title: Expanding Lipidome Coverage Using LC-MS/MS Data-Dependent Acquisition with Automated Exclusion List Generation.
abstract: Untargeted omics analyses aim to comprehensively characterize biomolecules within a biological system. Changes in the presence or quantity of these biomolecules can indicate important biological perturbations, such as those caused by disease. With current technological advancements, the entire genome can now be sequenced; however, in the burgeoning fields of lipidomics, only a subset of lipids can be identified. The recent emergence of high resolution tandem mass spectrometry (HR-MS/MS), in combination with ultra-high performance liquid chromatography, has resulted in an increased coverage of the lipidome. Nevertheless, identifications from MS/MS are generally limited by the number of precursors that can be selected for fragmentation during chromatographic elution. Therefore, we developed the software IE-Omics to automate iterative exclusion (IE), where selected precursors using data-dependent topN analyses are excluded in sequential injections. In each sequential injection, unique precursors are fragmented until HR-MS/MS spectra of all ions above a user-defined intensity threshold are acquired. IE-Omics was applied to lipidomic analyses in Red Cross plasma and substantia nigra tissue. Coverage of the lipidome was drastically improved using IE. When applying IE-Omics to Red Cross plasma and substantia nigra lipid extracts in positive ion mode, 69% and 40% more molecular identifications were obtained, respectively. In addition, applying IE-Omics to a lipidomics workflow increased the coverage of trace species, including odd-chained and short-chained diacylglycerides and oxidized lipid species. By increasing the coverage of the lipidome, applying IE to a lipidomics workflow increases the probability of finding biomarkers and provides additional information for determining etiology of disease. Graphical Abstract ᅟ.
authors.affiliation: Department of Chemistry, University of Florida, 214 Leigh Hall, Gainesville, FL, 32611, USA.
authors.forename: Jeremy P
authors.initials: JP
authors.lastname: Koelmel
authors.affiliation: College of Engineering, University of Florida, 412 Newell Drive, Gainesville, FL, 32611, USA.
authors.forename: Nicholas M
authors.initials: NM
authors.lastname: Kroeger
authors.affiliation: Department of Chemistry, University of Florida, 214 Leigh Hall, Gainesville, FL, 32611, USA.
authors.forename: Emily L
authors.initials: EL
authors.lastname: Gill
authors.affiliation: Department of Chemistry, University of Florida, 214 Leigh Hall, Gainesville, FL, 32611, USA.|Hollings Marine Laboratory, National Institute of Standards and Technology, 331 Ft. Johnson Rd., Charleston, SC, 29412, USA.
authors.forename: Candice Z
authors.initials: CZ
authors.lastname: Ulmer
authors.affiliation: Hollings Marine Laboratory, National Institute of Standards and Technology, 331 Ft. Johnson Rd., Charleston, SC, 29412, USA.
authors.forename: John A
authors.initials: JA
authors.lastname: Bowden
authors.affiliation: Department of Chemistry, University of Florida, 214 Leigh Hall, Gainesville, FL, 32611, USA.
authors.forename: Rainey E
authors.initials: RE
authors.lastname: Patterson
authors.affiliation: Department of Chemistry, University of Florida, 214 Leigh Hall, Gainesville, FL, 32611, USA.|Department of Pathology, Immunology, and Laboratory Medicine, College of Medicine, University of Florida, PO BOX 13303, 1395 Center Dr, Gainesville, FL, 32610, USA.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Yost
authors.affiliation: Department of Chemistry, University of Florida, 214 Leigh Hall, Gainesville, FL, 32611, USA. tgarrett@ufl.edu.|Department of Pathology, Immunology, and Laboratory Medicine, College of Medicine, University of Florida, PO BOX 13303, 1395 Center Dr, Gainesville, FL, 32610, USA. tgarrett@ufl.edu.
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Garrett
keywords: Data-dependent Analysis; Diglyceride; Lipidomics; Lipids; Mass spectrometry; Metabolomics; Tandem mass spectrometry