HPLC-ELSD法同时测定鲜地黄中3种低聚糖的含量

Simultaneous Determination of Three Oligosaccharides by HPLC-ELSD in the Fresh Root of Rehmannia glutinosa

目的建立同时测定中药鲜地黄中主要的3种低聚糖的HPLC-ELSD含量测定方法。方法采用HPLC-ELSD对鲜地黄中主要的低聚糖进行研究,采用Waters XBridgeTMAmide 3.5μm(4.6 mm×150 mm)色谱柱为固定相,乙腈-水-70∶30为流动相,流速为0.5 mL.min-1,柱温:25℃。ELSD检测器:漂移管温度40℃,雾化气体2.07 L.min-1,喷雾模式:冷却。结果该方法下,鲜地黄中3种低聚糖分离效果良好,具有良好的线性范围,分别为0.184～9.2μg(r=0.999 8),0.458～11.45μg(r=0.999 7)和0.924～11.55μg(r=0.999 2),平均加样回收率为96.94%(RSD,1.87%),98.87%(RSD,1.98%)和101.02%(RSD,3.23%)。结论该方法稳定、可靠、精密度高、重复性好,为有效控制鲜地黄药材的内在质量提供科学依据。

OBJECTIVE To establish a HPLC-ELSD method for simultaneous determination of Sucrose,Raffinose and Stachyose in the fresh root of Rehmannia glutinosa.METHODS Chromatographic separation was performed on a Waters XBridgeTM Amide 3.5 μm（4.6 mm×150 mm） with mobile phase of acetonitrile-water（70∶30） at a flow rate of 0.50 mL·min-1,the drift tube temperature of ELSD was set at 40 ℃,and nitrogen gas was 2.07 L·min-1.RESULTS A good resolution and linear range were obtaind under this HPLC-ELSD condition.The calibration of Sucrose,Raffinose and Stachyose were linear over the the ranges of 0.184-9.20 μg（r=0.999 8）,0.458-11.45 μg（r=0.999 7） and 0.924-11.55 μg（r=0.999 2）,respectively,and the average recoveries were 96.94%（RSD,1.87%）,98.87%（RSD,1.98%） and 101.02%（RSD,3.23%）,respectively.CONCLUSION The method is simple,reliable,accurate and reproducible for quality control of fresh root of Rehmannia glutinosa.