HPLC-CAD同时测定知母根茎与须根中5种呋甾皂苷的含量

Simultaneous determination of five furostanol saponins in rhizome and fibrous root of Anemarrhena asphodeloides Bge. by HPLC-CAD

目的建立高效液相色谱-电雾式检测器(HPLC-CAD)同时测定知母根茎与须根中5种呋甾皂苷含量的方法。方法采用Tech Mate C18-ST-Ⅱ色谱柱(250 mm×4.6 mm,5μm);流动相:乙腈-水(22∶78,V/V);流速1.0 ml/min;柱温30℃;CAD检测器参数:采集频率10 Hz;雾化温度55℃;filter:5 s。结果知母中5种主要呋甾皂苷:知母皂苷N、知母皂苷L、知母皂苷BⅡ、25R-知母皂苷BⅡ和25S-officinalisnin-Ⅰ分别在一定范围内呈良好的线性关系,相关系数为0.9992~0.9998,检测限分别为0.28、0.92、0.92、0.92和0.92 ng,定量限分别为0.92、2.77、2.77、2.77和2.76 ng,精密度、重复性、48 h稳定性的RSD均<3.0%,平均加样回收率分别为98.17%、101.37%、98.53%、97.63%和98.17%。结论本研究建立的HPLC-CAD法重复性好,结果准确可靠,可作为中药知母多成分含量测定的方法。

Objective To develop a method for the determination of five furostanol saponins（timosaponin N,timosaponin L,timosaponin BⅡ,25R-timosaponin BⅡ,and 25S-officinalisnin-Ⅰ）in rhizome and fibrous root of Anemarrhena asphodeloides Bge. by HPLC with the charged aerosol detector（CAD）. Methods The analysis was performed on Tech Mate C18-ST-II（250 mm×4.6 mm,5μm）with acetonitrile∶water（22∶78,V/V）,the flow rate of 1.0 ml/min and column temperature at 30 ℃. The Corona parameters were as follows：sampling rate 10 Hz,filter 5 s,and the nebulizer temperature 55℃. Results The approach showed good linearity for five saponins. The correlation coefficients（r2）for calibration curves varied from 0.9992 to 0.9998. The limits of detection（LOD）were0.28,0.92,0.92,0.92 and 0.92 ng for five steroidal saponins,respectively. The limits of quantitation（LOQ）were found to be 0.92,2.77,2.77,2.77 and 2.76 ng,respectively. RSD calculated from peak area of precision,repeatability and stability in 48 h were all less than 3.0%. The average recoveries of timosaponin N,timosaponin L,timosaponin BⅡ,25R-timosaponin BⅡ,and 25S-officinalisnin-Ⅰwere 98.17%,101.37%,98.53%,97.63%,and 98.17%,respectively. Conclusion The developed method is accurate,reliable,which could be applied to the quality control of multiple components in A. asphodeloides Bge.