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首页> 外文期刊>Journal of mass spectrometry: JMS >Development and validation of a liquid chromatography and ion spray tandem mass spectrometry method for the quantification of artesunate, artemether and their major metabolites dihydroartemisinin and dihydroartemisinin-glucuronide in sheep plasma
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Development and validation of a liquid chromatography and ion spray tandem mass spectrometry method for the quantification of artesunate, artemether and their major metabolites dihydroartemisinin and dihydroartemisinin-glucuronide in sheep plasma

机译：液相色谱和离子喷雾串联质谱法在绵羊血浆中青蒿琥酯，青蒿醚及其主要代谢物双氢青蒿素和双氢青蒿素-葡糖苷酸定量的开发和验证

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摘要

Recently, promising fasciocidal activities of artesunate and artemether were described in rats and sheep. Therefore, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to quantify artesunate, artemether and their metabolites dihydroartemisinin and dihydroartemisinin-glucuronide in sheep plasma. Protein precipitation with methanol was used for sample workup. Reversed-phase high-performance liquid chromatography (HPLC) was performed using an Atlantis C18 analytical column with a mobile phase gradient system of ammonium formate and acetonitrile. The analytes were detected by MS/MS using selected reaction monitoring (SRM) with electrospray ionisation in the positive mode (transition m/z 267.4 → 163.0). The analytical range for dihydroartemisinin, dihydroartemisinin-glucuronide and artesunate was 10-1000 ng/ml and for artemether 90-3000 ng/ml with a lower limit of quantification of 10 and 90 ng/ml, respectively. Inter- and intra-day accuracy and precision deviations were < 10%. Consistent relative recoveries (60-80%) were observed over the investigated calibration range for all analytes. All analytes were stable in the autosampler for at least 30 h (6 °C) and after three freeze and thaw cycles. The validation results demonstrated that the LC-MS/MS method is precise, accurate and selective and can be used for the determination of the artemisinins in sheep plasma. The method was applied successfully to determine the pharmacokinetic parameters of artesunate and its metabolites in plasma of intramuscularly treated sheep.
机译：最近，在大鼠和绵羊中描述了青蒿琥酯和蒿甲醚有前途的杀真菌活性。因此，开发了一种液相色谱-串联质谱（LC-MS / MS）方法来定量羊血浆中的青蒿琥酯，青蒿醚及其代谢产物双氢青蒿素和双氢青蒿素-葡萄糖醛酸。用甲醇沉淀蛋白质用于样品处理。反相高效液相色谱（HPLC）使用Atlantis C18分析柱和甲酸铵和乙腈的流动相梯度系统进行。使用选定的反应监测（SRM）通过MS / MS通过正离子模式下电喷雾电离（转换m / z 267.4→163.0）来检测分析物。双氢青蒿素，双氢青蒿素-葡糖苷酸和青蒿琥酯的分析范围为10-1000 ng / ml，蒿甲醚为90-3000 ng / ml，定量下限分别为10和90 ng / ml。日间和日内准确性和精确度偏差<10％。在所有分析物的标定研究范围内，观察到一致的相对回收率（60-80％）。在三个冷冻和融化循环后，所有分析物在自动进样器中稳定至少30 h（6°C）。验证结果表明，LC-MS / MS方法准确，准确且具有选择性，可用于测定绵羊血浆中的青蒿素。该方法已成功应用于肌肉注射绵羊的血浆中青蒿琥酯及其代谢产物的药代动力学参数测定。

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