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Validated assay for the evaluation of multiple glucuronidation activities in human liver microsomes via liquid chromatography-tandem mass spectrometry

机译:通过液相色谱-串联质谱法评估人肝微粒体中多种葡糖醛酸化活性的验证方法

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摘要

A sensitive and high-throughput liquid chromatography-tandem mass spectrometry system was developed and validated for the simultaneous determination of major human hepatic UDP-glucuronyltransferase forms in human liver microsomes. The analytes were detected using a triple-quadrupole mass spectrometer equipped with an electrospray ionization source in the negative ion and selected reaction monitoring modes. The method provided satisfactory linear concentration range, accuracy, precision, and stability. The developed method was successfully applied to the enzyme kinetic study of estradiol-3-O-glucuronidation, 4-methyl-umbelliferone-O-glucuronidation, propofol-O-glucuronidation, and 3-azido-3-deoxythymidine glucuronidation in human liver microsomes.
机译:开发了灵敏且高通量的液相色谱-串联质谱系统,并验证了该系统可同时测定人肝微粒体中主要的人肝UDP-葡萄糖醛酸转移酶形式。使用配备有电喷雾电离源的三重四极杆质谱仪以负离子和选定的反应监测模式检测分析物。该方法提供了令人满意的线性浓度范围,准确度,精密度和稳定性。该方法成功用于人体肝微粒体中雌二醇-3-O-葡萄糖醛酸化,4-甲基-伞形酮-O-葡萄糖醛酸化,丙泊酚-O-葡萄糖醛酸化和3-叠氮基-3-脱氧胸苷葡萄糖醛酸化的酶动力学研究。

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