RP-HPLC-RID法同时测定怀地黄中单糖、低聚糖的含量

Simultaneous determination of monosaccharide and oligosaccharides by RP-HPLC-RID in Huai-Radix Rehmanniae

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,5μm)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、41.35～144.73μg(生地黄),1.50～15.00、25.20～92.40、1.51～15.47、110.30～523.93μg(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,5μm）, 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 ob-tained 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 estab-lished 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.