In clinical lipidomics, it is a challenge to measure a large number of samples and to reproduce the quantitative results. We expanded the range of application of the tandem mass tag (TMT) method, which is widely used in proteomics, to lipidomic fields. There are various types of lipid molecule, for example, eicosanoids have a carboxyl group and phosphatidic acid has a phosphate group. We modified these functional groups simultaneously with TMT. This approach allows for a single analysis by mixing six samples and using one of the six samples as a bridging sample; the quantitative data can be easily normalized even if the number of measurements increases. To accommodate a large number of samples, we utilize a pooled serum sample of 300 individuals as a bridging sample. The stability of these lipid molecules in serum was examined as an analytical validation for the simultaneous TMT labeling. It was found that the stability of these lipid molecules in serum differs greatly depending on the lipid species. These findings reaffirmed the importance of proper sample preparation and storage to obtain reliable data. The TMT labeling method is expected to be a useful method for lipidomics with high-throughput and reliable reproducibility.

中文翻译：

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含羧基和磷酸盐基团的脂质分子串联质量标签标记方法的开发及其在人血清中的稳定性

在临床脂质组学中，测量大量样品并再现定量结果是一个挑战。我们将蛋白质组学中广泛使用的串联质谱标签（TMT）方法的应用范围扩展到脂质组学领域。脂质分子有各种类型，例如，类二十烷酸具有羧基，磷脂酸具有磷酸基。我们与TMT同时修改了这些功能组。这种方法可以通过混合六个样本并将六个样本之一用作桥接样本来进行单个分析。即使测量数量增加，定量数据也可以轻松归一化。为了容纳大量样本，我们将300个个体的合并血清样本用作桥接样本。检查这些脂质分子在血清中的稳定性，作为同时TMT标记的分析验证。已经发现，这些脂质分子在血清中的稳定性取决于脂质种类而有很大差异。这些发现再次证明了正确准备和存储样品以获得可靠数据的重要性。TMT标记方法有望成为一种具有高通量和可靠重现性的脂质组学方法。