LC-MS/MS测定大鼠血清中15种胆汁酸成分

Determination of Fifteen Bile Acids Concentration in Rats’Serum by LC-MS/MS

目的建立同时测定大鼠血清中15种胆汁酸浓度的液相色谱-串联质谱法(LC-MS/MS)。方法血清样本用乙腈蛋白沉淀后,采用Xterra RP18(4.6 mm×100 mm,3.5μm)色谱柱分离操作,以乙腈-10 mmol·L^-1醋酸铵水溶液为流动相,梯度洗脱,流速为1 mL·min^-1,用电喷雾离子源,负离子多反应监测。结果15种胆汁酸成分在定量范围内线性关系良好,日内和日间精密度均<10%,低、中、高浓度提取回收率在69.93%~88.83%。结论本研究建立的LC-MS/MS法简便、快速、专属性强,能够适用于大鼠血清中15种胆汁酸组分的检测。

OBJECTIVE To establish an LC-MS/MS method for simultaneous determination of 15 bile acids in rats’serum.METHODS Serum samples were protein precipitated with acetonitrile and the separation was performed on a Xterra RP18 column(4.6 mm×100 mm,3.5μm).All analytes were gradient eluted with acetonitrile and 10 mmol·L^-1 ammonium acetate in aqueous solution,of which flow rate was 1 mL·min^-1.Quantitative analysis of 15 bile acids was conducted by a triple-quadrupole mass spectrometer with negative-electrospray ionization(ESI)source,monitored under a multiple reaction monitoring(MRM)mode.RESULTS The calibration curves of 15 biles acids were linear in the quantitative range.The RSDs of the intra-day and inter-day precision results were all<10%.The average extraction recoveries of the control samples at low,middle and high concentrations were between 69.93%-88.83%.CONCLUSION Established LC-MS/MS method is simple,rapid and specific,and can be used for the determination of 15 bile acids in rat serum.