高效液相色谱-质谱串联法测定人血浆特拉唑嗪的浓度

HPLC-MS/MS Determination of Terazosin in Human Plasma

目的建立一种简便、灵敏的测定人体血浆中特拉唑嗪浓度的高效液相色谱-质谱串联(HPLC-MS/MS)法。方法血浆样品采用叔丁基甲醚进行液相萃取后,进样分析。采用Inertsil-C18色谱柱(2.1 mm×150 mm,3.0μm),流动相:甲醇-水(2 mmol.L-1醋酸铵)=90∶10,采用正离子、多反应监测方式测定样品浓度。检测离子为m/z 388.4μm/z 247.4(特拉唑嗪)和m/z 384.2μm/z 231.3(哌唑嗪,内标)。结果特拉唑嗪血浆样品在0.2～50.0 ng.mL-1浓度范围线性关系良好(r=0.997 3);最低定量浓度为0.2 ng.mL-1。日内与日间相对标准偏差(RSD)均<15%,回收率在98.83%～100.67%,低、中、高绝对回收率分别为75.40%,70.28%,67.39%,符合生物样品分析要求。结论该方法简便快速、灵敏准确,适用于特拉唑嗪在人体内的药动学研究。

Objective To establish a simple and sensitive high performance liquid chromatography tandem mass spectrometry（HPLC-MS/MS） method for determination of terazosin in human plasma. Methods The plasma samples were precipitated with the liqui=d extraction by tert-butyl ether and then were injected directly into the HPLC–MS/MS system.Inertsil C18 column（2.1 mm×150 mm,3.0 μm） was used,and methanol-2 mmol·L-1ammonium acetate（90：10） was taken as the mobile phase by gradient elution.Terazosin and the internal standard（prazosin） were separated by HPLC and quantitated by MS/MS by using electrospray ionization（ESI） and multiple reaction monitoring（MRM） in the positive ion mode.The most intense ＋ MRM transition of terazosin at m/z 388.4μm/z→m/z247.4 was used for quantitation,and the transition at m/z 384.2μm/z→m/z231.3 was used to monitor prazosin. Results The liner range of terazosin was 0.2-50 ng·mL-1（r= 0.9973）,while the minimal detectable plasma concentration was 0.2 ng·mL-1.The average recovery was between 98.83% and 100.67%.Intra-day and inter-day RSD were less than 15%.Low,medium and high absolute recoveries were 75.40%,70.28%,67.39%,in line with biological sample analysis requirements. Conclusion This method is simple,sensitive and accurate,which is suitable for the study of pharmacokinetics of terazosin in human plasma and urine.