Rapid identification of erythrocyte phospholipids in Sprague–Dawley rats by ultra high performance liquid chromatography with electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry

Abstract

A rapid, sensitive, and reliable approach for analyzing five kinds of erythrocyte phospholipids in Sprague-Dawley rats was provided by ultra high performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry with MassLynx(TM) MassFragment. Improving conventional high performance liquid chromatography techniques, ultra high performance liquid chromatography integrated with quadrupole time-of-flight tandem mass spectrometry offers high sensitivity and increased analytical speed by using columns packed with sub-2 μm particles (1.7 μm), which allows a faster separation to be achieved. Through this method, 83 phospholipids were tentatively characterized based on their mass spectra and tandem mass spectra, as well as by matching the in-house formula database within a mass error of 5 ppm, including 40 phosphatidylcholines, 24 phosphatidyl ethanolamines, three phosphatidylinositols, six phosphatidylserines, and ten sphingomyelins. Our present results proved that the established method could be used to qualitatively analyze complex erythrocyte phospholipids in Sprague-Dawley rats and provide a useful data base for pharmacology and phospholipidomics to seek potential biomarkers of disease prediction.

Fragmentation patterns of five types of phospholipids by ultra-high-performance liquid chromatography electrospray ionization quadrupole time-of-flight tandem mass spectrometry

“…The fragmentation patterns of ve major types of phospholipids (PE, PI, PC, PS and SM) were studied systematically in different ion modes at several elevated collision energies (10 to 50 eV). Combined with our previous research, 22,23 all the fragmentation patterns were summarized, including different characteristic ions of phospholipids in different ion modes, three patterns ([RC]O] + , [RCOO À ] À and R 0 CH]CO) of common fatty acyl chain ions of glycerophospholipids in different ion modes, and the variation trend of the ratios of sn-1/sn-2 carboxylate ions in ESI (À) mode at different collision energies (10 to 50 eV), which could be used to rapidly identify possible phospholipids according to their MS and MS/MS information.…”

“…In this paper, the fragmentation regularities of the ve types of phospholipids were studied systematically in different ion modes. Combined with our previous work, 22,23 all these fragmentation regularities were summarized, including three patterns ([RC]O] + , [RCOO À ] À and R 0 CH]CO) of common fatty acyl chain ions in different ion modes, the variation trend of the ratios of the sn-1/sn-2 carboxylate anions, and different characteristic ions at different collision energies (10 to 50 eV). These proposed methodologies have already been successfully applied for the high-throughput screening of phospholipid markers in phospholipidomics studies.…”

“…A gradient elution program was set as follows: 60% to 81.4% B at 0 to 4 min, 81.4% to 90% B at 4-20 min. 22…”

Quantitative analysis of N‐acylphosphatidylethanolamine molecular species in rat brain using solid‐phase extraction combined with reversed‐phase chromatography and tandem mass spectrometry

“…NAPE synthesis is catalyzed by N-acyltransferase that transfers a fatty acyl chain from the sn-1 position of phosphatidylcholine (PC) to the amino group of a PE molecule. The primary pathway Tissue levels of NAPE in mammals are in the range of a few nmol/g [1], and NAPEs are usually not covered by "comprehensive" HPLC, SFC, or shotgun mass spectrometric lipidomic approaches [12][13][14][15][16][17][18][19][20], but rather require specialized approaches in terms of sample preparation and identification. By hydrolysis of the sn-1 and sn-2 bonds of NAPE the resulting glycerophospho-NAE species can be determined either by LC-MS [9,21], GC-MS [22,23] or HPLC coupled to fluorescence detection [24].…”

Anti‐inflammatory activities and glycerophospholipids metabolism in KLA‐stimulated RAW 264.7 macrophage cells by diarylheptanoids from the rhizomes of Alpinia officinarum

“…Ultra‐high‐performance liquid chromatography with electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry/mass spectrometry (UPLC‐ESI‐Q‐TOF‐MS/MS) technique is widely used for glycerophospholipid extract analysis (Cífková et al, ; Li et al, ; Pi, Wu, Yang, Zeng, & Feng, ). In this study, UPLC‐ESI‐Q‐TOF‐MS/MS was applied to accurately analyze lipid components.…”