HPLC-MS/MS法测定癫痫患者体内普瑞巴林的血药浓度

Determination of pregabalin in epileptic patients by liquid chromatography tandem mass spectrometry

目的建立人血浆中普瑞巴林含量测定的液相色谱串联质谱方法,并用于癫痫患者的血药浓度测定。方法色谱柱:Diamonsil C_(18)柱(150 mm×4.6mm,5μm),流动相:甲醇-水(0.05%甲酸),梯度洗脱流量:1 mL·min^(-1),进样量:20μL,柱温:30℃。用电喷雾离子源进行正离子模式监测,用多反应监测模式(MRM)进行定量分析,源喷射电压为5500 V,离子源温度为650℃。考察该方法的专属性、标准曲线、检测限、精密度、回收率及稳定性。选取20名门诊癫痫患者,第1次口服普瑞巴林胶囊75 mg 1 h后,采静脉血,用HPLC-MS/MS法测定普瑞巴林的血药浓度。结果普瑞巴林在13.76~5270.00 ng·mL^(-1)内线性关系良好,最低定量下限为0.65 ng·mL^(-1)。平均回收率在94%以上,日间、日内精密度RSD值均小于15%,稳定性良好。用该方法测定20名癫痫患者血样,其血药浓度范围为1.76~2.78μg·mL^(-1)。结论本法简便、快速、灵敏度高、专属性好,可用于癫痫患者普瑞巴林血药浓度的测定。

Objective To develop a HPLC-MS/MS method for the determination of Pregabalin in epileptic patients.Methods Analysis was performed on a Diamonsil C（18） column（4.6 mm ×150 mm,5 μm）eluted with methanol and water（0.05% formic acid） in a gradient program.The flow rate was 1 mL · min^-1,the injection volume was 20μL,and the column temperature was 30℃.The multiple-reaction monitoring scanning（MRM） was employed for the quantification with switching electrospray ion source polarity in negative mode.The ion spray voltage was set at 5500 V and the turbo spray temperature was maintained at 650℃.The specificity,calibration curve,limit of detection,precision,recovery and stability of the method were investigated.20 patients with epilepsy were selected,and the serum concentration of pregabalin was determined by HPLC-MS/MS after oral administration of 75 mg pregabalin for one hour.Results Pregabalin showed a good linearity in the ranges of 13.76-5270.00 ng · mL^-1,the lower limit of quantification was 0.65 ng · mL^-1.The average recoveries varied from94.7%-106.2%.The inter and intra-day RSD were less than 15%.Pregabalin was proved to be stable under study station.The concentration of pregabalin in epileptic patients ranged from 1.76 to 2.78 μg · mL^-1.Conclusion The method is simple,rapid,and sensitive,which could be used for the determination of pregabalin in epileptic patients.