【摘要】 目的:建立人血清还原型谷胱甘肽浓度的LC-MS/MS检测方法,并研究甲磺酸帕拉德褔韦在慢性乙肝患者中潜在的谷胱甘肽耗竭风险。方法:以阿德福韦酯(10 mg·d^-1)和富马酸替诺福韦二吡呋酯(300 mg·d^-1)为对照,慢性乙型肝炎患者为受试者,监测多次口服甲磺酸帕拉德褔韦(90 mg·d^-1)的谷胱甘肽血清浓度。血清样品经N-乙基马来酰亚胺的维生素C水溶液衍生化及甲醇蛋白沉淀后检测。分析柱为Phenomenex Synergi Fusion-RP 80A觷column,流动相为2 mmol·L^-1甲酸铵和0.1%甲酸（A）、甲醇（B）,梯度洗脱。谷胱甘肽及内标（卡托普利）衍生物的离子对m/z为433.1→304.1、343.1→228.1。结果:该方法的线性关系（20²000）ng·m L^-1、精密度、稳定性、回收率等均符合方法学要求。甲磺酸帕拉德褔韦组的谷胱甘肽浓度为（126.47±34.75）ng·m L^-1（x±s）。结论:本方法符合方法学要求,可用于人体药动学研究。多次口服甲磺酸帕拉德褔韦(90 mg·d^-1)并无谷胱甘肽耗竭风险。更多还原

【Abstract】 Objective: To establish a sensitive LC-MS/MS method for monitoring the potential depletion of glutathione（GSH） in serum after repeated pradefovir mesylate（PRA） administration in HBV patients.Methods: The present study was conducted in HBV patients to examine the potential GSH depletion induced by PRA and evaluate the safety of oral multi-dosing of PRA, with the controls of adefovir dipivoxil（ADV） and tenofovir disoproxil fumarate（TDF）. Samples were derivatized with an N-ethylmaleimide（NEM）solution containing vitamin C to prevent GSH auto-oxidation more effectively, then deproteinized with methanol. After that, the analytes were eluted for HPLC on a Phenomenex Synergi Fusion-RP 80 A column.Captopril（CAP）, a common compound with thiol group, was the final choice of internal standard. The derivatives of GSH（GSH-NEM） and internal standard（CAP-NEM） were detected by positive multiple reaction monitoring（MRM） for their ion pairs m/z 433.1→304.1 and m/z 343.1→228.1, respectively. Results:A simple and sensitive method based on LC-MS/MS for measuring the serum concentrations of GSH has been developed and showed excellent selectivity, linearity(20-2000 ng·m L^-1), precision, accuracy, stability and recovery in this study. The GSH concentrations in serum of HBV patients after multiple-dose of 90 mg PRA was 126.47±34.75 ng·m L^-1. Conclusions: The GSH concentration in serum of HBV patients has not been significantly affected after multiple-dose of 90 mg PRA.更多还原