HPLC-ELSD法同时测定知母药材中5种成分的含量

Simultaneous determination of five components in Anemarrhenae Rhizoma by HPLC-ELSD

目的采用高效液相色谱-蒸发光散射检测方法(HPLC-ELSD)建立同时测定知母药材中新芒果苷、芒果苷、知母皂苷BⅡ、宝藿苷Ⅰ及知母皂苷AⅢ的含量测定方法。方法采用Agilent poroshell 120 EC-C_(18)柱,流动相采用乙腈-0.2%醋酸水系统,梯度洗脱;柱温为30℃,流速为0.7 ml/min;蒸发光散射检测器以氮气为雾化气,雾化气温度为40℃,漂移管温度为90℃,氮气体积流量为2.0 L/min;进样量为20μl。结果5种成分均能达到基线分离,新芒果苷24.1~386μg/ml(r=0.9993)、芒果苷23.2~371μg/ml(r=0.9986)、知母皂苷BⅡ54.2~867.2μg/ml(r=0.9956)、宝藿苷Ⅰ5.3~84.8μg/ml(r=0.9968)、知母皂苷AⅢ10~160μg/ml(r=0.9989)的浓度范围内呈现良好的线性关系。5种成分的平均加样回收率在101.8%~105.0%之间,重复性RSD小于2.4%,知母药材中上述5种成分含量分别为1.62%、0.82%、7.36%、0.07%、0.34%。结论该方法操作简便、准确、灵敏度高,可用于知母药材的多指标成分定量测定。

Objective To establish a method for the simultaneous determination of new mangiferin,mangiferin,artemisinin BⅡ,icariin and artemisinin A in Anemarrhenae Rhizoma by high performance liquid chromatography-evaporation light scattering detector(HPLC-ELSD).Methods The column was Agilent Poroshell 120 EC-C_(18).The mobile phase used acetonitrile-0.2%acetic acid water system with gradient elution.Column temperature was 30℃.Flow rate was 0.7 ml/min.Evaporative light scattering detector used nitrogen as atomizing gas.The atomizing gas temperature was 40℃and the drift tube temperature was 90℃.The nitrogen volume flow rate was 2.00 L/min and the sample volume was 20μl.Results The five components were able to achieve baseline separation.Neomangiferin,Mangiferin,Anemaponin BⅡ,Baohuoside I,Anemarrhena saponin AⅢwere determined as 24.1-386μg/ml(r=0.9993),23.2-371μg/ml(r=0.9986),54.2-867.2μg/ml(r=0.9956),5.3-84.8μg/ml(r=0.9968),10-160μg/ml(r=0.9989)respectively,which showed a good linear relationship within the concentration range.The average recovery rate of the five components was between 101.8%and 105.0%,and the repeatability RSD was less than 2.4%.The content of the above five components in Zhimu medicinal materials were 1.62%,0.82%,7.36%,0.07%,0.34%,respectively.Conclusion The method is simple,accurate,and highly sensitive,which could be used as the quantitative determination of multiple index components of Anemarrhenae Rhizoma.