超高效液相色谱紫外检测法测定吡仑帕奈在癫痫患者血浆中的药物浓度

Determination of the concentration of perampanel in the plasma of patients with epilepsy using ultra performance liquid chromatography-ultraviolet detection

目的:吡仑帕奈是我国新获批的新型抗癫痫药物。本研究旨在开发一种简单、快速的超高效液相色谱(ultra-high performance liquid chromatography,UPLC)法来测定癫痫患儿血浆中吡仑帕奈的药物浓度。方法:色谱柱为Waters ACQUITY UPLC BEH C柱(100 mm×2.1 mm,1.7μm),流动相为乙腈-1.0‰的甲酸水溶液(40∶60,v/v),流速0.2 mL·min^(-1),进样量8μL,检测波长320 nm,柱温30℃。采用内标法(内标为盐酸普萘洛尔)进行定量分析。结果:吡仑帕奈和内标溶液的保留时间分别为6.4和2.6 min^(-1)。吡仑帕奈在0.100~3.200μg·mL浓度范围内与峰面积(Y)的线性关系良好(Y=0.1099X+0.0023,r=0.999)。定量下限(LLOQ)、低浓度质控(LQC)、中浓度质控(MQC)和高浓度质控(HQC)的日内和日间精密度的相对标准偏差(RSD)值均<5.00%,准确度在理论浓度的±15.00%内,提取回收率为98.65%~101.18%,RSD值均<5.00%,室温、短期、长期和冻融冻存稳定性均较好(RSD<10.00%)。结论:该UPLC法前处理简单、分析灵敏度高、特异性强,能够满足临床吡仑帕奈治疗药物浓度监测的需求。

Objective:Perampanel is a newly approved antiepileptic drug in China.This study aimed to develop a simple and rapid ultra-high performance liquid chromatography(UPLC)method to determine the drug concentration of perampanel in the plasma of children with epilepsy.Methods:The separation was achieved by Waters ACQUITY UPLC BEH Ccolumn(100 mm×2.1 mm,1.7μm)using the mobile phase of acetonitrile-1.0‰formic acid aqueous solution(40∶60,v/v)at a flow rate of 0.2 mL·min^(-1).The injection volume was 8μL,the detection wavelength was set at 320 nm,and the column temperature at 30℃.The internal standard method(internal standard was propranolol hydrochloride)was used for quantitative analysis.Results:The retention times of perampanel and internal standard were 6.4 and 2.6 min^(-1),respectively.A good linear relationship between perampanel and peak area Y was obtained in the concentration range of 0.100~3.200μg·mL(Y=0.1099X+0.0023,r=0.999).The intra-day and inter-day precision’s relative standard deviation(RSD)values of LLOQ,LQC,MQC and HQC are all under 5.00%,the accuracy is within±15.00%of the theoretical concentration,the extraction recovery is 98.65%to 101.18%with the RSD values all under 5.00%.Room temperature,short-term,long-term and freeze-thaw storage stability are all satisfactory(RSD<10.00%).Conclusion:The UPLC method has the advantages of simple pretreatment,high sensitivity and specificity,and can meet the need of clinical drug concentration monitoring for perampanel.