Uncovering biologically significant lipid isomers with liquid chromatography, ion mobility spectrometry and mass spectrometry

Kyle, Jennifer; Zhang, Xing; Weitz, Karl K.; Monroe, Matthew E.; Ibrahim, Yehia; Moore, Ronald J.; Cha, Jeeyeon; Sun, Xiaofei; Lovelace, Erica S.; Wagoner, Jessica; Polyak, Stephen J.; Metz, Thomas O.; Dey, Sudhansu K.; Smith, Richard; Burnum-Johnson, Kristin; Baker, Erin

doi:10.1039/c5an02062j

Cited by 219 publications

(232 citation statements)

References 42 publications

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“…IM can also resolve additional lipid classes within each group on the basis of their different headgroups. There is strong consensus between the literature and our results that PCs have larger gas-phase conformations than other phospholipids (49,50). Within the PEs, we observe a clear separation between PEs with two ester linkages and PEs with one vinyl ether linkage (plasmalogen PEs, PEp), with the plasmalogen PEs having slightly larger CCS values than PEs with the same fatty acid compositions in positive mode (supplemental Table S1) and slightly smaller CCS values in negative mode (supplemental Table S2).…”

Section: Untargeted Lipidomics By Hilic-im-msmentioning

confidence: 69%

“…However, the positional isomers were not differentiated in this work. Previous work by Baker and coworkers (49) reported that positional isomers of phospholipids could be differentiated by IM separation, but the difference between the drift times of each pair of isomers is less than 1%, which is beyond the resolution of our instrument. In another study by Groessl et al (52), the effect of E/Z isomers on the CCS of PC(18:1/18:1) was also found to be less than 0.5%.…”

Section: Ccs Measurements Of Lipids By Twim-ms Calibrationmentioning

confidence: 93%

“…The decrease in CCS we observed between PEs and plasmalogen PEs in negative mode is consistent with the observations of Paglia et al (50), who reasoned that the decrease in CCS was the result of the additional double bond in the PEp. This reduction in CCS with the addition of double bonds is a common phenomenon in all lipid classes, with each double bond contributing a 1-5% decrease in CCS (28,49).…”

Section: Untargeted Lipidomics By Hilic-im-msmentioning

confidence: 99%

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Assessment of altered lipid homeostasis by HILIC-ion mobility-mass spectrometry-based lipidomics

Hines

Herron

2017

Journal of Lipid Research

117

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implicated in a number of human diseases, such as atherosclerosis (3), nonalcoholic fatty liver (4), diabetes (5), Alzheimer's disease (6), and cancer (7,8). Advances in lipidomic techniques and strategies, led by the LIPID MAPS consortium, have greatly enhanced our understanding of the distribution and biological roles of lipids (9-13). Modern mass spectrometry (MS), coupled with electrospray ionization (ESI), is the key to qualitative and quantitative lipidomic analysis. Typical MS-based lipidomic strategies are shotgun (i.e., direct infusion) lipidomics (9, 14) and liquid chromatography (LC)-MS lipidomics (11,15,16). Shotgun lipidomics relies on partial intrasource separation of lipid classes through varying the pH of the lipid solution and identification of lipid species by their characteristic fragmentation in tandem MS analysis (9,17). This approach has the advantage of being high-throughput, but it also has several disadvantages: a) suppression of low-abundant species by major polar lipids such as phosphatidylcholines; b) difficulty in analysis of lipid species that are poorly ionized by ESI; and c) inability to provide structural information on isobaric and isomeric species. The LC-MS-based strategy utilizes targeted analysis of each lipid class under conditions that are optimized for that particular class (11,15,16,18). This strategy has the advantages of being specific, sensitive, and comprehensive, but it also has limitations, such as being time consuming and less cost effective. To increase the throughput of the LC-MS lipidomics while maintaining the specificity and sensitivity, we desire improved chromatographic techniques and additional dimensions of separation that are orthogonal to LC and MS. Lipids play important roles in maintaining membrane structures and mediating signaling pathways (1, 2). Dysregulated lipid biosynthesis and metabolism have been Abstract Ion mobility-mass spectrometry (IM-MS) has proven to be a highly informative technique for the characterization of lipids from cells and tissues. We report the combination of hydrophilic-interaction liquid chromatography (HILIC) with traveling-wave IM-MS (TWIM-MS

