HPLC-MS／MS同时测定血浆中九种胆汁酸亚组分浓度

目的：建立高效液相色谱．串联质谱（HPLC．MS／MS）法测定胆酸、脱氧胆酸、鹅脱氧胆酸、石胆酸、甘氨胆酸、甘氨鹅脱氧胆酸、牛磺胆酸、牛磺脱氧胆酸、牛磺鹅脱氧胆酸九种胆汁酸亚组分浓度的方法。方法：采用色谱柱LunaC18 （2）柱（150×4．6mm，5ixm），柱温30℃，流动相以10mmol／L乙酸氨为流动相A、乙腈-甲醇（3：1）为流动相B梯度洗脱，流速：0．6mL／min，血浆样品经乙腈沉淀蛋白后进样，MRM模式定量。结果：HPLC-MS／MS法测定胆汁酸九种亚组分浓度的最低检测限为0．50-8．00ng／mL；日内和日间变异系数均小于10％；回收率在92．75％-110．07％之间。结论：所建HPLC-MS／MS法灵敏、准确，重现性好，适用于测定血浆中九种胆汁酸亚组分含量，能够为临床及科学研究提供可靠的数据参考。

Determination Nine Subcomponents of Bile Acids in Plasma by HPLC-MS/MS

Obiective： To establish a RP-HPLC-MS/MS method for determination of bile acids include cholic acid, deoxychol- ic acid, chenodeoxycholic acid, lithocholic acid, glycocholate acid, glycochenodeoxycholic acid, taurocholic acid, taurodeoxychoic acid, taurochenodeoxycholic acid in human plasma. Methods： The assay was conducted with an API-3000 LC-MS/MS system consis- ted of a Luna CIs （2） column （ 150 x4.6mm, 5μm）, column temperature was 30°C, the mobile phase A was 10 mmd/L ammoni- um acetate, and B was acetonitrile-methanol （3：1 ） for gradient elution. The flow rate was 0. 6 mL/min. Acetonitrile was used to pre- cipitate proteins from the plasma sample. Quantitated by MRM mode. The residues from the supernatant that were dissolved in metha- nol Results： The minimal detection htimit was O. 50 -8.00 ng/mL; The recovery rate of the HPLC assay was 92. 75 - 110.07% ; The RSD within-day and between-day were less 10% . Conclusion： This sensitive, accurate and simple assay was useful to detemina- tion the nine species of bile acid in human plasma.