LN MANDIBULAR ......................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 TABLE 8 MIN 313/023622 : 93 : (Microscopic pathology - continued) Print No: 0029 Main study animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- MISCELLANEOUS ......................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 PREPUTIAL GLANDS ...................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 PROSTATE .............................................. NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 SKIN .................................................. NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 STOMACH ............................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 TAIL .................................................. NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 THYMUS ................................................ NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 URETERS ............................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 UTERUS ................................................ NUMBER EXAMINED: 0 0 0 0 0 0 0 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 TABLE 8 MIN 313/023622 : 94 : (Microscopic pathology - continued) Print No: 0031 Recovery phase animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** TOP OF LIST ** HEART ................................................. NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 5 0 0 5 4 0 0 5 --MYOCARDITIS, VENTRICULAR REGION Total> 0 0 0 0 0 0 0 0 KIDNEYS ............................................... NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 0 0 0 2 2 0 0 4 --CORTICAL TUBULAR BASOPHILIA Minimal> 3 0 0 2 0 0 0 0 Slight> 1 0 0 1 0 0 0 0 Total> 4 0 0 3 0 0 0 0 --CORTICAL LYMPHOCYTIC INFILTRATION Minimal> 1 0 0 0 0 0 0 0 Slight> 1 0 0 0 0 0 0 0 Total> 2 0 0 0 0 0 0 0 --CORTICAL INTERSTITIAL FIBROBLAST PROLIFERATION/ FIBROSIS Total> 0 0 0 0 0 0 0 0 --UROTHELIAL HYPERPLASIA, UNILATERAL Total> 0 0 0 0 0 0 0 0 --PELVIC DILATATION Slight> 1 0 0 0 0 0 0 0 Total> 1 0 0 0 0 0 0 0 --MINERALISATION, CORTICOMEDULLARY Minimal> 0 0 0 0 2 0 0 1 Total> 0 0 0 0 2 0 0 1 --MEDULLARY MINERALISATION Minimal> 1 0 0 1 1 0 0 0 Total> 1 0 0 1 1 0 0 0 --FOCAL CAPSULAR CONGESTION Total> 0 0 0 0 0 0 0 0 TABLE 8 MIN 313/023622 : 95 : (Microscopic pathology - continued) Print No: 0031 Recovery phase animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** FROM PREVIOUS PAGE ** KIDNEYS ............................................... NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 0 0 0 2 2 0 0 4 --MEDULLARY TUBULAR BASOPHILIA Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 --MEDULLARY TUBULAR FIBROSIS Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 LARYNX ................................................ NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 5 4 4 1 4 4 4 2 --NECROSIS OF VENTRAL CARTILAGE Present> 0 0 0 4 0 0 0 3 Total> 0 0 0 4 0 0 0 3 --ARYTENOID - EPITHELIAL HYPERPLASIA Minimal> 0 1 1 0 0 0 0 0 Slight> 0 0 0 2 0 0 0 0 Total> 0 1 1 2 0 0 0 0 --ARYTENOID - EPITHELIAL HYPERKERATOSIS PARAKERATOSIS Minimal> 0 0 0 2 0 0 0 0 Total> 0 0 0 2 0 0 0 0 --ARYTENOID - SUBEPITHELIAL INFLAMMATORY CELL INFILTRATION Minimal> 0 1 0 0 0 0 0 1 Total> 0 1 0 0 0 0 0 1 --VENTROLATERAL EPITHELIAL HYPERPLASIA Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 --VENTROLATERAL EPITHELIAL SQUAMOUS METAPLASIA Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 TABLE 8 MIN 313/023622 : 96 : (Microscopic pathology - continued) Print No: 0031 Recovery phase animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** FROM PREVIOUS PAGE ** LARYNX ................................................ NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 5 4 4 1 4 4 4 2 --VENTRAL POUCH - EPITHELIAL HYPERPLASIA Minimal> 0 0 0 2 0 0 0 0 Total> 0 0 0 2 0 0 0 0 --VENTRAL POUCH - INFLAMMATORY CELL INFILTRATION Minimal> 0 0 0 2 0 0 0 0 Total> 0 0 0 2 0 0 0 0 --VENTRAL POUCH - FORIGN BODY IN LUMEN, RELATED TO EPITHELIAL HYPERPLASIA/ INFLAMMATORY CELL INFILTRATION Slight> 0 0 0 0 0 1 0 0 Moderate> 0 0 0 0 0 0 1 0 Total> 0 0 0 0 0 1 1 0 --PROMINENT DILATED ACINI VENTRAL GLAND Present> 0 0 0 0 0 0 0 1 Total> 0 0 0 0 0 0 0 1 LIVER ................................................. NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 5 1 0 0 2 5 4 1 --CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA Slight> 0 0 2 5 0 0 0 0 Total> 0 0 2 5 0 0 0 0 --CENTRILOBULAR HEPATOCYTE HYPERTROPHY Minimal> 0 1 3 0 0 0 0 4 Slight> 0 2 0 0 0 0 0 0 Total> 0 3 3 0 0 0 0 4 --CENTRILOBULAR HEPATOCYTE NECROSIS/INFLAMMATORY CELL INFILTRATION/SINUSOIDAL CONGESTION Moderate> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 --PROMINENT CENTRILOBULAR HEPATOCYTE PIGMENT Minimal> 0 0 0 0 0 0 0 2 Total> 0 0 0 0 0 0 0 2 TABLE 8 MIN 313/023622 : 97 : (Microscopic pathology - continued) Print No: 0031 Recovery phase animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** FROM PREVIOUS PAGE ** LIVER ................................................. NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 5 1 0 0 2 5 4 1 --FOCAL/MULTIFOCAL SINGLE CELL NECROSIS INFLAMMATORY CELL INFILTRATION Total> 0 0 0 0 0 0 0 0 --PARENCHYMAL INFLAMMATORY CELL INFILTRATION Minimal> 0 0 0 0 0 0 1 0 Total> 0 0 0 0 0 0 1 0 --PERIPORTAL LYMPHOCYTIC INFILTRATION Minimal> 0 0 1 0 1 0 0 0 Total> 0 0 1 0 1 0 0 0 --CENTRILOBULAR INFLAMMATORY CELL INFILTRATION Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 --EXTRAMEDULLARY HAEMOPOEISIS Minimal> 0 1 1 0 1 0 0 1 Total> 0 1 1 0 1 0 0 1 --PERIPORTAL HEPATOCYTE VACUOLATION Total> 0 0 0 0 0 0 0 0 --CENTRILOBULAR HEPATOCYTE VACUOLATION Slight> 0 1 0 0 0 0 0 0 Total> 0 1 0 0 0 0 0 0 --HEPATOCYTE VACUOLATION, MEDIAN CLEFT Slight> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 LUNGS & BRONCHI ....................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 2 4 3 0 2 4 4 1 --FOCAL BRONCHOALVEOLAR HYPERPLASIA Minimal> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 TABLE 8 MIN 313/023622 : 98 : (Microscopic pathology - continued) Print No: 0031 Recovery phase animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- ** FROM PREVIOUS PAGE ** LUNGS & BRONCHI ....................................... NUMBER EXAMINED: 5 5 5 5 4 5 5 5 NOT REMARKABLE: 2 4 3 0 2 4 4 1 --FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES Minimal> 1 1 1 2 0 1 0 2 Slight> 0 0 0 3 0 0 0 0 Total> 1 1 1 5 0 1 0 2 --AGGREGATIONS OF ALVEOLAR MACROPHAGES Minimal> 0 0 1 0 2 0 0 2 Total> 0 0 1 0 2 0 0 2 --ALVEOLITIS Minimal> 1 0 1 0 0 0 0 0 Total> 1 0 1 0 0 0 0 0 --ALVEOLAR HAEMORRHAGE Minimal> 0 0 0 0 0 0 1 0 Slight> 0 0 0 0 1 0 0 0 Total> 0 0 0 0 1 0 1 0 --PERIVASCULAR INFLAMMATORY CELL INFILTRATION Minimal> 2 0 0 1 1 0 0 1 Total> 2 0 0 1 1 0 0 1 --PROMINENT AGGREGATIONS OF LYMPHOCYTES Total> 0 0 0 0 0 0 0 0 --VASCULAR CONGESTION Total> 0 0 0 0 0 0 0 0 --FOCAL OSSEOUS DEPOSITION Minimal> 1 0 0 1 0 0 0 2 Total> 1 0 0 1 0 0 0 2 --EARLY AUTOLYSIS Total> 0 0 0 0 0 0 0 0 TABLE 8 MIN 313/023622 : 99 : (Microscopic pathology - continued) Print No: 0031 Recovery phase animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- TRACHEA ............................................... NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 5 0 0 4 4 0 0 5 --EPITHELIAL HYPERPLASIA Total> 0 0 0 0 0 0 0 0 --EPITHELIAL INFLAMMATORY CELL INFILTRATION Total> 0 0 0 0 0 0 0 0 --SUBEPITHELIAL INFLAMMATORY AGGREGATIONS Slight> 0 0 0 1 0 0 0 0 Total> 0 0 0 1 0 0 0 0 --SUBEPITHELIAL FIBROBLAST PROLIFERATION Total> 0 0 0 0 0 0 0 0 TRACHEAL BIFURC. ...................................... NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 5 0 0 5 4 0 0 5 URINARY BLADDER ....................................... NUMBER EXAMINED: 5 0 0 5 4 0 0 5 NOT REMARKABLE: 5 0 0 5 4 0 0 5 EPIDIDYMIDES .......................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 LN MANDIBULAR ......................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 MISCELLANEOUS ......................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 PREPUTIAL GLANDS ...................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 TABLE 8 MIN 313/023622 : 100 : (Microscopic pathology - continued) Print No: 0031 Recovery phase animals GROUP : 1 2 3 4 Printed: 18-SEP-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 5 5 4 5 5 5 ------------------------------------------------------------------------ -=- -=- -=- -=- -=- -=- -=- -=- PROSTATE .............................................. NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 SKIN .................................................. NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 STOMACH ............................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 TAIL .................................................. NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 THYMUS ................................................ NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 URETERS ............................................... NUMBER EXAMINED: 0 0 0 0 0 0 0 0 NOT REMARKABLE: 0 0 0 0 0 0 0 0 UTERUS ................................................ NUMBER EXAMINED: 0 0 0 0 0 0 0 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 APPENDIX 1 MIN 313/023622 : 101 : Weekly physical examination - individual findings Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 1M ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 1 7 14 SKIN ENCRUSTATION(S) HEAD STAINING BROWN HEAD 2-3 13-14 3 7 14 COAT HAIRLOSS FORELIMBS 3-14 41 8 18 COAT HAIRLOSS HEAD 13-18 43 8 18 EYES PROMINENT RIGHT 18 45 8 18 SKIN ENCRUSTATION(S) HEAD 13-15 APPENDIX 1 MIN 313/023622 : 102 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 2M ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 8 7 14 SKIN ENCRUSTATION(S) TAIL STAINING BROWN DORSAL BODY SURFACE 7-8 6-10 9 7 14 COAT HAIRLOSS DORSAL BODY SURFACE 2-3 10 7 14 COAT HAIRLOSS HEAD 13-14 46 8 18 COAT HAIRLOSS HEAD SKIN ENCRUSTATION(S) HEAD 4-5, 8-18 8-9 47 8 18 COAT HAIRLOSS DORSAL BODY SURFACE STAINING BROWN HEAD 2-4 10-18 APPENDIX 1 MIN 313/023622 : 103 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 2M ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 48 8 18 SKIN ENCRUSTATION(S) NOSE DORSAL BODY SURFACE 15-17 17-18 49 8 18 STAINING BROWN HEAD 18 50 8 18 COAT HAIRLOSS DORSAL BODY SURFACE SKIN ENCRUSTATION(S) DORSAL BODY SURFACE STAINING BROWN HEAD 9-18 8 12-18 APPENDIX 1 MIN 313/023622 : 104 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 3M ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 12 7 14 STAINING RED PENIS 4 13 7 14 COAT HAIRLOSS DORSAL BODY SURFACE 13-14 APPENDIX 1 MIN 313/023622 : 105 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 4M ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 16 7 14 STAINING BROWN HEAD DORSAL BODY SURFACE 10-14 5-14 17 7 14 STAINING BROWN HEAD 14 19 7 14 COAT HAIRLOSS HEAD DORSAL BODY SURFACE SKIN ENCRUSTATION(S) HEAD STAINING BROWN HEAD 4 2-3 4 6-14 20 7 14 STAINING BROWN HEAD 14 57 8 18 COAT HAIRLOSS HEAD 4-18 APPENDIX 1 MIN 313/023622 : 106 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 4M ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 58 8 18 COAT HAIRLOSS HEAD DORSAL BODY SURFACE STAINING BROWN DORSAL BODY SURFACE 7-9, 17-18 2-18 6-18 59 8 18 COAT HAIRLOSS HEAD 5-18 60 8 18 COAT HAIRLOSS FORELIMBS 5-18 APPENDIX 1 MIN 313/023622 : 107 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 1F ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 21 7 14 STAINING BROWN HEAD 8 22 7 14 STAINING BROWN VENTRAL BODY SURFACE 12-14 23 F 13 COAT HAIRLOSS VENTRAL BODY SURFACE STAINING BROWN HEAD 12-13 12-13 24 7 14 STAINING BROWN HEAD 3-8 61 8 18 COAT HAIRLOSS DORSAL BODY SURFACE 16-18 62 8 18 COAT HAIRLOSS HEAD DORSAL BODY SURFACE 6-11 16-18 APPENDIX 1 MIN 313/023622 : 108 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 1F ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 63 H 3 BUILD (DISPLACEMENT) LEFT HINDLIMB 3 RIGHT HINDLIMB 3 BODY TEMPERATURE REDUCED 3 COAT UNGROOMED 3 EXCRETA FAECES LOOSE 3 MUSCLE REACTION ABNORMAL GAIT SWAYING 3 POSTURE TILTED HEAD LEANS TO THE RIGHT 3 RIGHTING REFLEXES POOR 3 STAINING RED NOSE 3 64 8 18 STAINING BROWN HEAD 3-13, 17-18 65 8 18 COAT HAIRLOSS FORELIMBS 3-18 APPENDIX 1 MIN 313/023622 : 109 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 2F ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 27 7 14 STAINING BROWN HEAD DORSAL BODY SURFACE 3-6 11-14 66 8 18 STAINING BROWN VENTRAL BODY SURFACE DORSAL BODY SURFACE 12-17 18 67 8 18 COAT HAIRLOSS DORSAL BODY SURFACE STAINING BROWN HEAD 4-10 12-18 68 8 18 COAT HAIRLOSS FORELIMBS HEAD STAINING BROWN HEAD 15-18 10-18 12-15 69 8 18 COAT HAIRLOSS HEAD SKIN ENCRUSTATION(S) HEAD 8-18 8, 11-15, 17 APPENDIX 1 MIN 313/023622 : 110 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 2F ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 70 8 18 STAINING BROWN HEAD 5-6 APPENDIX 1 MIN 313/023622 : 111 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 3F ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 31 7 14 STAINING BROWN HEAD 5-14 33 7 14 COAT HAIRLOSS HEAD STAINING BROWN HEAD 7-14 5-14 34 7 14 STAINING BROWN HEAD DEATH CODE TERMINAL KILL 5-14 14 35 7 14 STAINING BROWN HEAD 12-14 71 8 18 STAINING BROWN HEAD 18 72 8 18 STAINING BROWN HEAD 12-18 APPENDIX 1 MIN 313/023622 : 112 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 3F ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 73 8 18 STAINING BROWN HEAD 5-18 75 8 18 COAT HAIRLOSS DORSAL BODY SURFACE 8-18 APPENDIX 1 MIN 313/023622 : 113 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 4F ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 36 7 14 COAT HAIRLOSS RIGHT FORELIMB STAINING BROWN HEAD 11-14 5-14 37 7 14 BUILD (DEFORMITY) PROMINENT AREA TAIL STAINING BROWN VENTRAL BODY SURFACE DORSAL BODY SURFACE 9-14 6-14 5-14 38 7 14 COAT UNGROOMED HAIRLOSS LEFT HINDLIMB STAINING BROWN VENTRAL BODY SURFACE DORSAL BODY SURFACE 8-14 10-14 4-14 4-14 39 7 14 STAINING BROWN HEAD 6-14 APPENDIX 1 MIN 313/023622 : 114 : (Weekly physical examination - continued) Print No: 0017 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 4F ANIMAL DEATH WK OF KEYWORD WEEKS 1-18 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 40 7 14 STAINING BROWN HEAD VENTRAL BODY SURFACE DORSAL BODY SURFACE 6-14 5-14 5-14 77 8 18 STAINING BROWN VENTRAL BODY SURFACE DORSAL BODY SURFACE 10-18 10-18 78 8 18 STAINING BROWN HEAD VENTRAL BODY SURFACE 6-18 6-18 79 8 18 STAINING BROWN HEAD 5-18 80 8 18 STAINING BROWN HEAD 5-15 APPENDIX 1 MIN 313/023622 : 115 : (Weekly physical examination - continued) Print No: 0008 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 1M ANIMAL DEATH WK OF KEYWORD WEEKS 1-4 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 81 7 4 SKIN ENCRUSTATION(S) HEAD 2-3 APPENDIX 1 MIN 313/023622 : 116 : (Weekly physical examination - continued) Print No: 0008 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 1F ANIMAL DEATH WK OF KEYWORD WEEKS 1-4 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 91 7 4 STAINING BROWN HEAD 4 93 7 4 STAINING BROWN UPPER DORSAL THORAX 3-4 94 7 4 DEATH CODE TERMINAL KILL 4 95 7 4 STAINING BROWN UPPER DORSAL THORAX 4 APPENDIX 1 MIN 313/023622 : 117 : (Weekly physical examination - continued) Print No: 0008 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ CATEGORY GROUP: 4F ANIMAL DEATH WK OF KEYWORD WEEKS 1-4 NUMBER CODE DEATH QUALIFIER ------------------------------------------------------------------------------------------------------------------------------------ 96 7 4 STAINING BROWN UPPER DORSAL THORAX 4 98 7 4 STAINING BROWN HEAD 4 100 7 4 STAINING BROWN HEAD 4 APPENDIX 2 Daily post - dose observation MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Main study animals Group Sign Animal No. 1 2 1M Red/brown staining around eyes 1 (Control) 2 1 3 2 4 3 5 Wet fur 1 35 2 25 3 45 4 25 5 25 Brown staining on head 1 2 3 4 5 Brown staining muzzle 4 Brown staining body 1 2M Red/brown staining around eyes 6 3 1 (Low dose) 7 8 41 9 11 10 3 Wet fur 6 44 7 34 8 44 9 34 10 3 4 Brown staining body 6 Brown staining on head 6 7 8 1 9 1 10 Animals exposed for 5 days a week for 13 weeks * Animals sent for necropsy over first 3 days of Week 14 : 118 : Week (Number of days in week signs observed) 3 4 5 6 7 8 9 10 11 12 1 1 1 1 1 2 2 1 1 1 2 2 4 1 2 1 1 3 1 13 14* 2 1 4 5 5 3 4 3 4 3 1 1 1 3 5 5 3 5 3 3 4 1 3 5 5 4 5 3 3 3 1 3 3 5 5 4 5 3 3 4 1 2 2 1 4 5 5 3 4 4 3 3 2 1 1 1 1 1 1 1 1 1 1 2 1 1 2 1 3 1 1 2 1 1 1 4 1 2 4 2 1 1 1 1 1 1 1 1 2 1 1 2 1 1 2 1 1 1 3 4 5 3 5 4 5 3 2 3 4 4 3 5 4 4 4 3 4 4 3 5 4 4 3 3 4 4 3 5 4 4 3 1 1 3 4 5 3 5 4 4 3 3 2 1 1 1 1 4 1 1 1 1 1 1 APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Main study animals Group Sign Animal No. Week (Number of days in week signs observed) 1 2 3 4 5 6 7 8 9 10 11 12 13 14* 3M Red/brown staining around eyes 11 1 1 1 (Inter. dose) 12 3 1 13 1 1 1 14 1 2 1 1 1 1 15 1 1 1 1 1 2 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 1 1 12 1 1 1 13 1 14 1 15 1 4M Red/brown staining around eyes 16 (High dose) 17 18 19 20 1 3 3 1 5 4 2 2 3 1 2 1 2 2 2 3 1 2 1 4 1 2 2 2 2 1 5 3 4 2 2 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 1 1 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 1 2 5 2 1 1 1 1 2 1 1 1 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. 