3 research outputs found
Fluorometric Sniff-Cam (Gas-Imaging System) Utilizing Alcohol Dehydrogenase for Imaging Concentration Distribution of Acetaldehyde in Breath and Transdermal Vapor after Drinking
Understanding concentration distributions,
release sites, and release
dynamics of volatile organic compounds (VOCs) from the human is expected
to lead to methods for noninvasive disease screening and assessment
of metabolisms. In this study, we developed a visualization system
(sniff-cam) that enabled one to identify a spatiotemporal change of
gaseous acetaldehyde (AcH) in real-time. AcH sniff-cam was composed
of a camera, a UV-LED array sheet, and an alcohol dehydrogenase (ADH)-immobilized
mesh. A reverse reaction of ADH was employed for detection of gaseous
AcH where a relationship between fluorescence intensity from nicotinamide
adenine dinucleotide and the concentration of AcH was inversely proportional;
thus, the concentration distribution of AcH was measured by detecting
the fluorescence decrease. Moreover, the image differentiation method
that calculated a fluorescence change rate was employed to visualize
a real-time change in the concentration distribution of AcH. The dynamic
range of the sniff-cam was 0.1–10 ppm which encompassed breath
AcH concentrations after drinking. Finally, the sniff-cam achieved
the visualization of the concentration distribution of AcH in breath
and skin gas. A clear difference of breath AcH concentration was observed
between aldehyde dehydrogenase type 2 active and inactive subjects,
which was attributed to metabolic capacities of AcH. AcH in skin gas
showed a similar time course of AcH concentration to the breath and
a variety of release concentration distribution. Using different NADH-dependent
dehydrogenases in the sniff-cam could lead to a versatile method for
noninvasive disease screening by acquiring spatiotemporal information
on various VOCs in breath or skin gas
Fiber-Optic Bio-sniffer (Biochemical Gas Sensor) Using Reverse Reaction of Alcohol Dehydrogenase for Exhaled Acetaldehyde
Volatile organic compounds (VOCs)
exhaled in breath have huge potential
as indicators of diseases and metabolisms. Application of breath analysis
for disease screening and metabolism assessment is expected since
breath samples can be noninvasively collected and measured. In this
research, a highly sensitive and selective biochemical gas sensor
(bio-sniffer) for gaseous acetaldehyde (AcH) was developed. In the
AcH bio-sniffer, a reverse reaction of alcohol dehydrogenase (ADH)
was employed for reducing AcH to ethanol and simultaneously consuming
a coenzyme, reduced form of nicotinamide adenine dinucleotide (NADH).
The concentration of AcH can be quantified by fluorescence detection
of NADH that was consumed by reverse reaction of ADH. The AcH bio-sniffer
was composed of an ultraviolet light-emitting diode (UV-LED) as an
excitation light source, a photomultiplier tube (PMT) as a fluorescence
detector, and an optical fiber probe, and these three components were
connected with a bifurcated optical fiber. A gas-sensing region of
the fiber probe was developed with a flow-cell and an ADH-immobilized
membrane. In the experiment, after optimization of the enzyme reaction
conditions, the selectivity and dynamic range of the AcH bio-sniffer
were investigated. The AcH bio-sniffer showed a short measurement
time (within 2 min) and a broad dynamic range for determination of
gaseous AcH, 0.02–10 ppm, which encompassed a typical AcH concentration
in exhaled breath (1.2–6.0 ppm). Also, the AcH bio-sniffer
exhibited a high selectivity to gaseous AcH based on the specificity
of ADH. The sensor outputs were observed only from AcH-contained standard
gaseous samples. Finally, the AcH bio-sniffer was applied to measure
the concentration of AcH in exhaled breath from healthy subjects after
ingestion of alcohol. As a result, a significant difference of AcH
concentration between subjects with different aldehyde dehydrogenase
type 2 (ALDH2) phenotypes was observed. The AcH bio-sniffer can be
used for breath measurement, and further, an application of breath
analysis-based disease screening or metabolism assessment can be expected
due to the versatility of its detection principle, which allows it
to measure other VOCs by using NADH-dependent dehydrogenases
Improved Sensitivity of Acetaldehyde Biosensor by Detecting ADH Reverse Reaction-Mediated NADH Fluoro-Quenching for Wine Evaluation
Acetaldehyde (AcH) is found in ambient
air, foods, and the living
body. This toxic substance is also contained in wine and known as
an important ingredient affecting the quality of wine. Herein, we
constructed and evaluated two different fiber-optic biosensors for
measurement of AcH in the liquid phase (AcH biosensor) using aldehyde
dehydrogenase (ALDH) or alcohol dehydrogenase (ADH). The AcH biosensor
measured a concentration of AcH using fluorescence intensity of a
reduced form of nicotinamide adenine dinucleotide (NADH) that was
produced or consumed via catalytic reaction of the respective enzyme.
In the AcH measurement system, an ultraviolet light emitting diode
(UV-LED) and photomultiplier tube (PMT) were connected to a bifurcated
optical fiber and were used to excite and detect NADH. A sensing region
was developed using an optical fiber probe and an enzyme-immobilized
membrane, buffer pH, and concentrations of a coenzyme in buffer solution
for ALDH forward reaction and ADH reverse reaction were optimized,
and the dynamic ranges were compared. ADH-mediated AcH biosensor showed
higher sensitivity, wider dynamic range (1–500 μM), and
capability of rapid measurement (less than 3 min) than ALDH-mediated
AcH biosensor (5–200 μM). ADH biosensor also presented
a high selectivity and allowed measurement of AcH in 9 different wine
samples (5 red and 4 white wines). The determined concentrations were
comparable to those measured by NADH absorbance method, which validated
the accuracy of the ADH biosensor in AcH measurement