LC-MS/MS测定大鼠血浆中16种胆汁酸及在甘草减毒研究中的应用

LC-MS / MS Method for Determination of Sixteen Bile Acids in Rat Plasma and Its Application in Liquorice Detoxification Research

目的 建立测定大鼠血浆中16种胆汁酸浓度的LC-MS/MS法。方法 鼠血浆样品经过甲醇沉淀蛋白后,上清液挥干,流动相复溶后,用Xtimate^TM C18柱分离,以甲醇-醋酸铵及甲酸水溶液为流动相进行梯度洗脱,流速0.25 m L·min^-1,进样5μL分析。结果 16种胆汁酸在0.005 6～3.800 0μmol·L^-1内,线性关系良好,定量下限浓度为0.005 6μmol·L^-1,低、中、高浓度的萃取回收率在70.3%～107.2%之间,日内、日间RSD均小于12.9%。结论 该方法准确可靠,灵敏度高,处理简便,适用于血浆中胆汁酸浓度的测定,为临床血浆样品中胆汁酸含量测定提供参考。

OBJECTIVE To establish an LC-MS/MS method for determination of the concentrations of 16 bile acids in rat plasma. METHODS The plasma protein was precipitated with methanol and then the supernatant was dried. Liquid chromatographic separation was performed on a XtimateTM Cl8 column with gradient elution of mobile phase consisting of methanol-ammonium acetate and formic acid buffer, the flow rate was 0. 25 mL · min^-1 , and the injection volume was 5 μL. RESULTS The calibration curves were linear in the concentration range of 0. 005 6 - 3. 800 0 μmol· L^-1. The lower limit of quantification was 0. 005 6 μmol · L ^-1. The recoveries of the samples at low, middle and high concentrations were 70. 3% - 107.2 %. The intra-day and inter-day RSDs were less than 12. 9%. CONCLUSION This method is accurate and sensitive for determination of bile acids in rat plasma, which provides reference for the determination of bile acids in clinical samples.