1 1 : 119 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Main study animals Group Sign Animal No. Week (Number of days in week signs observed) 1 2 3 4 5 6 7 8 9 10 11 12 13 14* 1F Red/brown staining around eyes 21 1 2 2 3 1 2 1 (Control) 22 1 23 1 1 2 24 25 1 Wet fur 21 4 5 3 5 5 5 5 5 5 5 5 5 5 2 22 4 5 5 5 5 5 5 4 5 5 4 5 5 2 23 4 5 3 5 5 2 4 3 5 4 4 5 5 1a 24 3 5 3 5 5 2 4 4 4 4 1 5 5 1 25 3 5 4 5 5 2 4 4 4 4 3 5 5 1 Brown staining on head 21 3 3 1 1 1 2 2 2 4 4 1 22 1 2 1 1 3 1 3 2 3 23 1 3 1 1 1 2 3 4 3 4 1 24 2 1 2 4 3 2 3 3 1 25 1 2 1 2 1 1 4 2F Red/brown staining around eyes 26 (Low dose) 27 28 29 30 1 2 2 3 2 3 5 1 2 1 1 1 1 1 2 3 1 2 1 1 1 1 2 2 1 1 Wet fur 26 3 4 3 4 5 3 5 4 5 3 5 5 4 2 27 4 4 3 4 5 3 5 5 5 5 5 5 4 2 28 5 4 3 4 5 4 5 5 5 4 5 4 4 2 29 3 4 3 4 5 4 5 4 5 5 5 5 3 2 30 3 5 3 4 5 4 5 5 5 3 5 4 5 2 Brown staining on head 26 1 1 2 3 27 1 1 1 1 1 1 28 1 2 1 3 1 1 2 2 3 2 29 2 1 1 1 1 1 1 1 30 1 1 4 2 1 Animals exposed for 5 days a week for 13 weeks * Animals sent for necropsy over first 3 days of Week 14. a Animal No. 23 died at bleed : 120 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Main study animals Group Sign Animal No. Week (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 31 (Inter. dose) 32 33 34 35 1 1 1 1 1 1 1 2 1 3 4 5 2 2 1 2 1 4 1 4 2 4 3 2 3 2 1 1 4 1 1 1 1 4 2 2 1 1 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 1 5 2 1 34 1 2 1 2 3 1 2 1 3 2 2 1 3 2 35 1 1 2 2 2 1 4F Red/brown staining around eyes 36 1 1 1 (High dose) 37 2 1 1 1 1 1 1 4 38 1 2 1 2 1 2 1 1 39 1 2 2 40 3 2 1 2 3 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 1 2 3 1 3 1 1 2 1 1 1 1 1 1 1 2 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 5 5 5 5 5 5 5 5 2 : 121 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------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 3 3 2 2 2 2 1 1 1 (Control) 42 5 5 1 2 1 1 1 43 2 1 2 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 3 1 42 3 5 5 5 5 3 4 3 3 3 2 43 3 5 3 5 5 3 4 3 4 3 3 1 44 3 5 4 5 5 4 4 3 4 3 2 1 45 3 5 3 5 5 3 4 4 3 3 1 Brown staining on head 41 42 1 43 44 45 3 1 2 2 1 1 1 1 1 1 1 2 1 1 1 1 2M Red/brown staining around eyes 46 1 2 (Low dose) 47 3 1 1 1 2 1 1 48 2 1 1 2 49 1 1 1 3 4 3 2 3 1 1 1 1 50 2 1 2 1 1 3 3 1 2 3 2 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 1 48 49 3 50 2 Animals exposed for 5 days a week for 13 weeks * Animals sent for necropsy over first 3 days of Week 14. 1 1 1 1 1 1 1 1 1 2 1 1 2 2 1 1 1 2 3 : 122 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Recovery phase animals Group Sign Animal No. Week (Number of days in week signs observed) 1 2 3 4 5 6 7 8 9 10 11 12 13 14* 3M Red/brown staining around eyes 51 (Inter. dose) 52 53 54 55 1 2 1 1 1 1 1 1 1 1 1 3 2 1 1 1 1 2 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 1 1 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 1 1 2 1 52 1 53 1 1 54 1 1 55 1 1 3 2 2 4M Red/brown staining around eyes 56 (High dose) 57 58 59 60 2 2 1 1 2 1 1 1 1 3 2 2 3 3 2 2 2 1 1 5 4 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 1 57 1 1 58 1 2 1 2 59 1 2 1 1 1 1 60 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 : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------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* 1F Red/brown staining around eyes 61 4 1 1 2 3 1 1 (Control) 62 1 1 1 1 1 4 2 2 63 1 1 64 2 1 1 1 4 65 1 1 1 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 1 2 1 3 3 4 2 4 2 1 3 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 (Low dose) 67 68 69 70 1 1 1 4 2 2 3 1 2 2 2 1 1 2 1 3 2 2 1 1 1 1 2 1 3 1 2 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 1 3 1 1 1 2 3 1 69 1 1 1 1 1 1 3 3 2 3 70 2 1 5 4 2 2 1 2 Brown staining body 70 1 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 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Recovery phase animals Group Sign Animal No. Week (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 (Inter. dose) 72 73 74 75 2 1 2 1 1 1 1 1 2 1 Wet fur 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 Brown staining on head 71 72 73 74 75 1 1 2 1 1 1 2 2 1 1 1 1 4 1 1 1 2 1 2 1 1 1 2 2 1 4F Red/brown staining around eyes 76 (High dose) 77 78 79 80 2 2 2 1 1 4 2 1 3 1 1 2 1 2 4 3 1 1 1 4 4 3 4 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 1 1 80 1 2 2 1 3 1 1 1 1 5 1 2 4 2 2 1 1 1 4 5 3 2 Brown staining body 76 3 78 3 Animals exposed for 5 days a week for 13 weeks * Animals sent for necropsy over first 3 days of Week 14. : 125 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Satellite study animals Group Sign 1M (Control) Red/brown staining around eyes Animal No. 81 82 83 84 85 Week (Number of days in week signs observed) 1 2 3 4 4 1 2 1 3 1 2 4 2 1 2 3 3 4 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 2 1 1 1 3 1 4 2 1F 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 91 92 1 93 94 95 Animals exposed for 5 days a week for 4 weeks 3 1 1 4 5 2 2 2 : 126 : APPENDIX 2 (Daily post - dose observation - continued) MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------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) 1 2 3 4 4 1 1 1 1 1 2 1 1 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 Red/brown staining around eyes 97 (High 98 dose) 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 MIN 313/023622 : 128 : Bodyweights - individual values (g) Print No: 0018 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ GROUP WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK ANIMAL -1 0 1 2 3 4 5 6 7 8 9 ------------------------------------------------------------------------------------------------------------------------------------ 1M 1 248 292 328 356 386 410 428 447 460 474 492 2 224 252 276 307 338 346 363 376 396 404 412 3 227 263 284 311 337 359 378 399 408 423 435 4 226 248 266 291 314 339 355 371 390 402 422 5 232 253 275 296 313 337 365 378 398 402 417 41 229 252 281 296 312 336 352 374 387 402 412 42 247 275 296 324 355 382 408 420 434 451 461 43 229 246 273 302 323 342 365 382 404 416 436 44 236 257 279 298 316 344 368 390 409 425 432 45 228 268 300 316 337 363 379 392 404 409 416 2M 6 241 278 304 337 365 393 409 437 452 469 490 7 224 239 254 272 287 302 319 332 352 366 380 8 232 279 328 342 372 392 416 412 430 451 466 9 237 276 292 319 344 364 374 393 408 416 426 10 235 257 265 293 321 336 352 368 391 404 419 46 244 282 311 336 365 391 409 427 442 459 472 47 225 240 253 271 283 299 309 324 335 344 351 48 245 282 308 323 343 361 375 398 422 434 442 49 221 251 275 289 307 323 336 354 366 373 376 50 246 284 305 321 343 356 371 380 392 408 409 3M 11 235 264 290 308 333 354 376 390 405 416 421 12 238 272 295 318 332 360 380 396 413 424 441 13 237 270 293 302 341 369 395 412 430 451 459 14 239 263 269 288 317 331 349 366 379 392 400 15 234 255 277 291 310 336 353 370 380 399 415 51 245 274 291 317 339 364 380 398 406 421 435 52 236 271 294 313 329 350 360 373 394 407 410 53 232 265 289 320 342 363 374 397 401 410 413 54 245 275 297 315 332 352 363 374 390 402 418 55 214 227 242 255 266 282 294 312 325 336 347 APPENDIX 3 MIN 313/023622 : 129 : (Bodyweights - continued) Print No: 0018 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ GROUP WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK ANIMAL -1 0 1 2 3 4 5 6 7 8 9 ------------------------------------------------------------------------------------------------------------------------------------ 4M 16 234 262 289 309 330 351 364 375 379 387 384 17 220 241 254 267 285 298 301 316 322 326 329 18 227 258 274 303 324 343 356 373 378 386 387 19 238 267 283 304 329 351 357 373 387 389 402 20 234 260 276 295 310 330 339 347 348 359 361 56 269 261 266 292 319 338 353 372 386 399 404 57 227 263 288 321 346 368 382 395 409 419 423 58 240 261 277 291 311 328 339 348 362 388 377 59 217 243 255 280 293 306 314 320 331 339 345 60 234 261 275 294 314 334 358 381 393 402 410 1F 21 171 184 190 205 217 230 230 250 259 263 258 22 170 185 199 206 208 225 239 237 238 245 255 23 173 188 198 208 229 238 239 251 254 257 259 24 177 193 202 207 225 234 246 240 257 261 265 25 177 191 206 217 224 229 241 252 260 258 268 61 195 207 217 237 246 262 268 276 282 287 294 62 174 188 201 213 213 216 231 241 251 246 263 63 189 196 214 231 A 64 186 194 196 210 211 223 216 227 239 253 243 65 172 185 197 197 211 227 232 243 252 256 266 2F 26 177 195 206 213 215 229 240 247 243 256 257 27 178 176 189 199 202 195 207 206 221 213 227 28 180 190 198 202 208 206 219 232 239 240 247 29 165 172 178 189 202 208 209 225 230 231 225 30 164 170 184 187 194 196 206 216 223 225 233 66 188 208 227 238 252 262 271 274 278 283 288 67 163 191 210 221 237 252 263 269 280 293 287 68 177 184 204 208 217 210 227 233 232 234 241 69 183 199 215 220 226 236 249 256 259 268 273 70 174 183 203 208 233 246 250 250 261 269 272 ------------------------------------------------------------------------------------------------------------------------------------ A Animal No. 63 sent for necropsy for humane reasons therefore no further bodyweights APPENDIX 3 MIN 313/023622 : 130 : (Bodyweights - continued) Print No: 0018 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ GROUP WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK ANIMAL -1 0 1 2 3 4 5 6 7 8 9 ------------------------------------------------------------------------------------------------------------------------------------ 3F 31 171 182 193 197 200 219 221 227 227 237 237 32 186 201 209 215 226 243 249 249 262 271 275 33 168 172 200 191 201 202 208 214 222 214 217 34 194 209 227 240 251 252 267 280 278 280 290 35 169 189 187 207 219 227 237 243 250 258 256 71 169 187 203 201 217 226 228 238 240 246 245 72 187 187 194 196 200 208 216 223 229 232 238 73 181 201 211 216 221 236 231 239 250 252 255 74 182 185 199 205 200 214 217 223 216 225 236 75 166 176 184 195 202 210 208 221 224 232 234 4F 36 185 193 213 208 207 219 228 234 230 238 243 37 176 180 202 211 218 219 228 240 244 240 246 38 178 198 211 209 225 231 239 239 244 244 256 39 167 178 192 196 204 205 215 218 223 221 225 40 184 203 190 226 234 244 252 256 259 263 265 76 163 173 186 184 188 192 200 199 205 211 213 77 174 181 195 190 189 195 192 205 208 216 219 78 190 206 211 211 215 215 224 226 231 229 234 79 188 207 228 238 244 249 263 268 271 271 277 80 182 185 202 191 202 204 212 224 238 235 235 APPENDIX 3 MIN 313/023622 : 131 : (Bodyweights - continued) Print No: 0018 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ GROUP WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK ANIMAL 10 11 12 13 14 15 16 17 ------------------------------------------------------------------------------------------------------------------------------------ 1M 1 500 510 516 506 2 426 433 436 429 3 456 464 470 462 4 427 444 449 437 5 433 440 453 455 41 421 433 449 464 473 485 499 506 42 465 476 492 488 500 516 529 542 43 444 454 461 466 487 500 515 531 44 447 461 471 477 489 502 518 533 45 418 424 430 439 446 455 466 472 2M 6 490 505 512 511 7 388 401 409 410 8 475 470 474 481 9 431 438 450 441 10 423 432 435 426 46 483 486 492 495 504 509 521 533 47 359 370 372 378 394 409 422 435 48 450 463 470 468 476 488 490 500 49 381 394 395 403 417 431 441 444 50 425 425 434 442 464 471 478 491 3M 11 431 435 442 436 12 451 458 469 464 13 448 453 455 454 14 403 412 415 402 15 432 450 461 458 51 446 455 468 460 475 501 505 514 52 418 426 440 434 446 459 468 487 53 406 434 442 454 478 492 501 509 54 427 440 451 467 481 493 499 504 55 362 366 375 380 397 409 407 413 APPENDIX 3 MIN 313/023622 : 132 : (Bodyweights - continued) Print No: 0018 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ GROUP WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK ANIMAL 10 11 12 13 14 15 16 17 ------------------------------------------------------------------------------------------------------------------------------------ 4M 16 389 390 394 378 17 336 341 349 337 18 395 401 397 392 19 408 416 422 416 20 372 375 379 357 56 413 416 421 429 444 461 466 478 57 433 439 442 444 461 478 501 515 58 378 384 393 395 406 415 426 436 59 358 358 363 366 386 399 424 430 60 422 429 429 431 431 433 452 466 1F 21 271 278 284 270 22 254 255 261 258 23 266 268 270 24 274 281 282 270 25 272 279 274 274 61 297 308 307 301 313 327 326 328 62 267 275 269 275 287 293 294 305 64 260 269 269 265 277 288 289 277 65 268 279 281 274 288 303 312 311 2F 26 262 264 269 261 27 234 233 230 226 28 251 251 245 243 29 232 243 239 232 30 239 247 245 243 66 286 289 294 293 300 312 320 324 67 301 307 310 301 317 328 330 330 68 242 246 240 234 278 265 274 275 69 273 276 282 287 297 295 303 310 70 266 278 283 281 282 303 303 299 3F 31 238 238 244 233 32 265 275 284 271 33 224 224 221 216 34 300 287 300 300 APPENDIX 3 MIN 313/023622 : 133 : (Bodyweights - continued) Print No: 0018 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ GROUP WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK ANIMAL 10 11 12 13 14 15 16 17 ------------------------------------------------------------------------------------------------------------------------------------ 3F 35 256 264 269 262 71 249 255 254 252 258 268 268 274 72 243 247 246 247 255 262 262 266 73 255 258 264 262 273 281 281 288 74 239 236 242 244 249 249 259 259 75 240 240 243 236 250 258 257 256 4F 36 244 243 248 242 37 256 258 253 240 38 248 257 254 242 39 227 228 224 217 40 269 269 275 259 76 215 217 221 216 223 229 237 242 77 222 226 227 229 241 251 258 262 78 237 238 242 230 255 267 268 278 79 279 280 280 279 291 302 303 318 80 241 241 248 250 257 268 273 272 APPENDIX 3 MIN 313/023622 : 134 : (Bodyweights - continued) Print No: 0009 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ GROUP WEEK WEEK WEEK WEEK WEEK ANIMAL -1 0 1 2 3 ------------------------------------------------------------------------------------------------------------------------------------ 1M 81 245 277 301 330 358 82 244 263 285 314 341 83 224 242 258 276 290 84 235 271 310 338 366 85 237 264 294 324 351 4M 86 232 270 301 319 339 87 232 272 293 321 342 88 235 262 273 290 305 89 249 277 295 321 343 90 238 278 296 316 331 1F 91 172 187 202 204 216 92 188 194 200 204 210 93 182 194 216 226 245 94 174 181 194 195 208 95 186 197 208 217 236 4F 96 183 190 196 193 200 97 182 192 195 194 202 98 177 190 192 193 200 99 169 180 192 196 200 100 167 183 191 203 217 APPENDIX 4 MIN 313/023622 : 135 : Food consumption - individual values (g/rat) Print No: 0019 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ GROUP WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK WEEK CAGE -1 1 2 3 4 5 6 7 8 9 10 ------------------------------------------------------------------------------------------------------------------------------------ 1M 1 181 180 196 205 213 213 215 214 216 212 216 9 183 163 176 186 197 198 194 192 195 193 190 2M 2 192 