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Section: Untargeted Lipidomics By Hilic-im-msmentioning

confidence: 69%

Section: Ccs Measurements Of Lipids By Twim-ms Calibrationmentioning

confidence: 93%

Section: Untargeted Lipidomics By Hilic-im-msmentioning

confidence: 99%

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Assessment of altered lipid homeostasis by HILIC-ion mobility-mass spectrometry-based lipidomics

Hines

Herron

2017

Journal of Lipid Research

117

View full text Add to dashboard Cite

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“…30 Coupling LC with ion-mobility has shown potential for facilitating multi-dimensional LC-IMS-MS measurements, permitting isomeric lipids within complex biological extract to be resolved. 31 Despite these significant advances in lipid isomer separation and analysis, these techniques do not consistently allow isomer identification (e.g. the determination of the location of double bonds, stereochemistry about the double bonds, or sn -positions) without requiring analysis of an exhaustive number of standards which may not be commercially or synthetically available.…”

Section: Introductionmentioning

confidence: 99%

Pinpointing Double Bond and sn-Positions in Glycerophospholipids via Hybrid 193 nm Ultraviolet Photodissociation (UVPD) Mass Spectrometry

et al. 2017

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Complete structural characterization of complex lipids, such as glycerophospholipids, by tandem mass spectrometry (MS/MS) continues to present a major challenge. Conventional activation methods do not generate fragmentation patterns that permit the simultaneous discernment of isomers which differ in both the positions of acyl chains on the glycerol backbone and the double bonds within the acyl chains. Herein we describe a hybrid collisional activation/UVPD workflow that yields near-complete structural information for glycerophospholipids. This hybrid MS3 strategy affords the lipid’s sum composition based on the accurate mass measured for the intact lipid as well as highly specific diagnostic product ions that reveal both the acyl chain assignment (i.e. sn-position) and the site-specific location of double bonds in the acyl chains. This approach is demonstrated to differentiate sn-positional and double bond positional isomers, such as the regioisomeric phosphatidylcholines PC 16:0/18:1(n-9) and PC 18:1(n-9)/16:0 and has been integrated into an LC-MS3 workflow.

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“…This method was originally developed for total lipid extractions 9,11 and more recently was amended for the simultaneous extraction of metabolites, proteins, and lipids from a single sample 12,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30, enabling less sample quantity and experimental variability 10 . In the MPLEx protocol, chloroform is not miscible with water, which provides the basis for the triphasic chemical separation of sample constituents into distinct fractions.…”

Section: Introductionmentioning

confidence: 99%

The MPLEx Protocol for Multi-omic Analyses of Soil Samples

Nicora

Nakayasu

Casey

et al. 2018

JoVE

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Mass spectrometry (MS)-based integrated metaproteomic, metabolomic, and lipidomic (multi-omic) studies are transforming our ability to understand and characterize microbial communities in environmental and biological systems. These measurements are even enabling enhanced analyses of complex soil microbial communities, which are the most complex microbial systems known to date. Multi-omic analyses, however, do have sample preparation challenges, since separate extractions are typically needed for each omic study, thereby greatly amplifying the preparation time and amount of sample required. To address this limitation, a 3-in-1 method for the simultaneous extraction of metabolites, proteins, and lipids (MPLEx) from the same soil sample was created by adapting a solvent-based approach. This MPLEx protocol has proven to be both simple and robust for many sample types, even when utilized for limited quantities of complex soil samples. The MPLEx method also greatly enabled the rapid multi-omic measurements needed to gain a better understanding of the members of each microbial community, while evaluating the changes taking place upon biological and environmental perturbations.

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Uncovering biologically significant lipid isomers with liquid chromatography, ion mobility spectrometry and mass spectrometry

Cited by 219 publications

References 42 publications

Assessment of altered lipid homeostasis by HILIC-ion mobility-mass spectrometry-based lipidomics

Assessment of altered lipid homeostasis by HILIC-ion mobility-mass spectrometry-based lipidomics

Pinpointing Double Bond and sn-Positions in Glycerophospholipids via Hybrid 193 nm Ultraviolet Photodissociation (UVPD) Mass Spectrometry

The MPLEx Protocol for Multi-omic Analyses of Soil Samples

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