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Amperometric biosensor for ethanol based on immobilization of alcohol dehydrogenase on a nickel hexacyanoferrate
1Department of Chemistry, Nanjing Normal University, Nanjing 210097. People's Republic of China.
Abstract:
Alcohol dehydrogenase (ADH) has been immobilized on a nickel hexacyanoferrate modified microband gold electrode surface by a glutaraldehyde/bovine serum albumin (BSA) cross-linking procedure to provide a new amperometric sensor for the assay of ethanol. The resulting enzyme electrode exhibits excellent electrocatalysis for the oxidation of reduced nicotinamide-adenine dinucleotide (NADH). The amperometric determination is based on the electrochemical detection of NADH which is generated in the enzymatic reaction of ethanol with NAD(+) under catalysis of ADH. The influence of various experimental conditions was examined for the determination of the optimum analytical performance. The sensor responds rapidly to ethanol with a detection limit of (5.0 +/- 0.3) x 10(-7) mol 1(-1). The response current increases linearly with ethanol concentration up to 5 mmol 1(-1). The sensor remains relatively stable for about 1 week.
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