文章编号:1008 - 9926(2005) 01 - 0034 - 04 中图分类号:R943 文献标识码:A

    地黄中寡糖的提取分离工艺研究①

    张汝学②,樊俊杰 ,贾正平 ,孔令媛 ,马慧萍 ,李茂星 ,任 俊 ,王 娟

    (中国人民解放军兰州军区兰州总医院 全军临床药理基地 甘肃 兰州 730050)

    摘 要:目的 研究地黄( Rehmannia glutinosa )中寡糖的提取分离工艺。方法 建立水苏糖的 HPLC测定法;以水苏糖含量为

    指标 ,采用正交试验优化地黄水提取物的提取工艺 ,对水提物进一步用离子交换树脂进一步纯化 ,再进行活性炭柱层析 ,以不

    同浓度梯度乙醇洗脱 ,分得寡糖的不同部位。结果 地黄水提取的最佳工艺为A2B3C2 ,即溶剂(水)体积10倍 ,提取 3 次 ,每次

    提取1h。水提物经活性炭柱层析得到4个部位 ,部位Ⅰ～Ⅳ,其中部位Ⅱ及部位Ⅲ中水苏糖的含量分别为 20. 99 %及 60. 51 %。

    结论 地黄水提取的较佳工艺条件为A2B3C2 ,活性炭柱层析可以较好地将地黄中寡糖进行分离 ,其中水苏糖的 HPLC测定法

    简便、稳定、可靠。

    关键词:地黄;寡糖;正交试验;活性炭层析;HPLC

    Extraction and Separation Process of Oligosacchrides from Rehmannia Glutinosa

    ZHANG Ru2Xue ,FAN Jun2 Jie ,J IA Zheng 2Ping ,KONGLing 2Yuan ,MA Hui2Ping ,LI Mao2Xing ,REN Jun ,WANGJuan

    (Base of Clinical Pharmacology ,Lanzhou General Hospital of PLA ,Lanzhou 730050 ,Gansu China)

    ABSTRACT:Aim To study extraction and separation process of oligosaccharides from Rehmannia glutinosa and HPLC

    determination of stachyose.Mehtods Orthogonal test was used to inspect the factors such as water volume ,times and time

    of extraction by boiled water method from R. glutinosa.Oligosacchrides of water extract from R. glutinosa were further sep2

    arated by charcoal column chromatography and the content of stachyose ,the main component of water extract from R. gluti 2

    nosa was determined by HPLC. Results According to the yields of water extract the influence factors were extraction time

    (B) > water volume (A) > time for extraction (C) in turn. The optimal extraction condition was found to be A2B3C2 ,that is ,with 10 volume of water for 1 hour ,3 times.Water extract was further separated by charcoal column chromatogra 2

    phy into four groups and identified by HPLC. It was found that the analysis of stachyose in water extract by HPLC was a

    stable and reliable method to determine the content of stachyose in the extract . Conclusion Oligosaccharides in R. gluti2

    nosa could be extracted at a fuller extent by boiled water method according to best procession of A2B3C2 ,that was with 10

    volume of water for 1 hour ,3 times.Determination of stachyose by HPLC was stable and reliable.

    KEY WORDS :Rehmannia glutinosa ;Oligosaccharides ;Orthogonal test ;Charcoal column chromatography ;HPLC

    地黄是一味重要的滋阴补肾中药 ,近年来 ,国内

    外对其进行了大量深入细致的化学、药理研究。日

    本学者对鲜地黄中水溶性组分进行的研究表明[1 ]

    ,含量最高的是寡糖成分 ,其中水苏糖( stachyose) 达

    48. 3 %。药理研究表明 ,地黄寡糖可增强造血功

    能[2、 3 ]

    ,促进免疫[4 ]

    ,降低血糖[5 ]

    ,水苏糖对亚临床

    肝性脑病具有治疗作用[6 ]

    ,但未见中国产地黄有关

    寡糖的提取、分离工艺研究报告。本文研究了国产

    地黄中寡糖的提取分离工艺流程 ,考察了提取次数、提取时间和溶媒用量对水提物得率的影响 ,确定了

    水苏糖得率高、适合大量生产的最佳提取条件;对地

    黄水提物中寡糖成分进行了活性炭柱层析分离 ,并

    采用高效液相色谱法 ,对分得的各个部位进行了水

    苏糖含量测定。

    1 仪器与试药

    1. 1 仪器 DHG- 90140 电热恒温鼓风干燥箱(上

    海医用恒温设备厂) ;TN - 100B扭力天平(上海第二

    天平仪器厂) ;LXJ - Ⅱ离心沉淀机(上海医用分析仪

    器厂) ;SCL - 6A高效液相色谱仪;RID26A 示差折光

    检测器;色谱柱(Spherisorb NH2 柱) (均为日本岛津

    制作所) 。

    · 43 · 解放军药学学报 第21卷 第1期 地黄中寡糖的提取分离工艺研究 张汝学

    ① ② 作者简介:张汝学(1963 - ) ,男,甘肃文县人,博士,副主任药师。研究方向:中药药理学。Tel : (0931) 8975884 ; E - mail :empriqqq @pub2

    lic. lz. gs. cn

    基金项目:国家自然科学基金,No. 30472186

    ? 1995-2005 Tsinghua Tongfang Optical Disc Co., Ltd. All rights reserved.1. 2 药材与试剂 地黄[兰州市医药公司 ,经兰州

    医学院药学系杨永健教授鉴定为怀庆地黄

    ( Rehmannia glutinosa Libosch. ) ] ; 732 型阳离子树

    脂、 717型阴离子树脂;活性炭颗粒(均为中国科学

    院兰州分院化物所试剂部) ;95 %乙醇、盐酸、氢氧化

    钠、醋酸(兰州军区总院药剂科提供) ;水苏糖标准品

    (Sigma 公司) ;乙腈色谱纯 ......