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Determination of 3-hydroxybutyrate in serum by flow-injection analysis using a co-immobilized 3-hydroxybutyrate dehydrogenase/NADH oxidase reactor and a chemiluminometer

✍ Scribed by Nobutoshi Kiba; Hidekazu Koemado; Junko Inagaki; Motohisa Furusawa

Publisher: Elsevier Science
Year: 1994
Tongue: English
Weight: 451 KB
Volume: 298
Category: Article
ISSN: 0003-2670
DOI: 10.1016/0003-2670(94)90051-5

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✦ Synopsis

A flow-injection system with a co-immobilized enzyme reactor is described for the determination of 3-hydroxybutyrate. 3-Hydroxybutyrate dehydrogenase and NADH oxidase were immobilized on aminated poly (vinyl alcohol) beads and packed into a stainless-steel column (5 cm X 4 mm i.d.). Serum was diluted IO-fold with borate buffer (pH 9.4) and ultrafiltered. Filtrate (20 ~1) was injected into the carrier stream. The hydrogen peroxide produced was detected chemiluminometrically via a luminol-hexacyanoferrate( III) reaction. The calibration graph was linear for 2 X lo-' M-5 X low4 M; the detection limit was 8 X 10e8 M and the sample throughput was 30 h-' without carryover.

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