169 184 197 202 204 201 208 208 207 207 10 195 173 184 190 196 194 196 198 196 193 198 3M 3 192 175 185 191 198 199 208 209 209 203 204 11 190 168 172 192 197 194 200 198 201 200 200 4M 4 186 157 173 179 189 181 190 180 189 180 180 12 186 150 171 179 188 185 189 191 193 187 193 1F 5 136 132 136 143 148 149 148 147 147 146 147 13 137 133 142 145 151 151 153 151 147 148 148 2F 6 131 126 126 132 136 138 138 137 136 139 134 14 150 147 149 157 158 157 156 154 158 151 148 3F 7 138 134 153 147 151 153 153 151 150 148 147 15 135 130 133 136 138 137 136 136 139 136 131 4F 8 144 137 137 140 141 144 144 143 140 143 140 16 140 128 122 125 121 126 127 125 126 125 122 APPENDIX 4 MIN 313/023622 : 136 : (Food consumption - continued) Print No: 0019 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ GROUP WEEK WEEK WEEK WEEK WEEK WEEK WEEK CAGE 11 12 13 14 15 16 17 ------------------------------------------------------------------------------------------------------------------------------------ 1M 1 218 215 190 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 11 202 199 199 205 205 206 205 4M 4 175 177 153 12 189 185 186 190 196 199 200 1F 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 APPENDIX 4 MIN 313/023622 : 137 : (Food consumption - continued) Print No: 0010 GROUP : 1 2 3 4 Printed: 24-JUN-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ GROUP WEEK WEEK WEEK WEEK CAGE -1 1 2 3 ------------------------------------------------------------------------------------------------------------------------------------ 1M 17 186 169 185 197 4M 18 196 161 177 188 1F 19 136 138 145 149 4F 20 137 122 131 136 MIN 313/023622 : 138 : APPENDIX 5 Haematology - individual values Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group Animal Hct / sex number L/L 1M 1 CTD 2 0.417 3 CTD 4 0.437 5 0.441 2M 6 7 8 9 10 0.451 0.438 0.455 0.437 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 3 4 ------POSF------ 30 100 300 Hb RBC Retic g/dL x1012/L % CTD 15.3 CTD 15.4 15.5 CTD 7.94 CTD 8.39 8.40 CTD 2.13 CTD 3.17 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 g/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 MIN 313/023622 : 139 : APPENDIX 5 (Haematology - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number L x109/L 1M 1 CTD 2 10.87 3 CTD 4 8.24 5 7.39 2M 6 8.81 7 8.43 8 13.10 9 7.87 10 9.83 3M 11 9.07 12 6.89 13 9.76 14 9.95 15 8.92 4M 16 10.15 17 8.72 18 11.40 19 11.33 20 10.71 CTD Clotted sample 2 3 4 ------POSF------ 30 100 300 E x109/L B x109/L M x109/L CTD 0.32 CTD 0.11 0.10 CTD 0.06 CTD 0.03 0.03 CTD 0.52 CTD 0.27 0.28 0.19 0.03 0.21 0.18 0.03 0.31 0.23 0.10 0.47 0.13 0.03 0.39 0.16 0.03 0.10 0.19 0.03 0.18 0.11 0.02 0.18 0.28 0.04 0.17 0.14 0.05 0.23 0.27 0.02 0.22 0.09 0.02 0.06 0.09 0.03 0.10 0.07 0.05 0.17 0.13 0.04 0.10 0.15 0.04 0.34 LUC x109/L CTD 0.25 CTD 0.19 0.08 0.06 0.06 0.15 0.12 0.04 0.05 0.04 0.04 0.06 0.07 0.04 0.03 0.06 0.05 0.11 Plt x109/L CTD 925 CTD 957 537 888 961 223 1108 909 580 869 1080 924 801 803 952 1107 1058 1416 PT sec CTD 12.8 CTD 13.2 CTD 13.5 14.0 CTD CTD 14.2 14.2 14.7 14.6 13.9 13.6 13.2 13.5 14.0 12.9 12.5 APTT sec CTD 14.9 CTD 23.1 CTD 15.9 16.9 CTD CTD 16.9 23.9 17.1 18.4 19.5 16.3 16.0 23.4 20.8 17.0 17.2 APPENDIX 5 (Haematology - continued) MIN 313/023622 : 140 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Anisocytosis 1M 1 CTD 2 - 3 CTD 4 - 5 - 2M 6 - 7 - 8 - 9 - 10 - 3M 11 - 12 - 13 - 14 - 15 - 4M 16 - 17 - 18 - 19 - 20 - CTD Clotted sample 2 3 4 ------POSF------ 30 100 300 Microcytosis Macrocytosis Hypochromasia CTD + CTD + + CTD - CTD - CTD - CTD - + - - + - - - - - + - - - - - - - - + - - + - - - - - - - - - - - - - - - - - - - - - - - Hyperchromasia CTD - CTD - MIN 313/023622 : 141 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group Animal Hct / sex number L/L 1F 21 CTD 22 0.435 23 NS 24 0.410 25 0.425 2F 26 0.440 27 0.411 28 0.432 29 0.423 30 0.420 3F 31 0.420 32 0.404 33 0.411 34 0.424 35 0.448 4F 36 0.437 37 0.437 38 0.443 39 0.417 40 0.423 CTD Clotted sample NS No sample obtained APPENDIX 5 (Haematology - continued) 2 3 4 ------POSF------ 30 100 300 Hb RBC Retic g/dL x1012/L % CTD 15.6 NS 14.9 15.0 CTD 7.90 NS 7.29 7.61 CTD 2.04 NS 2.89 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 pg 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 g/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 : 142 : APPENDIX 5 (Haematology - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number L x109/L 1F 21 CTD 22 4.84 23 NS 24 5.87 25 9.04 2F 26 5.77 27 7.17 28 6.53 29 6.70 30 8.40 3F 31 6.39 32 8.17 33 5.91 34 10.10 35 6.35 4F 36 8.74 37 6.79 38 6.68 39 8.39 40 8.72 CTD Clotted sample NS No sample obtained 2 3 4 ------POSF------ 30 100 300 E x109/L B x109/L M x109/L 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 x109/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 Plt 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 APPENDIX 5 (Haematology - continued) MIN 313/023622 : 143 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Anisocytosis 1F 21 CTD 22 - 23 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 2 3 4 ------POSF------ 30 100 300 Microcytosis Macrocytosis Hypochromasia CTD NS - CTD NS - CTD NS - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Hyperchromasia CTD NS - MIN 313/023622 : 144 : APPENDIX 5 (Haematology - continued) 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 4M 56 57 58 59 60 1 Control 0 Hct L/L 0.388 0.418 0.393 0.441 0.412 0.390 0.379 0.406 0.394 0.404 0.416 0.413 0.399 0.419 0.418 0.413 0.391 0.439 0.415 0.410 2 3 4 ------POSF------ 30 100 300 Hb RBC Retic g/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 pg 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 g/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 x109/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 : 145 : APPENDIX 5 (Haematology - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number L x109/L 1M 41 7.88 42 7.59 43 8.02 44 9.84 45 10.16 2M 46 5.51 47 6.83 48 10.06 49 5.61 50 8.26 3M 51 12.51 52 12.31 53 6.15 54 6.97 55 7.59 4M 56 8.52 57 7.20 58 9.00 59 8.81 60 8.31 CTD Clotted sample 2 3 4 ------POSF------ 30 100 300 E x109/L B x109/L M x109/L 0.10 0.01 0.30 0.13 0.03 0.32 0.20 0.04 0.27 0.18 0.03 0.22 0.13 0.06 0.15 0.13 0.02 0.10 0.08 0.02 0.16 0.07 0.04 0.36 0.07 0.01 0.21 0.07 0.03 0.16 0.11 0.06 0.15 0.30 0.08 0.48 0.09 0.02 0.11 0.10 0.02 0.16 0.14 0.04 0.22 0.28 0.05 0.29 0.15 0.03 0.14 0.07 0.03 0.07 0.29 0.03 0.24 0.09 0.02 0.19 LUC x109/L 0.23 0.21 0.28 0.26 0.09 0.06 0.11 0.15 0.11 0.11 0.06 0.31 0.06 0.10 0.09 0.19 0.09 0.08 0.10 0.13 Plt x109/L 1123 1045 1028 860 990 1021 1147 887 829 1028 799 910 879 789 960 1063 938 903 1098 925 PT sec 12.9 13.0 12.6 13.2 12.8 13.8 CTD 14.4 13.5 14.2 14.6 15.2 14.5 13.9 14.9 13.8 13.6 14.4 13.0 13.8 APTT sec 17.6 21.2 20.4 20.9 15.7 17.9 CTD 17.6 17.5 20.0 15.7 19.9 14.1 16.3 14.1 14.7 14.3 18.0 15.5 16.1 APPENDIX 5 (Haematology - continued) MIN 313/023622 : 146 : Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Anisocytosis 1M 41 - 42 - 43 - 44 - 45 - 2M 46 - 47 + 48 - 49 - 50 - 3M 51 - 52 - 53 - 54 - 55 - 4M 56 - 57 - 58 - 59 - 60 - 2 3 4 ------POSF------ 30 100 300 Microcytosis Macrocytosis Hypochromasia + - - + - - + - - - - - ++ - - + - - ++ - - + - - - - - + - - - - - - - - + - - - - - - - - + - - - - - - - - + - - - - - Hyperchromasia - MIN 313/023622 : 147 : APPENDIX 5 (Haematology - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group Animal Hct / sex number L/L 1F 61 0.377 62 0.395 64 0.392 65 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 3 4 ------POSF------ 30 100 300 Hb RBC Retic g/dL x1012/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 1.90 14.8 7.36 2.02 15.4 7.76 2.28 CTD CTD 1.88 14.6 7.32 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 x109/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 : 148 : APPENDIX 5 (Haematology - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number L x109/L 1F 61 7.44 62 4.84 64 5.56 65 7.35 2F 66 3.94 67 5.73 68 4.57 69 CTD 70 3.74 3F 71 5.41 72 3.90 73 2.95 74 2.69 75 5.69 4F 76 3.62 77 2.93 78 5.39 79 3.33 80 5.10 CTD Clotted sample 2 3 4 ------POSF------ 30 100 300 E x109/L B x109/L M x109/L 0.12 0.02 0.23 0.07 0.01 0.19 0.27 0.02 0.19 0.16 0.03 0.21 0.09 0.11 0.08 CTD 0.19 0.01 0.01 0.01 CTD 0.02 0.10 0.11 0.14 CTD 0.26 0.10 0.01 0.12 0.07 0.01 0.09 0.05 0.01 0.05 0.07 0.00 0.04 0.12 0.02 0.21 0.19 0.01 0.08 0.05 0.00 0.04 0.09 0.01 0.13 0.06 0.01 0.07 0.12 0.01 0.14 LUC x109/L 0.12 0.11 0.09 0.17 0.06 0.10 0.06 CTD 0.16 0.08 0.04 0.03 0.06 0.15 0.10 0.04 0.18 0.06 0.12 Plt x109/L 947 869 1107 877 685 877 1001 CTD 796 672 491 564 890 800 865 745 831 479 828 PT sec 13.7 13.3 13.1 12.6 13.7 CTD 13.9 13.1 13.4 14.6 13.8 CTD 14.7 14.2 14.8 12.9 13.3 CTD 13.8 APTT sec 18.8 17.4 17.1 15.0 16.7 CTD 18.7 16.8 16.9 17.7 15.7 CTD 19.3 18.4 23.4 15.6 18.6 CTD 17.5 APPENDIX 5 (Haematology - continued) MIN 313/023622 : 149 : Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Anisocytosis 1F 61 - 62 - 64 - 65 - 2F 66 - 67 - 68 - 69 CTD 70 - 3F 71 - 72 - 73 - 74 - 75 - 4F 76 - 77 - 78 - 79 - 80 - CTD Clotted sample 2 3 4 ------POSF------ 30 100 300 Microcytosis Macrocytosis Hypochromasia - - - - - - - - - - - - CTD - CTD - CTD - - - - - - - + - - - - - - - - - - - - - - - - - - - - - - - Hyperchromasia CTD - MIN 313/023622 : 150 : APPENDIX 6 Biochemistry - individual values Day 91 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 ALP U/L 1M 1 150 2 217 3 186 4 198 5 260 2M 6 290 7 212 8 213 9 218 10 354 3M 11 228 12 302 13 259 14 264 15 342 2 3 4 ------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 APPENDIX 6 (Biochemistry - continued) MIN 313/023622 : 151 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 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 2 3 4 ------POSF------ 30 100 300 SDH Bili Urea U/L mol/L mmol/L 17.2 2 5.90 18.8 2 8.02 14.3 1 5.61 12.0 1 5.47 14.4 1 5.26 14.3 1 5.86 12.9 2 5.87 10.7 1 7.59 16.6 2 6.12 11.2 1 7.37 15.5 1 6.07 12.4 2 5.55 11.5 1 6.02 9.7 1 5.53 8.6 2 5.82 Creat mol/L 47 55 51 55 56 54 54 53 51 49 53 51 46 46 45 Gluc 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 Chol 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 APPENDIX 6 (Biochemistry - continued) MIN 313/023622 : 152 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group Animal K Cl Ca Phos Total Prot Alb a1 a2 Beta / sex number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L g/L g/L 1M 1 2 3 4 5 4.0 107 5.1 106 3.7 105 3.6 106 3.8 106 2M 6 7 8 9 10 3.5 107 4.0 108 4.0 110 3.8 106 3.6 108 3M 11 12 13 14 15 4.5 110 3.9 106 4.4 107 4.0 108 3.9 106 INS Insufficient sample 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) MIN 313/023622 : 153 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Gamma g/L 2 3 4 ------POSF------ 30 100 300 A/G Alb a1 Ratio % % a2 Beta Gamma % % % 1M 1 4 0.91 47.7 20.1 5.8 19.5 6.8 2 INS INS INS INS INS INS INS 3 6 0.83 45.6 19.1 4.4 21.6 9.3 4 5 0.86 45.7 18.5 4.7 23.1 7.9 5 7 0.78 44.4 14.5 4.5 26.0 10.6 2M 6 7 7 6 8 6 9 10 10 5 0.89 46.7 17.2 0.86 45.8 16.0 0.74 42.6 18.9 0.74 42.9 15.9 1.00 49.6 17.1 4.4 21.5 10.2 5.6 23.0 9.5 4.7 24.4 9.5 3.8 23.1 14.3 4.7 20.7 8.0 3M 11 4 12 6 13 6 14 6 15 4 1.10 52.1 16.7 0.84 45.2 18.8 0.80 45.1 18.7 0.81 44.7 19.5 0.86 46.7 19.6 4.5 20.2 6.5 4.8 22.3 9.0 6.5 19.6 10.0 5.1 21.8 8.8 4.7 22.3 6.6 INS Insufficient samples MIN 313/023622 : 154 : APPENDIX 6 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 ALP U/L 4M 16 366 17 346 18 302 19 399 20 224 2 3 4 ------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) MIN 313/023622 : 155 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number CPK Total U/L 4M 16 258 17 175 18 137 19 124 20 147 2 3 4 ------POSF------ 30 100 300 SDH Bili Urea U/L mol/L mmol/L 13.7 1 6.59 12.2 1 6.17 12.7 1 6.68 11.4 1 7.34 10.1 1 7.27 Creat mol/L 45 52 46 48 48 Gluc mmol/L 6.47 5.26 5.07 5.88 5.79 Chol 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) MIN 313/023622 : 156 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group Animal K Cl Ca Phos Total Prot Alb a1 / sex number mmol/L mmol/L mmol/L mmol/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 a2 Beta g/L g/L 2 12 3 14 3 13 3 14 3 15 MIN 313/023622 : 157 : APPENDIX 6 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Gamma g/L 2 3 4 ------POSF------ 30 100 300 A/G Alb a1 Ratio % % 4M 16 4 17 6 18 6 19 7 20 4 1.17 53.1 18.6 1.03 50.0 16.9 1.06 50.8 18.0 0.95 48.2 18.8 0.82 45.6 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 MIN 313/023622 : 158 : APPENDIX 6 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group Animal ALP ALT AST LDH / sex number UL U/L U/L Total U/L 1F 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 LDH3 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 : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex Animal number CPK Total U/L SDH Bili Urea U/L mol/L mmol/L 1F 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 6.86 27 100 9.1 2 6.88 28 111 11.0 3 8.08 29 101 10.9 3 6.80 30 140 19.4 2 7.15 3F 31 107 7.9 2 7.01 32 116 11.1 2 5.39 33 100 9.2 2 5.90 34 155 14.8 2 6.36 35 182 10.7 2 6.38 NS No sample Animal 23 died at bleed Creat mol/L 74 69 NS 63 59 57 56 62 55 58 55 54 53 55 60 Gluc 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 Chol mmol/L 2.08 2.35 NS 2.33 1.86 2.50 1.64 2.52 2.42 2.79 1.91 1.96 2.28 2.99 1.50 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) MIN 313/023622 : 160 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group Animal K Cl Ca Phos Total Prot Alb a1 / sex number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L 1F 21 4.1 105 2.75 1.49 71 37 11 22 3.4 105 2.90 1.88 71 37 10 23 NS NS NS NS NS NS NS 24 3.2 106 2.80 1.79 68 35 10 25 3.4 106 2.79 1.24 71 37 10 2F 26 3.4 106 27 3.5 107 28 3.1 108 29 3.0 107 30 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 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 NS No sample Animal 23 died at bleed a2 Beta g/L g/L 4 13 3 14 NS NS 4 14 3 14 4 14 3 15 3 14 5 13 4 13 4 14 3 13 4 13 4 14 3 14 MIN 313/023622 : 161 : APPENDIX 6 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Gamma g/L 2 3 4 ------POSF------ 30 100 300 A/G Alb a1 Ratio % % 1F 21 7 1.09 51.8 15.4 22 6 1.09 52.1 14.2 23 NS NS NS NS 24 5 1.06 51.8 14.8 25 7 1.09 52.2 13.8 2F 26 5 27 9 28 5 29 9 30 6 1.34 57.2 13.5 0.92 47.5 14.8 1.24 54.8 15.6 1.08 52.5 13.4 1.15 54.0 14.6 3F 31 8 32 5 33 6 34 6 35 4 1.06 50.9 13.9 1.10 53.0 14.7 1.21 54.8 14.0 1.12 52.5 15.5 1.16 53.3 15.9 NS No sample Animal 23 died at bleed a2 Beta Gamma % % % 5.6 18.0 9.2 4.8 20.0 8.9 NS NS NS 5.2 20.9 7.2 4.1 19.9 10.0 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 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 MIN 313/023622 : 162 : APPENDIX 6 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 ALP UL 4F 36 92 37 142 38 124 39 94 40 162 2 3 4 ------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) MIN 313/023622 : 163 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number CPK Total U/L 4F 36 137 37 96 38 158 39 197 40 129 2 3 4 ------POSF------ 30 100 300 SDH Bili Urea U/L mol/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 mol/L 51 61 55 74 58 Gluc mmol/L 5.20 5.25 6.67 5.91 6.46 Chol 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 : 164 : APPENDIX 6 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group Animal K Cl Ca Phos Total Prot Alb a1 / sex number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L 4F 36 3.2 104 37 3.2 104 38 3.4 106 39 3.8 104 40 3.4 105 2.83 1.41 82 45 12 2.77 1.67 73 40 11 2.76 1.81 75 42 11 2.78 1.61 74 42 12 2.80 1.55 78 40 12 a2 Beta g/L g/L 4 13 3 14 4 13 3 11 3 16 MIN 313/023622 : 165 : APPENDIX 6 (Biochemistry - continued) Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Gamma g/L 2 3 4 ------POSF------ 30 100 300 A/G Alb a1 Ratio % % 4F 36 9 37 6 38 5 39 5 40 6 1.22 54.5 14.6 1.21 54.7 15.0 1.27 55.8 14.4 1.31 56.7 16.3 1.05 51.8 15.9 a2 Beta Gamma % % % 4.7 15.6 10.7 3.9 18.7 7.7 5.8 16.9 7.1 4.4 15.5 7.0 4.0 20.8 7.5 MIN 313/023622 : 166 : Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 