Division of Nutritional Sciences, Cornell University , Ithaca, New York 14853, United States.
Anal Chem. 2014 Feb 18;86(4):2175-84. doi: 10.1021/ac403845u. Epub 2014 Jan 28.
Recent advances in mass spectrometry have allowed for unprecedented characterization of human metabolism and its contribution to disease. Despite these advances, limitations in metabolomics technology remain. Here, we describe a metabolomics strategy that consolidates several recent improvements in mass spectrometry technology. The platform involves a high-resolution Orbitrap mass spectrometer coupled to faster scanning speeds, allowing for polarity switching and improved ion optics resulting in enhanced sensitivity. When coupled to HILIC chromatography, we are able to quantify over 339 metabolites from an extract of HCT8 cells with a linear range of over 4 orders of magnitude in a single chromatographic run. These metabolites include diverse chemical classes ranging from amino acids to polar lipids. In addition, we also detect over 3000 additional potential metabolites present in mammalian cells. We applied this platform to characterize the metabolome of eight colorectal cancer cell lines and observed both commonalities and heterogeneities across their metabolic profiles when cells are grown in identical conditions. Together these results demonstrate that simultaneous profiling and quantitation of the human metabolome is feasible.
近年来，质谱技术的进步使得对人类代谢及其对疾病的贡献进行前所未有的描述成为可能。尽管取得了这些进展，但代谢组学技术仍存在局限性。在这里，我们描述了一种代谢组学策略，该策略整合了质谱技术的几项最新改进。该平台涉及高分辨率轨道阱质谱仪与更快的扫描速度相结合，允许极性切换和改进的离子光学，从而提高了灵敏度。当与亲水相互作用色谱法结合使用时，我们能够从 HCT8 细胞提取物中定量超过 339 种代谢物，单个色谱运行的线性范围超过 4 个数量级。这些代谢物包括从氨基酸到极性脂质的多种化学类别。此外，我们还检测到哺乳动物细胞中存在的 3000 多种其他潜在代谢物。我们应用该平台来描述八种结直肠癌细胞系的代谢组，并观察到在相同条件下生长时，其代谢谱存在共性和异质性。这些结果共同表明，同时对人类代谢组进行分析和定量是可行的。
