An Enzymatic Assay of Reduced Glutathione Using Glutathione S-Aryltransferase with o-Dinitrobenzene as a Substrate1

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Abstract
A simple and sensitive enzymatic assay method for the determination of reduced glutathione (GSH) has been developed using glutathione S-aryltransferase with o-dinitrobenzene as a substrate. o-Dinitrobenzene is a good substrate for the enzyme and has low spontaneous reactivity with GSH at neutral pH. GSH can be determined by colorimetrically measuring nitrite released upon the enzymatic conjugation of GSH and o-dinitrobenzene by using a diazo-coupling method with N-(1-naphthyl)ethylenediamine dihydrochloride. This method is capable of quantitating 1 to 40 nmol of GSH, and can be applied to physiological samples containing deproteinizing reagents.

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