HPLC法测定绿豆芽中四种大豆异黄酮的含量

Determination of Four Isoflavone in Mung Bean Sprout by HPLC

建立同时测定绿豆芽中4种大豆异酮（大豆苷、染料木苷、大豆苷元和染料木素）含量的高效液相色谱方法,对从绿豆芽中提取4种大豆异黄酮的工艺进行优化,最佳工艺为：用60%的乙醇在55℃条件下超声提取60min,料液比为1∶12（g/mL）,同时探究绿豆发芽不同天数4种大豆异黄酮的含量。采用日本岛津LC-20A高效液相色谱仪进行测定的色谱条件为：色谱柱为phenomenex C18（150mm×4.6mm,5.0μm）,采用乙腈-0.2%甲酸为流动相进行梯度洗脱,流速为0.8mL/min,检测波长为260nm。大豆苷,染料木苷,大豆苷元,染料木素的线性范围分别是0.74μg/mL～100.00μg/mL,0.74μg/mL～200.00μg/mL,0.167μg/mL～500.00μg/mL,0.198μg/mL～160.00μg/mL（r〉0.9996）,线性关系良好;加样回收率（n=9）分别为100.02%,98.95%,99.28%,99.63%。

To develop an HPLC method for determination of four isoflavones,daidzin,genistin,daidzein,genistein in mung bean sprout.An phenomenex C18 column（150 mm×4.6 mm,5.0 μm）was applied,a gradient mobile phase composing of acetonitrile and 0.2% formic acid was used,and the flow rate was 0.8 mL/min,detective wavelength was set at 260 nm.The best condition of extracted four isoflavones of mung bean sprout was studied,the results showed the best condition was the ethanol concentration 60%,55℃,extraction for 60 min with the solid-liquid ratio 1：12（g/mL）.The results were obtained that quantitative linear range of four isoflavones was 0.74 μg/mL-100.00 μg/mL,0.74 μg/mL-200.00 μg/mL,0.167 μg/mL-500.00 μg/mL,0.198 μg/mL-160.00 μg/mL（r0.999 6）respectively,and the average recoveries（n=9）were 100.02 %,98.95 %,99.28 %,99.63 % respectively.A specific,sensitive,simple and reproducible HPLC method has been established for determination of four isoflavones in mung bean sprout.