RP-HPLC-RID法同时测定怀地黄中单糖、低聚糖的含量

目的: 建立同时测定怀地黄中主要的单糖和低聚糖(葡萄糖、蔗糖、棉籽糖、水苏糖)的RP-HPLC-RID含量测定方法。 方法: 采用Agilent Zorbax NH₂(4.6 mm×250 mm,5 μm)色谱柱,以乙腈-水(70∶30)为流动相,流速1 mL·min^-1,柱温30 ℃。RID检测器内部温度30 ℃。 结果: 葡萄糖、蔗糖、棉籽糖和水苏糖的线性范围分别为1.50~15.00、25.20~92.40、8.30~49.80、41.35~144.73 μg(生地黄),1.50~15.00、25.20~92.40、1.51~15.47、110.30~523.93 μg(鲜地黄);加样回收率(n=5)分别为96.6%、97.5%、99.4%、100.1%(生地黄),96.4%、98.4%、97.3%、99.1%(鲜地黄)。 结论: 本方法经过了系统的方法学考察,能够准确测定出怀地黄鲜品及生品中4种糖类成分的含量分布情况,专属性强,可为怀地黄药材的内在质量控制提供科学依据。

Objective: To develop a RP-HPLC-RID method for simultaneous determination of monosaccharide and oligosaccharide(including glucose,sucrose,raffinose,stachyose)contents in Huai-Radix Rehmanniae from genuine producing area. Methods: Chromatographic separation was performed on an Agilent Zorbax NH₂ column(4.6 mm×250 mm,5 μm),a mobile phase of acetonitrile-water(70∶30)was adopted with a flow-rate of 1.0 mL·min^-1.A RID detector was applied,and the inner temperature was set at 30 ℃. Results: The results were obtained that quantitative linear range of glucose,sucrose,raffinose and stachyose in unprocessed Rehmannia root was 1.50-15.00,25.20-92.40,8.30-49.80,and 41.35-144.73 μg,respectively,and that in fresh Rehmannia root was 1.50-15.00,25.20-92.40,1.51-15.47,and 110.30-523.93 μg,respectively.The average recovery(n=5)in unprocessed Rehmannia root was 96.6%,97.5%,99.4% and 100.1%,and that in fresh Rehmannia root was 96.4%,98.4%,97.3% and 99.1%. Conclusion: System methodological study results suggest the established method can accurately determine the four kinds of oligosaccharides in Huai-Radix Rehmanniae for both fresh and dried samples with strong specificity,which can be applied to quality control of Huai-Radix Rehmanniae.