High-Throughput Quantitative Lipidomics Analysis of Nonesterified Fatty Acids in Human Plasma
High-Throughput Quantitative Lipidomics Analysis of Nonesterified Fatty Acids in Human Plasma
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DOI：
10.1021/acs.jproteome.6b00198
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发表时间：
2016-07-01
影响因子：
4.4
通讯作者：
Masoodi, Mojgan
中科院分区：
文献类型：
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作者：
Christinat, Nicolas;Morin-Rivron, Delphine;Masoodi, Mojgan
我们提出了一种高通量、非靶向脂质组学方法，使用液相色谱与高分辨率质谱联用来定量分析非酯化脂肪酸。我们应用这种方法筛选人血浆样本中从中链到超长链（8 至 24 个碳原子）的多种脂肪酸。该方法使我们能够通过色谱法将支链物质与其直链异构体分离，以及分离生物学上重要的 omega-3 和 omega-6 多不饱和脂肪酸。我们使用 51 种脂肪酸来证明该方法的定量能力，定量限度在纳摩尔范围内；然而，该方法不仅限于这些脂肪酸种类。高通量样品制备是在机器人平台上开发和进行的，可在 3 小时内同时提取 96 个样品。该高通量平台用于评估不同类型的人血浆采集和制备对健康供体非酯化脂肪酸谱的影响。已将抗凝剂 EDTA 和肝素的使用与简单凝血进行了比较，并且在大多数非酯化脂肪酸浓度中仅检测到有限的变化。
We present a high-throughput, nontargeted lipidomics approach using liquid chromatography coupled to high-resolution mass spectrometry for quantitative analysis of nonesterified fatty acids. We applied this method to screen a wide range of fatty acids from medium-chain to very long-chain (8 to 24 carbon atoms) in human plasma samples. The method enables us to chromatographically separate branched-chain species from their straight-chain isomers as well as separate biologically important omega-3 and omega-6 polyunsaturated fatty acids. We used 51 fatty acid species to demonstrate the quantitative capability of this method with quantification limits in the nanomolar range; however, this method is not limited only to these fatty acid species. High-throughput sample preparation was developed and carried out on a robotic platform that allows extraction of 96 samples simultaneously within 3 h. This high-throughput platform was used to assess the influence of different types of human plasma collection and preparation on the nonesterified fatty acid profile of healthy donors. Use of the anticoagulants EDTA and heparin has been compared with simple clotting, and only limited changes have been detected in most nonesterified fatty acid concentrations.