High-throughput Lipidomics Analysis Research Articles

Comprehensive lipid profile analysis of three fish roe by untargeted lipidomics using ultraperformance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry

Fish roe is a significant nutrition source from marine byproducts, containing rich contents of long-chain omega-3 fatty acids (n 3-LC PUFAs). However, investigations focused on the nutritional values and lipid composition of various fish roe species are limited. This research aims to perform a high-throughput lipidomics analysis to comprehensively evaluate and compare the lipid profiles of three consumed marine fish roe using ultraperformance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC Q-TOF-MS/MS). Results showed notable differences in three fish roe species in terms of sphingolipids (SLs), phospholipids (PLs), and glycerolipids (GLs) compositions. Furthermore, the high percentages of n-3 PUFA-PLs in yellow croaker and mackerel roe indicate their significant nutritional value. In total, 449 lipid molecular species of 13 lipid subclasses were identified; of them, 31 lipid molecular species could serve as potential biomarkers for the differentiation of three fish roe. This study provides comprehensive lipid profiles of three fish roe, demonstrating their potential values in functional food industries.

Alveolar type II epithelial cell FASN maintains lipid homeostasis in experimental COPD.

Alveolar epithelial type II (AEC2) cells strictly regulate lipid metabolism to maintain surfactant synthesis. Loss of AEC2 cell function and surfactant production are implicated in the pathogenesis of the smoking-related lung disease chronic obstructive pulmonary disease (COPD). Whether smoking alters lipid synthesis in AEC2 cells and whether altering lipid metabolism in AEC2 cells contributes to COPD development are unclear. In this study, high-throughput lipidomic analysis revealed increased lipid biosynthesis in AEC2 cells isolated from mice chronically exposed to cigarette smoke (CS). Mice with a targeted deletion of the de novo lipogenesis enzyme, fatty acid synthase (FASN), in AEC2 cells (FasniΔAEC2) exposed to CS exhibited higher bronchoalveolar lavage fluid (BALF) neutrophils, higher BALF protein, and more severe airspace enlargement. FasniΔAEC2 mice exposed to CS had lower levels of key surfactant phospholipids but higher levels of BALF ether phospholipids, sphingomyelins, and polyunsaturated fatty acid-containing phospholipids, as well as increased BALF surface tension. FasniΔAEC2 mice exposed to CS also had higher levels of protective ferroptosis markers in the lung. These data suggest that AEC2 cell FASN modulates the response of the lung to smoke by regulating the composition of the surfactant phospholipidome.

High-throughput lipidomics analysis to discover lipid biomarkers and profiles as potential targets for evaluating efficacy of Kai-Xin-San against APP/PS1 transgenic mice based on UPLC-Q/TOF-MS.

Lipid metabolism has a significant function in the central nervous system and Alzheimer's disease (AD) is an age-related senile disease characterized by central nerve degeneration. The pathological development of AD is closely related to lipid metabolism disorders. To reveal the influence of Kai-Xin-San (KXS) on lipid metabolism in APP/PSI transgenic mice and potential therapeutic targets for treating AD, brain tissue samples were collected and analyzed by high-throughput lipidomics based on UPLC-Q/TOF-MS. The collected raw data were processed by multivariate data analysis to discover the potential biomarkers and lipid metabolic profiles. Compared with the control wild-type mouse group, nine potential lipid biomarkers were found in the AD model group, of which seven were up-regulated and two were down-regulated. Orally administrated KXS can reverse the changes in these potential biomarkers. Compared with the model group, a total of six differential metabolites showed a recovery trend and may be potential targets for KXS to treat AD. This study showed that high-throughput lipidomics can be used to discover the perturbed pathways and lipid biomarkers as potential targets to reveal the therapeutic effects of KXS.

High-Throughput Screening of Lipidomic Adaptations in Cultured Cells.

High-throughput screening of biologically active substances in cell cultures remains challenging despite great progress in contemporary lipidomic techniques. These experiments generate large amounts of data that are translated into lipid fingerprints. The subsequent visualization of lipidomic changes is key to meaningful interpretation of experimental results. As a demonstration of a rapid and versatile pipeline for lipidomic analysis, we cultured HeLa cells in 96-well format for four days in the presence or absence of various inhibitors of lipid metabolic pathways. Visualization of the data by principle component analysis revealed a high reproducibility of the method, as well as drug specific changes to the lipidome. Construction of heatmaps and networks revealed the similarities and differences between the effects of different drugs at the lipid species level. Clusters of related lipid species that might represent distinct membrane domains emerged after correlation analysis of the complete dataset. Taken together, we present a lipidomic platform for high-throughput lipidomic analysis of cultured cell lines.

Identification of Double Bond Position Isomers in Unsaturated Lipids by m-CPBA Epoxidation and Mass Spectrometry Fragmentation.

