Flow injection procedures for the determination of ethanol and alcohol dehydrogenase using co-immobilised bacterial luciferase and oxidoreductase

Abstract

Bacterial luciferase and oxidoreductase extracted from Vibrio harveyi were co-immobilised on cyanogen bromide-activated Sepharose 4B and used in a flow injection manifold for the rapid and sensitive determination of ethanol and alcohol dehydrogenase. The detection limits were 30 pmol for ethanol and 0.03 pmol for alcohol dehydrogenase. The relative standard deviations were 0–1.8%(n= 5) for ethanol over the range 1 × 10^–6–1 × 10^–1M and 0–2.0%(n= 5) for alcohol dehydrogenase over the range 0.03–30 pmol; the sample throughput was 60 h^–1. For the determination of ethanol, alcohol dehydrogenase was immobilised on cyanogen bromide-activated Sepharose 4B and incorporated upstream of the co-immobilised bacterial luciferase and oxidoreductase in the reaction coil.