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Chromium (VI) is carcinogenic through intermediates formed in the cellular milieu by reduction with small reductants like glutathione (GSH), ascorbate, cysteine, and NADPH. Although the reduction of chromate by thiols has been investigated, the participation of Cr(IV) intermediates has been inferred only indirectly due to the Cr(IV) refractive behavior towards EPR spectroscopy. Biological data from numerous reports indicate that Cr(IV) is the species most likely responsible for the carcinogenicity of Cr(VI). Our kinetic studies suggested that in acidic solutions, glycine buffer at pH 2.8, the reduction of chromate with GSH involves mostly a chromium(IV) intermediate. As a step towards the full characterization of the paramagnetic species involved in the reduction of chromate by thiols at neutral pH, we embarked on an investigation of the reduction of chromate with GSH in glycine buffer at pH 2.8 using a Superconducting QUantum Interference Device (SQUID) magnetometer. Our results indicate a strong influence of temperature and confirm the presence of Cr(IV). At 2 K, the saturation magnetization method was applied to the frozen reaction when it reached the peak of formation of intermediates and the contributions were calculated to be 30% of Cr(IV) and 69% of Cr(V). When the Curie–Weiss method was applied to determine the effective magnetic moment, the use of the linear portion of the curve, 100–200 K, yielded 58% Cr(IV) and 42% Cr(V); when data from the region below the temperature of liquid N2 (77 K) is employed, the intermediate is exclusively Cr(IV).

Magnetic Characterization of Chromium Intermediates in the Reduction of Chromium (VI) by Glutathione in Acidic Solutions