Callus Formation and Its Isoflavonoid Accumulation in Maackia amurensis

    LUO Jian2ping , WU Li2 fu , J IANG Shao2tong

    (Department of Biotechnology and Food Engineering , Hefei University of Technology , Hefei 230009 , Anhui , China)

    [ Abstract] Objective : To obtain Maackia amurensis callus and investigate some factors influencing total isoflavonoid produc2

    tion in callus. Method : Different media were used for callus induction f rom cotyledonary and hypocotyl explants. Total isoflavonoid

    content was determined by UV spect rophotometer. Result : Medium types and hormone combinations influenced both callus formation

    and their developmental states. MS medium su pplemented with NAA BA or containing 2 ,42D was suitable for callus induction f rom

    cotyledonary explants. Among the basal media tested , MS , N6 and B5 were suitable for cotyledonary callus formation and SH for

    hypocotyl callus formation. The formed callus could synthesize isoflavones. Media containing NAA were suitable for isoflavonoid pro2

    duction in cotyledonary callus and media containing 2 ,42D BA were suitable for hypocotyl callus , but 2 ,42D KT at high concen2

    t ration inhibited the isoflavonoid accumulation in hypocotyl and coytledonary calli . According to the developmentally morphological

    characters , the formed callus could be classified into four types. Ⅱ 2type callus with yellow in color and f riability in texture showed the

    highest accumulation of isoflavones. Conclusion : The influences of medium type and hormone combinations on Maackia amurensis cal 2

    lus formation are reflected in both developmentally morphological characters and isoflavonoid accumulation of calli . Yellow , f riable cal 2

    lus induced f rom cotyledonary and hypocotyl explants in MS or N6 medium su pplemented with 2 ,42D and BA exhibit the optimum

    growth and isoflavonoid production.

    [ Key words] Maackia amurensis ; callus formation ; morphological character ; isoflavone

    [责任编辑 袁桂京]

    葛根药材 HPLC指纹图谱研究

    何春年1

    , 李 敏2

    , 曹志高3

    , 郭红英3

    , 王春兰1

    , 余世春2

    (1.中国医学科学院 中国协和医科大学 药用植物研究所 ,北京 100094 ;

    2.安徽安泰医药生物技术有限公司 ,安徽 合肥 230088 ;

    3.安徽大东方药业有限公司 ,安徽 合肥 230088 )

    [摘要] 目的:对不同来源的葛根药材指纹图谱比较研究。方法:应用 RP2HPLC法、 Kromasil C18分析柱、乙

    腈2水二元梯度洗脱 ,流速为 1. 0 mL· min - 1

    ,检测波长 250 nm。结果与结论:不同来源的葛根药材所含黄酮类成分

    均得到很好的分离 ,该法可用于鉴别葛根的品种 ,并为葛根注射液及含葛根药材的中药复方注射液指纹图谱研究

    奠定基础。

    [关键词] 葛根;指纹图谱;高效液相色谱;梯度洗脱

    [中图分类号] R 282. 5 [文献标识码] A [文章编号] 100125302 (2003) 1221140205

    豆 科 植 物 葛 根 包 括 野 葛 Pueraria lobata

    ( Iwild. ) Ohwi 与甘藤葛 P. thomsoni i Benth. ,均收

    [收稿日期] 2002209220

    [通讯作者] 何春年 Tel : (010) 62899734 E2mail :chunnianhe

    @163. com

    载于《中国药典》各版 ,广泛应用于临床。目前尚未

    见文献报道有关葛根药材及其制剂的指纹图谱研

    究。按照国家药品监督管理局关于“中药注射剂指

    纹图谱研究的技术要求(暂行)”的通知 [1 ]

    ,作者参

    照相关文献[225 ]

    ,对不同产地的葛根药材进行了指

    纹图谱研究。

    · 1411 ·

    第28 卷第12 期

    2003 年12 月 中 国 中 药 杂 志

    China Journal of Chinese Materia Medica

    Vol . 28 , Issue 12

    December , 2003

    ? 1995-2005 Tsinghua Tongfang Optical Disc Co., Ltd. All rights reserved.1 仪器与试剂

    岛津 LC210AT 液相色谱仪 ,包括 LC210AT 色

    谱泵、 SPD210A 紫外检测器、混合器。江申J S23030

    色谱工作站。葛根素对照品(批号 7522200108 中国

    药品生物制品检定所提供) ,乙腈为色谱纯 ,水为高

    纯水 ,其他试剂均为分析纯。药材:野葛与粉葛 ,经

    安徽中医学院生药教研室周建理教授鉴定为 Puer2

    aria lobata 或 P. thomsoni i ,见表 1。

    表 1 葛根药材样品来源及名称

    编号 来源 采集时间

    A 安徽霍山县城郊 2001210

    B 安徽岳西县 2000211

    C 安徽岳西县 2000211

    D 安徽金寨县 2001209

    E 安徽太湖县 2000212

    F 安徽霍山县(粉葛) 2000211

    G 四川宜宾 2002201

    2 实验方法与结果

    2. 1 色谱分析条件

    流动相为乙腈2水二元梯度洗脱 ,流速 1. 0 mL·

    min - 1

    ,检测波长 250 nm ,色谱柱 Kromasil52C18 (4. 6

    mm×150 mm , 5μm)江苏汉邦科技有限公司 ,柱温

    30 ℃ ......