Potentiometric Determination of NAD(P)H Coenzymes Using a Nitrite-Selective Membrane Electrode and a Nitrate Reductase Enzyme and the Application to Glucose Assay in Human Serum.

KATSU Takashi, HASHIMOTO Takeshi, HIRAMAKI Shima

doi:10.2116/analsci.15.135

Potentiometric Determination of NAD(P)H Coenzymes Using a Nitrite-Selective Membrane Electrode and a Nitrate Reductase Enzyme and the Application to Glucose Assay in Human Serum.

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KATSU Takashi

Faculty of Pharmaceutical Sciences, Okayama University
HASHIMOTO Takeshi

Faculty of Pharmaceutical Sciences, Okayama University
HIRAMAKI Shima

Faculty of Pharmaceutical Sciences, Okayama University

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NAD(P)H coenzyme concentrations were determined with the combination of a nitrite-selective membrane electrode and a nitrate reductase enzyme. The method was based on the detection of nitrite formed by the enzymatic reaction of NAD(P)H with nitrate catalyzed by nitrate reductase. The lower detection limits for both these coenzymes were around 100 µM. This system was applied to detect glucose by combining an additional glucose dehydrogenase enzyme. Serum glucose levels could be determined through the detection of nitrite.