Potentiation of sodium chromate(VI)-induced chromosomal aberrations and mutation by vitamin B2 in Chinese hamster V79 cells

Electron spin resonance (ESR) measurements on solutions and isolated powders provide direct evidence for the involvement of Cr(V) species in the reduction of Cr(VI) by NAD(P)H. ESR analysis of an isolated Cr(V)-NAD(P)H solid yields g parallel = 1.9831 and g perpendicular = 1.9772, indicating that the unpaired electron occupies the dz2 orbital of the Cr(V) ion, with square-pyramidal geometry. Addition of hydrogen peroxide (H2O2) to the NAD(P)H-Cr(VI) reaction mixtures suppresses the Cr(V) species and generates hydroxyl (.OH) radicals. The .OH radicals were detected via ESR spin trapping, employing 5,5-dimethyl-1-pyrroline-N-oxide and alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone as spin traps. The dependence of Cr(V) and .OH radical formation on the H2O2 and Cr(VI) concentrations indicates that the Cr(V) species react with H2O2 to generate the .OH radicals. Similar results were obtained by using various diols (arabinose, cellobiose, FAD, fructose, glyceraldehyde, ribose, and tartaric acid), alpha-hydroxycarboxylic acids, and glutathione. Investigations with superoxide dismutase showed no significant participation of O2- in the generation of .OH radicals. These results thus indicate that the Cr(V) complexes, produced in the reduction of Cr(VI) by cellular reductants, react with H2O2 to generate .OH radicals, which might be initiators of the primary events in the Cr(VI) cytotoxicity.