High Throughput UHPLC-MS-Based Lipidomics Using Vacuum Jacketed Columns



Plumb, Robert S, Isaac, Giorgis, Rainville, Paul D, Hill, Jason, Gethings, Lee A, Johnson, Kelly A, Lauterbach, Joshua and Wilson, Ian D ORCID: 0000-0002-8558-7394
(2022) High Throughput UHPLC-MS-Based Lipidomics Using Vacuum Jacketed Columns. JOURNAL OF PROTEOME RESEARCH, 21 (3). pp. 691-701.

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Abstract

Reversed-phase UHPLC-MS is extensively employed for both the profiling of biological fluids and tissues to characterize lipid dysregulation in disease and toxicological studies. With conventional LC-MS systems the chromatographic performance and throughput are limited due to dispersion from the fluidic connections as well as radial and longitudinal thermal gradients in the LC column. In this study vacuum jacketed columns (VJC), positioned at the source of the mass spectrometer, were applied to the lipidomic analysis of plasma extracts. Compared to conventional UHPLC, the VJC-based methods offered greater resolution, faster analysis, and improved peak intensity. For a 5 min VJC analysis, the peak capacity increased by 66%, peak tailing reduced by up to 34%, and the number of lipids detected increased by 30% compared to conventional UHPLC. The narrower peaks, and thus increased resolution, compared to the conventional system resulted in a 2-fold increase in peak intensity as well a significant improvement in MS and MS/MS spectral quality resulting in a 22% increase in the number of lipids identified. When applied to mouse plasma samples, reproducibility of the lipid intensities in the pooled QC ranged from 1.8-12%, with no related drift in <i>t</i><sub>R</sub> observed.

Item Type: Article
Uncontrolled Keywords: high-throughput, lipidomics, NIST 1950 plasma, mouse plasma, gefitinib
Divisions: Faculty of Health and Life Sciences
Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology
Depositing User: Symplectic Admin
Date Deposited: 07 Feb 2023 12:36
Last Modified: 07 Feb 2023 12:37
DOI: 10.1021/acs.jproteome.1c00836
Open Access URL: https://doi.org/10.1021/acs.jproteome.1c00836
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3168253