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衍生化LC-MS/MS法测定大鼠血浆中内源性谷胱甘肽的含量

徐鹏遥, 杨漾, 苏梦翔, 狄斌

徐鹏遥, 杨漾, 苏梦翔, 狄斌. 衍生化LC-MS/MS法测定大鼠血浆中内源性谷胱甘肽的含量[J]. 中国药科大学学报, 2018, 49(2): 209-214. DOI: 10.11665/j.issn.1000-5048.20180211
引用本文: 徐鹏遥, 杨漾, 苏梦翔, 狄斌. 衍生化LC-MS/MS法测定大鼠血浆中内源性谷胱甘肽的含量[J]. 中国药科大学学报, 2018, 49(2): 209-214. DOI: 10.11665/j.issn.1000-5048.20180211
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XU Pengyao, YANG Yang, SU Mengxiang, DI Bin. Determination of endogenous glutathione in rat plasma by a new derivative LC-MS/MS method[J]. Journal of China Pharmaceutical University, 2018, 49(2): 209-214. DOI: 10.11665/j.issn.1000-5048.20180211
Citation: XU Pengyao, YANG Yang, SU Mengxiang, DI Bin. Determination of endogenous glutathione in rat plasma by a new derivative LC-MS/MS method[J]. Journal of China Pharmaceutical University, 2018, 49(2): 209-214. DOI: 10.11665/j.issn.1000-5048.20180211
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衍生化LC-MS/MS法测定大鼠血浆中内源性谷胱甘肽的含量

  • 1.

    中国药科大学江苏省药物分子设计与成药性优化重点实验室

  • 2.

    中国药科大学蛋白质化学与结构生物学重点实验室

  • 3.

    中国药科大学药物分析系

Determination of endogenous glutathione in rat plasma by a new derivative LC-MS/MS method

摘要

    摘要
  • 摘要: 建立一种巯基衍生化LC-MS/MS法测定大鼠血浆中内源性谷胱甘肽含量的方法。血浆样品经衍生化处理后采用电喷雾离子源(ESI)-三重四极杆质谱仪于正离子模式下进行选择反应检测(SRM)。色谱柱为Zorbax HILIC PLUS(4.6 mm×100 mm，3.5 μm)；流动相为乙腈-水(含0.1%甲酸)(75∶25)；流速为1 mL/min；柱温为35 ℃。甘氨酰酪氨酸作为内标物，4-(N-马来酰亚胺)苯基三甲基碘化铵(MPTA)作为衍生化试剂。方法学验证结果表明:衍生化后的谷胱甘肽在3.000~2 000 ng/mL范围内线性良好，r=0.997 1，最终定量下限约为10 pmol/L。准确度、提取回收率和基质效应均符合生物样本分析的要求。该方法对生物体内谷胱甘肽准确的定量、定性分析，以及其存在状态的正确评价有着重要的临床意义。
    Abstract: To develop a rapid and sensitive liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the determination of endogenous glutathione in rat plasma. Glycyltyrosine was used as the internal standard(IS)and 4-(N-maleimido)phenyl trimethylammonium iodide(MPTA)was used as the derivation reagent. Chromatographic separation was achieved on a Zorbax HILIC PLUS column(4. 6 mm×100 mm, 3. 5 μm)and the mobile phase consisted of acetonitrile and 0. 1% formic acid(75 ∶25)pumped at a flow rate of 1. 0 mL/min. Detection was carried out on a triple quadrupole tandem mass spectrometer by selected reaction monitoring(SRM)in the can meet positive ion mode. The linearity ranged from 3. 000 to 2 000 ng/mL(r=0. 997 1); and the limit of detection of glutathione in rat plasma was 10 pmol/L. Matrix effect, stability, precision and accuracy of the method met the requirements. The proposed method was proved to be selective and sensitive, which is suitable for the quantification of endogenous glutathione in rat plasma.
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  • 刊出日期:  2018-04-24

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