A method was developed for the analysis of reducing sugars extracted with 80% aqueous ethanol from fish muscle. It was based on the reaction of the reducing sugars with the chromophoric reagent, 4-N, N-dimethylamino-4'-azobenzene sulfonyl hydrazide, and their separation by reversed-phase high-performance liquid chromatography (HPLC) or high-performance silical gel thin-layer chromatography. The absorbance at 485 nm provided a sensitivity of the picomole level. This method was applied to the reducing sugar analysis of 20 mg of fish muscle. The change of reducing sugar contents in fish muscle of various stages of freshness was compared with that of K values measured by ion-pair reversed-phase HPLC.
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