Highly sensitive voltammetric determination of NADH based on N-CQDs decorated SnO <sub>2</sub>/ionic liquid/carbon paste electrode

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Abstract

A highly sensitive and selective modified electrode was successfully developed for the monitoring of nicotinamide adenine dinucleotide (NADH) in the presence of folic acid. In this regard, a carbon paste electrode (CPE) was functionalized by the nitrogen-doped carbon quantum dots/tin oxide (N-CQDs/SnO ₂) nanocomposite and 1-butyl-2,3-dimethyl imidazolium hexafluorophosphate ([C ₄DMIM][PF ₆]) ionic liquid (IL). The structure and surface morphology of the nanocomposite were characterized by various methods, including field emission scanning electron microscopy, energy dispersive spectroscopy (EDS), high-resolution transmission electron microscopy (HR-TEM), and x-ray diffraction (XRD). The modified electrode displayed powerful and long-lasting electron mediating activity, with well-separated NADH and folic acid oxidation peaks. The sensing response of the developed [C ₄DMIM][PF ₆]/N-CQDs/SnO ₂/CPE platform was evaluated by determining NADH via the voltammetric technique under the optimized operating conditions. The current peaks of the square wave voltammograms of NADH and folic acid increased linearly with enhancing its concentrations within the ranges of 0.003–275 μM NADH and 0.4–380 μM folic acid. The detection limits for NADH and folic acid were obtained at 0.8 nM and 0.1 μM, respectively. Interference species such as glucose, urea, tryptophan, glycine, methionine, and vitamin B ₁₂ had no influence on the ability of the fabricated modified electrode to detect the target species. The low detection limit, high sensitivity, excellent selectivity, superior stability, and cost-effectiveness made it suitable for the quantification of NADH in the real biological samples with the recovery percent values in the range of 97.5%–103%.

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Ionic Liquids for Clean Technology

Kenneth Seddon (1997)

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Quantitative flux analysis reveals folate-dependent NADPH production

Jing-Jing Fan, Jiangbin Ye, Jurre Kamphorst … (2014)

ATP is the dominant energy source in animals for mechanical and electrical work (e.g., muscle contraction, neuronal firing). For chemical work, there is an equally important role for NADPH, which powers redox defense and reductive biosynthesis 1 . The most direct route to produce NADPH from glucose is the oxidative pentose phosphate pathway (oxPPP), with malic enzyme sometimes also important. While the relative contribution of glycolysis and oxidative phosphorylation to ATP production has been extensively analyzed, similar analysis of NADPH metabolism has been lacking. Here we demonstrate the ability to directly track, by liquid chromatography-mass spectrometry, the passage of deuterium from labeled substrates into NADPH, and combine this approach with carbon labeling and mathematical modeling to measure cytosolic NADPH fluxes. In proliferating cells, the largest contributor to cytosolic NADPH is the oxPPP. Surprisingly a nearly comparable contribution comes from serine-driven one-carbon metabolism, where oxidation of methylene tetrahydrofolate to 10-formyl-tetrahydrofolate is coupled to reduction of NADP+ to NADPH. Moreover, tracing of mitochondrial one-carbon metabolism revealed complete oxidation of 10-formyl-tetrahydrofolate to make NADPH. Since folate metabolism has not previously been considered an NADPH producer, confirmation of its functional significance was undertaken through knockdown of methylenetetrahydrofolate dehydrogenase (MTHFD) genes. Depletion of either the cytosolic or mitochondrial MTHFD isozyme resulted in decreased cellular NADPH/NADP+ and GSH/GSSG ratios and increased cell sensitivity to oxidative stress. Thus, while the importance of folate metabolism for proliferating cells has been long recognized and attributed to its function of producing one carbon units for nucleic acid synthesis, another crucial function of this pathway is generating reducing power.