ALP U/L 1M 41 172 42 208 43 143 44 198 45 187 2M 46 178 47 240 48 268 49 231 50 254 3M 51 171 52 239 53 200 54 126 55 240 APPENDIX 6 (Biochemistry - continued) 2 3 4 ------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 LDH4 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 : 167 : Day 121 Group : Compound : Exposure level (ppm) : 1 Control 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 3 4 ------POSF------ 30 100 300 SDH Bili Urea U/L mol/L mmol/L 17.1 2 5.10 9.6 1 5.07 10.8 2 5.10 10.6 1 4.52 13.5 2 4.73 14.9 2 6.87 15.0 1 5.33 14.4 2 5.20 11.1 1 6.23 12.7 2 6.03 7.6 1 5.49 13.0 2 4.74 12.2 1 6.20 11.1 0 5.39 20.6 2 6.75 Creat mol/L 48 49 50 46 51 54 45 46 50 49 49 49 46 50 52 Gluc 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 Chol 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 : 168 : APPENDIX 6 (Biochemistry - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group Animal K Cl Ca Phos Total Prot Alb a1 / sex number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L 1M 41 42 43 44 45 2M 46 47 48 49 50 3M 51 52 53 54 55 4.1 102 3.6 101 3.5 102 4.2 103 3.8 102 3.8 104 3.9 103 4.1 102 3.8 103 3.8 102 4.1 102 3.5 103 3.5 102 4.1 104 3.9 103 2.62 1.80 66 32 11 2.58 1.74 66 32 13 2.61 1.90 65 29 12 2.64 1.84 63 32 11 2.54 1.66 68 32 11 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 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 a2 Beta g/L g/L 5 15 4 14 5 15 5 13 5 14 5 16 4 15 8 16 5 13 5 14 4 14 3 16 4 14 4 14 4 13 APPENDIX 6 (Biochemistry - continued) MIN 313/023622 : 169 : Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 Gamma g/L 2 3 4 ------POSF------ 30 100 300 A/G Alb a1 Ratio % % a2 Beta Gamma % % % 1M 41 3 42 4 43 4 44 2 45 6 2M 46 5 47 4 48 6 49 4 50 5 3M 51 5 52 8 53 5 54 4 55 4 0.94 48.1 16.3 7.7 22.7 5.2 0.94 47.8 19.4 5.9 21.4 5.5 0.81 44.6 18.4 8.0 22.7 6.3 1.03 50.2 17.5 7.6 21.0 3.7 0.89 47.5 16.1 7.2 20.8 8.4 0.81 44.0 16.5 7.3 24.8 7.3 0.97 48.8 14.3 6.7 23.3 6.9 0.67 39.4 14.8 12.3 24.5 9.1 0.88 46.9 17.3 8.4 21.5 6.0 0.88 46.5 14.4 8.5 21.9 8.7 0.88 46.9 16.5 0.76 43.6 14.9 0.87 46.2 15.8 0.80 45.2 19.7 0.90 48.2 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 : 170 : 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 APPENDIX 6 (Biochemistry - continued) 2 3 4 ------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 15 10 17 13 29 LDH2 U/L 13 11 15 14 20 LDH3 U/L 8 8 7 8 11 LDH4 U/L 19 14 15 15 32 LDH5 U/L 174 103 136 116 174 APPENDIX 6 (Biochemistry - continued) MIN 313/023622 : 171 : Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number CPK Total U/L 4M 56 163 57 309 58 214 59 304 60 475 2 3 4 ------POSF------ 30 100 300 SDH Bili Urea U/L mol/L mmol/L 31.6 1 4.66 26.1 1 5.18 8.4 2 5.25 12.6 1 6.30 16.4 1 5.52 Creat mol/L 48 51 44 50 46 Gluc mmol/L 5.33 6.04 5.60 5.19 5.13 Chol 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) MIN 313/023622 : 172 : Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group Animal K Cl Ca Phos Total Prot Alb a1 / sex number mmol/L mmol/L mmol/L mmol/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 a2 Beta g/L g/L 5 15 5 14 5 13 4 14 5 13 MIN 313/023622 : 173 : Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 Gamma g/L 4M 56 6 57 5 58 4 59 5 60 4 APPENDIX 6 (Biochemistry - continued) 2 3 4 ------POSF------ 30 100 300 A/G Alb a1 Ratio % % 0.78 44.4 17.1 0.94 48.0 16.2 0.85 46.5 18.7 0.92 47.8 19.4 0.94 48.7 17.2 a2 Beta Gamma % % % 7.0 22.4 9.0 6.9 21.6 7.3 7.3 21.3 6.2 5.2 20.9 6.8 7.6 20.6 5.9 MIN 313/023622 : 174 : Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 ALP U/L 1F 61 106 62 157 64 105 65 100 2F 66 112 67 65 68 129 69 105 70 65 3F 71 90 72 107 73 101 74 96 75 125 4F 76 81 77 104 78 123 79 113 80 141 APPENDIX 6 (Biochemistry - continued) 2 3 4 ------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 LDH2 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 APPENDIX 6 (Biochemistry - continued) MIN 313/023622 : 175 : Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number CPK Total U/L 1F 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 3 4 ------POSF------ 30 100 300 SDH Bili Urea U/L mol/L mmol/L 15.1 2 7.86 10.7 2 5.94 34.7 3 6.31 15.8 3 7.29 13.2 2 6.68 26.7 1 5.77 10.8 2 7.35 17.9 2 5.47 16.0 1 5.88 14.6 2 6.45 15.3 2 5.62 12.8 2 5.83 8.9 2 6.38 13.9 2 6.35 18.9 2 8.86 11.5 1 5.68 57.6 1 6.66 16.2 1 7.05 10.9 1 6.12 Creat mol/L 64 58 58 57 56 54 55 62 59 59 64 55 51 55 58 55 69 59 54 Gluc 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 Chol 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 : 176 : APPENDIX 6 (Biochemistry - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group Animal K Cl Ca Phos Total Prot Alb a1 / sex number mmol/L mmol/L mmol/L mmol/L g/L g/L g/L 1F 61 3.2 100 62 3.5 102 64 3.3 100 65 3.2 100 2F 66 3.6 103 67 3.3 101 68 2.9 101 69 3.7 104 70 3.5 102 3F 71 3.3 103 72 3.7 101 73 3.2 103 74 3.2 103 75 3.2 103 4F 76 3.5 104 77 3.4 104 78 3.5 102 79 3.9 107 80 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 a2 Beta g/L g/L 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 MIN 313/023622 : 177 : Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 Gamma g/L 1F 61 4 62 5 64 6 65 5 2F 66 4 67 7 68 4 69 5 70 6 3F 71 5 72 5 73 5 74 5 75 10 4F 76 5 77 3 78 5 79 5 80 7 APPENDIX 6 (Biochemistry - continued) 2 3 4 ------POSF------ 30 100 300 A/G Alb a1 Ratio % % 1.34 57.7 12.8 1.28 56.6 12.0 1.06 51.3 13.1 1.47 59.2 10.4 1.27 55.5 12.0 1.03 50.6 11.9 1.26 55.1 12.9 1.26 55.8 13.4 1.17 54.2 14.7 1.20 55.2 12.6 1.23 54.5 13.4 1.06 51.3 15.3 1.25 55.4 15.2 1.05 51.2 13.2 1.19 54.9 13.1 1.31 56.0 11.1 1.19 54.3 11.2 1.21 54.3 12.8 1.15 53.4 14.4 a2 Beta Gamma % % % 5.6 18.9 5.0 5.0 19.4 7.0 5.9 20.8 9.0 5.2 18.7 6.5 6.9 19.3 6.2 7.0 20.6 9.9 5.8 20.3 5.9 5.8 17.9 7.2 4.9 19.2 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 7.6 6.7 21.7 4.6 7.1 20.9 6.5 6.6 19.4 6.8 4.8 18.0 9.4 MIN 313/023622 : 178 : APPENDIX 7 Urinalysis - individual values Day 29 Group Compound Exposure level (ppm) Group / sex Animal number : 1 : Control : 0 App. 2 3 4 ------POSF------ 30 100 300 Vol pH SG ml g/L Prot Gluc Keto 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 1F 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 1.01 0 0 1.29 0 TR 1.41 0 TR 1.19 0 TR 0.78 0 0 1.05 0 TR 0.93 0 TR 0.97 0 TR 1.44 0 TR 1.20 0 TR 0.56 0 0 0.42 0 0 0.32 0 0 0.40 0 0 0.45 0 0 0.46 0 0 0.46 0 0 0.69 0 0 0.30 0 0 0.44 0 0 Bili Blood 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 MIN 313/023622 : 179 : APPENDIX 7 (Urinalysis - continued) Day 29 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group Animal Epi / sex number Leuc RBC 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 1F 91 0 1 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 APPENDIX 7 (Urinalysis - continued) Day 29 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex Animal number Cl Total mmol U-Na mmol/L U-K mmol/L U-Cl mmol/L MIN 313/023622 : 180 : 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 1F 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 MIN 313/023622 : 181 : APPENDIX 7 (Urinalysis - continued) Day 91 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 App. 1M 1 CPY 2 CMY 3 CMY 4 CMY 5 PY 2M 6 CPY 7 CMY 8 CPY 9 CPY 10 CMY 3M 11 CPY 12 CPY 13 CPY 14 CPY 15 CPY 2 3 4 ------POSF------ 30 100 300 Vol pH SG ml g/L 9.0 7.0 1031 4.2 7.7 1037 6.2 6.8 1043 4.0 7.4 1041 12.4 7.8 1022 6.8 7.2 1036 2.8 7.1 1062 7.8 7.5 1032 4.8 7.1 1042 4.2 7.7 1049 8.0 7.2 1031 9.0 7.3 1030 7.4 6.9 1039 6.2 6.5 1036 5.2 6.7 1044 Prot Gluc Keto g/L 0.53 0 0 0.98 0 TR 1.17 0 TR 1.40 0 TR 0.64 0 0 0.62 0 TR 1.12 0 TR 0.90 0 TR 1.47 0 TR 0.91 0 TR 0.82 0 TR 0.70 0 TR 0.91 0 TR 0.89 0 + 0.94 0 TR Bili Blood 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 MIN 313/023622 : 182 : APPENDIX 7 (Urinalysis - continued) Day 91 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 Epi 1M 1 0 2 0 3 0 4 0 5 0 2M 6 1 7 1 8 0 9 0 10 0 3M 11 0 12 0 13 0 14 0 15 0 2 3 4 ------POSF------ 30 100 300 Leuc RBC Cryst 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 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) MIN 313/023622 : 183 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Cl Total mmol 2 3 4 ------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 : 184 : APPENDIX 7 (Urinalysis - continued) Day 91 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 App. 4M 16 CMY 17 CMY 18 CPY 19 CPY 20 CMY 2 3 4 ------POSF------ 30 100 300 Vol pH SG ml g/L 2.8 6.8 1066 2.8 6.7 1068 5.2 6.7 1043 6.2 6.6 1043 3.0 6.4 1070 Prot Gluc Keto g/L 0.90 0 TR 1.60 0 TR 0.72 0 TR 1.11 0 TR 1.44 0 TR Bili Blood 0 0 0 0 0 0 0 0 0 0 MIN 313/023622 : 185 : APPENDIX 7 (Urinalysis - continued) Day 91 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 Epi 2 3 4 ------POSF------ 30 100 300 Leuc RBC Cryst Sperm 4M 16 0 0 0 0 0 17 0 0 0 0 0 18 0 0 0 0 1 19 0 0 0 0 1 20 0 0 0 0 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 : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Cl Total mmol 2 3 4 ------POSF------ 30 100 300 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 : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 App. 1F 21 PY 22 MY 23 PY 24 PY 25 PY 2F 26 PY 27 PY 28 PY 29 PY 30 CPY 3F 31 MY 32 PY 33 MY 34 MY 35 PY 2 3 4 ------POSF------ 30 100 300 Vol pH SG ml g/L 3.2 6.6 1033 2.6 6.4 1050 1.8 6.9 1029 8.0 6.6 1031 5.8 6.6 1037 4.0 6.6 1040 2.6 6.4 1044 6.0 6.8 1032 3.8 7.2 1029 4.8 6.6 1034 2.0 6.2 1049 2.6 6.5 1050 2.0 6.6 1064 3.0 6.4 1064 4.2 6.3 1042 Prot Gluc Keto g/L 0.14 0 0 0.28 0 0 0.16 0 0 0.14 0 0 0.22 0 0 0.23 0 0 0.29 0 0 0.18 0 0 0.15 0 0 0.20 0 0 0.26 0 0 0.31 0 0 0.25 0 0 0.23 0 0 0.20 0 0 Bili Blood 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 MIN 313/023622 : 188 : APPENDIX 7 (Urinalysis - continued) Day 91 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 Epi 1F 21 0 22 0 23 0 24 0 25 0 2F 26 0 27 0 28 0 29 0 30 0 3F 31 0 32 0 33 0 34 0 35 0 2 3 4 ------POSF------ 30 100 300 Leuc RBC Cryst 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 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 APPENDIX 7 (Urinalysis - continued) MIN 313/023622 : 189 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Cl Total mmol 2 3 4 ------POSF------ 30 100 300 U-Na mmol/L U-K mmol/L U-Cl mmol/L 1F 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 : 190 : APPENDIX 7 (Urinalysis - continued) Day 91 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 App. 4F 36 CMY 37 MY 38 MY 39 MY 40 MY 2 3 4 ------POSF------ 30 100 300 Vol pH SG ml g/L 3.2 6.4 1043 2.6 7.2 1045 1.8 6.5 1084 1.8 6.6 1078 2.0 6.2 1090 Prot Gluc Keto g/L 0.39 0 0 0.23 0 0 0.30 0 0 0.23 0 0 0.40 0 0 Bili Blood 0 0 0 0 0 0 0 0 0 0 MIN 313/023622 : 191 : APPENDIX 7 (Urinalysis - continued) Day 91 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 Epi 4F 36 0 37 0 38 0 39 0 40 0 2 3 4 ------POSF------ 30 100 300 Leuc RBC Cryst 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Sperm 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 APPENDIX 7 (Urinalysis - continued) MIN 313/023622 : 192 : Day 91 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Group / sex Animal number Cl Total mmol U-Na mmol/L U-K mmol/L U-Cl mmol/L 4F 36 0.33 88.2 170.5 102.1 37 0.17 59.6 254.5 63.5 38 0.21 57.8 406.5 115.9 39 0.24 137.0 376.5 134.7 40 0.29 121.8 342.0 144.9 MIN 313/023622 : 193 : APPENDIX 7 (Urinalysis - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 App. 1M 41 CPY 42 PY 43 CPY 44 CPY 45 CPY 2M 46 CPY 47 CPY 48 CMY 49 CPY 50 CPY 3M 51 CPY 52 CPY 53 MY 54 PY 55 MY 2 3 4 ------POSF------ 30 100 300 Vol pH SG ml g/L 3.2 6.8 1033 8.0 6.7 1036 12.0 7.6 1025 10.2 7.2 1026 2.0 7.2 1032 8.8 7.0 1033 5.4 6.9 1043 4.8 7.0 1058 5.8 7.0 1040 6.4 7.1 1037 9.2 6.9 1032 5.0 6.7 1044 2.6 6.5 1058 9.0 6.9 1036 3.2 6.6 1068 Prot Gluc Keto g/L 1.07 0 TR 0.94 0 TR 0.55 0 TR 0.77 0 + 1.03 0 + 2.18 0 + 1.08 0 TR 1.01 0 TR 1.22 0 TR 1.02 0 TR 0.82 0 TR 0.92 0 TR 1.41 0 TR 1.46 0 TR 1.12 0 TR Bili Blood 0 0 0 0 0 0 0 0 0 + 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 MIN 313/023622 : 194 : APPENDIX 7 (Urinalysis - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 Epi 1M 41 0 42 1 43 0 44 0 45 0 2M 46 0 47 0 48 1 49 0 50 0 3M 51 0 52 0 53 0 54 0 55 0 2 3 4 ------POSF------ 30 100 300 Leuc RBC Cryst 0 0 0 1 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 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 APPENDIX 7 (Urinalysis - continued) MIN 313/023622 : 195 : Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Cl Total mmol 2 3 4 ------POSF------ 30 100 300 U-Na mmol/L U-K mmol/L U-Cl 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 : 196 : APPENDIX 7 (Urinalysis - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 App. 4M 56 PY 57 PY 58 MY 59 PY 60 MY 2 3 4 ------POSF------ 30 100 300 Vol 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 Gluc Keto g/L 0.98 0 TR 0.76 0 TR 1.18 0 TR 1.10 0 TR 1.07 0 TR Bili Blood 0 0 0 0 0 0 0 0 0 0 MIN 313/023622 : 197 : APPENDIX 7 (Urinalysis - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 Epi 4M 56 0 57 0 58 0 59 0 60 0 2 3 4 ------POSF------ 30 100 300 Leuc RBC Cryst 1 0 0 0 0 0 0 0 0 0 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 APPENDIX 7 (Urinalysis - continued) MIN 313/023622 : 198 : Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Cl Total mmol 2 3 4 ------POSF------ 30 100 300 U-Na mmol/L U-K mmol/L U-Cl mmol/L 4M 56 57 58 59 60 0.38 59.5 210.5 66.0 0.55 57.3 192.0 64.9 0.31 68.6 285.0 76.9 0.31 89.0 163.5 50.1 0.25 70.5 306.0 79.3 MIN 313/023622 : 199 : APPENDIX 7 (Urinalysis - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 App. 2 3 4 ------POSF------ 30 100 300 Vol pH SG ml g/L Prot Gluc Keto g/L 1F 61 PY 5.4 6.2 1039 0.27 0 0 62 PY 1.6 6.0 1047 0.30 0 0 64 PY 6.6 6.2 1033 0.18 0 0 65 MY 2.4 5.9 1043 0.31 0 0 2F 66 MY 4.2 6.0 1039 0.17 0 0 67 PY 5.0 6.2 1037 0.28 0 0 68 PY 4.2 6.4 1041 0.20 0 0 69 PY 4.2 6.4 1040 0.19 0 0 70 PY 6.0 6.0 1037 0.23 0 0 3F 71 PY 2.4 6.0 1060 0.22 0 0 72 PY 3.2 6.0 1042 0.15 0 0 73 MY 1.2 6.2 1060 0.34 0 0 74 MY 2.6 6.1 1047 0.18 0 0 75 PY 4.2 6.3 1036 0.29 0 0 4F 76 MY 3.0 6.1 1045 0.19 0 0 77 MY 3.8 6.2 1047 0.28 0 0 78 MY 2.0 6.2 1060 0.26 0 0 79 PY 4.8 6.2 1033 0.23 0 0 80 PY 3.2 6.4 1047 0.37 0 0 Bili Blood 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 MIN 313/023622 : 200 : APPENDIX 7 (Urinalysis - continued) Day 121 Group : Compound : Exposure level (ppm) : Group / sex Animal number 1 Control 0 Epi 2 3 4 ------POSF------ 30 100 300 Leuc RBC Cryst Sperm 1F 61 0 0 0 0 0 62 0 0 0 0 0 64 0 0 0 0 0 65 0 1 0 0 0 2F 66 0 0 0 0 0 67 0 0 0 0 0 68 0 0 0 0 0 69 1 0 0 0 0 70 0 0 0 0 0 3F 71 0 0 0 0 0 72 0 0 0 0 0 73 0 0 0 0 0 74 0 0 0 0 0 75 0 0 0 0 0 4F 76 0 0 0 0 0 77 0 0 0 0 0 78 0 0 0 0 0 79 0 0 0 0 0 80 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 APPENDIX 7 (Urinalysis - continued) Day 121 Group : Compound : Exposure level (ppm) : 1 Control 0 Group / sex Animal number Cl Total mmol 2 3 4 ------POSF------ 30 100 300 U-Na mmol/L U-K mmol/L U-Cl mmol/L MIN 313/023622 : 201 : 1F 61 0.31 57.5 162.5 56.5 62 0.07 51.5 225.5 45.9 64 0.26 53.9 146.5 40.0 65 0.13 72.2 176.0 54.6 2F 66 0.11 43.9 140.5 27.3 67 0.52 72.7 218.0 103.5 68 0.35 95.0 189.5 83.8 69 0.24 62.3 206.5 56.3 70 0.38 65.6 151.5 63.3 3F 71 0.10 34.3 260.5 40.8 72 0.09 19.1 197.5 28.9 73 0.05 56.9 365.5 45.4 74 0.20 38.4 243.5 75.6 75 0.34 87.8 128.0 80.0 4F 76 0.20 62.7 211.0 68.1 77 0.21 75.7 196.5 55.1 78 0.12 80.1 316.0 60.8 79 0.23 49.9 162.0 47.1 80 0.18 92.3 207.0 55.4 APPENDIX 8 MIN 313/023622 : 202 : Absolute organ weights - individual values (g) Print No: 0026 GROUP : 1 2 3 4 Printed: 23-JUL-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR ------------------------------------------------------------------------------------------------------------------------------------ 1M 1 500.7 2.14 0.086 1.803 3.00 17.89 1.673 2 421.8 2.03 0.050 1.227 2.46 13.39 1.384 3 457.3 1.98 0.069 1.383 2.55 15.49 1.467 4 433.8 1.99 0.045 1.406 2.44 14.55 1.601 5 449.6 1.94 0.054 1.481 2.52 12.76 1.474 ------------------------------------------------------------------------------------------------------------------------------------ 