HPLC-MS/MS法同时测定人血浆中利培酮和帕利哌酮的浓度

Simultaneous determination of risperidone and paliperidone in human plasma by HPLC-MS/MS

目的建立高效液相色谱联用质谱法同时测定人血浆中利培酮和帕利哌酮的浓度。方法内标选用利培酮-d4和帕利哌酮-d4,血浆样品采用甲醇直接沉淀蛋白处理,色谱柱为Waters Atlantis T3(2.1 mm×50.0 mm,5.0μm),流动相A为含0.1%甲醇和5.00 mmol·L^(-1)醋酸铵的水溶液,流动相B为含0.1%甲醇的乙腈,梯度洗脱;应用电喷雾离子化,正离子模式下进行多反应监测利培酮(m/z 411.2→191.1),帕利哌酮(m/z 427.0→207.0)和内标利培酮-d4(m/z 415.2→195.1),帕利哌酮-d4(m/z 431.2→211.1)。结果利培酮和帕利哌酮分别在0.05~30 ng·mL^(-1)、0.05~15 ng·mL^(-1)内与峰面积线性关系良好,提取回收率在90.66%~96.82%,日内及日间RSD均小于15%,稳定性考察结果良好。结论该方法灵敏、准确、简单、快速,可用于临床血药浓度检测和药动学研究。

Objective To determine the plasma concentrations of risperidone and paliperidone by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Methods Plasma samples were treated by the protein precipitation method with methanol and the internal standard was risperidone-d4 and paliperidone-d4.Waters Atlantis T3(2.1 mm×50.0 mm,5.0μm)was used.Gradient elution was conducted with mobile phase A and B.The mobile phase A was an aqueous solution containing 0.1%methanol and 5.00 mmol·L^(-1) ammonium acetate,while B was acetonitrile containing 0.1%methanol.MS detection was used with an electrospray ionization source operated in a multi-reaction monitoring mode.The ion transitions were m/z 411.2→191.1(risperidone),m/z 427.0→207.0(paliperidone),m/z 415.2→195.1(risperidone-d4),and m/z 431.2→211.1(paliperidone-d4).Results The linearity of risperidone was 0.05~30 ng·mL^(-1) and that of pariperidone was 0.05~15 ng·mL^(-1).The inter-day and intra-day RSDs were all lower than 15%,and the method recoveries were at 90.66%~96.82%,with good stability.Conclusion This method is fast,simple,accurate and sensitive,which is suitable for plasma concentration determination and pharmacokinetic study.