高效液相色谱法分离纯化微生物油脂中的花生四烯酸

Isolation and Purification of Arachidonic Acid in Microbial Lipids by High Performance Liquid Chromatography

为满足花生四烯酸样品制备的需要,将微生物油脂中制得的脂肪酸先经尿素包合(脲包法)和甲酯化后,再利用反相高效液相色谱法分离纯化其中的花生四烯酸。在C18反相柱上,以甲醇 水(体积比为95∶5)为流动相,采用示差折光检测器检测,流速为5mL/min时,目标产物的分离度较佳。用气相色谱/质谱法(GC/MS)进行分析的结果表明,花生四烯酸甲酯在纯化后的样品中的质量分数由38%提高到99%以上,基本达到标准物质的质量要求。

A simple isolation and purification method for arachidonic acid (5,8,11,14eicosatetraenoic acid, AA) from microbial lipids is presented. At first, AA was concentrated by urea inclusion reaction from mixed fatty acids made from microbial lipids, and then esterified with BF3methanol. AA was successfully isolated and purified with reversedphase high performance liquid chromatography on a μBondapakTM C18 preparative column (30 mm id×300 mm, 15 μm), using methanolwater (95∶5 in volume ratio) as the mobile phase at a flow rate of 5 mL/min and detected by a 410 Waters differential refractometer. After urea inclusion reaction the sample was refined and the content of AA was greatly increased. Finally, the purity of AA reached 99% from the initial content of 38%. The analysis was carried out by gas chromatography/mass spectrometry (GC/MS).