HPLC-MS/MS法快速测定人血浆中氟哌噻吨浓度及生物等效性研究

目的建立测定人血浆中氟哌噻吨浓度的高效液相串联质谱法（HPLC-MS/MS）,用于氟哌噻吨美利曲辛片的生物等效性研究。方法以Agilent ZORBAX Eclipse Plus C18（4.6mm×150mm,5μm）为色谱柱,流动相为乙腈（含1%甲酸）：0.02mol/L甲酸铵水溶液（80：20,V：V）,流速：0.8ml/min;柱温：40℃,以醋酸乙酯：二氯甲烷（4：1,V：V）为提取剂。样品经电喷雾离子源正离子化后,通过三重四级杆串联质谱仪,采用选择性反应监测（SRM）对氟哌噻吨（m/z435.2→305.1）和氯普噻吨（m/z316.2→231.1）进行测定。结果氟哌噻吨的高（1μg/L）、中（0.5μg/L）、低（0.05μg/L）3个浓度的平均回收率分别为101.01%、96.19%和106.63%,日内（n=5）、日间（n=3）RSD均小于15%;分析方法的最低定量限为0.025μg/L。线性范围为：0.025～2.5μg/L,回归方程为：F=0.6625ρ＋0.0049,r=0.997（n=7）,权重为1/ρ2。结论该方法灵敏、准确、简单、快速,可用于临床血浓监测和药动学研究。

Rapid determination of flupentixol in plasma by HPLC-MS/MS and its bioequivalence

Objective To develop a HPLC-MS/MS method for flupentixol assay in plasma,and to determine its bioequivalence. Methods Flupentixol was extracted with ethyl acetate-dichloromethane.The residues were analyzed with a HPLC-MS/MS system （Agilent ZORBAX Eclipse Plus C18,4.6mm-150mm,5μm） with the mobile phase consisted of aeetonitrile （1% formic acid） with 0.02mol/L ammonium formate （80：20,V：V）.Agilent 6410 triple quad mass spectrometer system equipped with an eleetrospray ionization ion-trap source was used as the detector and operated in the positive ion mode.Selected reaction monitoring （SRM） using the precursor to product ion combinations of m/z 435.2→305.1 and m/z 316.2→231.1 was performed to detect flupentixol and the internal standard, respectively. Results The average recoveries for flupentixol were 101.01%,96.19% and 106.63%,respectively.The within-day（n=5） and between-day（n=3） precision of variation was less than 15%.The calibration curves for flupentixol had good linearity,r=0.997 （n=7）,over the range of 0.025-2.5μg/L.The limits of quantitation for flupentixol was 0.025μg/L. Conclusion The method provides a sensitive,accurate,precise and reliable analytical procedure for clinical monitoring of flupentixol plasma and its phamacokinetic studies.