A Bio-Fluorometric Acetone Gas Imaging System for the Dynamic Analysis of Lipid Metabolism in Human Breath

Abstract

We constructed an imaging system to measure the concentration of acetone gas by acetone reduction using secondary alcohol dehydrogenase (S-ADH). Reduced nicotinamide adenine dinucleotide (NADH) was used as an electron donor, and acetone was imaged by fluorescence detection of the decrease in the autofluorescence of NADH. In this system, S-ADH–immobilized membranes wetted with buffer solution containing NADH were placed in a dark box, and UV-LED excitation sheets and a high-sensitivity camera were installed on both sides of the optical axis to enable loading of acetone gas. A hydrophilic polytetrafluoroethylene (H-PTFE) membrane with low autofluorescence was used as a substrate, and honeycomb-like through-hole structures were fabricated using a CO2 laser device. After loading the enzyme membrane with acetone gas standards, a decrease in fluorescence intensity was observed in accordance with the concentration of acetone gas. The degree of decrease in fluorescence intensity was calculated using image analysis software; it was possible to quantify acetone gas at concentrations of 50–2000 ppb, a range that includes the exhaled breath concentration of acetone in healthy subjects. We applied this imaging system to measure the acetone gas in the air exhaled by a healthy individual during fasting.