Division of Metabolic and Nutritional Medicine, Dr. von Hauner Children's Hospital, University of Munich Medical Center, Munich, Germany.
PLoS One. 2010 Aug 9;5(8):e12045. doi: 10.1371/journal.pone.0012045.
Plasma fatty acid (FA) composition reflects dietary intake and endogenous turnover and is associated with health outcomes on a short and long term basis. The total plasma FA pool represents the composition of all FA containing lipid fractions. We developed a simplified and affordable high-throughput method for the analysis of total plasma FA composition, suitable for large studies.
METHODOLOGY/PRINCIPAL FINDINGS: The total lipid FA from 100 microl plasma is transferred in situ into methyl esters, avoiding initial extraction and drying steps. The fatty acid methyl esters are extracted once and analyzed by gas chromatography. For the new direct in situ transesterification method optimal, reaction parameters were determined. Intra-assay analysis (n=8) revealed coefficients of variation below 4% for FA contributing more than 1% to total FA.
CONCLUSIONS/SIGNIFICANCE: The results show good agreement with FA concentrations obtained by a reference method. The new direct in situ transesterification method is robust and simple. Sample preparation time and analysis costs are reduced to a minimum. This method is an economically and ecologically superior alternative to conventional methods for assessing plasma FA status in large studies.
血浆脂肪酸 (FA) 组成反映了饮食摄入和内源性代谢，并与短期和长期的健康结果相关。总血浆 FA 池代表了所有含脂类 FA 的组成部分。我们开发了一种简化且经济实惠的高通量方法，用于分析总血浆 FA 组成，适用于大型研究。
方法/主要发现：从 100 微升血浆中提取的总脂质 FA 就地转化为甲酯，避免了初始提取和干燥步骤。脂肪酸甲酯提取一次，并通过气相色谱法进行分析。对于新的直接原位酯交换方法，确定了最佳的反应参数。对于总 FA 贡献超过 1%的 FA，日内分析（n=8）显示变异系数低于 4%。
结论/意义：结果与参考方法得到的 FA 浓度具有良好的一致性。新的直接原位酯交换方法具有较强的稳健性和简单性。样品制备时间和分析成本降低到最低。与传统方法相比，这种方法在大型研究中评估血浆 FA 状态具有经济和生态上的优势。
