RP-HPLC法测定大豆异黄酮的含量

Determination the Content of the Soybean Isoflavone by RP-HPLC

目的:采用反相高效液相色谱法,建立大豆提取物中6种大豆异黄酮—大豆苷,黄豆黄苷,染料木苷,大豆苷元,黄豆黄素,染料木素的含量测定方法。方法:色谱柱:welchrom-C_(18)柱(250 mm×4.6 mm;5μm),流动相:甲醇-水-乙腈=35∶65∶8(磷酸调节pH到2.6-3.0);流速:0.8 ml·min^(-1);检测波长:254 nm,柱温:35℃。结果:线性范围:大豆苷1.48～29.54 mg·L^(-1)(r=0.9998),黄豆黄苷1.58～31.64 mg·L^(-1)(r=0.9998),染料木苷1.09～21.84 mg·L^(-1)(r=0.9998),大豆苷元0.97～19.46 mg·L^(-1)(r=0.9998),黄豆黄素1.20～23.96 mg·L^(-1)(r=0.9999),染料木素1.31～26.26 mg·L^(-1)(r=0.9999)。大豆苷,黄豆黄苷,染料木苷,大豆苷元,黄豆黄素,染料木素平均回收率分别为101.56%,100.45%,102.75%,100.83%,100.92%,101.53%,RSD分别为1.66%,1.10%,1.89%,1.10%,1.52%,1.35%(n=6)。结论:本方法便捷、灵敏、准确、重复性好,适用于大豆产品中大豆异黄酮含量检测。

Objective ： To establish a RP-HPLC method for determination of the six main isoflavone constituents, daidzin, glycitin, genistin, daidzein, glycitein and genistein from extract of soybean. Method ： The welchrom-Cls column （ 250 mm ×4.6 mm;5 μm） was used. The mobile phase consisted of methanol-water-aeetonitrile （ 35： 65：8 ） （ adjust pH to 2. 6-3.0 with phosphoric acid）. The flow rate was 0. 8 ml·min^-1 ;The detective wavelength was 254 nm. Result： There was a good linear relationship within the range of 1.48-29.54 mg·L-1 of daidzin （r =0. 999 8） ,1.58-31.64 mg.L-1 of glycitin （r =0. 999 8） ,1.09-21.84mg·L^-1 of genistin（r =0. 999 8） ,0. 97- 19. 46 mg. L - 1 of daidzein （ r = 0. 999 8 ）, 1. 20-23.96 mg·L ^- 1of glycitein （ r = 0. 999 9）, 1. 31-26. 26 mg·L -1 of genistein （ r = 0. 999 9）. The reeoverises of daidzin, glyeitin, genistin, daidzein, glycitein and genistein were 101.56%, 100. 45% , 102.75%, 100. 92%, 101.53% ,and RSD were 1.66% ,1.10% ,1.89% ,1.10% ,1.52% ,1.35% （n =6） ,respectively. Conclusion： The method is proved to be simple, accurate and reproducible for the determination of isoflavone in extract of soybean.