超高效液相色谱法测定癫痫患儿拉考沙胺血药浓度

Determination of Blood Concentration of Lacosamide in Children with Epilepsy by Ultra-High Performance Liquid Chromatography

目的:建立测定癫痫患儿拉考沙胺(LCM)血药浓度的超高效液相色谱(UPLC)方法。方法:内标为盐酸普萘洛尔;色谱柱为Waters ACQUITY UPLC BEH C_(18)柱(100 mm×2.1 mm,1.7μm);流动相为甲醇-10 mmol/L磷酸二氢铵溶液(45:55,用甲酸调至pH 4.0),流速0.2 mL/min,进样量2μL;检测波长240 nm;柱温30℃。结果:LCM和普萘洛尔的保留时间分别为2.468 min和5.475 min。LCM(X)在1.0~40.0μg/mL浓度范围内与峰面积(Y)的线性关系良好(Y=0.0494X+0.0222,r=0.9998),检测下限为0.50μg/mL(S/N=4),定量下限为1.00μg/mL(S/N=12)。日内精密度1.88%~3.29%,日间精密度2.33%~5.68%,准确度88.75%~105.29%,加样回收率为96.58%~101.76%。LCM血清样品的室温稳定性、长期稳定性和冻融稳定性均较好(偏差<15%)。结论:该方法预前处理简单,分析灵敏度高、特异性强,稳定可靠,能够满足临床LCM治疗药血浓度监测的需求。

Objective:To establish the ultra-high performance liquid chromatography(UPLC)method for determination of blood concentration of lacosamide(LCM)in children with epilepsy.Methods:Phenytoin sodium was used as the internal standard.The Waters ACQUITY UPLC BEH C_(18) column(100 mm×2.1 mm,1.7μm)was used,the mobile phase was methanol-10 mmol/L ammonium dihydrogen phosphate solution(45∶55,adjusted with formic acid to pH 4.0),the flow rate was 0.2 mL/min,the injection volume was 2μL,the detection wavelength was 240 nm,and the column temperature was 30℃.Results:The retention time of LCM and propranolol was 2.468 min and 5.475 min,respectively.LCM(X)showed good linearity with peak area(Y)in the concentration range from 1.0 to 40.0μg/mL(Y=0.049,4X+0.022,2,r=0.999,8),the lower limit of detection was 0.50μg/mL(S/N=4)and the lower limit of quantification was 1.00μg/mL(S/N=12).The intra-day precision was from 1.88%to 3.29%,the inter-day precision was from 2.33%to 5.68%,the accuracy was from 88.75%to 105.29%,and the extraction recovery rate was from 96.58%to 101.76%.The room temperature stability,long-term stability and freeze-thaw stability of LCM samples were good(deviation<15%).Conclusion:The UPLC method has the advantages of simple pre-treatment,high analytical sensitivity,specificity,stability and reliability,and can meet the needs of clinical drug concentration monitoring for LCM treatment.