Generation of a Collision Cross Section Library for Multi-Dimensional Plant Metabolomics Using UHPLC-Trapped Ion Mobility-MS/MS.

Mark Schroeder,Heino M Heyman,Aiko Barsch,Sven W Meyer,Lloyd W Sumner

doi:10.3390/metabo10010013

Mark Schroeder, Heino M Heyman + Show 3 more

Open Access

https://doi.org/10.3390/metabo10010013

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Abstract

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.

Highlights

Metabolomics is a revolutionary systems biology tool for understanding plant metabolism and elucidating gene function [1,2,3]
Balston Model N2-80A membrane based nitrogen generation system were used to record Trapped ion mobility spectrometry (TIMS) CCSN2 values for 146 authentic plant natural products mostly consisting of flavonoids, glycosylated flavonoids, isoflavonoids, triterpenes, and glycosylated triterpenes
We report here a collision cross section (CCS) library focused on plant natural products and specialized metabolites that contains 146 compounds, 343 CCS values, and 29 isomers annotated

Summary

Introduction

Metabolomics is a revolutionary systems biology tool for understanding plant metabolism and elucidating gene function [1,2,3]. The continual refinement, increasing scope and larger-scale have led to the evolution of modern plant metabolomics, which has matured as a valuable tool for advancing our understanding of plant biology and physiology [2,4,5,6]. The vast utility of metabolomics is well documented in the literature, its full scientific promise has not yet been realized due to multiple technical challenges. These challenges have been reviewed by the metabolomics community and a set of grand challenges identified by the Plant, Algae, and Microbial Metabolomics Research Coordination. We propose a multi-dimensional, combinatorial solution for both using UHPLC-trapped ion mobility coupled to mass spectrometry (UHPLC-TIMS-TOF-MS)

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