Comprehensive mass spectrometry lipidomics of human biofluids and ocular tissues

Journal of Lipid Research · 8 authors, 5 centres

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TL;DR

The two methods are complementary, with each providing distinct advantages for different lipid classes, enabling annotation of approximately 500 lipid species across 15 classes.

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Lipid annotation was achieved from MS2 scans in positive and negative ionization modes, combining LipidSearch database results with manual spectra elucidation. This dual approach annotated 15 lipid classes and approximately 500 lipid species. The authors demonstrated a positive correlation between automatic peak extraction via R and manual extraction from Freestyle software. HILIC was found to be preferable for polar lipids such as glycerophospholipids, which are major classes in ocular tissues like the retina. A limitation noted is that CEs tend to fragment in ESI sources, reducing CE detection. The study provides a methodological framework for comprehensive lipid profiling of human ocular and biofluid samples, relevant to future AMD research.

STUDY TYPE

other

SPECIALTY

ophthalmology

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AI pipeline

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ORIGINAL PAPER

PMC10027555

Comprehensive mass spectrometry lipidomics of human biofluids and ocular tissues

Journal of Lipid Research

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ABSTRACT

Abstract Evaluating lipid profiles in human tissues and biofluids is critical in identifying lipid metabolites in dysregulated metabolic pathways. Due to various chemical characteristics, single-run lipid analysis has not yet been documented. Such approach is essential for analyzing pathology-related lipid metabolites. Age-related macular degeneration, the leading cause of vision loss in western countries, is emblematic of this limitation. Several studies have identified alterations in individual lipids but the majority are based on targeted approaches. In this study, we analyzed and identified approximately 500 lipid species in human biofluids (plasma and erythrocytes) and ocular tissues (retina and retinal pigment epithelium) using the complementarity of hydrophilic interaction liquid chromatography (HILIC) and reversed-phase chromatography (RPC), coupled to high-resolution mass spectrometry. For that, lipids were extracted from human eye globes and blood from 10 subjects and lipidomic analysis was carried out through analysis in HILIC and RPC, alternately. Furthermore, we illustrate the advantages and disadvantages of both techniques for lipid characterization. RPC showed greater sensitivity in hydrophobicity-based lipid separation, detecting diglycerides, triglycerides, cholesterol, and cholesteryl esters, whereas no signal of these molecules was obtained in HILIC. However, due to coelution, RPC was less effective in separating polar lipids like phospholipids, which were separated effectively in HILIC in both ionization modes. The complementary nature of these analytical approaches was essential for the detection and identification of lipid classes/subclasses, which can then provide distinct insights into lipid metabolism, a determinant of the pathophysiology of several diseases involving lipids, notably age-related macular degeneration. Supplementary key words: lipidomic analysis, eye/retina, RPE/Choroid, erythrocytes, plasma, mass spectrometry, age-related macular degeneration Abbreviations: AMD, age-related macular degeneration; Cer, ceramide; chol, cholesterol; DAG, diglyceride; FFA, free fatty acyls; GPh, glycerophospholipids; Hex2Cer, dihexosylceramide; HexCer, hexosylceramide; HILIC, hydrophilic interaction liquid chromatography; HRMS, high-resolution mass spectrometry; IS, internal standard; LPC, lysophosphatidylcholine; LPE, lysophosphatidylethanolamine; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PG, phosphatidylglycerol; PI, phosphatidylinositol; PR, photoreceptor; PS, phosphatidylserine; PUFA, polyunsaturated fatty acids; QC, quality control; RPC, reversed-phase chromatography; RPE, retinal pigment epithelium; SM, sphingomyelin; TAG, triglyceride; VLC, very long chain; VLC-PC, PC species containing VLC-PUFA

CITATION EXPORT

Glenda Vasku, Caroline Peltier, Zhiguo He et al.. Comprehensive mass spectrometry lipidomics of human biofluids and ocular tissues. Journal of Lipid Research. (2023). https://doi.org/10.1016/j.jlr.2023.100343

AUTHORS

8 · 5 CENTRES

Glenda Vasku, Caroline Peltier, Zhiguo He, Gilles Thuret, Philippe Gain, Pierre-Henry Gabrielle, Niyazi Acar, Olivier Berdeaux

1Eye and Nutrition Research Group, Centre des Sciences du Goût et de l'Alimentation, CNRS, INRAE, Institut Agro, Université de Bourgogne Franche-Comté, Dijon, France
2ChemoSens Platform, Centre des Sciences du Goût et de l’Alimentation, Institut Agro, CNRS, INRAE, Université Bourgogne Franche-Comté, Dijon, France
3INRAE, PROBE Research infrastructure, ChemoSens facility, Dijon, France
4Department of Ophthalmology, Biology, Imaging, and Engineering of Corneal Grafts, Faculty of Medicine, Saint Etienne, France
5Department of Ophthalmology, University Hospital, Dijon, France