Akoto Lawrence, Vreuls René J J, Irth Hubertus, Pel Roel, Stellaard Frans
Vrije Universiteit, Department of Analytical Chemistry and Applied Spectroscopy, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands.
J Chromatogr A. 2008 Apr 4;1186(1-2):365-71. doi: 10.1016/j.chroma.2007.08.080. Epub 2007 Sep 11.
Gas chromatography (GC) has in recent times become an important tool for the fatty acid profiling of human blood and plasma. An at-line procedure used in the fatty acid profiling of whole/intact aquatic micro-organisms without any sample preparation was adapted for this work. A direct thermal desorption (DTD) interface was used to profile the fatty acid composition of human plasma and whole human blood of eight volunteers in a procedure omitting the usual lipid extraction steps that precede sample methylation in the traditional (off-line) protocols. Trimethylsulfonium hydroxide (TMSH) was used as reagent for thermally assisted methylation. In a fully automated manner, the liner of the GC injector is used as a sample-and-reaction container with the aid of the DTD interface. The fatty acid methyl ester (FAME) profiles obtained using this novel approach, were very identical to those obtained when the traditional off-line protocol was applied. FAME yields obtained in the at-line DTD method were found to be very similar for saturated fatty acids, but significantly higher for polyunsaturated fatty acids compared to off-line yields. As a result of the contribution of circulating cell membranes in blood, substantial differences were observed when the amount of FAMEs obtained in whole human blood and human plasma samples were compared after their analysis. Thanks to the fully automated operation of this novel procedure, large series of analyses can easily be performed.
气相色谱法(GC)近来已成为分析人体血液和血浆中脂肪酸谱的重要工具。本研究采用了一种用于完整水生微生物脂肪酸谱分析的在线程序,无需任何样品制备。使用直接热脱附(DTD)接口,在一个省略了传统(离线)方案中样品甲基化之前通常的脂质提取步骤的程序中,对八名志愿者的人体血浆和全血中的脂肪酸组成进行分析。使用氢氧化三甲锍(TMSH)作为热辅助甲基化试剂。借助DTD接口,以全自动方式将气相色谱进样器的衬管用作样品和反应容器。使用这种新方法获得的脂肪酸甲酯(FAME)谱与应用传统离线方案时获得的谱非常相似。在线DTD方法中获得的FAME产率对于饱和脂肪酸而言非常相似,但与离线产率相比,多不饱和脂肪酸的产率显著更高。由于血液中循环细胞膜的作用,在分析后比较全血和人体血浆样品中获得的FAME量时,观察到了显著差异。得益于这种新方法的全自动操作,可以轻松进行大量分析。