高纯度亚麻木酚素的分离纯化和分析

Preparation and Characterization of Highly Pure Flax Lignan from Flaxseeds

开展高纯度亚麻木酚素的分离纯化和分析方法研究,选取亚麻籽为研究对象,通过正己烷脱脂、乙醇提取、AB-8大孔吸附树脂分离、高速逆流色谱(high-speed countercurrent chromatography,HSCCC)纯化制备得到高纯度亚麻木酚素。其中,大孔吸附树脂的洗脱溶剂为80%乙醇溶液,HSCCC溶剂体系为叔丁基甲醚-正丁醇-乙腈-水(1∶3∶1∶5,V/V)。实验分离得到的亚麻木酚素经薄层色谱、高效液相色谱-二极管阵列检测器-蒸发光散射检测器联用和高效液相色谱-质谱联用进行纯度分析,结果显示杂质质量分数低于0.5%。通过采用多种高效液相色谱条件分析,峰面积归一化法进行纯度计算,其纯度在99.3%～99.5%之间。最后采用紫外光谱、红外光谱、高分辨质谱、核磁共振波谱和元素分析等方法鉴定其结构为亚麻木酚素。结果表明,该分离纯化方法简单可行,纯度分析方法准确可靠,制备得到的样品具有纯度高的优点,可以应用于相关产品的质量控制和方法验证。

Highly pure flax lignan was prepared from flaxseeds by n-hexane defatting, ethanol extraction, AB-8 macroporous resin separation and high-speed countercurrent chromatography (HSCCC) purification. Flax lignan was eluted with 80% ethanol from AB-8, and the HSCCC solvent system was composed of tert-butyl methyl ether, n-butanol, acetonitrile and water (1:3:1:5, V/V). The results showed that the impurity content was less than 0.5%, as detected by thin layer chromatography (TLC), high performance liquid chromatography (HPLC), HPLC-DAD-ELSD and HPLC-MS. The purity of flax lignan was between 99.3%-99.5%, as calculated by peak area normalization under various HPLC conditions. The structure was identified by ultraviolet spectroscopy (UV), infrared spectroscopy (IR), high-resolution mass spectrometry (HRMS), nuclear magnetic resonance (NMR) and elemental analysis. The purification method proved to be simple and feasible, and the methods used for purity analysis were accurate and reliable. The flax lignin prepared in this study is enough pure to be used in method validation and quality control.