Fluorometric gas-imaging system (sniff-cam), using the extinction of NADH with an ADH reverse reaction, for acetaldehyde in the gas phase
| Autor: | Kohji Mitsubayashi, Munire Naisierding, Toshiyuki Sato, Yuuki Hayakawa, Kenta Iitani, Takahiro Arakawa, Koji Toma |
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| Rok vydání: | 2017 |
| Předmět: |
Tris
Acetaldehyde 02 engineering and technology Nicotinamide adenine dinucleotide Photochemistry 01 natural sciences Biochemistry Reversible reaction Analytical Chemistry chemistry.chemical_compound Electrochemistry Ultraviolet light Environmental Chemistry Spectroscopy Alcohol dehydrogenase Chromatography Ethanol biology 010401 analytical chemistry Alcohol Dehydrogenase NAD 021001 nanoscience & nanotechnology Fluorescence 0104 chemical sciences chemistry biology.protein 0210 nano-technology Oxidation-Reduction |
| Zdroj: | The Analyst. 142:3830-3836 |
| ISSN: | 1364-5528 0003-2654 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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