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The quantity of L-malate was determined using apparatus comprised of a reactor with immobilized malate dehydrogenase (MDH) and aspartate amino- transferase (AST) in a flow line. NADH formed by an enzymatic reaction was fluorometrically detected. The optimal concentration of NAD+ in the carrier containing 0.1M glutamate was determined. The maximum peak areas due to NADH were observed at pH 8.0 when the pH of the carrier consisting of Tris buffer ranged from 7.0 to 8.5. Various buffer types were also examined as carrier media at pH 8.0 and Tris buffer showe dt hemaximum peak area. When the carrier composed of Tris buffer (0.1M, pH 8.0) was used, the calibration curve for malate was linear in the range of 0.05-50µ M( r = 1.000). The detec- tion limit (S/N = 3) was 0.03µM. Relative standard deviations of the peak area at 1µ Ma nd 10µ Mw ere 1.5% (n = 7) and 0.36% (n = 7), respectively. Thirty samples of malate (10µM) were analyzed for 1hr. This method was applied to the analysis of malate in several beverages, and malate content determined by this method agreed with that determined by a com- mercially available test-kit method.

journal of health science

Determination of L-Malate Using Immobilized Malate Dehydrogenase and Aspartate Aminotransferase in a Flow System and its Application to Analyze the L-Malate Content of Beverages