多酶体系中谷胱甘肽HPLC分析方法研究

Study on HPLC analysis method of glutathione in multienzyme system

为了建立一种快速有效的方法来满足体外酶催化法生产谷胱甘肽中各复杂成分的定性及定量检测。依据反应液各组分的性质与特性,利用HPLC研究建立了一种新的分离方法。结果表明,通过此方法可实现L-半胱氨酸、L-谷氨酸钠、ATP、AMP、ADP、还原型谷胱甘肽、氧化型谷胱甘肽各组分的分离,分离度均>1.5,L-半胱氨酸与甘氨酸分离度>1.2;底物及产物曲线线性为R^(2)>0.99,回收率为94.6%~110.0%,连续检测6次RSD均<2.0%;且单个样品检测时间为15 min,与药典片剂检测方法45 min相比,缩短了检测时间,可快速出具检测结果,更有利于日常分析检测。

In order to establish a rapid and effective method for the qualitative and quantitative determination of complex components of glutathione produced by enzyme catalysis in vitro.According to the properties and characteristics of each component of the reaction liquid,a new separation method was established by HPLC.The results showed that the separation degree of L-cysteine,L-sodium glutamate,ATP,AMP,ADP,reduced glutathione and oxidized glutathione was greater than 1.5,and the separation degree of L-cysteine and glycine was greater than 1.2;The linear R^(2) of the substrate and product curves was greater than 0.99,the recoveries were between 94.6% and 110.0%,and the RSDS were all less than 2.0% for 6 consecutive tests;Moreover,the detection time of a single sample is 15 min,compared with the pharmacopeia tablet detection method of 45 min,the detection time is greatly shortened,the test results can be quickly issued,and it is more conducive to daily analysis and detection.