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To develop a novel and automated assay determining plasma thiol/disulphide homeostasis, which consists of thiol–disulphide exchanges.

Native thiol and total thiol concentrations were synchronously measured as a paired test. In the first vessel, the amount of native thiol groups was measured by a modified Ellman reagent. At the parallel run, first, dynamic disulphide bonds were reduced to free thiol groups by NaBH₄. The unused reductant remnants were completely removed by formaldehyde. Thus, the total thiol amount could be accurately measured. Mercaptoethanol solutions were used as calibrators. The half value of the difference between total thiol and native thiol amounts gave the disulphide bond amount.

No separation step for the assay was needed. All processes were performed using an automated analyser within about 10 min. Plasma disulphide levels were 17.29 …