饲料中大豆异黄酮检测方法的建立

Establishment of a method for detecting the soy isoflavones in feeds

[目的]建立用于检测饲料中大豆提取物的高效液相色谱方法(HPLC),为大豆异黄酮在畜牧业的合理使用提供支持。[方法]通过查阅文献,采用HPLC色谱技术,样品经甲醇溶液超声提取后,以乙腈-0.1%醋酸水为流动相梯度洗脱,260 nm波长检测。[结果]建立的HPLC检测方法,线性范围为0.2~20μg/mL,定量限可达0.197μg/mL,以2、15μg/mL的质量浓度进行添加回收,添加回收率为97.45%~102.54%。[结论]按建立的HPLC方法检测5%纯度的大豆异黄酮,可检测6种组分的含量分别为:大豆苷1.13%、大豆黄苷0.22%、染料木苷2.79%、大豆苷元0.55%、大豆黄素0.06%和染料木素0.29%,总黄酮含量为5.04%。

[Objectives]A high-performance liquid chromatography(HPLC)method for detecting soybean extracts in feeds was established to provide support for the rational use of soy isoflavones in animal husbandry.[Methods]By reviewing literature and using HPLC chromatography technology,the sample was extracted by ultrasound from methanol solution and eluted with acetonitrile 0.1%acetic acid-water as the mobile phase gradient.The detection was performed at a wavelength of 260 nm.[Results]The detection method established with HPLC had a linear range of 0.2-20μg/mL,with a quantification limit of 0.197μg/mL.The mass concentration of 2μg/mL and 15μg/mL was added for recovery,with a recovery rate of 97.45%-102.54%.[Conclusions]According to the established HPLC method for detecting 5%purity of soy isoflavones,the content of six components including daidzin 1.13%,daidzin 0.22%,genistein 2.79%,daidzein 0.55%,daidzin 0.06%,and genistein 0.29%were detected.The total flavonoid content was 5.04%.