Method for the determination of the reduced form of nicotinamide adenine dinucleotide
申请人:Kyowa Medex Co. Ltd.
公开号:EP0153872A2
公开(公告)日:1985-09-04
A method is disclosed for the determination of NAD(P)H which comprises reacting the NAD(P)H with (1) peroxidase or thiol reductase and (2) diaphorase or an electron carrier, in the presence of a chromogen and determining the pigment thus formed. The chromogens used are compounds of the formula(l):
wherein Y is hydrogen or hydroxyl; R1, R2 and R3 are each
groups wherein Z is hydroxyl, amino, substituted amino, alkyl, substituted alkyl, alkenyl, aryl, substituted aryl, acyl, halogen, nitro, sulfo, carboxyl, or alkoxy; n is 0, 1, 2 or 3; provided that at least one Z in R1, R2 and R3 is hydroxyl, amino, or substituted amino.
Also disclosed are reagents comprising the chromogen in admixture with the peroxidase or thiol oxide reductase and the diaphorase or electron carrier.
本发明公开了一种测定 NAD(P)H 的方法,该方法包括使 NAD(P)H 与(1)过氧化物酶或硫醇还原酶和(2)二磷酸酶或电子载体在显色剂存在下发生反应,并测定由此形成的色素。所用的显色剂为式(l)化合物:
其中 Y 为氢或羟基;R1、R2 和 R3 分别为以下基团
基团,其中 Z 是羟基、氨基、取代氨基、烷基、取代烷基、烯基、芳基、取代芳基、酰基、卤素、硝基、磺基、羧基或烷氧基;n 是 0、1、2 或 3;条件是 R1、R2 和 R3 中至少有一个 Z 是羟基、氨基或取代氨基。
另外还公开了一些试剂,包括与过氧化物酶或氧化硫还原酶和二磷酸盐酶或电子载体混合的发色剂。