A direct method for fatty acid methyl ester synthesis: Application to wet meat tissues, oils, and feedstuffs
A direct method for fatty acid methyl ester synthesis: Application to wet meat tissues, oils, and feedstuffs
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DOI：
10.2527/jas.2006-491
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发表时间：
2007-06-01
影响因子：
3.3
通讯作者：
Gaskins, C. T.
中科院分区：
文献类型：
--
作者：
O'Fallon, J. V.;Busboom, J. R.;Gaskins, C. T.
一个简化的协议，以获得脂肪酸甲酯（FAME）直接从新鲜的组织，油，或饲料，没有事先的有机溶剂萃取，提出。利用该方案，FAME合成在高达33%水的存在下进行。将湿组织或其它样品在含有C13：0作为内标的1 N KOH/MeOH中在55 ℃下透化并水解1.5小时。中和KOH，并在55 ℃下通过H2SO 4催化将FFA甲基化1.5小时。然后将己烷加入反应管中，涡旋混合并离心。将己烷移液至气相色谱小瓶中，用于随后的气相色谱分析。为方便起见，所有反应均在单个螺旋盖Pyrex管中进行。该方法符合脂肪酸分析的许多标准，包括不异构化CLA或引入脂肪酸伪影。它适用于新鲜、冷冻或冻干的组织样品，以及油、蜡和饲料。与其他方法相比，该方法省时省力，经济实惠。其独特的性能，包括容易的样品制备，是因为水包括在FAME反应混合物中，而不是消除。
A simplified protocol to obtain fatty acid methyl esters (FAME) directly from fresh tissue, oils, or feedstuffs, without prior organic solvent extraction, is presented. With this protocol, FAME synthesis is conducted in the presence of up to 33% water. Wet tissues, or other samples, are permeabilized and hydrolyzed for 1.5 h at 55 degrees C in 1 N KOH in MeOH containing C13:0 as the internal standard. The KOH is neutralized, and the FFA are methylated by H2SO4 catalysis for 1.5 h at 55 degrees C. Hexane is then added to the reaction tube, which is vortex-mixed and centrifuged. The hexane is pipetted into a gas chromatography vial for subsequent gas chromatography. All reactions are conducted in a single screw-cap Pyrex tube for convenience. The method meets many criteria for fatty acid analysis, including not isomerizing CLA or introducing fatty acid artifacts. It is applicable to fresh, frozen, or lyophilized tissue samples, in addition to oils, waxes, and feedstuffs. The method saves time and effort and is economical when compared with other methods. Its unique performance, including easy sample preparation, is achieved because water is included rather than eliminated in the FAME reaction mixtures.