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

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.

Unraveling mosquito metabolism with mass spectrometry-based metabolomics

“…Ultrahigh-resolution mass spectrometry (UHRMS; Rs >30 000) instruments feature QTOFMS-based options that include a GC-HRT+ system, and a timsTOF system that couples a trapped ion mobility spectrometry (TIMS) device to a QTOFMS. The TIMS device expands the capability of this instrument to resolve coeluting or isobaric substances in the gas phase prior to TOF analysis [64]. The OrbiTrap (OT)-class of MS systems boast maximum mass resolutions from 120 000 up to 1 000 000 in a simple-to-use platform, and these instruments allow for fine isotopolog peak distributions that reveal and improve the fidelity of molecular/product ion formula assignment [65].…”

Comprehensive multicomponent characterization and fingerprinting analysis of Lanqin Oral Liquid by ultra‐high‐performance liquid chromatography coupled with ion mobility‐quadrupole time‐of‐flight mass spectrometry

“…It was noted that CCS could play complementary role in characterizing the multicomponents from LOL. Three currently available natural product‐specific CCS libraries were fully searched to validate the identifications of LOL ingredients [26–28]. As a result, seven groups of molecules (together with the isomers) could receive hits in the database as given in Table S6.…”

Plants Metabolome Study: Emerging Tools and Techniques

“…McCullagh et al [130] used the TW CCSN 2 library to screen the steviol glycosides in 55 food commodities. Schroeder et al [131] identified 146 plant natural compounds, 343 CCS values, and 29 isomers annotated (various flavonoids and isoflavonoids) in Medicago truncatula based on CCS, retention time, accurate mass, and molecular formula. The combination of a large-scale CCS database and different MS/MS spectra will assist in the discovery of new metabolites.…”