The utility of metabolomics is well documented; however, its full scientific promise has not yet been realized due to multiple technical challenges. These grand challenges include accurate chemical identification of all observable metabolites and the limiting depth-of-coverage of current metabolomics methods. Here, we report a combinatorial solution to aid in both grand challenges using UHPLC-trapped ion mobility spectrometry coupled to tandem mass spectrometry (UHPLC-TIMS-TOF-MS). TIMS offers additional depth-of-coverage through increased peak capacities realized with the multi-dimensional UHPLC-TIMS separations. Metabolite identification confidence is simultaneously enhanced by incorporating orthogonal collision cross section (CCS) data matching. To facilitate metabolite identifications, we created a CCS library of 146 plant natural products. This library was generated using TIMS with N2 drift gas to record the TIMSCCSN2 of plant natural products with a high degree of reproducibility; i.e., average RSD = 0.10%. The robustness of TIMSCCSN2 data matching was tested using authentic standards spiked into complex plant extracts, and the precision of CCS measurements were determined to be independent of matrix affects. The utility of the UHPLC-TIMS-TOF-MS/MS in metabolomics was then demonstrated using extracts from the model legume Medicago truncatula and metabolites were confidently identified based on retention time, accurate mass, molecular formula, and CCS.

中文翻译：

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使用UHPLC捕获的离子淌度MS / MS为多维植物代谢组学生成碰撞截面库。

代谢组学的效用已被充分证明。然而，由于多重技术挑战，其完整的科学承诺尚未实现。这些重大挑战包括对所有可观察到的代谢物进行准确的化学鉴定以及当前代谢组学方法的有限覆盖范围。在这里，我们报告了一种组合解决方案，以解决使用UHPLC捕获的离子淌度质谱与串联质谱（UHPLC-TIMS-TOF-MS）结合的两个挑战。TIMS通过增加多维UHPLC-TIMS分离所实现的峰容量，提供了更大的覆盖深度。通过合并正交碰撞截面（CCS）数据匹配，可以同时提高代谢物识别的可信度。为便于代谢物鉴定，我们创建了一个包含146种植物天然产物的CCS库。该文库是使用TIMS和N2漂移气生成的，以高重复性记录了植物天然产物的TIMSCCSN2。即，平均RSD = 0.10％。使用掺入复杂植物提取物中的真实标准品测试了TIMSCCSN2数据匹配的鲁棒性，并且确定了CCS测量的精度与基质影响无关。然后使用豆科植物苜蓿模型提取物证明了UHPLC-TIMS-TOF-MS / MS在代谢组学中的效用，并根据保留时间，准确的质量，分子式和CCS可靠地鉴定了代谢物。使用加标到复杂植物提取物中的真实标准品测试了TIMSCCSN2数据匹配的鲁棒性，并且确定了CCS测量的精度与基质影响无关。然后使用豆科植物苜蓿模型提取物证明了UHPLC-TIMS-TOF-MS / MS在代谢组学中的效用，并根据保留时间，准确的质量，分子式和CCS可靠地鉴定了代谢物。使用掺入复杂植物提取物中的真实标准品测试了TIMSCCSN2数据匹配的鲁棒性，并且确定了CCS测量的精度与基质影响无关。然后使用豆科植物苜蓿模型提取物证明了UHPLC-TIMS-TOF-MS / MS在代谢组学中的效用，并根据保留时间，准确的质量，分子式和CCS可靠地鉴定了代谢物。