Issue 20, 2017

Fluorometric gas-imaging system (sniff-cam), using the extinction of NADH with an ADH reverse reaction, for acetaldehyde in the gas phase

Abstract

A gas-imaging system (sniff-cam) that allows fluorometric visualization of a two-dimensional (2-D) distribution of gaseous acetaldehyde (AcH) was developed. It employed a reverse reaction of a nicotinamide adenine dinucleotide (NADH) dependent enzyme that led to consumption of NADH in that reaction. The system was constructed with a highly sensitive camera, an ultraviolet light emitting diode array sheet, two band pass filters and an alcohol dehydrogenase (ADH)-immobilized mesh that was used for AcH detection. The reverse reaction of the ADH catalyzed the reduction of AcH to ethanol and the oxidation of NADH to NAD+, which occurred when gaseous AcH was applied to the ADH immobilized mesh that was wetted with a slightly acidic NADH solution. As NADH has an autofluorescence property [emission (λem) at 490 nm; excitation (λex) at 340 nm], the presence of gaseous AcH was visualized by a decrease of fluorescence of the NADH at the ADH immobilized mesh. After constructing the gaseous AcH imaging system, optimizations of pH, and concentration of the NADH solution were performed. As a result of the optimizations (500 μM of NADH in 0.1 M of Tris hydrochloride (Tris-HCl) buffer at pH 6.5), the AcH sniff-cam showed a wide dynamic range (0.1–10 ppm) for gaseous AcH with a high correlation coefficient (R = 0.999). Furthermore, a fluorescence gradient with a rounded shape centered in a gas outlet was observed. These results demonstrated that the AcH sniff-cam utilizing the fluorescence decrease of NADH could be used to quantitatively evaluate the 2-D distribution of gaseous AcH.

Graphical abstract: Fluorometric gas-imaging system (sniff-cam), using the extinction of NADH with an ADH reverse reaction, for acetaldehyde in the gas phase

Supplementary files

Article information

Article type
Paper
Submitted
27 Mar 2017
Accepted
17 Jul 2017
First published
19 Jul 2017

Analyst, 2017,142, 3830-3836

Fluorometric gas-imaging system (sniff-cam), using the extinction of NADH with an ADH reverse reaction, for acetaldehyde in the gas phase

K. Iitani, T. Sato, M. Naisierding, Y. Hayakawa, K. Toma, T. Arakawa and K. Mitsubayashi, Analyst, 2017, 142, 3830 DOI: 10.1039/C7AN00524E

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