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基于磷酸化组蛋白H2AX质谱定量分析的药品基因毒性杂质的毒效应导向高通量筛查方法研究

瞿敏敏 ,  陈佳 ,  徐斌 ,  张雅姣 ,  徐华 ,  谢剑炜

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引用本文: 瞿敏敏, 陈佳, 徐斌, 张雅姣, 徐华, 谢剑炜. 基于磷酸化组蛋白H2AX质谱定量分析的药品基因毒性杂质的毒效应导向高通量筛查方法研究[J]. 分析化学, 2021, 49(9): 1531-1539. doi: 10.19756/j.issn.0253-3820.211096 shu
Citation:  QU Min-Min,  CHEN Jia,  XU Bin,  ZHANG Ya-Jiao,  XU Hua,  XIE Jian-Wei. A Toxic Effect-Directed Screening Method for Genotoxic Impurities in Drugs Based on Mass Spectrometry Quantitative Analysis of Phosphorylated Histone H2AX[J]. Chinese Journal of Analytical Chemistry, 2021, 49(9): 1531-1539. doi: 10.19756/j.issn.0253-3820.211096 shu

基于磷酸化组蛋白H2AX质谱定量分析的药品基因毒性杂质的毒效应导向高通量筛查方法研究

  • 军事科学院军事医学研究院毒物药物研究所, 抗毒药物与毒理学国家重点实验室, 北京 100850

    • 通讯作者: 徐华,E-mail:huarxu@163.com
  • 基金项目:

    国家重点研发计划项目（No.2018YFC1602600）和国家自然科学基金项目（No.21974151）资助。

摘要: 细胞中组蛋白H2AX磷酸化水平的变化能灵敏指示DNA断裂的毒性反应，基于此，本研究建立了药品中基因毒性杂质的同位素稀释高效液相色谱-串联质谱高通量筛查方法。H2AX及其磷酸化特异肽段的检出限分别为1和2 ng/mL，准确度和精密度均符合方法学要求。以两种重要的基因毒性杂质甲磺酸乙酯（EMS）及N-亚硝基二甲胺（NDMA）为代表，考察了两种具有不同药物代谢酶体系的受试细胞HepG2和HeLa，结果表明，EMS在两种受试细胞中均呈现基因毒性特点，而NDMA仅在具有相应药物代谢酶的HepG2细胞中呈现基因毒性。EMS与NDMA的检测灵敏度分别为0.06和0.03 μg/g，满足国际限量检出标准要求（EMS：0.6 μg/g，NDMA：0.3 μg/g）。将本方法应用于5种市售药片中基因毒性杂质的筛查，药片用水溶解后即直接与细胞孵育，药物活性成分及赋形剂不干扰测定，结果表明，本方法具有简便、灵敏度高和准确度好的特点，可用于药品中基因毒性杂质的快速高通量筛查。

English


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  • 收稿日期:  2021-01-31
  • 修回日期:  2021-04-06
通讯作者: 陈斌, bchen63@163.com
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