液相色谱-串联质谱法测定人血浆中泮托拉唑的浓度

目的: 建立LC-MS/MS法测定人血浆中泮托拉唑的浓度。 方法: 人血浆样本以乙腈沉淀蛋白后,选用Zorbax SB-C₁₈ Narrow-Bore色谱柱(150 mm×2.1 mm,5 μm),以甲醇-10 mmol·L^-1乙酸铵(65:35)为流动相,流速为0.40 mL·min^-1;选用API3200型三重四极杆串联质谱仪的多重反应监测(MRM)扫描方式进行监测,电喷雾离子化源,正离子方式,选择监测离子反应分别为m/z 384.1→m/z 200.2(泮托拉唑)和m/z 370.1→m/z 252.0(内标兰索拉唑)。 结果: 泮托拉唑和兰索拉唑的保留时间分别为1.75 min和2.36 min;血浆中泮托拉唑的线性范围为0.0100～6.00 mg·L^-1(r>0.99),定量下限为0.0100 mg·L^-1;日内、日间相对标准差(RSD)均小于5%;相对偏差(RE)均在±4%的范围以内;平均提取回收率为(97.2±5.4)%;稳定性试验中,在各种贮存条件下血浆中泮托拉唑均较稳定。 结论: 该方法快速、灵敏、准确、专属性强、重现性好,适用于人血浆中泮托拉唑浓度的测定,可应用于泮托拉唑钠肠溶片的人体生物等效性研究。

Objective: To develop an LC-MS/MS method for determination of pantoprazole in human plasma. Methods: After protein precipitation with acetonitrile,the analyte and internal standard,lansoprazole,were separated on a Zorbax SB-C₁₈ Narrow-Bore analytical column(150 mm×2.1 mm,5 μm) with the mobile phase of methanol and 10 mmol·L^-1 ammonium acetate (65:35) at a flow rate of 0.40 mL·min^-1.Detection was carried out by electrospray positive ionization mass spectrometry in the multiple reaction monitoring (MRM) mode.The MRM transitions of m/z 384.1→m/z 200.2 and m/z 370.1→m/z 252.0 were used to quantify pantoprazole and IS,respectively. Results: Pantoprazole and internal standard were eluted at 1.75 and 2.36 min,respectively.The calibration curve was linear over the concentration range of 0.0100-6.00 mg·L^-1 with the lower limit of quantitation (LLOQ) 0.0100 mg·L^-1.Intra-day and inter-day relative standard deviations were both less than 5%,and the relative errors were within±4%.The mean extract recoveries were (97.2±5.4)%.In the stability studies,pantoprazole in plasma was found to be stable under various storage conditions. Conclusion: It is a rapid,sensitive,selective and reliable method for the determination of pantoprazole in human plasma.The method was successfully applied to a bioequivalence study of pantoprazole sodium enteric-coated tablet in healthy volunteers.