用毛细管电泳-电致化学发光法定量测定药品中盐酸倍他司汀

Determination of Betahistine Hydrochloride in Pharmaceuticals by Pre-column Derivatization Capillary Electrophoresis Coupled with Electrochemiluminescence

本文选用L-精氨酸为内标物,并采用纳米粒子/交联壳聚糖涂饰开管柱为分析柱,成功建立了用柱前衍生毛细管电泳-电致化学发光法(CE-ECL)定量测定药品中盐酸倍他司汀的新方法。盐酸倍他司汀经N-甲基化衍生反应后,它与联吡啶钌检测试剂的ECL共发光强度能增长14倍,可极大提高分析检测的灵敏度。再采用内标标准曲线法进行定量时,又可以彻底消除既有CE-ECL法发光试剂消耗造成的响应信号衰减及其衍生反应条件波动导致工作曲线弯曲的缺陷。在实验优化的分析条件下,待测物盐酸倍他司汀与内标物L-精氨酸衍生物的电泳峰高比值与标样浓度在5~800μmol/L范围内呈现良好的线性关系(R^(2)=0.9995),检出限为0.79μmol/L(S/N=3),相对标准偏差(RSD)小于0.9%。同时,本法对三种药品制剂中的盐酸倍他司汀进行了实样测定,加标回收率能达到96.2%~104%范围内。

In this paper,a new method for the quantitative determination of betahistine hydrochloride by pre-column derivatization reaction capillary electrophoresis coupled with electrochemiluminescence(CE-ECL)was successfully established using a nanoparticles/cross-linked chitosan coating open-tube column as the internal standard.After the N-methylation derivatization reaction of betahistine hydrochloride,its ECL co-luminescence intensity with bipyridyl ruthenium detection reagent can be increased by 14 times,which can greatly improve the sensitivity of the analytical detection.When the internal standard curve method is used for quantification,the defects of the working curve caused by the depletion of luminescent reagents of the existing CE-ECL method and the fluctuation of the derivatization reaction conditions can be completely eliminated.Under the experimentally optimized analytical conditions,the electrophoretic peak intensity ratios of betahistine hydrochloride and L-arginine derivatives of the internal standard showed good linearity(R^(2)=0.9995)in the range of 5~800μmol/L with the detection limit of 0.79μmol/L(S/N=3)and the relative standard deviation(RSD)less than 0.9%.Meanwhile,the method was applied to the determination of betahistine hydrochloride in three pharmaceutical preparations with the spiked recoveries in the range of 96.2%~104%.