Abstract
Shotgun lipidomics has evolved into a myriad of multi-dimensional strategies for molecular lipid characterization, including bioinformatics tools for mass spectrum interpretation and quantitative measurements to study systems-lipidomics in complex biological extracts. Taking advantage of spectral mass accuracy, scan speed and sensitivity of improved quadrupole linked time-of-flight mass analyzers, we developed a bias-free global lipid profiling acquisition technique of sequential precursor ion fragmentation called MS/MSALL. This generic information-independent tandem mass spectrometry (MS) technique consists of a Q1 stepped mass isolation window through a set mass range in small increments, fragmenting and recording all product ions and neutral losses. Through the accurate MS and MS/MS information, the molecular lipid species are resolved, including distinction of isobaric and isomeric species, and composed into more precise lipidomic outputs. The method demonstrates good reproducibility and at least 3 orders of dynamic quantification range for isomeric ceramides in human plasma. More than 400 molecular lipids in human plasma were uncovered and quantified in less than 12 min, including acquisitions in both positive and negative polarity modes. We anticipate that the performance of sequential precursor ion fragmentation both in quality and throughput will lead to the uncovering of new avenues throughout the biomedical research community, enhance biomarker discovery and provide novel information target discovery programs as it will prospectively shed new insight into affected metabolic and signaling pathways.
Highlights
The unprecedented advances in mass spectrometry (MS), combined with the need for comprehensive lipid measurements from the research and medical communities, have led to the rapid evolution of lipidomics
Shotgun lipidomics workflow was developed consisting of the ordered acquisition of high resolution, accurate mass time-of-flight detection of all precursor and product ions: an acquisition technique termed MS/MSALL
We demonstrated that the sequential precursor ion fragmentation (MS/MSALL) technique successfully reveals the make-up of the human plasma lipidome
Summary
The unprecedented advances in mass spectrometry (MS), combined with the need for comprehensive lipid measurements from the research and medical communities, have led to the rapid evolution of lipidomics. Lipid classes such as glycerophospholids, glycerolipids, glycosphingolipids and sterol lipids can be identified by distinguishing their characteristic headgroup ions, long-chain bases, fatty acid acyl ions and corresponding neutral losses These types of ions have been used in precursor ion scanning (PIS), neutral loss scanning (NLS) and multiple reaction monitoring (MRM) experiments, primarily carried out on triple quadrupole (QqQ) instruments due to the high sensitivity, favorable quantitative capabilities, acquisition speed and selectivity of this technology [8,9,10,11]. MS/MSALL represents a novel scanning technique amenable to the analysis of biological lipid extracts and automated untargeted high throughput molecular lipidomics
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