Novel cytotoxicity test based on menadione-catalyzed H2O2 productivity for food safety evaluation.

Menadione-catalyzed H(2)O(2) production by viable cells was proportional to viable cell number, and the assay of this H(2)O(2) production was applied to the cytotoxicity test of 17 substances which were used for international validation of fixed-dose procedure as an alternative to the classical LD(50) test. The cytotoxicity of substances tested was observed 4 h after the incubation with animal cells, and the viability was determined in 10 min according to menadione-catalyzed H(2)O(2) production assay. IC(50) of each substance required for 50% inhibition of menadione-catalyzed H(2)O(2) production was similar among HepG2, HuH-6KK, HUVE, Vero, Intestine407, NIH/3T3 and Neuro-2a cells. Twelve substances, 3 substances and 2 substances showed the difference of one, two and three orders in the magnitude between LD(50) and IC(50), respectively. These results show that menadione-catalyzed H(2)O(2) production assay is useful for the rapid detection of toxic compounds having the basal cytotoxicity common to various cells, but is unfit for the detection of organ-specific toxic compounds.