霍山石斛与美花石斛多糖柱前衍生HPLC特征图谱比较  

Comparative Study on Pre-column Derivation HPLC Characteristic Spectrum of Polysaccharide from Dendrobium Huoshanense and Dendrobium Loddigesii

作　　者：黄月纯[1] 叶家宏[2] 戴亚峰 任晋[2] 顺庆生[4] 魏刚[2] 

机构地区：[1]广州中医药大学第一附属医院,广东广州510405 [2]广州中医药大学,广东广州510006 [3]九仙尊霍山石斛股份有限公司,安徽六安237003 [4]上海健康职业技术学院,上海200237

出　　处：《中药新药与临床药理》2014年第6期725-730,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省产业技术研究与开发专项资金(2013B060400022)

摘　　要：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,5μm）;流动相为乙腈-0.02 mol·L^-1的乙酸铵溶液（冰乙酸调p H=6.7）,梯度洗脱;检测波长为250 nm;柱温为30℃;流速为1 m L·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 huoshane nse and De ndrob ium 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 C18column（250 mm ×4.6 mm, 5 μm）, the acetonitrile-0.02 mol·L-1ammonium acetatesolution（p H=6.7）（gradient elution）was employed as a mobile phase,the detection was carried out at 250 nm with flow rate being of 1 m L·min^-1at 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 De ndrob ium huoshane nse,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 loddige sii,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 De ndrobium huoshane nse and Dendrobium loddige sii were 0.994-1.000,0.934-1.000 respectively,and the similarity of the mutual modes between De ndrob ioum huoshane ns e and De ndrob ium loddiges ic 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 De ndrob ium huoshane nse and De ndrobium loddigesii.

关 键 词：霍山石斛 美花石斛 多糖 柱前衍生化 特征图谱 高效液相色谱法 

分 类 号：R284.1[医药卫生—中药学]