Development and validation of a UHPLC-ESI-MS/MS method for the simultaneous quantification of mammal lysophosphatidylcholines and lysophosphatidylethanolamines in serum

Abstract

Recent investigations based on non-targeted metabolomics have proposed lysophospholipids (Lyso-PLs) as biomarkers of different diseases. In particular, lysophosphatidylcholines (Lyso-PCs) and lysophosphatidylethanolamines (Lyso-PEs) have been associated with serious lipid pathologies. Methods to determine the different molecular species in a biological sample and to quantify even less abundant species are required for the evaluation of the Lyso-PL pattern as a novel comprehensive biomarker of dyslipidemia. This study describes the development and validation of an ultra-high-performance liquid chromatography coupled to tandem mass spectrometry assay for the determination of a large number of Lyso-PCs and Lyso-PEs in biological samples. The method was validated in rat serum using two simple methanol-based extractions with low sample volumes (5-50μL) that covered the wide concentration range of these metabolites. In total, thirty-one Lyso-PLs were separated and quantified with low method limits of detection and quantification, reaching values of 0.2 and 0.8nM, respectively. The method was subsequently applied in the identification of Lyso-PL-related changes produced by the chronic intake of a cafeteria diet. The results showed alterations in the majority of Lyso-PCs and Lyso-PEs in rat serum. Furthermore, multivariate analysis indicated that the comprehensive evaluation of serum Lyso-PLs could be an excellent indicator of the nutritional phenotype associated with an increased risk of lipid disorders.

Serum lysophospholipid levels are altered in dyslipidemic hamsters

“…Based on the exhaustive study of the fragmentation data, we concluded that both patterns represented Lyso-PL species. In addition, all these Lyso-PLs identified were molecular species containing the acyl chain in sn1 -position, which generally is the most common circulating isomer for these metabolites 38 . When collision energies of 10 V were applied in the positive ionization mode, several daughter ions were generated: a) the loss of the phosphorylethanolamine head group ([M + H] + −141), b) the loss of water ([M + H] + −18), c) the ion corresponding to the ethanolamine ([M + H] + = 62) and its fragment, d) the ethylamine ([M] + = 44), e) the phosphocholine ([M – H + 2 H] + = 184), and f) choline ions ([M + H] + = 104).…”

Variations in hepatic lipid species of age-matched male mice fed a methionine-choline-deficient diet and housed in different animal facilities

“…A separate analysis nevertheless identified strongly elevated LPC in the liver of MCD diet and high fat diet fed mice [52]. This suggests that hepatic LPC levels were modified by diet, genetics and / or housing conditions [53] but were not mandatory for NASH pathogenesis.…”

Serum metabolic profiling using small molecular water‐soluble metabolites in individuals with schizophrenia: A longitudinal study using a pre–post‐treatment design

“…For all the identified LysoPC in this study, LysoPC(14:0) and LysoPC(15:0) were upregulated in post‐treatment patients compared to pre‐treatment patients after adjusting clinical medication therapies. LysoPC, a series of abundant polar lipids, can participate in peroxidation and biosynthetic dysfunction of membrane phospholipids . LysoPC are also components of oxidized low‐density lipoprotein and regulate multiple biological processes, such as apoptosis control .…”