FETAX INTERLABORATORY VALIDATION-STUDY - PHASE-II TESTING

The Frog Embryo Teratogenesis Assay-Xenopus (FETAX) is a 96-h whole embryo developmental toxicity screening assay that can be used in ecotoxicology and in detecting mammalian developmental toxicants when an in vitro metabolic activation system is employed. A standardized American Society for Testing and Materials (ASTM) guide for the conduct of FETAX has been published along with a companion atlas that helps in embryo staging and identifying malformations. As part of the ASTM process, an interlaboratory validation study was undertaken to evaluate the repeatability and reliability of FETAX. Six different laboratories participated in the study. Each laboratory utilized one technician with the exception of one laboratory, which utilized two independent technicians. In Phase I, FETAX proved to be more repeatable and reliable than many other bioassays. However, some excessive variation was observed in a few laboratories. Some of this variation may have been due to an initial lack of experience with the assay by some technicians. Phase II, which is reported here, showed far less intralaboratory and interlaboratory variability than did Phase I. Nonteratogens such as saccharin and sodium cyclamate showed the most consistent results, whereas more variability was observed for the teratogens caffeine and 5-fluorouracil. Interlaboratory coefficient of variation values for all FETAX end points ranged from 7.3 to 54.7%. The minimum concentration to inhibit growth proved to be the most variable end point for three of the four test chemicals, whereas the LC50 and EC50 (malformation) proved to be less variable.