高效液相色谱-串联质谱法测定血浆中熊去氧胆酸的浓度

Determination of ursodeoxycholic acid in human plasma by liquid chromatography tandem mass spectrometry

目的：建立高效液相色谱-质谱联用的方法测定人血浆内的熊去氧胆酸浓度的方法。方法：采用Shimadzu VP-ODS(4．6 mm×150 mm,5μm)色谱柱；柱温25℃；流动相为(A)乙腈-水(含5 mmol·L^(-1)的乙酸铵)(35：65,V/V),(B)乙情；梯度程序为0～2min,A：B=94：6；2～4．5min,A：B= 80：20：4．5～7min,A：B=94：6；流速为1．0 mL·min^(-1)；通过液相串联质谱,电喷雾离子源(ESI),以选择反应监测(SRM)方式进行检测；离子极性监测负离子(-)；检测离子为熊去氧胆酸m/z 391．1 [M-H]^-→391．1[M-H]^-,盐酸西替利嗪(内标)m/z 387．1[M-H]^-→387．1[M-H]^-。结果：熊去氧胆酸的最低定量限为40．144μg·L^(-1),线性范围为40．14～12 043μg·L^(-1)(r=0．995)。结论：该方法简便、灵敏度高,可以用来进行熊去氧胆酸的人体药动学和生物等效性研究。

AIM： To develope a high-performance liquid chromatographic tandem mass spectrometric method for the determination of ursodexycholic acid （UDCA） in human plasma. METHODS： Shimadzu VP-ODS （4.6 mm×150 mm, 5μm） chromatographic column was used with 25 ℃ column temperature. The mobile phase consisted of A： acetonitrile/5 mmol·L^-1 ammonium acetate （35 ： 65, V/V） and B： acetonitrile at a flowrate of 1.0mL·min^-1, with gradient procedure： 0-2min, A：B=94：6; 2-4.5min, A：B=80：20; 4.5- 7 min, A ： B = 94 ： 6. A Finnigan TSQ tandem mass spectrometer equipped with electronic spray ionization source was used as detector and operated in the negative ion mode and selected reaction monitoring （SRM）. The ion of UDCA to detect was m/z 391.1 [M-H]^-→391.1 [M-H]^-, and the cetirizine ion （internal standard） was m/z 386.9 [M-H]^-→386.9 [M-H]^-. RESULTS： The lower limit of qualification was 40.144μg·L^-1. The standard curve was in linear over a working range of 40.14 - 12 043μg·L^-1 with an standard correlation coefficient of 0.995. CONCLUSION： This method is and bioequivalence studies. simple, sensitive, and suitable for clinical pharmacokinetics