Fiber-Optic Bio-sniffer (Biochemical Gas Sensor) Using Reverse Reaction of Alcohol Dehydrogenase for Exhaled Acetaldehyde
Fiber-Optic Bio-sniffer (Biochemical Gas Sensor) Using Reverse Reaction of Alcohol Dehydrogenase for Exhaled Acetaldehyde
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DOI：
10.1021/acssensors.7b00865
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发表时间：
2018-02-01
期刊：
影响因子：
8.9
通讯作者：
Mitsubayashi, Kohji
中科院分区：
文献类型：
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作者：
Iitani, Kenta;Chien, Po-Jen;Mitsubayashi, Kohji
呼吸中呼出的挥发性有机化合物（VOC）作为疾病和代谢的指标具有巨大的潜力。由于呼吸样本可以非侵入性地收集和测量，因此呼吸分析在疾病筛查和代谢评估中的应用是预期的。本研究开发了一种针对气态乙醛（AcH）的高灵敏度和选择性生化气体传感器（生物嗅探器）。在AcH生物嗅探器中，采用醇脱氢酶（ADH）的逆反应将AcH还原为乙醇，同时消耗辅酶，还原形式的烟酰胺腺嘌呤二核苷酸（NADH）。AcH的浓度可以通过荧光检测ADH逆反应消耗的NADH来定量。乙酰胆碱生物嗅探器由紫外发光二极管（UV-LED）作为激发光源、光电倍增管（PMT）作为荧光检测器和光纤探针组成，这3个部件通过分叉光纤连接。利用流动池和ADH固定膜构建了光纤探针的气敏区。在实验中，通过对酶反应条件的优化，考察了乙酰胆碱生物嗅探器的选择性和动态范围。AcH生物嗅探器显示出短的测量时间（在2分钟内）和宽的动态范围，用于测定气态AcH，0.02-10 ppm，其包括呼出气中的典型AcH浓度（1.2-6.0 ppm）。此外，基于ADH的特异性，AcH生物嗅探器对气态AcH表现出高选择性。传感器输出仅从含乙酰胆碱的标准气体样品中观察到。最后，乙酰胆碱生物嗅探器被应用于测量呼气中乙酰胆碱浓度从健康受试者摄入酒精后。结果，观察到具有不同醛脱氢酶2型（ALDH 2）表型的受试者之间AcH浓度的显著差异。AcH生物嗅探器可以用于呼吸测量，并且由于其检测原理的多功能性，可以预期基于呼吸分析的疾病筛查或代谢评估的应用，这允许其通过使用依赖于NADH的酶来测量其他VOC。
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.