玫瑰多糖的酶法提取及柱前衍生HPLC分析.docVIP

玫瑰多糖的酶法提取及柱前衍生HPLC分析 　　摘要：通过酶法提取玫瑰多糖，用柱前衍生高效液相色谱法分析含量。结果表明，非酶法对干花和玫瑰花渣的多糖提取率分别为9.865、5.087 g/100 g；酶法对其的多糖提取率为12.487、7.599 g/100 g。用柱前衍生高效液相色谱法分析玫瑰多糖，发现该多糖是由甘露糖、鼠李糖、葡萄糖醛酸、葡萄糖、半乳糖、阿拉伯糖6种单糖组成的，其物质量比为0.934 4∶0.981 8∶1.156 6∶0.976 3∶0.974 0∶0.974 5。酶法提取和不加酶提取的玫瑰多糖一级结构的糖组成相同。 　　关键词：玫瑰多糖；酶法提取；柱前衍生；一级结构 　　中图分类号：O657.7 文献标识码：A 文章编号：0439-8114（2013）20-5031-03 　　Analysis of Rose Polysaccharide by Enzyme Extraction and Pre-column 　　Derivatization HPLC 　　ZHANG Yue-hui1，HAN Li-wen2，ZHENG Lan2，SHI Jian-guo2，YANG Jun-hui2，MA Yao-hong1 　　（1.Shandong Institute of Light Industry， Jinan 250353， China； 2. Biology Institute / Key Laboratory for Biosensor of Shandong Province，Shandong Academy of Sciences， Jinan 250014， China） 　　Abstract： Polysaccharide of rose was extracted by enzyme method and analyzed by HPLC with pre-column derivatization. The results showed that the yield rate of rose polysaccharide from dried flowers and rose dregs by normal method were 9.865 g/100 g， 5.087 g/100 g， respectively. The yield rate of rose polysaccharide by enzyme method were 12.487 g/100 g， 7.599 g/100 g， respectively. The results of monosaccharide composition of rose polysaccharide by HPLC with pre-column derivatization showed that it was composed of mannose， rhamnose， glucuronic acid， glucose， galactose and arabinose with a molar ratio of 0.934 4∶0.981 8∶1.156 6∶0.976 3∶0.974 0∶0.974 5. The sugar composition of primary structure of rose polysaccharide extracted by enzyme method was the same as those extracted by normal method . 　　Key words： rose polysaccharide； enzyme method extraction； pre-column derivatization； primary structure 　　多糖作为一种生命活性物质，具有清除自由基和抗肿瘤作用[1，2]。孔心渭等[3]通过肿瘤坏死因子（TNF）单克隆抗体的ELISA法，发现野玫瑰多糖通过调节肿瘤坏死因子的分泌而对急性肾炎有一定的治疗作用。白伟芳[4]测定了玫瑰花多糖在体外对羟基自由基和超氧阴离子自由基的清除作用，以及玫瑰花多糖的抗肿瘤活性。玫瑰花中的总糖质量浓度高达29.5 g/100 g，其中可溶性糖8.8 g/100 g。马猛华等[5]提取玫瑰花中多糖，提取率为0.6 g/100 g。陈晓梅等[6]分析了铁皮石斛原球茎多糖的结构和功能。本试验分析了玫瑰花提取精油后废水废渣中多糖的提取方法，并以1-苯基-3甲基-5吡唑啉酮（PMP）作为柱前衍生剂[7，8]，采用HPLC分析其单糖及糖醛酸的组成，并鉴定酶法对单糖及糖醛酸的组成有无影响，以期为多糖结构及功能研究