UPLC-MS/MS法测定人血浆中丙硫氧嘧啶的含量

Determination of propylthiouracil in human plasma by UPLC-MS/MS

目的建立UPLC-MS/MS法测定人血浆中丙硫氧嘧啶的含量,为临床治疗药物监测(TDM)和生物等效性试验(BE)提供方法学基础。方法采用Agilent SB-C18柱(4.6 mm×150 mm,5μm),流动相为甲醇-0.1%甲酸水溶液(80∶20,V/V),等度洗脱。质谱采用AJS-ESI源,多反应监测(MRM)正离子模式,检测离子对为:丙硫氧嘧啶m/z 171.1→112.1、内标(丙硫氧嘧啶-D5)m/z 176.1→117.0。结果血浆中丙硫氧嘧啶在10~5 000 ng/ml范围内线性关系良好,r=0.999 3;批内和批间的精密度和准确度良好(RSD<10%,RE<±10%);不同浓度的基质效应均<110%,变异系数<5%;不同浓度的平均回收率为101.60%~113.56%,符合方法学要求。结论该方法快速简便、灵敏准确,适用于血浆中丙硫氧嘧啶的含量测定。

Objective To establish a method for the determination of propylthiouracil in human plasma by UPLCMS/MS and provide methodological basis for therapeutic drug monitoring(TDM) and bioequivalence test(BE) in clinical. Methods The chromatographic separation was performed on an Agilent SB-C18 column(4.6 mm×150 mm, 5 μm), the mobile phase was methanol and water containing 0.1% formic acid(80∶20, V/V), isocratic elution. MS condition was optimized in the positive ion detection mode by multiple reaction monitoring(MRM), along with the Agilent JetStream electrospray source interface(AJS-ESI).The precursors to the product ion transitions were m/z 171.1→112.1 for propylthiouracil and m/z 176.1→117.0 for the internal standard(IS). Results The calibration curve was linear in the range of 10-5 000 ng/ml for propylthiouracil in human plasma,r=0.999 3. The intra-day and inter-day precision and accuracy were good(RSD<10%,RE<±10%). The matrix effect of different concentrations was less than 110% and the coefficient of variation was less than 5%. The average recovery of different concentrations was 101.60%-113.56%, which conformed with the requirement of methodological validation. Conclusion The method is rapid, sensitive and accurate, which can be used for the determination of propylthiouracil in human plasma.