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DL-LACTONE CAS N°:79-50-5 - page 99 / 113

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OECD SIDS 5. TOXICITY

DL-LACTONE

ID: 79-50-5 DATE: 18.01.2006

monitoring system, Pearson Technical Services, Debenham, Stowmarket, England), during the motor activity test, males were caged individually and females were caged with their offspring. The assigned males were tested during week 4 of treatment and the assigned females were tested during lactation (all before blood sampling). In order to avoid hypothermia of pups, dams were removed from the pups for not more than 30-40 minutes. Body weights: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females were weighed on days 0, 7, 14 and 21 of gestation and during lactation on days 1 and 4. Food consumption: Weekly, for males and females. During the mating period analysis of food consumption was suspended. Food consumption of mated females was measured on gestation days 0, 7, 14 and 21 and during lactation on days 1 and 4. Water consumption: Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effect was suspected. Reproduction processes: Male number paired with, mating date, confirmation of pregnancy and delivery day were recorded.

Observations offspring Each litter was examined to determine the following if practically possible: - the numbers of live and dead pups at the First Litter Check (= check at day 1 of lactation) and daily thereafter (if possible, defects or cause of death were evaluated); - the individual weight of all live pups on days 1 and 4 of lactation; - sex of all pups (by assessment of the ano-genital distance); - the number of pups with physical or behavioural abnormalities, daily. Clinical labooratory investigations F0 animals Blood samples were collected from 5 males and 5 females randomly selected from each group under isoflurane anaesthesia immediately prior to scheduled post mortem examination, between 07:30 and 09:30 am. The animals were fasted overnight (with a maximum of 20 hours) before blood sampling, but water was provided. Blood samples were drawn from the retro-orbital sinus of all rats/sex/group and collected into tubes prepared with EDTA for haematological parameters (0.25 ml), with citrate for clotting tests (1.0 ml) and Li-heparin-treated tubes for clinical biochemistry parameters (1.0 ml). The following parameters were determined. Haematology: Erythrocytes count (RBC); Haemoglobin (HB); Haematocrit (HCT); Mean corpuscular volume (MCV); Mean corpuscular haemoglobin (MCH); Mean corpuscular haemoglobin concentration (MCHC); Platelet count; Red cell distribution width; Total leucocytes count (WBC); Differential leucocyte count; Clotting Potential; Prothrombin time (PT); Partial thromboplastin time (APTT). Clinical Biochemistry: Alanine aminotransferase (ALAT); Alkaline phosphatase (ALP); Aspartate aminotransferase (ASAT); Bilirubin, total; Chloride; Cholesterol, total; Creatinine; Glucose; Phosphorus (inorganic); Protein, total; Protein, albumin; Urea; Calcium; Potassium; Sodium.

Pathology, F0 animals Termination: All animals surviving to the end of the observation period and all moribund animals were anaesthetised using iso-flurane and subsequently exsanguinated. All animals

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