High-Throughput Analysis of Total Plasma Fatty Acid Composition with Direct In Situ Transesterification
High-Throughput Analysis of Total Plasma Fatty Acid Composition with Direct In Situ Transesterification
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DOI：
10.1371/journal.pone.0012045
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发表时间：
2010-08-09
期刊：
影响因子：
3.7
通讯作者：
Koletzko, Berthold
中科院分区：
文献类型：
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作者：
Glaser, Claudia;Demmelmair, Hans;Koletzko, Berthold
背景资料：血浆脂肪酸（FA）组成反映了膳食摄入和内源性周转，并与短期和长期的健康结果相关。总血浆FA池代表所有含FA脂质组分的组成。我们开发了一种简化的和负担得起的高通量的方法，用于分析总血浆FA的组成，适合大型studies.Methodology/主要调查结果：总脂质FA从100亩血浆原位转移到甲酯，避免了最初的提取和干燥步骤。提取一次脂肪酸甲酯，并通过气相色谱法进行分析。为新型直接原位酯交换法的优化，确定了反应参数。批内分析（n = 8）显示FA的变异系数低于4%，贡献超过1%的总FA.Conclusions/显着性：结果显示出良好的一致性与FA浓度通过参考方法获得。新的直接原位酯交换法是稳健和简单的。将样品制备时间和分析成本降至最低。在大型研究中，该方法是评估血浆FA状态的传统方法的经济和生态上级替代方法。
Background: 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 mu l 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.