LC-MS测定黑豆中异黄酮和花色苷的含量

Determination of Isoflavone and Anthocyanin in Black Soybean Seed( Glycine max) by LC-MS

采用高效液相色谱质谱联用技术(HPLC-MS),建立黑豆异黄酮和花色苷含量测定方法。应用Waters Xselect HSS T3色谱柱,以乙腈-0.1%乙酸水溶液为流动相梯度洗脱黑豆异黄酮;应用X-Terra MS C18色谱柱,以10%甲酸水溶液-甲醇/乙腈/水/甲酸(22.5/22.5/40/10,V/V)混合液为流动相梯度洗脱黑豆花色苷;以电喷雾离子源(ESI)选择离子监控模式(SIM)检测目标化合物,并通过标准品对照比较,实现黑豆12个异黄酮及8个花色苷单体化合物的定量测定。该方法操作简便、稳定、耗时少,具有良好的灵敏度、重现性和回收率,可用于黑豆种质资源的化学评价及质量控制。

In this study,an efficient high performance liquid chromatography-mass spectrometry（ LC-MS） method was established to quantitatively determine the isoflavone and anthocyanin concentrations in black soybean seed. Each sample was injected into Waters Xselect HSS T3 C18 column for isoflavone analysis,the mobile phase consisted of acetonitrile and 0. 1% acetic acid in HPLC water. For anthocyanin,X-Terra MS C18 column was used,the mobile phase consisted of10% formic acid and methanol/acetonitrile/water/formic acid（ 22. 5/22. 5/40/10,V/V）. Selected ion monitoring（SIM） mode was used in the mass spectral acquisition. Identification of the isoflavones and anthocyanins were achieved by comparing the sample retention times and mass spectra with those of standard compounds. In this study,twelve isoflavones and eight anthocyanins were quantified in absolute terms via linear regression of their corresponding standards. The established analysis method was simple,stable and rapid. It can be used for the identification and quantitative determination of isoflavone and anthocyanin of black soybean seed.