中药天冬中甾体皂苷25-epi-officinalisninⅡ的液相色谱含量测定方法的建立

Establishment of a liquid chromatographic method for determination of the content of steroidal saponin 25-epi-officinalisninⅡ in Asparagi Radix

目的建立中药天冬中甾体皂苷25-epi-officinalisninⅡ的含量测定方法,为天冬药材的质量控制奠定基础。方法采用超高效液相色谱-电雾式检测器(UHPLC-CAD)检测,检测器工作气压59.2 psi(1 psi=6.895 kPa),雾化温度35℃。采用Kinetex?C_(18)(100 mm×4.6 mm,2.6μm)色谱柱,乙腈-0.1%乙酸水(19∶81,V/V)等度洗脱,流速1.2 ml/min,进样量5μl。结果该皂苷与其他成分实现良好分离。在0.0519～1.0380 mg/ml范围内呈良好的线性关系(r=0.9990)。检测限为12.5 ng,定量限为25.0 ng,精密度、重复性、24 h稳定性的RSD均<3.0%,平均加样回收率为98.63%。收集的11批不同产地天冬样本含量测定结果为0.36～6.86 mg/g。结论本研究建立的UHPLC-CAD方法灵敏、稳定,可用于中药天冬中25-epi-officinalisninⅡ的含量测定。

Objective To develop a method for the determination of 25-epi-officinalisninⅡand for the future use in quality control of Asparagi Radix.Methods Ultra performance liquid chromatography system with charged aerosol detector(UHPLC-CAD)was used.The CAD working pressure was 59.2 psi(1 psi=6.895 kPa)and the nebulizer chamber temperature was 35℃.The analysis was performed on a Kinetex?C18(100 mm×4.6 mm,2.6μm)column using acetonitrile-water with 0.1%acetic acid(19∶81,V/V)at a flow rate of 1.2 ml/min,and the injection volume of sample solution was 5μl.Results The 25-epi-officinalisninⅡwas well separated from other components and had a good linear relationship(r=0.9990)in the range of 0.0519-1.0380 mg/ml.The limit of detection and quantification was 12.5 and 25.0 ng,respectively.The precision,repeatability and stability were all good as judged by the RSD values(all below 3.0%).The average recovery rate was 98.63%.The contents of 25-epi-officinalisninⅡin the 11 batches of Asparagi Radix samples from different producing area were between 0.36 and 6.86 mg/g.Conclusion The established UHPLC-CAD method showed a well sensitivity and stability,which might be potentially used for the quality control of Asparagi Radix by quantitative determination of 25-epi-officinalisninⅡ.