快速检测大豆籽粒中十二种异黄酮组分的HPLC方法  

A Rapid HPLC Method for Determination of 12 Isoflavone Components in Soybean Seeds

作　　者：孙君明[1] 孙宝利[2] 韩粉霞[1] 闫淑荣[1] 杨华[1] 菊池彰夫 

机构地区：[1]中国农业科学院作物科学研究所/国家农作物基因资源与基因改良重大科学工程,中国北京100081 [2]中国农业科学院环境与可持续发展研究所,中国北京100081 [3]日本东北农业研究中心,日本秋田县019-2112

出　　处：《中国农业科学》2009年第7期2491-2498,共8页Scientia Agricultura Sinica

基　　金：国家转基因重大专项(2008ZX08004-003);国家“948”项目(2006-G1);国家“863”计划项目(2006AA100104);科技支撑计划项目(2006BAD13B05和2006BAD01A04);院所基金项目(2060302-2);国家自然科学基金项目(30000107)

摘　　要：【目的】利用高效液相色谱(HPLC)技术快速检测大豆异黄酮组分,为大豆的异黄酮标记辅助育种提供重要依据。【方法】以大豆异黄酮12种组分为标准样品,利用HPLC技术结合吸收波峰鉴定的方法进行异黄酮组分的分析。【结果】不同异黄酮苷元组分黄豆苷元(daidzein)、大豆黄素(glycitein)和染料木素(genistein)分别具有不同的最大紫外光谱吸收波长,分别位于250、257和260nm左右。采用YMC-C18反相色谱柱,以含0.1%(v/v)乙酸13%～30%(v/v)的乙腈为流动相,在35℃柱温条件下,经梯度洗脱,结合紫外吸收检测,可以准确地定性和定量鉴定出12种不同异黄酮组分。【结论】该方法样品用量少,检测异黄酮含量准确、快速,适合大量样本分析。[Objective] It is highly important and has utilization values to determine the isoflavone components by HPLC as it can provide bases for molecular assistant selection ofisoflavone in soybean breeding. [Method] Based on the standard samples of 12 isoflavone components, the isoflavone components were analyzed using the determination of absorbance peaks method by HPLC. [Result] The results showed that there were different maximum absorbance wavelengths for the daidzein, glycitein, and genistein, which were 250 nm, 257 nm and 260 nm, respectively. A linear gradient elution of acetonitrile （13%-30%） containing 0,1% acetic acid using a mobile phase was applied on a YMC-Cl8 column at 35 ℃. The 12 isoflavone components were determined using the UV detector by HPLC. [Conclusion] It is concluded that this is a rapid, precise method which adapted to determine the large numbers of samples with microanalysis.

关 键 词：大豆(Glycine MAX L.Merri) 异黄酮 HPLC 

分 类 号：S565.1[农业科学—作物学]