柱前衍生化HPLC分析不同来源、不同生长年限铁皮石斛多糖的组成和含量

 目的 建立柱前衍生化HPLC分析不同种植基地、不同生长年限铁皮石斛多糖中单糖的组成和主要单糖的含量，并根据含量水平进行聚类分析及判别分析。方法 铁皮石斛药材用乙醇脱脂，沸水提取得到的铁皮石斛多糖，用3.0 mol·L-1盐酸水解，水解产物加入1-苯基-3-甲基-5-吡唑啉酮(PMP进行衍生化，采用HPLC测定单糖衍生物；并利用SPSS16.0进行聚类分析及判别分析。结果 铁皮石斛多糖主要由甘露糖和葡萄糖组成，不同生长年限、9个种植基地的13批铁皮石斛样品甘露糖的含量范围为19.65%~34.27%，均符合药典标准；并随着生长年限的增加，甘露糖含量降低。不同生长年限的铁皮石斛甘露糖与葡萄糖峰面积比差异较大。聚类分析将13批样品按生长年限聚成3类；并建立了判别函数，回判准确率100％。结论 铁皮石斛多糖中单糖的组成和主要单糖的含量结合化学计量学方法判别铁皮石斛的生长年限是切实可行的，该研究为铁皮石斛的质量评价和开发利用提供了实验依据。

Analysis of Monosaccharide Compositions and Contents in Polysaccharide of Dendrobium officinale from Different Sources and Growth Years by Pre-column Derivatization-HPLC

OBJECTIVE To develop a method based on pre-column derivation-HPLC combined with cluster analysis and discriminant analysis to determine the monosaccharide compositions and contents in polysaccharide of Dendrobium officinale and to identify growth years. METHODS After medicinal Dendrobium was degreazed by ethanol， the polysaccharides were extracted with hot water and hydrolyzed with 3.0 mol·L-1HCl. Then， the monosaccharides were derivated by PMP. The monosaccharide derivatives were assayed by HPLC. Cluster analysis and discriminant analysis(DA were performed by SPSS16.0. RESULTS The polysaccharide of Dendrobium candidum was composed of mannose and glucose. The ranges of the mannose contents were 19.65％-34.27％， and all met the standard of Chinese Pharmacopoeia. However， the mannose content decreased with growth years. The peak area ratio between mannose and glucose was significantly different. The samples analyzed can be clustered into three classes and the same ages were clustered into one class， meanwhile the recognition function was established and the accuracy of classification testing was 100％. CONCLUSION The monosaccharide compositions and contents in polysaccharide of Dendrobium officinale， combined with chemometrics， can accurately identify growth year. The study provided experimental evidence for quality evaluation and development of Dendrobium officinale.