Untargeted Comprehensive Two-Dimensional Liquid Chromatography Coupled with High-Resolution Mass Spectrometry...

In this work, a new strategy for the chemometric analysis of two-dimensional liquid chromatography-high-resolution mass spectrometry (LC × LC-HRMS) data is proposed. This approach consists of a preliminary compression step along the mass spectrometry (MS) spectral dimension based on the selection of the regions of interest (ROI), followed by a further data compression along the chromatographic dimension by wavelet transforms. In a secondary step, the multivariate curve resolution alternating least squares (MCR-ALS) method is applied to previously compressed data sets obtained in the simultaneous analysis of multiple LC × LC-HRMS chromatographic runs from multiple samples. The feasibility of the proposed approach is demonstrated by its application to a large experimental data set obtained in the untargeted LC × LC-HRMS study of the effects of different environmental conditions (watering and harvesting time) on the metabolism of multiple rice samples. An untargeted chromatographic setup coupling two different liquid chromatography (LC) columns [hydrophilic interaction liquid chromatography (HILIC) and reversed-phase liquid chromatography (RPLC)] together with an HRMS detector was developed and applied to analyze the metabolites extracted from rice samples at the different experimental conditions. In the case of the metabolomics study taken as example in this work, a total number of 154 metabolites from 15 different families were properly resolved after the application of MCR-ALS. A total of 139 of these metabolites could be identified by their HRMS spectra. Statistical analysis of their concentration changes showed that both watering and harvest time experimental factors had significant effects on rice metabolism. The biochemical insight of the effects of watering and harvesting experimental factors on the changes in concentration of these detected metabolites in the investigated rice samples is attempted.