2M 6 509.2 2.06 0.063 1.554 2.92 15.57 1.668 7 403.3 1.90 0.052 1.282 2.23 12.30 1.264 8 478.1 2.16 0.047 1.428 2.74 17.33 1.510 9 433.0 2.01 0.060 1.514 2.66 13.52 1.498 10 424.9 2.01 0.046 1.105 2.48 14.12 1.397 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 8 MIN 313/023622 : 203 : (Absolute organ weights - continued) Print No: 0026 GROUP : 1 2 3 4 Printed: 23-JUL-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR ------------------------------------------------------------------------------------------------------------------------------------ 3M 11 430.6 1.83 0.055 1.557 2.40 14.24 1.450 12 465.0 2.03 0.050 1.492 3.18 17.99 1.540 13 448.0 2.01 0.065 1.270 2.82 17.26 1.561 14 401.6 1.96 0.047 1.166 2.48 14.91 1.366 15 457.4 2.07 0.060 1.415 3.04 16.94 2.043 ------------------------------------------------------------------------------------------------------------------------------------ 4M 16 377.6 2.03 0.069 1.395 2.72 16.91 1.878 17 332.9 1.88 0.055 1.025 2.15 16.77 1.765 18 385.0 2.06 0.045 1.318 2.63 19.38 2.044 19 403.3 2.04 0.065 1.625 2.51 20.97 2.131 20 358.4 1.97 0.058 1.270 2.23 18.03 2.077 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 8 MIN 313/023622 : 204 : (Absolute organ weights - continued) Print No: 0026 GROUP : 1 2 3 4 Printed: 23-JUL-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR ------------------------------------------------------------------------------------------------------------------------------------ 1F 21 269.8 1.84 0.049 1.028 1.53 8.71 1.125 22 253.9 2.04 0.060 0.913 1.41 8.30 1.142 24 269.9 1.94 0.053 1.018 1.70 8.53 1.236 25 267.8 1.85 0.073 0.979 1.82 9.98 1.177 ------------------------------------------------------------------------------------------------------------------------------------ 2F 26 265.1 1.88 0.066 1.067 1.70 9.09 1.171 27 226.0 1.81 0.066 0.908 1.57 7.23 1.037 28 244.5 1.86 0.063 1.023 1.86 8.81 1.118 29 235.2 1.89 0.051 0.899 1.70 8.41 1.074 30 243.0 1.86 0.085 0.960 1.61 9.15 1.090 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 8 MIN 313/023622 : 205 : (Absolute organ weights - continued) Print No: 0026 GROUP : 1 2 3 4 Printed: 23-JUL-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR ------------------------------------------------------------------------------------------------------------------------------------ 3F 31 235.3 1.79 0.069 1.032 1.61 8.90 1.063 32 270.1 1.93 0.076 0.946 1.91 10.00 1.220 33 218.2 1.86 0.057 0.843 1.78 8.61 1.070 34 299.8 1.98 0.061 0.952 1.84 11.96 1.410 35 260.1 1.84 0.071 0.943 1.67 9.82 1.309 ------------------------------------------------------------------------------------------------------------------------------------ 4F 36 243.4 1.88 0.063 1.203 1.98 11.69 1.703 37 242.6 1.93 0.054 0.893 1.41 11.08 1.322 38 241.7 1.97 0.070 0.880 1.65 14.47 1.598 39 217.3 1.79 0.057 0.757 1.46 11.14 1.647 40 260.2 2.03 0.076 0.865 1.71 12.85 1.714 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 8 MIN 313/023622 : 206 : (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 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR ------------------------------------------------------------------------------------------------------------------------------------ 1M 41 466.6 2.17 0.049 1.320 2.76 13.02 1.398 42 506.1 2.09 0.075 1.417 2.75 11.85 1.599 43 496.4 1.97 0.053 1.413 2.78 12.36 1.519 44 493.2 2.13 0.057 1.718 2.91 12.24 1.389 45 450.3 2.21 0.051 1.484 2.80 11.22 1.455 ------------------------------------------------------------------------------------------------------------------------------------ 2M 46 498.8 2.23 0.045 1.405 2.98 14.24 1.549 47 409.2 1.99 0.047 1.685 2.75 11.50 1.422 48 485.8 2.13 0.050 1.481 3.34 13.44 1.487 49 423.7 2.17 0.054 1.254 2.73 10.70 1.406 50 470.8 2.14 0.065 1.417 3.06 12.50 1.470 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 8 MIN 313/023622 : 207 : (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 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR ------------------------------------------------------------------------------------------------------------------------------------ 3M 51 502.1 2.09 0.049 1.568 2.83 13.72 1.555 52 465.1 2.09 0.050 1.596 2.99 14.91 1.512 53 497.0 2.11 0.053 1.378 3.07 14.63 1.457 54 487.9 2.18 0.058 1.677 3.33 15.37 1.499 55 396.1 1.96 0.048 1.348 2.24 11.05 1.201 ------------------------------------------------------------------------------------------------------------------------------------ 4M 56 451.5 1.94 0.058 1.735 2.92 16.08 1.514 57 491.2 2.11 0.054 1.771 3.26 16.25 1.831 58 414.0 1.97 0.054 1.555 2.76 13.44 1.457 59 407.7 2.00 0.042 1.218 2.55 16.07 1.637 60 449.9 2.22 0.044 1.449 2.79 15.79 1.642 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 8 MIN 313/023622 : 208 : (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 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR ------------------------------------------------------------------------------------------------------------------------------------ 1F 61 313.0 2.05 0.078 1.196 1.86 9.45 1.365 62 285.2 1.94 0.078 1.083 1.92 9.62 1.276 64 270.2 1.94 0.073 1.164 1.75 7.58 1.206 65 297.2 1.95 0.064 1.125 1.97 9.29 1.314 ------------------------------------------------------------------------------------------------------------------------------------ 2F 66 304.1 2.17 0.065 0.864 1.83 8.35 1.242 67 316.9 2.01 0.078 1.223 2.27 9.64 1.279 68 258.3 1.95 0.055 0.908 1.77 7.93 1.128 69 286.4 2.12 0.072 1.136 1.78 8.97 UNSUITABLE 70 291.7 2.01 0.083 1.020 1.84 9.12 1.150 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 8 MIN 313/023622 : 209 : (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 Xybion protocol number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL GROUP ANIMAL BODY WT (g) BRAIN ADRENALS HEART KIDNEYS LIVER LUNGS & BR ------------------------------------------------------------------------------------------------------------------------------------ 3F 71 260.4 1.89 0.065 0.889 1.66 8.15 1.187 72 251.6 1.88 0.075 0.940 1.73 8.59 1.132 73 276.7 2.00 0.080 1.137 1.96 8.95 1.079 74 251.5 1.85 0.062 1.023 1.73 7.81 1.046 75 249.9 2.00 0.060 0.944 1.76 8.71 1.003 ------------------------------------------------------------------------------------------------------------------------------------ 4F 76 232.2 1.90 0.055 0.849 1.43 8.63 1.082 77 250.8 1.87 0.057 0.988 1.75 9.02 1.142 78 259.0 2.03 0.063 0.918 1.53 9.49 1.259 79 297.6 1.94 0.071 0.950 1.74 11.37 1.242 80 265.0 1.94 0.071 0.918 1.88 10.40 1.212 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 8 MIN 313/023622 : 210 : (Absolute organ weights - continued) Print No: 0013 GROUP : 1 2 3 4 Printed: 23-JUL-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL GROUP ANIMAL BODY WT (g) ADRENALS BRAIN HEART KIDNEYS LIVER LUNGS & BR ------------------------------------------------------------------------------------------------------------------------------------ 1M 81 353.0 0.076 1.88 1.510 2.42 10.74 1.424 82 334.2 0.045 2.10 1.214 2.41 9.47 1.126 83 280.2 0.037 1.79 0.933 1.91 7.83 1.005 84 350.5 0.054 2.01 1.408 2.47 10.31 1.379 85 346.2 0.051 1.91 1.353 2.41 9.78 1.319 ------------------------------------------------------------------------------------------------------------------------------------ 4M 86 333.4 0.063 1.98 1.481 2.37 12.42 1.852 87 325.3 0.059 1.89 1.391 2.28 13.17 1.596 88 281.2 0.040 2.02 1.141 2.03 11.18 1.699 89 335.2 0.061 2.03 1.361 2.53 13.72 1.620 90 314.4 0.057 1.92 1.144 2.43 13.05 1.757 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 8 MIN 313/023622 : 211 : (Absolute organ weights - continued) Print No: 0013 GROUP : 1 2 3 4 Printed: 23-JUL-02 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Xybion protocol number: MIN 313A ------------------------------------------------------------------------------------------------------------------------------------ TERMINAL GROUP ANIMAL BODY WT (g) ADRENALS BRAIN HEART KIDNEYS LIVER LUNGS & BR ------------------------------------------------------------------------------------------------------------------------------------ 1F 91 206.8 0.061 1.84 0.795 1.62 6.72 1.021 92 204.5 0.053 1.80 0.923 1.41 6.76 1.088 93 234.1 0.084 1.78 1.061 1.49 7.19 0.979 94 194.3 0.061 1.81 0.850 1.52 7.10 0.948 95 224.5 0.072 1.98 0.914 1.68 8.24 1.161 ------------------------------------------------------------------------------------------------------------------------------------ 4F 96 191.9 0.064 1.69 0.835 1.42 7.96 1.310 97 193.2 0.079 1.82 0.828 1.55 8.33 1.467 98 193.2 0.070 1.80 0.842 1.39 7.59 1.286 99 188.0 0.056 1.78 0.781 1.46 7.90 1.201 100 206.9 0.078 1.87 0.849 1.56 7.87 1.407 ------------------------------------------------------------------------------------------------------------------------------------ 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 MIN 313/023622 : 213 : Individual pathological findings Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0001 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: 500.7 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : LIVER : -MEDIAN CLEFT PALE AREA(S); ONE, SUBCAPSULAR, 1MM. >NO SIGNIFICANT LESION LUNGS & BRONCHI : -ALVEOLITIS,-MINIMAL -ALVEOLAR HAEMORRHAGE,-MINIMAL STOMACH : -ANTRUM WHITE NODULE(S); RIDGE, 1MM. MUCOSA, ONE, NEAR TO LIMITING ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 214 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0002 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: 421.8 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- HEART : -MYOCARDITIS, VENTRICULAR REGION,-MINIMAL, MULTI-FOCAL LN MANDIBULAR : -ENLARGED; 10MM. -CONGESTED, MINIMAL LUNGS & BRONCHI : -ALVEOLAR HAEMORRHAGE,-MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 215 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0003 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: 457.3 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- HEART : -MYOCARDITIS, VENTRICULAR REGION,-MINIMAL, FOCAL KIDNEYS : -MINERALISATION, CORTICOMEDULLARY,-MINIMAL LARYNX : -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL LN MANDIBULAR : -CONGESTED LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 216 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0004 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: 433.8 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY 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 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 217 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 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 ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- HEART : -MYOCARDITIS, VENTRICULAR REGION,-MINIMAL, FOCAL KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL LIVER : -PERIPORTAL LYMPHOCYTIC INFILTRATION,-MINIMAL LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-MINIMAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 218 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0041 SEX: MALE DOSE GROUP: 1 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: 466.6 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL -MEDULLARY MINERALISATION,-MINIMAL LUNGS & BRONCHI : -PERIVASCULAR INFLAMMATORY CELL INFILTRATION,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; LEFT SIDE OF FACE, DIFFUSE. ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 219 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0042 SEX: MALE DOSE GROUP: 1 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: 506.1 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-SLIGHT, FOCAL LUNGS & BRONCHI : -PERIVASCULAR INFLAMMATORY CELL INFILTRATION,-MINIMAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 220 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0043 SEX: MALE DOSE GROUP: 1 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: 496.4 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, MULTI-FOCAL -CORTICAL LYMPHOCYTIC INFILTRATION,-SLIGHT, MULTI-FOCAL LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL ALVEOLAR MACROPHAGES,-MINIMAL -ALVEOLITIS,-MINIMAL -FOCAL OSSEOUS DEPOSITION,-MINIMAL THICKENING/ SCATTERED SKIN : -HAIRLOSS, MINIMAL; RIGHT SIDE OF FACE, DIFFUSE. ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 221 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0044 SEX: MALE DOSE GROUP: 1 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 ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL -CORTICAL LYMPHOCYTIC INFILTRATION,-MINIMAL, MULTI-FOCAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 222 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0045 SEX: MALE DOSE GROUP: 1 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: 450.3 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -PELVIC DILATATION,-SLIGHT - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 223 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0006 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 509.2 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : LIVER : -MEDIAN CLEFT PALE AREA(S); ONE, SUBCAPSULAR, 1MM. -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-SLIGHT -PARENCHYMAL INFLAMMATORY CELL INFILTRATION,-MINIMAL, FOCAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 224 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0007 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 403.3 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT LN MANDIBULAR : -CONGESTED, MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 225 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0008 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 478.1 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT LUNGS & BRONCHI : -ALVEOLITIS,-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 : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0009 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 433.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT LN MANDIBULAR : -ENLARGED; UP TO 13MM. LUNGS & BRONCHI : -ALVEOLAR HAEMORRHAGE,-MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 227 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0010 SEX: MALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 424.9 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY 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. ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 228 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0046 SEX: MALE DOSE GROUP: 2 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: 498.8 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- SKIN : -HAIRLOSS, MINIMAL; SIDES OF FACE, DIFFUSE. - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 229 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0047 SEX: MALE DOSE GROUP: 2 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: 409.2 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE VACUOLATION,-SLIGHT LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL STOMACH : -ANTRUM WHITE NODULE(S); RIDGE, 1MM. MUCOSA, ONE, NEAR TO LIMITING ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 230 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0048 SEX: MALE DOSE GROUP: 2 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: 485.8 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-SLIGHT SKIN : -SCAB(S); RIGHT DORSAL LUMBAR REGION, ONE, 3X1MM. STOMACH : -ANTRUM WHITE NODULE(S); RIDGE, 1MM. MUCOSA, ONE, NEAR TO LIMITING ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 231 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0049 SEX: MALE DOSE GROUP: 2 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: 423.7 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY 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 : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0050 SEX: MALE DOSE GROUP: 2 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: 470.8 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY 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 MIN 313/023622 : 233 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0011 SEX: MALE DOSE GROUP: 3 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 ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-MODERATE -CENTRILOBULAR HEPATOCYTE VACUOLATION,-MINIMAL MISCELLANEOUS : -ANIMAL THIN, MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 234 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0012 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 465.