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


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1

Beijing Key Laboratory of Bioprocess, Beijing University of Chemical Technology, Beijing 100029, China

2

Engineering Research Center for Semiconductor Integrated Technology, Institute of Semiconductors, Chinese Academy of Sciences, Beijing 100083, China

3

Analytical and Testing Center, Beijing Normal University, Beijing 100875, China





*

Authors to whom correspondence should be addressed.



Academic Editor: Vittorio M. N. Passaro

Abstract 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





Autor: Haiyan Teng 1, Mingyang Lv 1, Luo Liu 1, Xin Zhang 1,* , Yongmei Zhao 2, Zhenglong Wu 3 and Haijun Xu 1,*

Fuente: http://mdpi.com/



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