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Sensors 2017, 17(4), 788;

Quantitative Detection of NADH Using a Novel Enzyme-Assisted Method Based on Surface-Enhanced Raman Scattering

Beijing Key Laboratory of Bioprocess, Beijing University of Chemical Technology, Beijing 100029, China
Engineering Research Center for Semiconductor Integrated Technology, Institute of Semiconductors, Chinese Academy of Sciences, Beijing 100083, China
Analytical and Testing Center, Beijing Normal University, Beijing 100875, China
Authors to whom correspondence should be addressed.
Academic Editor: Vittorio M. N. Passaro
Received: 19 January 2017 / Revised: 30 March 2017 / Accepted: 30 March 2017 / Published: 7 April 2017
(This article belongs to the Section Physical Sensors)
Full-Text   |   PDF [2407 KB, uploaded 7 April 2017]   |  


An enzymatic method for quantitative detection of the reduced form of nicotinamide-adenine dinucleotide (NADH) using surface-enhanced Raman scattering was developed. Under the action of NADH oxidase and horseradish peroxidase, NADH can generate hydrogen peroxide (H2O2) in a 1:1 molar ratio, and the H2O2 can oxidize a chromogen into pigment with a 1:1 molar ratio. Therefore, the concentration of NADH can be determined by detecting the generated pigment. In our experiments, eight chromogens were studied, and o-tolidine (OT) was selected because of the unique Raman peaks displayed by its corresponding pigment. The optimal OT concentration was 2 × 10−3 M, and this gave the best linear relationship and the widest linear range between the logarithmic H2O2 concentration and the logarithmic integrated SERS intensity of the peak centered at 1448 cm−1. Under this condition, the limit of detection for NADH was as low as 4 × 10−7 M. Two NADH samples with concentrations of 2 × 10−4 and 2 × 10−5 M were used to validate the linear relationship, and the logarithmic deviations were less than 3%. View Full-Text
Keywords: surface enhanced Raman scattering; quantitative detection; NADH; enzyme-assisted surface enhanced Raman scattering; quantitative detection; NADH; enzyme-assisted

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Teng, H.; Lv, M.; Liu, L.; Zhang, X.; Zhao, Y.; Wu, Z.; Xu, H. Quantitative Detection of NADH Using a Novel Enzyme-Assisted Method Based on Surface-Enhanced Raman Scattering. Sensors 2017, 17, 788.

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