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-MODERATE -CENTRILOBULAR HEPATOCYTE VACUOLATION,-MINIMAL LN MANDIBULAR : -CONGESTED, MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 235 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0013 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 448.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-MODERATE -CENTRILOBULAR HEPATOCYTE VACUOLATION,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; REGION, DIFFUSE. SIDES OF FACE AND RIGHT DORSAL THORACIC ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 236 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0014 SEX: MALE DOSE GROUP: 3 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 ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : LIVER : -MEDIAN CLEFT PALE AREA(S); ONE, SUBCAPSULAR, 1MM. -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-MODERATE -CENTRILOBULAR HEPATOCYTE VACUOLATION,-SLIGHT LUNGS & BRONCHI : -PROMINENT AGGREGATIONS OF LYMPHOCYTES,-MINIMAL -VASCULAR CONGESTION,-SLIGHT ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 237 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0015 SEX: MALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 457.4 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -ARYTENOID - SUBEPITHELIAL INFLAMMATORY CELL INFILTRATION,- MINIMAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-MODERATE -EXTRAMEDULLARY HAEMOPOEISIS,-MINIMAL LUNGS & BRONCHI : -CONGESTED, MINIMAL LUNGS & BRONCHI : -ALVEOLAR HAEMORRHAGE,-MARKED ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 238 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0051 SEX: MALE DOSE GROUP: 3 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: 502.1 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-MINIMAL LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-MINIMAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 239 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0052 SEX: MALE DOSE GROUP: 3 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: 465.1 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-MINIMAL -EXTRAMEDULLARY HAEMOPOEISIS,-MINIMAL LN MANDIBULAR : -ENLARGED; RIGHT, ONE, 12MM. LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL -ALVEOLITIS,-MINIMAL STOMACH : -ANTRUM WHITE NODULE(S); RIDGE, PUNCTATE. MUCOSA, ONE, NEAR TO LIMITING ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 240 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0053 SEX: MALE DOSE GROUP: 3 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: 497.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT -PERIPORTAL LYMPHOCYTIC INFILTRATION,-MINIMAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 241 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0054 SEX: MALE DOSE GROUP: 3 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: 487.9 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-SLIGHT - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 242 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0055 SEX: MALE DOSE GROUP: 3 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: 396.1 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- EPIDIDYMIDES : -SWELLING(S); RIGHT CAUDA, ONE, 3MM. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 243 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0016 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 377.6 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-MODERATE LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MODERATE TRACHEA : -EPITHELIAL HYPERPLASIA,-MINIMAL, FOCAL -EPITHELIAL INFLAMMATORY CELL INFILTRATION,-MINIMAL, FOCAL -SUBEPITHELIAL INFLAMMATORY AGGREGATIONS,-MINIMAL, FOCAL -SUBEPITHELIAL FIBROBLAST PROLIFERATION,-MINIMAL, FOCAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 244 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0017 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 332.9 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-MODERATE LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-SLIGHT -ALVEOLITIS,-MINIMAL MISCELLANEOUS : -ANIMAL THIN STOMACH : -CORPUS WHITE NODULE(S); 1MM. MUCOSA, ONE NEAR TO LIMITING RIDGE, ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 245 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0018 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 385.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -FOCAL CAPSULAR CONGESTION,-MINIMAL LIVER : -ENLARGED; 19.38G. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-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) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0019 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 403.3 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LIVER : -MEDIAN CLEFT PALE AREA(S); -ENLARGED; 20.97G. ONE, SUBCAPSULAR, 2MM. 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 MIN 313/023622 : 247 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0020 SEX: MALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 358.4 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL -ARYTENOID - SUBEPITHELIAL INFLAMMATORY CELL INFILTRATION,- MINIMAL LIVER : -ENLARGED; 18.03G. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-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); RIDGE, 1MM. MUCOSA, ONE, NEAR TO LIMITING TRACHEA : -EPITHELIAL HYPERPLASIA,-MINIMAL, FOCAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 248 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0056 SEX: MALE DOSE GROUP: 4 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: 451.5 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL 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 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 249 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0057 SEX: MALE DOSE GROUP: 4 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: 491.2 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS 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. ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 250 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0058 SEX: MALE DOSE GROUP: 4 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: 414.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-SLIGHT, MULTI-FOCAL -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; REGION. RIGHT SCAPULAR REGION AND CRANIAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 251 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0059 SEX: MALE DOSE GROUP: 4 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: 407.7 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS 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 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 252 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0060 SEX: MALE DOSE GROUP: 4 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: 449.9 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -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 MIN 313/023622 : 253 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0021 SEX: FEMALE 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: 269.8 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL LYMPHOCYTIC INFILTRATION,-MINIMAL, FOCAL LARYNX : -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL LIVER : -PERIPORTAL LYMPHOCYTIC INFILTRATION,-SLIGHT - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 254 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0022 SEX: FEMALE 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: 253.9 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- MISCELLANEOUS : -FUR STAINING; DORSUM, BROWN. - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 255 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0023 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: UNSCHEDULED (F) DATE OF DEATH: 20-MAR-02 STUDY DAY OF DEATH: 91 STUDY WEEK OF DEATH: 13 TERMINAL BODY WEIGHT: 242.4 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- 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 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 256 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0024 SEX: FEMALE 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: 269.9 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-SLIGHT, FOCAL -CORTICAL INTERSTITIAL FIBROBLAST PROLIFERATION/ FIBROSIS,- SLIGHT, FOCAL LIVER : -EXTRAMEDULLARY HAEMOPOEISIS,-MINIMAL LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-SLIGHT -ALVEOLAR HAEMORRHAGE,-MINIMAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 257 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0025 SEX: FEMALE 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: 267.8 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL SKIN : -HAIRLOSS, MINIMAL; DIFFUSE. VENTRAL SURFACE AND RIGHT HINDLIMB, ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 258 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0061 SEX: FEMALE DOSE GROUP: 1 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: 313.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-MINIMAL -PERIVASCULAR INFLAMMATORY CELL INFILTRATION,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; REGION, DIFFUSE. RIGHT SCAPULAR AND RIGHT DORSAL THORACIC ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 259 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0062 SEX: FEMALE DOSE GROUP: 1 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: 285.2 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -MINERALISATION, CORTICOMEDULLARY,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; DORSAL THORACIC REGION, DIFFUSE. ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 260 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0063 SEX: FEMALE DOSE GROUP: 1 SACRIFICE STATUS: UNSCHEDULED (H) DATE OF DEATH: 03-JAN-02 STUDY DAY OF DEATH: 15 STUDY WEEK OF DEATH: 3 TERMINAL BODY WEIGHT: 213.3 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -CORTICAL TUBULAR BASOPHILIA,-MINIMAL, FOCAL -CORTICAL INTERSTITIAL FIBROBLAST PROLIFERATION/ FIBROSIS,- MINIMAL, FOCAL -MEDULLARY MINERALISATION,-MINIMAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 261 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0064 SEX: FEMALE DOSE GROUP: 1 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: 270.2 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -PERIPORTAL LYMPHOCYTIC INFILTRATION,-MINIMAL LN MANDIBULAR : -ENLARGED; 9MM. LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-MINIMAL -ALVEOLAR HAEMORRHAGE,-SLIGHT ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 262 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0065 SEX: FEMALE DOSE GROUP: 1 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: 297.2 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -MINERALISATION, CORTICOMEDULLARY,-MINIMAL -MEDULLARY MINERALISATION,-MINIMAL LIVER : -EXTRAMEDULLARY HAEMOPOEISIS,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; FORELIMBS, DIFFUSE. ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 263 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0026 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 265.1 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -PELVIC DILATION, MINIMAL; RIGHT. LUNGS & BRONCHI : -FOCAL OSSEOUS DEPOSITION,-MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 264 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0027 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 226.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- SKIN : -HAIRLOSS, MINIMAL; DORSAL THORACIC REGION, DIFFUSE. UTERUS : -FLUID DISTENSION, MODERATE - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 265 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0028 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 244.5 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- UTERUS : -FLUID DISTENSION, MODERATE - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 266 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0029 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 235.2 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LN MANDIBULAR : -ENLARGED; ONE, 12MM. LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-MINIMAL -PERIVASCULAR INFLAMMATORY CELL INFILTRATION,-MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 267 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0030 SEX: FEMALE DOSE GROUP: 2 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 243.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -VENTRAL POUCH - EPITHELIAL HYPERPLASIA,-MINIMAL LIVER : -PERIPORTAL LYMPHOCYTIC INFILTRATION,-MINIMAL STOMACH : -ANTRUM WHITE NODULE(S); RIDGE, 1MM. MUCOSA, ONE, NEAR TO LIMITING THYMUS : -DARK UTERUS : -FLUID DISTENSION, MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 268 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0066 SEX: FEMALE DOSE GROUP: 2 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: 304.1 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 269 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0067 SEX: FEMALE DOSE GROUP: 2 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: 316.9 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : -PELVIC DILATION, MODERATE; RIGHT. LARYNX : -VENTRAL POUCH - FORIGN BODY IN LUMEN, RELATED TO EPITHELIAL HYPERPLASIA/ INFLAMMATORY CELL INFILTRATION,-SLIGHT URETERS : -DISTENDED; RIGHT, 1MM. ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 270 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0068 SEX: FEMALE DOSE GROUP: 2 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: 258.3 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; RIGHT, DIFFUSE. UTERUS : -FLUID DISTENSION, MODERATE ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 271 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0069 SEX: FEMALE DOSE GROUP: 2 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: 286.4 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- SKIN : -HAIRLOSS, MINIMAL; RIGHT SIDE IF FACE, DIFFUSE. UTERUS : -FLUID DISTENSION, MODERATE - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 272 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0070 SEX: FEMALE DOSE GROUP: 2 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: 291.7 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LN MANDIBULAR : -ENLARGED; 9MM. - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 273 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0031 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 235.3 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LN MANDIBULAR : -ENLARGED; UP TO 10MM. - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 274 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0032 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 270.1 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -PERIPORTAL HEPATOCYTE VACUOLATION,-MINIMAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 275 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0033 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 218.2 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS 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 ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 276 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0034 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 299.8 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- STOMACH : -ANTRUM WHITE NODULE(S); MUCOSA, TWO, NEAR TO LIMITING RIDGE, PUNCTATE. - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 277 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0035 SEX: FEMALE DOSE GROUP: 3 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 260.1 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -VENTRAL POUCH - EPITHELIAL HYPERPLASIA,-MINIMAL LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-MINIMAL MISCELLANEOUS : -ANIMAL THIN, MINIMAL ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 278 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0071 SEX: FEMALE DOSE GROUP: 3 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: 260.4 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -VENTRAL POUCH - FORIGN BODY IN LUMEN, RELATED TO EPITHELIAL HYPERPLASIA/ INFLAMMATORY CELL INFILTRATION,-MODERATE LIVER : -MEDIAN CLEFT PALE AREA(S); ONE, SUBCAPSULAR. LIVER : >NO SIGNIFICANT LESION STOMACH : -ANTRUM WHITE NODULE(S); PUNCTATE. MUCOSA, TWO, NEAR TO LIMTING RIDGE, ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 279 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0072 SEX: FEMALE DOSE GROUP: 3 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: 251.6 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -PARENCHYMAL INFLAMMATORY CELL INFILTRATION,-MINIMAL, FOCAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 280 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0073 SEX: FEMALE DOSE GROUP: 3 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: 276.7 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- STOMACH : -ANTRUM WHITE NODULE(S); MUCOSA, THREE, NEAR TO LIMITING RIDGE, 1MM. UTERUS : -FLUID DISTENSION, MINIMAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 281 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0074 SEX: FEMALE DOSE GROUP: 3 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: 251.5 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LUNGS & BRONCHI : -ALVEOLAR HAEMORRHAGE,-MINIMAL UTERUS : -FLUID DISTENSION, MODERATE ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 282 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0075 SEX: FEMALE DOSE GROUP: 3 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: 249.9 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- SKIN : -HAIRLOSS, MINIMAL; LEFT DORSUM, DIFFUSE. - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO MICROSCOPIC FINDINGS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 283 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0036 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 243.4 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- KIDNEYS : KIDNEYS : -PELVIC DILATION, MINIMAL; RIGHT. -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; 9MM. -CONGESTED; RIGHT, ONE. LUNGS & BRONCHI : -INCOMPLETE COLLAPSE, MINIMAL LUNGS & BRONCHI : -FOCAL OSSEOUS DEPOSITION,-MINIMAL SKIN : -HAIRLOSS, MINIMAL; RIGHT FORELIMB, DIFFUSE. ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 284 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0037 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 242.6 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY 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) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0038 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 241.7 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT -ARYTENOID - EPITHELIAL HYPERPLASIA,-MINIMAL, FOCAL LIVER : -ENLARGED; 14.47G. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-MODERATE -PROMINENT CENTRILOBULAR HEPATOCYTE PIGMENT,-MINIMAL LUNGS & BRONCHI : -INCOMPLETE COLLAPSE, MINIMAL LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL ALVEOLAR MACROPHAGES,-SLIGHT -ALVEOLAR HAEMORRHAGE,-MINIMAL -FOCAL OSSEOUS DEPOSITION,-MINIMAL THICKENING/ SCATTERED MISCELLANEOUS : -ANIMAL THIN SKIN : -HAIRLOSS, MINIMAL; DIFFUSE. HINDLIMBS AND LEFT CERVICAL REGION, ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 286 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0039 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 217.3 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LIVER : -MEDIAN CLEFT PALE AREA(S); ONE, SUBCAPSULAR, 3MM. LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY EXTENDING INTO MIDZONAL AREA,-MODERATE -HEPATOCYTE VACUOLATION, MEDIAN CLEFT,-MINIMAL LUNGS & BRONCHI : -FOAMY ALVEOLAR MACROPHAGES/SEPTAL THICKENING/ SCATTERED ALVEOLAR MACROPHAGES,-SLIGHT MISCELLANEOUS : -ANIMAL THIN SKIN : -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) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0040 SEX: FEMALE DOSE GROUP: 4 SACRIFICE STATUS: SCHEDULED, TERMINAL SACRIFICE DATE OF DEATH: 21-MAR-02 STUDY DAY OF DEATH: 92 STUDY WEEK OF DEATH: 14 TERMINAL BODY WEIGHT: 260.2 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- 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 MIN 313/023622 : 288 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0076 SEX: FEMALE DOSE GROUP: 4 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: 232.2 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY 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) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0077 SEX: FEMALE DOSE GROUP: 4 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: 250.8 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY 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) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0078 SEX: FEMALE DOSE GROUP: 4 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: 259.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LARYNX : -NECROSIS OF VENTRAL CARTILAGE,-PRESENT LUNGS & BRONCHI : -AGGREGATIONS OF ALVEOLAR MACROPHAGES,-MINIMAL -FOCAL OSSEOUS DEPOSITION,-MINIMAL UTERUS : -FLUID DISTENSION, MODERATE ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 291 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0079 SEX: FEMALE DOSE GROUP: 4 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: 297.6 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY 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 *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 9 MIN 313/023622 : 292 : (Individual pathological findings - continued) Print No: 0034 GROUP : 1 2 3 4 Printed: 09-SEP-03 COMPOUND : CONTROL --------POSF-------- EXPOSURE LEVEL(PPM): 0 30 100 300 Protocol Number: MIN 313 ------------------------------------------------------------------------------------------------------------------------------------ ANIMAL NUMBER: 0080 SEX: FEMALE DOSE GROUP: 4 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: 265.0 GRAMS ------------------------------------------------------------------------------------------------------------------------------------ PATHOLOGY OBSERVATIONS NECROPSY HISTOPATHOLOGY ---------------------------------------------------------------- ---------------------------------------------------------------- LIVER : -CENTRILOBULAR HEPATOCYTE HYPERTROPHY,-MINIMAL LUNGS & BRONCHI : -PERIVASCULAR INFLAMMATORY CELL INFILTRATION,-MINIMAL -FOCAL OSSEOUS DEPOSITION,-MINIMAL - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ------------------------------------------------------------------------------------------------------------------------------------ *** ANIMAL HAS NO GROSS OBSERVATIONS RECORDED *** ------------------------------------------------------------------------------------------------------------------------------------ APPENDIX 10 pH values for terminal urine - individual values MIN 313/023622 Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------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 7.2a 15 8.4 35 7.3 4 16 8.1 36 8.1 17 - 37 - 18 - 38 8.4 19 - 39 8.0 20 8.1 40 8.2 a Sample taken from bladder at necropsy - Sample volume < 8 l 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 2 3 4 ------POSF------ 30 100 300 Recovery study animals Group Males Animal pH number 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 l 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 2 3 4 ------POSF------ 30 100 300 Satellite animals Group Males Animal pH number Females Animal pH number 1 81a 7.1 91 - 82 - 92 7.6 83a 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 l required to obtain accurate pH reading a Sample pH'd approximately 1.5 hours following sample collection due to failure of pH hardware : 295 : APPENDIX 11 Cell proliferation findings MIN 313/023622 : 296 : Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Satellite animals Group Animal Level 1 No. Positive Negative Total 1M 81 60 940 1000 82 34 966 1000 83 31 969 1000 84 15 985 1000 85 46 954 1000 4M 86 70 930 1000 87 46 954 1000 88 27 973 1000 89 31 969 1000 90 25 975 1000 1F 91 49 951 1000 92 41 959 1000 93 51 949 1000 94 35 965 1000 95 64 936 1000 4F 96 63 937 1000 97 35 965 1000 98 42 958 1000 99 40 960 1000 100 41 959 1000 * Cell proliferation index (%positive cells) C.P.I.* 6.0 3.4 3.1 1.5 4.6 7.0 4.6 2.7 3.1 2.5 4.9 4.1 5.1 3.5 6.4 6.3 3.5 4.2 4.0 4.1 Positive 57 40 26 24 47 64 60 36 34 31 54 60 46 37 59 55 50 51 45 55 Level Negative 943 960 974 976 953 936 940 964 966 969 946 940 954 963 941 945 950 949 955 945 2 Total 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 C.P.I.* 5.7 4.0 2.6 2.4 4.7 6.4 6.0 3.6 3.4 3.1 5.4 6.0 4.6 3.7 5.9 5.5 5.0 5.1 4.5 5.5 Positive 59 33 34 23 73 81 49 38 27 39 46 32 53 36 56 61 46 53 38 45 Level Negative 941 967 966 977 927 919 951 962 973 961 954 968 947 964 944 939 954 947 962 955 3 Total 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 1000 Group Mean cell proliferation index (%positive cells) Male Female 1 4.0 4.8 4 4.4 4.8 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 MIN 313/023622 : 297 : APPENDIX 11 (Cell proliferation findings - continued) Group : Compound : Exposure level (ppm) : 1 Control 0 2 3 4 ------POSF------ 30 100 300 Main group animals Group Animal Level 1 No. Positive Negative Total 1M 1 NR NR NR 2 NR NR NR 3 NR NR NR 4 NR NR NR 5 NR NR NR 4M 16 NR NR NR 17 NR NR NR 18 NR NR NR 19 NR NR NR 20 NR NR NR 1F 21 NR NR NR 22 NR NR NR 23 NR NR NR 24 NR NR NR 25 NR NR NR 4F 36 NR NR NR 37 NR NR NR 38 NR NR NR 39 NR NR NR 40 NR NR NR * Cell proliferation index (%positive cells) 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 Negative NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 2 Total 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 Negative NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR 3 Total NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR NR Group Mean cell proliferation index (%positive cells) Male Female 1 NR NR 4 NR NR NR Not recorded 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 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 the sampling 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 l/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-IR) 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 l/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 l/minute was used as extract for each Group. For the dose groups this comprised 29 l/minute from the base of the chamber and 1 l/minute drawn for analytical sampling. A 1 l/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 l/minute and a calibrated flow of 30 l/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 l/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 t99 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 l/minute was used for the nitrogen and ethylene in nitrogen cylinders, the analyser drew 1 l/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 of 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 l/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 the 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 of 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 100 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 are presented in Table D. The chamber temperatures were similar for all groups on each day of the study. DISCUSSION Control of the POSF vapour delivery 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 l Filtration m Air extract : 308 : FIGURE B MIN 313/023622 Schematic of a Fourier Transform Infrared spectrophotometer and the sampling system Key a Nitrogen cylinder b Ethylene in nitrogen cylinder c Flowmeter regulated to 1 l/minute d Test compound sample line from chamber e 10 cm IR cell f FT-IR spectrophotometer g Flowmeter regulated to 2 l/minute h Diaphragm pump i Sample line return to waste j Toggle Valves k Time-controlled chamber switching system : 309 : TABLE A Operating conditions for the inhalation exposure system MIN 313/023622 Parameter Target concentration of POSF (ppm) Chamber airflows (l/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 2 (Low dose) 3 (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 Analyser Concentration (ppm) Actual Concentration (ppm) no. Gp2 Gp3 Gp4 Gp2 Gp3 Gp4 1 35 114 327 22 87 261 2 33 117 350 21 89 279 3 36 132 302 23 101 240 4 1 5 2,3 6 1 7 42 147 393 28 114 315 8 22 127 363 12 97 290 9 1 10 21 130 359 11 100 287 11 41 147 380 27 114 304 12 41 132 370 27 101 296 13 43 132 369 29 102 295 14 42 124 365 28 95 292 15 35 140 378 22 108 303 16 46 146 390 31 112 313 17 45 123 376 30 94 301 18 42 125 371 28 96 297 19 43 125 381 29 95 305 20 44 126 376 29 96 301 21 4 40 117 354 26 89 283 22 40 120 385 26 91 308 23 42 129 386 28 99 310 24 58 143 393 41 111 315 25 38 123 364 25 94 291 26 41 135 365 27 104 292 27 41 136 363 27 105 290 28 42 133 373 28 102 299 29 42 134 378 27 103 303 30 44 137 371 29 105 297 31 41 140 382 27 108 306 32 39 135 368 25 104 295 33 40 139 371 26 107 297 34 1 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 1 No data collected due to computer problem with data capture programme 2 Problem with ethylene in nitrogen calibration cylinder, no data analytical data collected 3 Ethylene in nitrogen calibration cylinder ran out 4 New Ethylene in nitrogen calibration cylinder : 311 : TABLE B MIN 313/023622 (Chamber concentration of POSF (ppm) - daily mean values - continued) Exposure no. 41 42 43 44 45 46 47 48 3 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 Overall Mean sd CV (%) Mean (1-65) sd 3 CV (%) sd CV (%) Analyser Gp2 37 38 41 36 40 40 40 39 37 37 40 42 40 40 39 39 42 41 36 39 48 44 44 46 43 42 40 Concentration Gp3 127 134 139 131 138 138 141 129 133 136 141 142 146 144 140 142 145 139 140 145 147 138 141 138 137 139 146 (ppm) Gp4 352 342 365 359 358 360 362 364 357 367 374 373 405 380 374 378 371 375 383 393 422 378 413 411 406 415 403 40 135 375 4.9 8.1 20.2 12.1 6.0 5.4 40 135 374 5.0 8.1 19.4 12.3 6.0 5.2 Ethylene in nitrogen calibration cylinder Standard deviation Coefficient of variation (sd 100/mean) Actual Gp2 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 26 4.1 15.5 ran out Concentration Gp3 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 6.4 104 6.6 6.4 (ppm) Gp4 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 5.5 299 15.9 5.3 : 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: 1 No data collected due to computer problem with data capture programme 2 Problem with ethylene in nitrogen calibration cylinder, no data analytical data collected 5 Nominal concentrations were calculated from the following equations: Concentration (ppm) = V 106 Va + V V = W R T 760 mm Hg M Atm where V = W = M = R = T = Atm = Va = the exposure gaseous volume of POSF mass of POSF molecular weight POSF (502.14 g/mole) gas constant (0.08205 l atm mol-1 K-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 100/mean) : 313 : TABLE C 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 1 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 : TABLE C 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 1 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 : 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) 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 1 2 1 315 290 1 287 304 296 295 292 303 313 301 297 305 301 283 308 310 315 291 292 290 299 303 297 306 295 297 1 310 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 : 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) 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 : Exposure no. