An automated shotgun lipidomics platform for high throughput, comprehensive, and quantitative analysis of blood plasma intact lipids

Abstract

Blood plasma has gained protagonism in lipidomics studies due to its availability, uncomplicated collection and preparation, and informative readout of physiological status. At the same time, it is also technically challenging to analyze due to its complex lipid composition affected by many factors, which can hamper the throughput and/or lipidomics coverage. To tackle these issues, we developed a comprehensive, high throughput, and quantitative mass spectrometry-based shotgun lipidomics platform for blood plasma lipid analyses. The main hallmarks of this technology are (i) it is comprehensive, covering 22 quantifiable different lipid classes encompassing more than 200 lipid species; (ii) it is amenable to high-throughput, with less than 5 min acquisition time allowing the complete analysis of 200 plasma samples per day; (iii) it achieves absolute quantification, by inclusion of internal standards for every lipid class measured; (iv) it is highly reproducible, achieving an average coefficient of variation of <10% (intra-day), approx. 10% (inter-day), and approx. 15% (inter-site) for most lipid species; (v) it is easily transferable allowing the direct comparison of data acquired in different sites. Moreover, we thoroughly assessed the influence of blood stabilization with different anticoagulants and freeze-thaw cycles to exclude artifacts generated by sample preparation.Practical applications: This shotgun lipidomics platform can be implemented in different laboratories without compromising reproducibility, allowing multi-site studies and inter-laboratory comparisons. This possibility combined with the high-throughput, broad lipidomic coverage and absolute quantification are important aspects for clinical applications and biomarker research.

The BUME method: a new rapid and simple chloroform-free method for total lipid extraction of animal tissue

“…Using this method for lipid extraction of plasma and tissue, the authors show that the procedure is associated with high recoveries of a wide range of plasma lipids and fully comparable with the Folch method. Further development by Surma et al ., with some modifications in initial sample to solvent ratio, enables the method to be scaled down and automated extraction of plasma lipids could be performed in the 96-well format25. Whether this method could be applied to tissue needs to be further investigated and lies outside the scope of this study.…”

Plasma Lipid Profiling of Patients with Chronic Ocular Complications Caused by Stevens-Johnson Syndrome/Toxic Epidermal Necrolysis

“…Full MS data in the negative ion mode were used for quantification. The estimated concentrations of PCs were determined using a curation curve constructed using one PC standard (PC[12:0/12:0] in the present study), as frequently applied in the previous studies [25,26]. The slope and goodness-of-fit values for the linear regression (R 2 ) of the PC standard (PC[12:0/12:0]) were 1.001 and 0.998, respectively.…”

Role of n-3 Polyunsaturated Fatty Acids in Ameliorating the Obesity-Induced Metabolic Syndrome in Animal Models and Humans

“…The plasma lipid profile plays an important role in metabolic syndrome incidence and severity [67,68,200]. Techniques for high-throughput and sensitive analysis of blood plasma lipids should be further developed [201]. Thus, routine plasma examination could be applied for monitoring health status.…”