Flow-injection determination of 3-hydroxybutyrate in serum with an immobilized 3-hydroxybutyrate dehydrogenase reactor and fluorescence detection.
A flow-injection system with an immobilized enzyme reactor is proposed for the determination of 3-hydroxybutyrate. 3-Hydroxybutyrate dehydrogenase is immobilized on aminated poly(vinyl alcohol) beads and packed into a stainless-steel column (4 cm x 4 mm I.D.). Serum is diluted and filtered. Sample solution (20 mul) is injected into the carrier stream [4mM NAD(+) in glycine buffer (pH 9.3)]. The NADH formed is detected at 465 nm (excitation at 340 nm). The calibration graph is linear for 0.7-500muM 3-hydroxybutyrate; the detection limit is 0.5muM.
Kiba
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N
Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Yamanashi University, Kofu 400, Japan.
Koemado
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H
Furusawa
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M