目的 研究比较霍山石斛与美花石斛多糖柱前衍生高效液相色谱（HPLC）特征图谱的差异。方法 以盐酸水解石斛多糖，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法分析单糖的衍生物。色谱柱为Kromasil 100-5 C18（250 mm×4.6 mm，5 μm）；流动相为乙腈-0.02 mol·L-1的乙酸铵溶液（冰乙酸调pH=6.7），梯度洗脱；检测波长为250 nm；柱温为30 ℃；流速为1 mL·min-1。采用国家药典委员会中药色谱指纹图谱相似度评价系统软件（2004A版）生成对照图谱。结果 10批霍山石斛多糖柱前衍生特征图谱标示出6个特征峰，主要由D-甘露糖（71.8 %）与D-葡萄糖（27.2 %）组成，D-甘露糖与D-葡萄糖峰面积比值为2.05～3.71，此外含微量的D-半乳糖醛酸、D-半乳糖、D-木糖、D-阿拉伯糖。10批美花石斛多糖柱前衍生特征图谱亦标示出6个特征峰，主要为D-甘露糖（42.9 %）与D-葡萄糖（54.0 %）组成，D-甘露糖与D-葡萄糖峰面积比值为0.67～0.98，此外含微量的D-半乳糖醛酸、D-半乳糖、D-木糖、D-阿拉伯糖。与对照图谱比较，10批霍山石斛、美花石斛的相似度分别为0.994～1.000，0.934～1.000，霍山石斛与美花石斛共有模式之间的相似度为0.871。结论 该方法准确可靠，重复性好；两种石斛多糖构成差异较大，不宜混用。

Objective To compare the differences of pre-column derivation HPLC characteristic spectrum of the polysaccharide from Dendrobium huoshanense and Dendrobium loddigesii. Methods Dendrobium polysaccharide samples were hydrolyzed with hydrochloric acid and derivated by 1-phenyl-3-methyl-5-pyrazolone（PMP）. HPLC was performed on Kromasil 100-5 C18 column（250 mm×4.6 mm，5 μm），the acetonitrile -0.02 mol·L-1 ammonium acetatesolution（pH=6.7）（gradient elution） was employed as a mobile phase，the detection was carried out at 250 nm with flow rate being of 1 mL·min-1 at column temperature 30 ℃. The common mode was generated by the State Pharmacopoeia Commission specific chromatogram similarity evaluation system software（2004A version）. Results Six common peaks were separated from 10 batches of Dendrobium huoshanense，mainly composed of D-mannose（71.8 %） and D-glucose（27.2 %）. The ratio of peak area of D-mannose to that of D-glucose was 2.05~3.71. The common peaks also contained traces of galacturonic acid，D-galactose，D-xylose and D-arabia sugar. Six common peaks were separated from 10 batches of Dendrobium loddigesii，mainly composed of D-mannose（42.9 %） and D-glucose（54.0 %）. The ratio of peak area of D-mannose to that of D-glucose was 0.67~0.98. The common peaks also contained traces of D-galacturonic acid，D-galactose，D-xylose and D-arabia sugar. The similarities of 10 batches of Dendrobium huoshanense and Dendrobium loddigesii were 0.994~1.000，0.934~1.000 respectively，and the similarity of the mutual modes between Dendrobioum huoshanense and Dendrobium loddigesic was 0.871. Conclusion The components of the two kinds of Dendrobium polysaccharide are quite different and can not be confused. The established method is accurate and reproducible，which will provide reference for the quality control of Dendrobium huoshanense and Dendrobium loddigesii.

R284.1

广东省产业技术研究与开发专项资金（2013B060400022）。