Abstract
Nicotinamide adenine dinucleotide (NADH) is an important cofactor involved in metabolic redox reactions in living cells. The detection of NADH in living animal cells is a challenge. We developed a one-step monitoring method for NADH via an electrocatalytic reaction that uses a surface-modified, screen-printed electrode (SPE) having a redox active monolayer 4′-mercapto-N-phenlyquinone diamine (NPQD) formed by a self-assembled monolayer (SAM) of an aromatic thiol, 4-aminothiophenol (4-ATP). This electrode has a limit of detection (LOD) of 0.49 μM and a sensitivity of 0.0076 ± 0.0006 μM/μA in cell culture media, which indicates that it retains its selectivity. The applicability of this NADH sensor was demonstrated for the first time by cell viability monitoring via NADH-sensing in cell culture supernatants.
Highlights
Nicotinamide adenine dinucleotide (NADH) is the most well-known biomarker of the redox state of a cell
To oxidize NADH to NAD+, a potential is required as the driving force, and we found that the minimum potential for oxidation is 400 mV
We have demonstrated that changes in the NADH concentration, as determined by our electrocatalytic sensor, in live cells can be used to monitor cytotoxic events caused by the addition of toxic compounds such as Polyhexamethylene guanidine-phosphate (PHMG-p), which provides a framework for toxicity studies for the first time
Summary
Nicotinamide adenine dinucleotide (NADH) is the most well-known biomarker of the redox state of a cell. Mitochondrial dysfunction characterized by inefficient ATP production contributes to neurodegenerative diseases, such as Alzheimer’s, Parkinson’s, and Huntington’s diseases, cardiovascular disease [1,2], diabetes and metabolic syndrome [3,4,5], and lung diseases [6,7]. Many studies present various quantification methods for NADH because it exists in cells at a higher concentration than other coenzymes, and its quantification is not affected significantly by blood [8]. The classical method for the determination of NADH is an optical assay using absorbance [10]
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