HPLC法研究还原型谷胱甘肽降解产物

目的研究还原型谷胱甘肽(GSH)在不同破坏条件下的降解产物;在此研究基础上,建立了氧化型谷胱甘肽(GSSG)杂质测定方法。方法采用CenturySIL AQ C18色谱柱(4.6 mm×200 mm,5μm),50 mmol.L-1磷酸二氢钾溶液(含10 mmol.L-1庚烷磺酸钠,用磷酸调节pH值至3.0)-甲醇(体积比97∶3)为流动相;检测波长为210 nm。结果通过对比HPLC保留时间,确定破坏样品中主要降解产物为GSSG、二肽半胱氨酰甘氨酸、谷氨酸、半胱氨酸、甘氨酸和胱氨酸。按外标法计算GSH原料中GSSG的含量为0.32%。结论建立的GSSG测定方法结果准确,重现性好,可用于GSH原料中GSSG的控制。

Study on the degradation products in reduced glutathione by HPLC

Objective To study the degradation products of reduced glutathione(GSH)in various destruction tests and establish an HPLC method for assaying glutathione oxidized(GSSG)based on above-mentioned study.Methods Century SIL AQ C18 column(4.6 mm×200 mm,5 μm)was used,the mobile phase was 10 mmol·L-1 sodium heptanesulfonate in 50 mmol·L-1 potassium dihydrogen phosphate solution(pH 3.0)-methanol(V∶V=97∶3),the detection wavelength was set at 210 nm.Results By comparison with the retention time of the standards,the major degradation products from samples of destruction tests were identified,including cysteinylglycine,GSSG and amino acids:glutamic,cysteine,glycine and cystine.The content of GSSG in GSH substances was 0.32% by external standard method.Conclusions The method of GSSG assaying is accurate,specific and can be used in the quality standard of the drug.