HPLC测定中药渣中人参皂苷Rg1、Re与Rb1含量

Determination of Ginsenosides Rg_1,Re and Rb_1 in Chinese Herbal Residues by HPLC

建立了一种利用反相高效液相色谱检测中药渣中人参皂苷Rg_1、Re和Rb_1含量的分析方法。考察了提取试剂、提取温度、提取溶剂倍量、提取时间对提取效果的影响,最终将样品以10倍体积的70%甲醇于40℃水浴中超声提取0.5 h。采用Zorbax SB-C_(18)(4.6 mm×150 mm,5μm)柱分离,以乙腈和水为流动相进行梯度洗脱,流速为1.0 m L/min,进样量为20μL,柱温为20℃,检测波长为203 nm,采用外标法定量。在该条件下,人参皂苷Rg_1、Re、Rb_1分别在18.74~257.61、37.48~468.54、3.88~96.93 mg/L范围内线性关系良好(r^2>0.999)。人参皂苷Rg_1、Re和Rb_1的定量下限(S/N=10)分别为17.69、13.94、1.14 mg/L。3个不同浓度的加标回收率为89.1%~107%,相对标准偏差(RSD,n=3)为0.6%~4.1%。该方法具有操作简便快捷、检测灵敏度高等特点。

A fast and sensitive reversed high performance liquid chromatographic(HPLC)method was developed for the simultaneous detection of three ginsenosides,i.e.Rg 1,Re and Rb 1 in Chinese herbal residues.Effects of solvent,temperature,solvent-matrix ratio and time on extraction efficiency were investigated.The sample was ultrasonically extracted with 10 times liquid of 70%methanol at 40℃for 0.5 h.Chromatographic separation was performed on a Zorbax SB-C 18(4.6 mm×150 mm,5μm)column by gradient elution using water-acetonitrile as mobile phase at a flow rate of 1.0 mL/min.The injection volume was 20μL,the column temperature was 20℃and the detection wavelength was 203 nm.External calibration of peak area versus concentration was used for the quantitation.There were excellent linearities for ginsenosides Rg 1,Re and Rb 1 in the ranges of 18.74-257.61,37.48-468.54 and 3.88-96.93 mg/L,respectively,with their correlation coefficients all above 0.999.The limits of quantitation(LOQ,S/N=10)for Rg 1,Re and Rb 1 were 17.69,13.94 and 1.14 mg/L,respectively.The average recoveries at three spiked levels were in the range of 89.1%-107%,with relative standard deviations(RSD,n=3)of 0.6%-4.1%.The method was applied in the detection of ginsenosides Rg 1,Re and Rg 1 in Chinese herbal samples with simplicity,rapidness and high sensitivity.