90-DAY INHALATION STUDY IN RATS, USING G7-25 CIGARETTES STUDY PROTOCOL This inhalation study will examine the toxicological responses observed in rats exposed to smoke from cigarettes formulated with 2 different inclusions of an additive ("test cigarettes"). Responses will be compared with those in animals exposed to smoke from cigarettes without the additive ("reference cigarettes"). The study will use 7 groups each containing 32 male and 32 female Sprague-Dawley rats, plus a sentinel group comprising 50 animals. Three concentrations of smoke from the each of the test and reference cigarettes will be used. The comparisons of test and reference will be made on the basis of the amounts of wet total particulate matter (WTPM) presented to the animals. There will also be a sham-exposure group ("controls") and a group of sentinels. Animals will be exposed one hour per day, 5 days per week for 14 consecutive weeks (total of 65 exposures). The experiment will include a study of the reversibility of any changes observed. End-points will include histopathology, plasma nicotine and cotinine, organ and body weights, feed consumption, CO-oximetry, and measurements of respiratory physiology. OBJECTIVE by Xybion Medical Systems (Cedar Knolls, The objective of this study (TOX-43) is NJ) will be used for scheduling and for data to compare toxicological responses of rats acquisition and analysis. exposed to mainstream smoke from cigarettes Any records that would be required to containing 0 or 30% of G7-25 sheet, in a reconstruct the experiment will be maintained. sub-chrönic nose-only inhalation study using This study will not be listed as a regulated Sprague-Dawley rats. study and the results will not be submitted to any regulatory agency. In general, the spirit of EXPERIMENTAL DESIGN Good Laboratory Practice (21 CFR, Part 58, The experimental design is based on the 1987) will be followed. Data will be reviewed OECD guideline No. 413, "Sub-chronic for accuracy of reporting, using staff Inhalation Toxicity: 90-day study", adopted 12 experienced in Quality Assurance (QA) review May 1981. of toxicology studies. Original laboratory Three groups of animals will be exposed notebooks and specimens will be stored at the to smoke from the test cigarette (30% G7-25) evaluation facilities. and three groups to smoke from the reference cigarette (0% G7-25). The actual dilutions of EXPERIMENTAL ANIMALS the smoke will be adjusted to provide for each This section was written with reference comparison a smoke with comparable made to the Format for preparation of concentrations (low, medium, high) of WTPM protocols and ACUP' S, prepared by the at the animal ports. In addition there will be a Chairman of the RJRT Institutional Animal sham-exposure group, placed in restraint tubes Care and Use Committee (IACUC) on July 26, but exposed only to dilution air. A group of 1991. non-exposed animals will serve as sentinels for Animal Selection and Justification for Test disease. Each inhalation group will consist of System 32 animals per sex; there will be 25 animals The study requires the use of animals, as per sex in the sentinel group. there are no alternative systems for examining Animals will be exposed to smoke for the inhalation toxicology (if any) of the one hour per day, 5 days per week for 14 addition of G7-25 sheet into cigarettes. consecutive weeks (excluding holidays), to The Sprague-Dawley rat (Crl: CD/BR, give a total of 65 exposure days. Sub-groups VAF/Plus) is chosen as the experimental of surviving animals groups will be kept for an animal, because it has frequently been used in additional 14 weeks without treatment, as a inhalation studies and there is a large amount reversibility study. of background data available in the scientific literature. DATA MANAGEMENT / QUALITY The number of animals to be used is the ASSURANCE minimum associated with meaningful Specialist computer software produced statistical analyses of the data, based on the DRAFT: DO NOT QUOTE. Page 1 Source:https://wwwindustrydocumentsucsf.edu/docs/ffbb0103