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辅酶ⅡNADP(H)广泛存在于动物、植物、微生物及培养细胞中。NADP⁺和NADPH含量的测定可获得NADP(H)总量（NADPH + NADP⁺）及NADPH/NADP⁺比值，其变化与磷酸戊糖途径（PPP）、生物合成及抗氧化反应密切相关。NADPH/NADP⁺比值不仅是细胞氧化还原状态的重要标志之一，而且在PPP途径、生物合成和抗氧化代谢中具有重要调控作用。 本试剂盒采用酸性和碱性提取液分别提取样品中的NADP⁺和NADPH；NADPH通过PMS递氢作用还原氧化型噻唑蓝（MTT）生成甲臜（formazan），于570 nm处检测吸光度，从而定量NADPH含量；NADP⁺则先经6-磷酸葡萄糖脱氢酶（G6PDH）还原为NADPH，再通过上述MTT还原体系进行测定，从而实现NADP⁺和NADPH的分别定量。
Coenzyme II NADP(H) is widely present in animals, plants, microorganisms, and cultured cells. The determination of NADP⁺ and NADPH contents yields the total NADP(H) (NADPH + NADP⁺) and the NADPH/NADP⁺ ratio. Changes in these parameters are closely associated with the pentose phosphate pathway (PPP), biosynthesis, and antioxidant reactions. The NADPH/NADP⁺ ratio is not only an important indicator of cellular redox status but also plays a significant regulatory role in the PPP pathway, biosynthesis, and antioxidant metabolism. This kit employs acidic and alkaline extraction buffers to separately extract NADP⁺ and NADPH from samples. NADPH reduces oxidized thiazolyl blue tetrazolium bromide (MTT) to formazan via the electron-carrying action of phenazine methosulfate (PMS), and the absorbance is measured at 570 nm for NADPH quantification. NADP⁺ is first reduced to NADPH by glucose-6-phosphate dehydrogenase (G6PDH), followed by quantification through the same MTT reduction system, thereby enabling the separate quantification of NADP⁺ and NADPH.
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