Optical isopropanol biosensor using NADH-dependent secondary alcohol dehydrogenase (S-ADH)

TALANTA

卷:159

Article

Chien, Po-Jen¹  Ye, Ming²  Suzuki, Takuma¹  Toma, Koji²  Arakawa, Takahiro²  Iwasaki, Yasuhiko³  Mitsubayashi, Kohji^1,2 

[1] Tokyo Med & Dent Univ, Grad Sch Med & Dent Sci, Bunkyo Ku, 1-5-45 Yushima, Tokyo 1138549, Japan

[2] Tokyo Med & Dent Univ, Inst Biomat & Bioengn, Dept Biomed Devices & Instrumentat, Chiyoda Ku, 2-3-10 Kanda Surugadai, Tokyo 1010062, Japan

[3] Kansai Univ, Fac Chem Mat & Bioengn, 3-3-35 Yamate Cho, Suita, Osaka 5640836, Japan

关键词: Biosensor;    Isopropanol;    NADH;    Secondary alcohol dehydrogenase;    UV-LED;    Fiber-optic;   

DOI  :  10.1016/j.talanta.2016.06.036

来源: Elsevier

【 摘 要 】

Isopropanol (IPA) is an important solvent used in industrial activity often found in hospitals as antiseptic alcohol rub. Also, IPA may have the potential to be a biomarker of diabetic ketoacidosis. In this study, an optical biosensor using NADH-dependent secondary alcohol dehydrogenase (S-ADH) for IPA measurement was constructed and evaluated. An ultraviolet light emitting diode (UV-LED, lambda=340 nm) was employed as the excitation light to excite nicotinamide adenine dinucleotide (NADH). A photomultiplier tube (PMT) was connected to a two-way branch optical fiber for measuring the fluorescence emitted from the NADH. S-ADH was immobilized on the membrane to catalyze IPA to acetone and reduce NAD(+) to be NADH. This IPA biosensor shows highly sensitivity and selectivity, the calibration range is from 500 nmol L-1 to 1 mmol L-1. The optimization of buffer pH, temperature, and the enzyme-immobilized method were also evaluated. The detection of IPA in nail related cosmetic using our IPA biosensor was also carried out. The results showed that large amounts of IPA were used in these kinds of cosmetics. This IPA biosensor comes with the advantages of rapid reaction, good reproducibility, and wide dynamic range, and is also expected to use for clinical IPA detections in serum or other medical and health related applications. (C) 2016 Elsevier B.V. All rights reserved.

【 授权许可】

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