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

【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.