Reduction of hexavalent chromium by mitochondria: methodological implications and possible mechanisms.

The reduction of hexavalent chromium by rat liver mitochondria was studied under various experimental conditions. The occurrence of possible artifacts arising from inadequate methods for Cr(VI) detection was also tested. Erroneous results due to the presence of NAD(P)H (above 0.1 mM) could be demonstrated when the diphenylcarbazide method is employed. Modifications of this colorimetric procedure are proposed, in order to minimize artifacts occurring when Cr(VI) is detected in NAD(P)H-added mixtures. Although no interfering substances that can affect colorimetric methods are normally present in mitochondrial preparations, some methodological precautions are recommended. Mitochondria can reduce hexavalent chromium using either succinate or glutamate as electron donors. The succinate-supported reduction of Cr(VI) is active especially in the presence of ADP or an uncoupler, while reduction induced by glutamate occurs only in the presence of respiratory-chain inhibitors. In mitochondria supplemented by NAD-linked or FAD-linked substrates, different mechanisms in Cr(VI) reduction can therefore take place.