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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。
Aim To study the degradation products of reduced glutathione (GSH) under different destructive conditions. On the basis of this study, a method for the determination of glutathione (GSSG) impurities was established. Methods CenturySIL AQ C18 column (4.6 mm × 200 mm, 5 μm), 50 mmol.L-1 potassium dihydrogen phosphate solution (containing 10 mmol.L-1 sodium heptanesulfonate, pH adjusted to 3.0 with phosphoric acid) - methanol (97: 3 by volume) as mobile phase. The detection wavelength was 210 nm. Results By comparing HPLC retention times, the main degradation products in the disrupted sample were identified as GSSG, dipeptide cysteinylglycine, glutamic acid, cysteine, glycine and cystine. Calculated by external standard GSH GSH GSH content of 0.32%. Conclusion The established method of GSSG determination is accurate and reproducible. It can be used for the control of GSSG in GSH raw materials.