title: Interlaboratory validation of the in vitro eye irritation tests for cosmetic ingredients (9). Evaluation of cytotoxicity test on HeLa cells.
abstract: The cytotoxicity test using MTT on HeLa cells (HeLa-MTT) was evaluated as an alternative method to the Draize eye irritation test (Draize test) by six to eight laboratories. The 50% inhibition concentration (EC(50)) for MTT reduction was calculated. A total of thirty-nine test chemicals were examined. The average interlaboratory coefficient of variation (CV) of the present assay was 25%. Comparison of the in vitro test results with those of the Draize test revealed a correlation coefficient of -0.799 between the log(EC(50)) value and the maximum average total score (MAS) for a 10% solution or suspension. The following characteristics of HeLa-MTT have become apparent through this validation: (1) HeLa-MTT could be applied to all substances including water-insoluble substances, esters, a colour additive, and a substance which is known to directly reduce MTT; (2) there is good interlaboratory reproducibility and strong correlation between HeLa-MTT and Draize MAS; (3) results for strong acids, alkanolamines and alcohols (lower mono-ol) clearly deviate from other samples with respect to the correlation between HeLa-MTT and Draize MAS. These results suggest that HeLa-MTT may be useful for predicting the Draize MAS if definite criteria can be established for applicable compounds.
authors.affiliation: Japan Cosmetic Industry Association, Hatsumei Bldg, 9-14, Toranomon 2-chome, Minato-ku, Tokyo 105Japan; Yakult Central Institute for Microbiological Research, 1796 Yaho, Kunitachi-shi, Tokyo 186-8650Japan.
authors.forename: K
authors.initials: K
authors.lastname: Chiba
authors.forename: I
authors.initials: I
authors.lastname: Makino
authors.forename: J
authors.initials: J
authors.lastname: Ohuchi
authors.forename: Y
authors.initials: Y
authors.lastname: Kasai
authors.forename: H
authors.initials: H
authors.lastname: Kakishima
authors.forename: K
authors.initials: K
authors.lastname: Tsukumo
authors.forename: T
authors.initials: T
authors.lastname: Uchiyama
authors.forename: E
authors.initials: E
authors.lastname: Miyai
authors.forename: J
authors.initials: J
authors.lastname: Akiyama
authors.forename: Y
authors.initials: Y
authors.lastname: Okamoto
authors.forename: H
authors.initials: H
authors.lastname: Kojima
authors.forename: H
authors.initials: H
authors.lastname: Okumura
authors.forename: Y
authors.initials: Y
authors.lastname: Tsurumi
authors.forename: M
authors.initials: M
authors.lastname: Usami
authors.forename: K
authors.initials: K
authors.lastname: Katoh
authors.forename: S
authors.initials: S
authors.lastname: Sugiura
authors.forename: A
authors.initials: A
authors.lastname: Kurishita
authors.forename: M
authors.initials: M
authors.lastname: Sunouchi
authors.forename: A
authors.initials: A
authors.lastname: Miyajima
authors.forename: M
authors.initials: M
authors.lastname: Hayashi
authors.forename: Y
authors.initials: Y
authors.lastname: Ohno