Separation and quantitation of free fatty acids and fatty acid methyl esters by reverse phase high pressure liquid chromatography.
Separation and quantitation of free fatty acids and fatty acid methyl esters by reverse phase high pressure liquid chromatography.
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发表时间：
1983
影响因子：
6.5
通讯作者：
M. Aveldaño;M. Vanrollins;L. Horrocks
中科院分区：
文献类型：
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作者：
M. Aveldaño;M. Vanrollins;L. Horrocks
十八烷基甲硅烷基柱上的反相高压液相色谱法（HPLC）可分离由哺乳动物组织磷脂制备的游离脂肪酸或脂肪酸甲酯的混合物。乙腈-水混合物用于洗脱酯。用磷酸水溶液代替水来分离游离酸。不饱和化合物通过其在192 nm处的吸收进行检测和定量。在205 nm处更好地检测饱和物。复杂混合物中脂肪酸的洗脱顺序随乙腈浓度的变化而变化。在任何给定的浓度下，一些化合物重叠。然而，通过改变溶剂强度，可以解析任何感兴趣的脂肪酸，包括许多几何异构体和位置异构体。通过银化薄层色谱法（TLC）根据不饱和度进行预分馏的甲基酯通过单独用CH 3CN洗脱而快速、完全分离。TLC-反相HPLC可用作分析和制备程序。辛基硅烷基柱用于快速分离和改进馏分中微量或低紫外吸收组分的检测。例如，在牛脑甘油磷脂中已检测到具有多达32个碳的单烯脂肪酸。测量了3H-和14C-标记的脂肪酸的比放射性和复杂脂质的酰基之间的放射性分布。该方法不推荐用于完全成分分析，但可用于在标记脂肪酸的周转和代谢转化研究期间测定特定放射性。
Reverse phase high pressure liquid chromatography (HPLC) on octadecylsilyl columns separates mixtures of either free fatty acids or fatty acid methyl esters prepared from mammalian tissue phospholipids. Acetonitrile-water mixtures are used for the elution of esters. Aqueous phosphoric acid is substituted for water for the separation of the free acids. Unsaturated compounds are detected and quantitated by their absorption at 192 nm. Saturates are detected better at 205 nm. The order of elution of fatty acids in complex mixtures varies as a function of acetonitrile concentration. At any given concentration, some compounds overlap. However, by varying the solvent strength, any fatty acid of interest can be resolved including many geometrical and positional isomers. Methyl esters prefractionated according to unsaturation by argentation thin-layer chromatography (TLC) are rapidly and completely separated by elution with CH3CN alone. Argentation TLC-reverse phase HPLC can be used as an analytical as well as a preparative procedure. Octylsilyl columns are used for rapid resolution and improved detection of minor or low ultraviolet-absorbing components in the fractions. For example, monoenoic fatty acids with up to 32 carbons have been detected in bovine brain glycerophospholipids. Specific radioactivities of 3H- and 14C-labeled fatty acids and the distribution of radioactivity among acyl groups from complex lipids are measured. The method is not recommended for complete compositional analysis, but is useful for determinations of specific radioactivities during studies on turnover and metabolic conversions of labeled fatty acids.