Image Sensing of Gaseous Acetone Using Secondary Alcohol Dehydrogenase-Immobilized Mesh for Exhaled Air

• Published in: Analytical Chemistry Vol. 96; no. 28; pp. 11549 - 11556
• Main Authors: Iitani, Kenta, Suzuki, Mika, Ichikawa, Kenta, Toma, Koji, Arakawa, Takahiro, Mitsubayashi, Kohji
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 16.07.2024; American Chemical Society (ACS)
• Subjects: Acetone; Acetone - analysis; Acetone - chemistry; Adenine; air; Alcohol dehydrogenase; Alcohol Oxidoreductases - chemistry; Alcohol Oxidoreductases - metabolism; alcohols; analytical chemistry; Biosensing Techniques; Biosensors; Breath Tests; Buffer solutions; catalytic activity; Cotton; Crosslinking; Dehydrogenase; Dehydrogenases; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - metabolism; Exhalation; fasting; fluorescence; Gas chromatography; gas chromatography-mass spectrometry; Gases - analysis; Gases - chemistry; glutaraldehyde; Humans; Immobilization; immobilized enzymes; Lipid metabolism; Lipids; Mass spectrometry; Mass spectroscopy; NAD; NAD (coenzyme); NAD - analysis; NAD - chemistry; NAD - metabolism; Nicotinamide; Nicotinamide adenine dinucleotide; people; Potassium phosphate; Potassium phosphates; solvents
• ISSN: 0003-2700; 1520-6882; 1520-6882
• DOI: 10.1021/acs.analchem.4c02251

Human-borne acetone is a potent marker of lipid metabolism. Here, an enzyme immobilization method for secondary alcohol dehydrogenase (S-ADH), which is suitable for highly sensitive and selective biosensing of acetone, was developed, and then its applicability was demonstrated for spatiotemporal imaging of concentration distribution. After various investigations, S-ADH-immobilized meshes could be prepared with less than 5% variation by cross-linking S-ADH with glutaraldehyde on a cotton mesh at 40 °C for 15 min. Furthermore, high activity was obtained by adjusting the concentration of the coenzyme nicotinamide adenine dinucleotide (NADH) solution added to the S-ADH-immobilized mesh to 500 μM and the solvent to a potassium phosphate buffer solution at pH 6.5. The gas imaging system using the S-ADH-immobilized mesh was able to image the decrease in NADH fluorescence (ex 340 nm, fl 490 nm) caused by the catalytic reaction of S-ADH and the acetone distribution in the concentration range of 0.1–10 ppm-v, including the breath concentration of healthy people at rest. The exhaled breath of two healthy subjects at 6 h of fasting was quantified as 377 and 673 ppb-v, which were consistent with the values quantified by gas chromatography–mass spectrometry.