Continuous comprehensive two-dimensional liquid chromatography–electrospray ionization mass spectrometry of complex lipidomic samples
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A new continuous comprehensive two-dimensional liquid chromatography–electrospray ionization mass spectrometry method has been developed for the lipidomic characterization of complex biological samples. The reversed-phase ultra-high-performance liquid chromatography with a C18 column (150 mm × 1 mm, 1.7 μm) used in the first dimension makes the separation of numerous lipid species differing in their hydrophobic part of the molecule, mainly fatty acyl chain lengths and the number and positions of double bonds, possible. Coeluted lipid species in the first dimension are resolved by the fast hydrophilic interaction liquid chromatography separation (50 mm × 3 mm, 2.7 μm, core–shell particles) of lipid classes according to their different polarities in the second dimension. Retention times in both dimensions, accurate m/z values, and tandem mass spectra provide high confidence in the identification of lipid species. The retention behavior of individual lipids in reversed-phase mode follows the equivalent carbon number pattern, which provides an additional tool for unambiguous identification. This analytical method is applied for the lipidomic characterization of total lipid extracts of human plasma and porcine brain samples, which resulted in the identification of 143 lipid species from four lipid categories and ten lipid classes.
KeywordsLipids Lipidomics Comprehensive 2D liquid chromatography Mass spectrometry Plasma Brain
This work was supported by ERC CZ project no. LL1302 sponsored by the Ministry of Education, Youth, and Sports of the Czech Republic. E.C. acknowledges the support of project no. CZ.1.07/2.3.00/30.0021 sponsored by the Ministry of Education, Youth, and Sports of the Czech Republic.
- 1.LIPID MAPS (2014) LIPID MAPS Lipidomics Gateway. http://www.lipidmaps.org/. Accessed 2 Dec 2014
- 2.Fahy E, Subramaniam S, Brown HA, Glass CK, Merrill AH, Murphy RC, Raetz CRH, Russell DW, Seyama Y, Shaw W, Shimizu T, Spener F, van Meer G, VanNieuwenhze MS, White SH, Witztum JL, Dennis EA (2005) A comprehensive classification system for lipids. Eur J Lipid Sci Technol 107:337–364CrossRefGoogle Scholar
- 12.Holčapek M, Cífková E, Červená B, Lísa M, Vostálová J, Galuszka J (2015) Determination of nonpolar and polar lipid classes in human plasma, erythrocytes and plasma lipoprotein fractions using ultrahigh-performance liquid chromatography-mass spectrometry. J Chromatogr A 1377:85–91CrossRefGoogle Scholar
- 13.Sokol E, Almeida R, Hannibal-Bach HK, Kotowska D, Vogt J, Baumgart J, Kristiansen K, Nitsch R, Knudsen J, Ejsing CS (2013) Profiling of lipid species by normal-phase liquid chromatography, nanoelectrospray ionization, and ion trap–orbitrap mass spectrometry. Anal Biochem 443:88–96CrossRefGoogle Scholar
- 14.Cífková E, Holčapek M, Lísa M, Ovčačíková M, Lyčka A, Lynen F, Sandra P (2012) Nontargeted quantitation of lipid classes using hydrophilic interaction liquid chromatography-electrospray ionization mass spectrometry with single internal standard and response factor approach. Anal Chem 84:10064–10070CrossRefGoogle Scholar
- 16.Lísa M, Netušilová K, Franěk L, Dvořáková H, Vrkoslav V, Holčapek M (2011) Characterization of fatty acid and triacylglycerol composition in animal fats using silver-ion and non-aqueous reversed-phase high-performance liquid chromatography/mass spectrometry and gas chromatography/flame ionization detection. J Chromatogr A 1218:7499–7510CrossRefGoogle Scholar
- 18.Holčapek M, Dvořáková H, Lísa M, Girón AJ, Sandra P, Cvačka J (2010) Regioisomeric analysis of triacylglycerols using silver-ion liquid chromatography–atmospheric pressure chemical ionization mass spectrometry: comparison of five different mass analyzers. J Chromatogr A 1217:8186–8194CrossRefGoogle Scholar
- 25.D'Attoma A, Grivel C, Heinisch S (2012) On-line comprehensive two-dimensional separations of charged compounds using reversed-phase high performance liquid chromatography and hydrophilic interaction chromatography. Part I: orthogonality and practical peak capacity considerations. J Chromatogr A 1262:148–159CrossRefGoogle Scholar
- 38.Folch J, Lees M, Stanley GHS (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226:497–509Google Scholar
- 42.Berdeaux O, Juaneda P, Martine L, Cabaret S, Bretillon L, Acar N (2010) Identification and quantification of phosphatidylcholines containing very-long-chain polyunsaturated fatty acid in bovine and human retina using liquid chromatography/tandem mass spectrometry. J Chromatogr A 1217:7738–7748CrossRefGoogle Scholar