Polette A, Durand P, Floccard B, Blache D
INSERM U. 63-22, Bron, France.
Anal Biochem. 1992 Nov 1;206(2):241-5. doi: 10.1016/0003-2697(92)90360-j.
The present report describes a simple method to selectively extract free fatty acids and analyze them by capillary gas-liquid chromatography. The procedure is based on the use of fumed silicon dioxide. In the presence of plasma, this material induces a rapid rise in the viscosity of the mixture and presents the ability to trap large particles such as emulsified lipids and lipoproteins. Albumin-bound fatty acids are thus left in the aqueous media. We present applications of our procedure for the analysis of free fatty acids in 0.2 ml of plasma from rat or human. By comparison with the method utilizing thin-layer chromatography for the separation of fatty acids and gas chromatography analysis, the present method has been found to be reliable and simple. The recovery of linoleic acid was 92.1 +/- 8.2%, a value which is about twice better than that obtained with the procedure using thin-layer chromatography. In particular, long-chain polyunsaturated fatty acids were better preserved. Our procedure does not require the use of organic solvents and its simplicity and reproducibility make it suitable for routine specific determination of the composition of free fatty acids in biological samples.
本报告描述了一种选择性提取游离脂肪酸并通过毛细管气液色谱法进行分析的简单方法。该方法基于使用气相二氧化硅。在血浆存在的情况下,这种材料会使混合物的粘度迅速升高,并具有捕获大颗粒(如乳化脂质和脂蛋白)的能力。因此,与白蛋白结合的脂肪酸会留在水性介质中。我们展示了该方法在分析大鼠或人类0.2ml血浆中游离脂肪酸的应用。与利用薄层色谱法分离脂肪酸和气相色谱分析的方法相比,本方法已被证明是可靠且简单的。亚油酸的回收率为92.1±8.2%,该值约为使用薄层色谱法获得的值的两倍。特别是,长链多不饱和脂肪酸得到了更好的保留。我们的方法不需要使用有机溶剂,其简单性和可重复性使其适用于生物样品中游离脂肪酸组成的常规特定测定。