A New Fluorometric Determination of Disulfides in Urine, Blood and Some Mammalian Tissues by KCN-NAM Method

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Abstract
Disulfides, oxidized glutathione (GSSG) and cystine [(Cys)2], were determined with a new fluorescent reagent N-(9-acridinyl)maleimide via corresponding thiols which were derived by the reduction with potassium cyanide. Linear relations were obtained between the fluorescence intensities and the disulfides concentrations ranged from 5 pmol/ml to 5 nmol/ml of GSSG and 5 pmol/ml to 2.7 nmol/ml of (Cys)2, respectively. This method is 100-times more sensitive than the conventional colorimetric method. It enables determinations of disulfides in 2 µl of urine, 6 µl of blood and 0.6 µg of tissues.