Department of Nutrition and Food Science, University of Maryland, College Park, MD 20742, USA.
Food Chem. 2012 Dec 15;135(4):2789-95. doi: 10.1016/j.foodchem.2012.06.043. Epub 2012 Jul 13.
A saponification/extraction procedure and high performance liquid chromatography (HPLC) analysis method were developed and validated for simultaneous analysis of phytosterols, tocopherols and lutein (a carotenoid) in soybeans. Separation was achieved on a phenyl column with a ternary, isocratic solvent system of acetonitrile, methanol and water (48:22.5:29.5, v/v/v). Evaporative light scattering detection (ELSD) was used to quantify β-sitosterol, stigmasterol, campesterol, and α-, δ- and γ-tocopherols, while lutein was quantified with visible light absorption at 450 nm. Peak identification was verified by retention times and spikes with external standards. Standard curves were constructed (R(2)>0.99) to allow for sample quantification. Recovery of the saponification and extraction was demonstrated via analysis of spiked samples. Also, the accuracy of results of four soybeans using the described saponification and HPLC analytical method was validated against existing methods. This method offers a more efficient alternative to individual methods for quantifying lutein, tocopherols and sterols in soybeans.
建立并验证了一种用于同时分析大豆中植物甾醇、生育酚和叶黄素（一种类胡萝卜素）的皂化/提取程序和高效液相色谱（HPLC）分析方法。在苯基柱上，采用乙腈、甲醇和水（48:22.5:29.5，v/v/v）的三元等度溶剂系统实现分离。蒸发光散射检测（ELSD）用于定量β-谷甾醇、豆甾醇、菜油甾醇和α-、δ-和γ-生育酚，而叶黄素则通过可见光在 450nm 处的吸收进行定量。通过保留时间和外部标准品的添加来验证峰的识别。通过分析加标样品，构建了标准曲线（R²>0.99）以允许对样品进行定量。通过分析加标样品，验证了皂化和提取的回收率。此外，还针对现有的方法，对描述的皂化和 HPLC 分析方法对四种大豆的结果的准确性进行了验证。与单个方法相比，该方法为定量分析大豆中的叶黄素、生育酚和甾醇提供了一种更有效的替代方法。
