Institute of Systems Biomedicine, Department of Pathology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking-Tsinghua Center for Life Sciences, Peking University Health Science Center, Beijing 100191, China.
Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education and Beijing Key Laboratory of Cardiovascular Receptors Research, Beijing 100191, China.
J Pharm Biomed Anal. 2017 Nov 30;146:53-58. doi: 10.1016/j.jpba.2017.07.020. Epub 2017 Aug 12.
A simple, sensitive and rapid method using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed and validated for quantification of glycine-β-muricholic acid (Gly-MCA) in mouse plasma for the first time. Plasma samples were pretreated with protein precipitation. The analyte and internal standard were separated on a Shimadzu Shim-pack XR-ODS column (4.6×50mm, 2.2μm) using 0.1% formic acid-water-methanol as mobile phase, with a runtime of 5min. Detection was performed using negative ion electrospray tandem mass spectrometry via multiple reaction monitoring (MRM) scan mode. The linear range was 5ng-2μg/ml (correlation coefficient >0.995) for Gly-MCA with a lower limit of quantitation of 5ng/ml. The intra-day and inter-day precision were less than 9.2% for the analyte and accuracy was from 0.4% to 7.0%. This analytical method was then successfully applied to a pharmacokinetic study of Gly-MCA following oral administration and intraperitoneal injection in mice.
首次建立并验证了一种采用高效液相色谱-串联质谱（HPLC-MS/MS）的简单、灵敏、快速的方法，用于定量检测小鼠血浆中的甘氨熊去氧胆酸（Gly-MCA）。血浆样品经蛋白沉淀预处理后，采用 Shimadzu Shim-pack XR-ODS 柱（4.6×50mm，2.2μm）进行分析，以 0.1%甲酸-水-甲醇为流动相，运行时间为 5min。采用负离子电喷雾串联质谱法，通过多反应监测（MRM）扫描模式进行检测。Gly-MCA 的线性范围为 5ng-2μg/ml（相关系数>0.995），定量下限为 5ng/ml。分析物的日内和日间精密度均小于 9.2%，准确度为 0.4%-7.0%。该分析方法随后成功应用于小鼠口服和腹腔注射后甘氨熊去氧胆酸的药代动力学研究。
