Quantification of L-Tartrate in Wine by Stopped-Flow Injection Analysis Using Immobilized D-Malate Dehydrogenase and Fluorescence Detection.

TSUKATANI Tadayuki, MATSUMOTO Kiyoshi

doi:10.2116/analsci.16.265

Quantification of L-Tartrate in Wine by Stopped-Flow Injection Analysis Using Immobilized D-Malate Dehydrogenase and Fluorescence Detection.

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TSUKATANI Tadayuki

Biotechnology and Food Research Institute, Fukuoka Industrial Technology Center
MATSUMOTO Kiyoshi

Institute of Food Biotechnology, Department of Bioscience and Biotechnology, Division of Bioresource and Bioenvironmental Sciences, Graduate School Kyushu University

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A stopped-flow injection system with an immobilized enzyme reactor is proposed for the quantification of L-tartrate in wine. L-Tartrate was oxidized using the secondary activity of D-malate dehydrogenase (D-MDH), which was immobilized on aminopropyl-controlled pore glass beads with glutaraldehyde. NADH produced by D-MDH reaction was detected fluorometrically at 460 nm (excitation at 340 nm). The calibration graph was linear over the range 0.01 - 1.20 mM with a correlation coefficient of 0.999 when the stopped-time was 180 s. The relative standard deviation for ten measurements was 0.81% at the 1.0 mM level. The sample throughput was 12 h^-1. The results obtained by the proposed method and an HPLC method are in good agreement with each other. The D-MDH reactor was stable for one month when used daily under the optimal conditions.