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 TABLE D Chamber temperature - exposure mean values MIN 313/023622 Gp1 (Control) 20.8 20.3 20.5 20.7 20.4 20.6 22.0 21.7 21.3 21.8 22.3 22.0 21.3 21.3 21.3 21.3 21.3 21.3 21.4 21.3 20.9 20.8 21.2 21.0 20.9 21.0 21.0 21.3 21.7 20.8 20.8 20.8 20.8 20.8 20.8 20.9 20.0 20.2 20.0 20.0 Temperature (oC) Gp2 (Low dose) Gp3 (Inter dose) 20.5 20.4 20.9 20.4 20.6 20.2 20.4 20.3 20.5 20.3 20.3 20.0 20.3 20.3 20.4 20.4 20.2 20.3 20.2 20.4 20.5 20.3 20.8 20.2 21.0 20.4 21.0 20.3 20.8 20.0 20.0 20.3 20.0 20.4 20.0 20.0 20.9 20.4 20.0 19.9 20.0 20.0 20.0 20.0 20.8 20.3 20.0 20.0 20.6 20.0 20.5 20.0 20.8 20.3 20.7 20.0 20.7 20.2 20.8 20.2 20.7 20.1 20.7 19.9 20.5 20.0 20.3 19.9 20.0 20.0 20.4 19.9 20.2 20.0 20.4 20.4 20.3 20.0 20.2 20.0 Gp4 (High dose) 21.3 21.3 20.9 21.1 21.2 20.9 21.3 21.2 21.5 21.0 21.2 20.7 21.0 21.0 20.9 21.2 20.9 21.0 21.2 20.9 20.8 20.8 20.8 20.9 20.8 20.9 20.8 20.8 20.9 20.9 20.6 20.5 20.4 20.0 20.5 20.4 20.4 20.8 20.5 20.4 : 320 : TABLE D MIN 313/023622 (Chamber temperature - exposure mean values - continued) Exposure Temperature (oC) no. Gp1 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 sd CV (%) 20.6 0.71 3.4 20.4 0.33 1.6 20.1 0.23 1.1 Mean (1-65) sd CV (%) 20.6 0.67 3.2 20.4 0.31 1.5 20.1 0.21 1.0 sd standard deviation CV (%) Coefficient of variation (sd 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 l/minute using a laboratory pump. The chamber atmosphere was drawn through the IR cell 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-IR 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. Vapour Concentration (ppm) = AutoQuant Concentration - 8.2414 (1) 1.2219 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 = V 1,000,000 ppm (2) Va + V V = W R T 760 mm Hg (3) M Atm V = W = M = R = T = Atm = Va = gaseous volume of POSF (ml) mass of POSF (g) molecular weight of POSF (502.14 g/mole) Gas constant (0.08205 ml atm mmol-1 K-1) temperature (K) atmospheric pressure (mmHg) volume of air (ml) : 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 MIDAC 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 I2001 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 l/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 l/minute directly through the IR cell. Production of a Background Scan A flowrate of 1l/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 Amount( ppm ) = a A b A = absorbance at a given frequency of POSF in the sample spectrum a = absorption coefficient (absorptivity) of POSF in the sample spectrum, which is derived from the reference calibration transfer standard spectrum b = 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 l/minute is drawn through the IR cell. After a period of 3-5 minutes, a FT-IR 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 hour periods showed that the final calibration gave the path length values differed by less than 2% from the initial calibration value of 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 I2000 series FT-IR 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 IR 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 IR cell is 1 l/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-IR system. The analyser draws 1 l/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-IR 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-IR 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 conc (ppm) = (AutoQuant Conc (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 = 2000ppm 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 Mean 6 34.81 34.80 115.07 114.25 329.33 327.13 sd 1.396 0.833 6.249 6 sd CV (%) CV (%) 4.0 0.7 1.9 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation Coefficient of variation (sd 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 15 7 16 7 32.34 35.15 35.74 95.68 117.75 119.89 31.57 299.11 321.14 Mean 17 7 18 7 35.44 36.03 36.54 118.82 122.00 121.99 310.13 360.78 364.65 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 Mean 8 33.48 33.33 117.32 349.74 sd 1.573 2.416 14.502 7 8 sd CV (%) 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 Standard deviation Coefficient of variation (sd 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) 3 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 Mean 6 37.45 36.39 128.14 132.25 352.90 302.07 sd 1.201 7.136 124.866 CV (%) 3.3 5.4 41.3 7 42 -0.33 9 -0.30 9 -0.32 9 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 Mean 6 44.45 42.16 148.59 147.12 396.89 392.66 sd 1.347 1.817 14.728 6 9 sd CV (%) 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 Standard deviation Coefficient of variation (sd 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 54 122.93 10 55 140.34 10 56 154.99 10 57 170.37 10 58 183.67 10 59 195.64 10 60 25.27 105.90 207.48 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 70 71 Mean 23.31 23.59 a 22.09 a 22.33 a 128.38 126.39 128.83 126.77 6 360.28 362.33 362.78 362.76 sd 1.213 1.682 5.156 6 10 sd CV (%) a 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 Standard deviation Coefficient of variation (sd 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 11 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 Mean 6 23.19 21.41 129.88 130.22 365.15 358.66 sd 0.896 1.083 10.307 6 11 sd CV (%) 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. Standard deviation Coefficient of variation (sd 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 Mean 6 41.11 41.15 123.08 147.34 356.98 380.17 sd 0.654 18.751 20.016 CV (%) 1.6 12.7 5.3 12 96 39.05 84.53 12 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 Mean 6 40.82 41.12 131.64 131.95 373.78 369.53 sd 0.443 1.676 15.366 6 12 sd CV (%) 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 Standard deviation Coefficient of variation (sd 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 Mean 6 44.16 43.18 140.35 132.33 381.80 368.73 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 Mean 6 44.12 42.34 136.03 123.98 373.60 364.93 sd 7.257 34.546 13.011 6 sd CV (%) CV (%) 17.1 27.9 3.6 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 13 113.67 311.48 133 12.34 13 137.61 367.35 134 6.86 13 143.16 382.52 135 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 Mean 6 42.17 35.41 140.46 140.32 375.38 378.16 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 Mean 6 47.96 46.13 146.32 145.65 399.36 390.49 sd 0.843 1.066 9.293 6 13 sd CV (%) 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 Standard deviation Coefficient of variation (sd 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 Mean 6 46.34 44.79 128.27 122.97 389.64 375.96 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 Mean 6 42.80 42.15 129.60 125.34 398.75 371.45 sd 0.838 2.653 15.016 6 sd CV (%) CV (%) 2.0 2.1 4.0 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 43.55 43.165 126.59 124.54 388.45 381.43 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 Mean 6 41.47 43.77 130.40 125.78 374.77 376.21 sd 9.918 2.861 11.188 6 14 sd CV (%) 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 Standard deviation Coefficient of variation (sd 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 Mean 6 40.06 39.70 121.05 117.16 372.45 353.96 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 Mean 6 41.64 39.77 124.38 119.90 391.23 384.86 sd 0.820 2.012 11.067 6 sd CV (%) CV (%) 2.1 1.7 2.9 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 42.81 41.90 130.97 128.78 382.97 386.46 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 Mean 6 56.62 57.84 141.56 143.38 393.35 393.19 sd 2.564 2.979 7.571 6 sd CV (%) CV (%) 4.4 2.1 1.9 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 40.73 38.46 126.59 122.73 369.85 363.65 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 Mean 6 41.16 40.76 137.71 135.12 369.57 365.25 sd 0.811 2.141 6.783 6 sd CV (%) CV (%) 2.0 1.6 1.9 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 40.85 40.81 137.40 135.99 370.96 363.09 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 Mean 6 42.72 41.98 132.04 132.59 375.06 373.11 sd 0.838 1.384 3.019 6 sd CV (%) CV (%) 2.0 1.0 0.8 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 43.69 41.79 136.46 134.05 384.56 378.23 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 Mean 6 43.94 44.24 138.59 136.98 377.14 371.26 sd 0.584 3.702 10.786 6 sd CV (%) CV (%) 1.3 2.7 2.9 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 42.54 41.45 141.35 139.93 390.94 381.53 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 Mean 6 41.64 39.36 139.90 135.39 375.60 368.42 sd 1.346 2.964 13.520 6 sd CV (%) CV (%) 3.4 2.2 3.7 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 41.44 40.48 139.56 138.51 376.69 370.77 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 6 sd CV (%) CV (%) 1.8 0.9 3.45 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 40.13 39.46 136.51 134.01 370.27 369.11 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 Mean 6 40.24 39.72 128.53 134.28 390.12 382.36 sd 0.503 4.365 9.985 6 sd CV (%) CV (%) 1.3 3.3 2.6 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 100/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 Mean 6 43.64 42.01 138.35 136.59 394.35 388.63 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 Mean 6 44.12 41.45 145.97 145.78 394.04 395.87 sd 7.009 1.172 3.188 6 sd CV (%) CV (%) 16.9 0.8 0.8 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 42.64 42.31 136.10 137.21 393.04 385.59 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 Mean 6 38.26 37.39 124.90 126.67 353.65 351.97 sd 0.678 3.335 3.677 6 sd CV (%) CV (%) 1.8 2.6 1.0 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 40.63 38.43 135.89 134.40 350.37 341.51 sd 0.947 1.382 21.440 CV (%) 2.5 1.0 6.3 43 456 15 457 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 Mean 6 41.40 41.44 141.33 138.766 374.68 364.92 sd 0.5670 1.4997 9.862 6 15 sd CV (%) 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 Standard deviation Coefficient of variation (sd 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 17 145.81 17 469 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 Mean 6 36.42 35.77 131.71 131.12 363.46 358.66 sd 0.736 8.257 4.031 CV (%) 2.1 6.3 1.1 45 480 36.88 98.70 16 481 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 Mean 6 39.65 138.63 137.63 364.80 357.74 sd 0.477 1.674 12.013 6 16 17 sd CV (%) 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 Standard deviation Coefficient of variation (sd 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 Mean 6 42.03 40.38 139.07 138.47 365.07 360.18 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 Mean 6 41.83 40.09 132.85 140.93 367.19 362.41 sd 0.987 7.080 2.574 6 12 sd CV (%) 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 Standard deviation Coefficient of variation (sd 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 Mean 6 40.26 39.15 134.75 128.91 369.94 363.63 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 Mean 6 37.33 36.97 129.82 132.59 359.01 356.58 sd 0.442 5.883 6.876 6 sd CV (%) CV (%) 1.2 4.4 1.9 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 38.22 36.82 137.31 135.91 384.60 366.66 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 Mean 6 40.53 39.50 142.62 140.90 383.57 373.61 sd 0.880 1.715 11.222 6 sd CV (%) CV (%) 2.2 1.2 3.0 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 43.14 41.59 138.80 141.53 376.45 373.19 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 Mean 6 45.38 39.84 145.88 146.44 399.94 404.67 sd 2.098 4.160 13.442 6 sd CV (%) CV (%) 5.3 2.8 3.3 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 43.01 39.83 141.63 143.59 381.31 380.04 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 Mean 6 40.19 38.59 142.25 140.32 381.92 373.73 sd 1.075 1.804 15.800 6 sd CV (%) CV (%) 2.8 1.3 4.2 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 40.50 39.47 142.01 142.37 382.02 378.33 sd 1.104 2.406 13.373 CV (%) 2.8 1.7 3.5 57 624 39.14 12 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 Mean 6 43.75 42.33 148.50 145.15 391.78 370.58 sd 0.899 2.387 39.677 6 12 sd CV (%) 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 Standard deviation Coefficient of variation (sd 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 Mean 6 42.22 41.48 127.41 138.93 384.83 374.83 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 Mean 6 44.51 36.26 142.54 140.14 397.84 382.59 sd 12.155 1.418 27.317 6 18 sd CV (%) 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 Standard deviation Coefficient of variation (sd 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 Mean 6 39.87 38.85 144.33 144.65 402.71 392.71 sd 1.262 1.973 11.516 6 sd CV (%) CV (%) 3.2 1.4 2.9 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 10 96.65 673 10 115.11 674 10 132.16 675 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 10 115.67 686 14 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 10 14 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 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 Mean 6 40.82 43.93 138.14 137.93 405.98 377.69 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 Mean 6 17.15 43.54 142.20 141.15 420.97 412.80 sd 8.844 1.679 11.212 6 sd CV (%) CV (%) 20.3 1.2 2.7 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 Mean 6 47.13 46.37 135.42 138.36 416.88 410.79 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 Mean 6 37.61 42.63 137.77 137.44 415.27 406.02 sd 2.208 1.277 13.789 6 sd CV (%) CV (%) 5.2 0.9 3.4 The initial concentration measurement of each exposure was excluded from all calculations Standard deviation. Coefficient of variation (sd 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 14 14 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 12 14 sd CV (%) 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 Standard deviation Coefficient of variation (sd 100/mean) : 365 : 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 2 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 2 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 366 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................................................................................................ 11 STUDY SCHEDULE AND STRUCTURE .............................................................................. 11 TEST SUBSTANCE AND FORMULATION .......................................................................... 12 ANIMAL MANAGEMENT ..................................................................................................... 13 SERIAL OBSERVATIONS...................................................................................................... 15 NECROPSY AND HISTOLOGY............................................................................................. 19 PATHOLOGY .......................................................................................................................... 22 DATA TREATMENT............................................................................................................... 22 QUALITY ASSURANCE AND ARCHIVING PROCEDURES ............................................. 26 RESULTS...................................................................................................................................... 27 DISCUSSION ............................................................................................................................... 35 REFERENCES.............................................................................................................................. 36 FIGURES 1. Bodyweights - group mean values.......................................................................................... 37 : 367 : TABLES MIN 313/023622 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 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: MIN 313/023622 : 733 : MIN 313/023622 : 734 : MIN 313/023622 HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2001 : 735 : MIN 313/023622 HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2003 : 736 : MIN 313/023622 HUNTINGDON RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2005 : 737 : MIN 313/023622 EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2001 : 738 : MIN 313/023622 EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2003 : 739 : MIN 313/023622 EYE RESEARCH CENTRE GLP COMPLIANCE STATEMENT 2005 : 740 :