Determination of Significance：

Coenzyme I include both reduced and oxidized forms and plays a role in hydrogen transfer in biological oxidation. Oxidized coenzyme I also called nicotinamide adenine dinucleotide (NAD+) which is the coenzyme of dehydrogenase. It is an important role in glycolysis, gluconeogenesis, tricarboxylic acid cycle and respiratory chain. Intermediate product will transfer hydrogen to NAD make it become NADH(reduced coenzyme I). NADH acts as a carrier for hydrogen and synthesizes ATP by chemosmosis coupling in the respiratory chain. NADH has important physiological significance in the body. It is closely related to substance metabolism, energy metabolism, anti-cell aging, anti-oxidation and the occurrence of some diseases. A decrease in coenzyme I levels in the body can lead to cell damage or death.

Measurement Principle：

Extract the sample of NAD+ and NADH with acidic and alkaline extract solution respectively, NADH reduces the oxidized Thiazolyl Blue Tetrazolium Blue (MTT) to form formazan by hydrogen transfer from PMS, formazan has characteristic absorption at 570nm. NAD could be reduced to NADH by alcohol dehydrogenase. Further, MTT reduction method was used to detect NAD+.

Self Provided：

Microplate reader/Spectrophotometer, desk centrifuge, desk centrifuge, transferpettor, micro glass cuvette/ 96 well flat-bottom plate, mortar/ homogenizer, ice and distilled water.

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