Lipids are highly diverse biomolecules associated with several biological functions including structural constituent, energy storage, and signal transduction. It is essential to characterize lipid structural isomers and further understand their biological roles. Unsaturated lipids contain one or multiple carbon-carbon double bonds. Identifying double bond position presents a major challenge in unsaturated lipid characterization. Recently, several advancements have been made for double bond localization by mass spectrometry (MS) analysis. However, many of these studies require complex chemical reactions or advanced mass spectrometers with special fragmentation techniques, which limits the application in lipidomics study. Here, an innovative meta-chloroperoxybenzoic acid ( m-CPBA) epoxidation reaction coupling with collision-induced dissociation (CID)-MS/MS strategy provides a new tool for unsaturated lipidomics analysis. The rapid epoxidation reaction was carried out by m-CPBA with high specificity. Complete derivatization was achieved in minutes without overoxidized byproduct. Moreover, diagnostic ion pair with 16 Da mass difference indicated localization of carbon-carbon double bond in MS/MS spectra. Multiple lipid classes were evaluated with this strategy and generated abundant fragments for structural analysis. Unsaturated lipid analysis of yeast extract using this strategy took less than 30 min, demonstrating the potential for high-throughput lipidomics analysis by this approach. This study opens a door for high throughput unsaturated lipid analysis with minimal requirement for instrumentation, which could be widely applied in lipidomics analysis.

Dynamic Assessment of Functional Lipidomic Analysis in Human Urine.

The development of enabling mass spectrometry platforms for the quantification of diverse lipid species in human urine is of paramount importance for understanding metabolic homeostasis in normal and pathophysiological conditions. Urine represents a non-invasive biofluid that can capture distinct differences in an individual's physiological status. However, currently there is a lack of quantitative workflows to engage in high throughput lipidomic analysis. This study describes the development of a MS/MS(ALL) shotgun lipidomic workflow and a micro liquid chromatography-high resolutiontandem mass spectrometry (LC-MS/MS) workflow for urine structural and mediator lipid analysis, respectively. This workflow was deployed to understand biofluid sample handling and collection, extraction efficiency, and natural human variation over time. Utilization of 0.5mL of urine for structural lipidomic analysis resulted in reproducible quantification of more than 600 lipid molecular species from over 20 lipid classes. Analysis of 1mL of urine routinely quantified in excess of 55 mediator lipid metabolites comprised of octadecanoids, eicosanoids, and docosanoids generated by lipoxygenase, cyclooxygenase, and cytochrome P450 activities. In summary, the high-throughput functional lipidomics workflow described in this study demonstrates an impressive robustness and reproducibility that can be utilized for population health and precision medicine applications.

Discovering lipid phenotypic changes of sepsis-induced lung injury using high-throughput lipidomic analysis

The aim of this study was to use lipidomics to identify lipid molecules that could predict patients with sepsis-induced lung injury.

An Improved High‐Throughput Lipid Extraction Method for the Analysis of Human Brain Lipids

We have developed a protocol suitable for high-throughput lipidomic analysis of human brain samples. The traditional Folch extraction (using chloroform and glass-glass homogenization) was compared to a high-throughput method combining methyl-tert-butyl ether (MTBE) extraction with mechanical homogenization utilizing ceramic beads. This high-throughput method significantly reduced sample handling time and increased efficiency compared to glass-glass homogenizing. Furthermore, replacing chloroform with MTBE is safer (less carcinogenic/toxic), with lipids dissolving in the upper phase, allowing for easier pipetting and the potential for automation (i.e., robotics). Both methods were applied to the analysis of human occipital cortex. Lipid species (including ceramides, sphingomyelins, choline glycerophospholipids, ethanolamine glycerophospholipids and phosphatidylserines) were analyzed via electrospray ionization mass spectrometry and sterol species were analyzed using gas chromatography mass spectrometry. No differences in lipid species composition were evident when the lipid extraction protocols were compared, indicating that MTBE extraction with mechanical bead homogenization provides an improved method for the lipidomic profiling of human brain tissue.

High-throughput lipidomic analysis of fatty acid derived eicosanoids and N-acylethanolamines

Fatty acid-derived eicosanoids and N-acylethanolamines (NAE) are important bioactive lipid mediators involved in numerous biological processes including cell signaling and disease progression. To facilitate research on these lipid mediators, we have developed a targeted high-throughput mass spectrometric based methodology to monitor and quantitate both eicosanoids and NAEs, and can be analyzed separately or together in series. Each methodology utilizes scheduled multiple reaction monitoring (sMRM) pairs in conjunction with a 25 min reverse-phase HPLC separation. The eicosanoid methodology monitors 141 unique metabolites and quantitative amounts can be determined for over 100 of these metabolites against standards. The analysis covers eicosanoids generated from cycloxygenase, lipoxygenase, cytochrome P450 enzymes, and those generated from non-enzymatic pathways. The NAE analysis monitors 36 metabolites and quantitative amounts can be determined for 33 of these metabolites against standards. The NAE method contains metabolites derived from saturated fatty acids, unsaturated fatty acids, and eicosanoids. The lower limit of detection for eicosanoids ranges from 0.1 pg to 1 pg, while NAEs ranges from 0.1 pg to 1000 pg. The rationale and design of the methodology is discussed. This article is part of a Special Issue entitled Lipodomics and Imaging Mass Spectrometry.