Masood M Athar, Salem Norman
Laboratory of Membrane Biochemistry and Biophysics, National Institutes on Alcohol Abuse and Alcoholism, National Institutes of Health, Rockville, MD, USA.
Lipids. 2008 Feb;43(2):171-80. doi: 10.1007/s11745-007-3130-9. Epub 2007 Dec 15.
Fatty acid analysis is an important research tool, and indices derived from essential fatty acid contents serve as useful biomarkers related to cardiovascular and other chronic disease risk. Both clinical and basic studies of essential fatty acid composition are becoming ever larger in magnitude leading to delays while the rather laborious lipid analyses are performed. A robotic transesterification procedure has been developed for high-throughput analysis of plasma fatty acid methyl esters. In this approach, robots perform most steps including plasma and reagent transfer, transesterification reaction via heating at 80 degrees C in open tubes with multiple reagent additions, followed by two-phase extraction and transfer of lipid extracts to GC vials. The vials are then placed directly onto a GC autosampler carousel for robotic sample injection. An improved fast GC method is presented in which the peaks of interest are eluted within 6 min. This method is readily scalable to prepare and analyze 200 samples per day (1,000 samples per week) so that large clinical trials can be accommodated.
脂肪酸分析是一种重要的研究工具，从必需脂肪酸含量得出的指标可作为与心血管疾病及其他慢性疾病风险相关的有用生物标志物。对必需脂肪酸组成的临床和基础研究规模越来越大，导致在进行相当繁琐的脂质分析时出现延迟。已开发出一种用于高通量分析血浆脂肪酸甲酯的机器人酯交换程序。在这种方法中，机器人执行大多数步骤，包括血浆和试剂转移、在开放式试管中于80摄氏度加热并多次添加试剂进行酯交换反应，随后进行两相萃取并将脂质提取物转移至气相色谱小瓶中。然后将小瓶直接放置在气相色谱自动进样器转盘上进行机器人进样。本文介绍了一种改进的快速气相色谱方法，其中感兴趣的峰在6分钟内洗脱。该方法易于扩展，每天可制备和分析200个样品（每周1000个样品），从而能够满足大型临床试验的需求。
