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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 method for the determination of six soy isoflavones-daidzin, glycitein, genistin, daidzein, glycitein and genistein in soybean extract by reversed-phase high performance liquid chromatography. Methods: Column: Welchrom-C 18 column (250 mm × 4.6 mm; 5 μm) with mobile phase of methanol-water-acetonitrile = 35:65:8 · Min ~ (-1); detection wavelength: 254 nm, column temperature: 35 ℃. Results: The linear range was 1.48-29.54 mg · L -1 for daidzin (r = 0.9998), 1.58-31.64 mg · L -1 for glycitein (r = 0.9998), 1.09-21.84 for genistin (r = 0.9998), daidzein 0.97 ~ 19.46 mg · L -1 (r = 0.9998), and the peak values of glycitein 1.20 ~ 23.96 mg · L -1 (r = 0.9999), genistein 1.31 ~ 26.26 mg · L -1 (r = 0.9999). The average recoveries of daidzin, glycinin, daidzein, daidzein, glycitein and genistein were 101.56%, 100.45%, 102.75%, 100.83%, 100.92% and 101.53%, respectively, with RSDs of 1.66% , 1.10%, 1.89%, 1.10%, 1.52%, 1.35% (n = 6). Conclusion: The method is convenient, sensitive, accurate and reproducible. It is suitable for the determination of soybean isoflavones content in soybean products.