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Updated: Jan 25, 2026

Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases
Published on: February 24, 2018
Simple aminophenol-based electrochemical probes for non-enzymatic, dual amperometric detection of NADH and hydrogen
Murugan Thiruppathi1, Pei-Ying Lin1, Yi-Te Chou1
1Bioanalytical Chemistry and Nanobiomedicine Laboratory, Department of Biochemical Science and Technology, National Taiwan University, Taipei 10617, Taiwan.
Abstract:
Non enzymatic detection of NADH and H2O2 is of practical significance for both environmental and biological prospective. However, there is no simple, straight forward electrochemical sensor available for sensing of them in real samples. Addressing this challenge, we report a simple stimuli responsive aminophenol, pre-anodized screen printed carbon electrode (SPCE*/AP) based electrochemical probes for dual detection of NADH and H2O2. Aminophenol prepared and adsorbed on the electrode from aminophenylboronic acid via boronic acid deprotection with H2O2. The SPCE*/AP fabricated with this process was characterized by cyclic voltammetry (CV), scanning electron microscope (SEM), Raman spectroscopy, UV-visible spectroscopy, and X-ray photoelectron spectroscopy (XPS). Amperometric detection results showed that SPCE*/AP electrodes exhibited linearity from 50 µM to 500 µM and from 200 µM to 2 mM with a detection limit (S/N = 3) of 4.2 µM and 28.9 µM for NADH and H2O2, respectively. Excellent reproducibility and selectivity for NADH and H2O2 were observed for this electrochemical platform. In addition, the matrix effect was investigated further using the same technique to analyze NADH and H2O2 in human urine samples, human serum samples, cell culture medium (containing 10% fetal bovine serum, FBS), and environmental water samples (tap water and rain water). Also, the present sensor demonstrated promising outcomes with living cells (normal cells and cancer cells).
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