Minor lipids profiling in subcutaneous and epicardial fat tissue using LC/MS with an optimized preanalytical phase

. 2019 Apr 15 ; 1113 () : 50-59. [epub] 20190312

Jazyk angličtina Země Nizozemsko Médium print-electronic

Typ dokumentu časopisecké články

Perzistentní odkaz   https://www.medvik.cz/link/pmid30897405
Odkazy

PubMed 30897405
DOI 10.1016/j.jchromb.2019.03.006
PII: S1570-0232(18)31552-6
Knihovny.cz E-zdroje

Analysis of bioactive lipids in adipose tissue could lead to better understanding of the pathogenesis of obesity and its complications. However, current MS methods are limited by a high content of triacylglycerols (TAGs), which markedly surpasses the amount of other lipids and suppresses their ionization. The aim of our study was thus to optimize the preanalytical phase of lipid analysis in adipose tissue, focusing in particular on less-abundant lipids. Next, the optimized method was used to describe the differences between epicardial and subcutaneous adipose tissues obtained from patients undergoing cardiac surgery. Lipids were extracted using a modified Folch method with subsequent detachment of TAGs by thin layer chromatography (TLC). The extracts with/without TAGs were analyzed by tandem LC/MS. The repeatability of the presented method expressed by the median of the coefficients of variation was 12/5% for analysis with/without TAGs separation, respectively. The difference in the relative abundance of TAGs gained with/without TLC was, on average, 19% and did not reach significance (p value > 0.05) for any identified TAG. The novel preanalytical step allowed us to detect 37 lipids, which could not have been detected without TAG separation, because their signal to noise ratio is <5 in current methods of untargeted lipidomics. These lipids belong predominately to ceramides, glycerophosphatidylserines, glycerophosphatidylinsitols, sphingomyelins, glycerophosphatidylcholines, glycerophosphatidylethanolamines, diacylglycerols. The two adipose tissue depots differed mainly in the following lipid classes: glycerophosphatidylcholines, glycerophosphatidylinositols, glycerophosphatidylethanolamine, and sphingomyelins. Moreover, other major lipids showed distinctly different distributions between the two adipose tissues. Among these, the changes in TAGs were the most striking, which correspond to previously published data describing the differences between omental and subcutaneous adipose tissue. Implementation of the TLC step for the elimination of TAGs was crucial for enhancing the MS detection limit of minor lipids in adipose tissue. The differences between the overall lipid profiles of subcutaneous and epicardial tissue reflect their different functions arising from their location.

2nd Department of Surgery Department of Cardiovascular Surgery 1st Faculty of Medicine Charles University and General University Hospital Prague Czech Republic

4th Medical Department 1st Faculty of Medicine Charles University and General Faculty Hospital Prague U Nemocnice 2 128 08 Praha 2 Czech Republic

Department of Anaesthesiology Resuscitation and Intensive Care 1st Faculty of Medicine Charles University and General University Hospital Prague Czech Republic

Faculty of Applied Sciences University of West Bohemia Univerzitní 8 306 14 Plzeň Czech Republic; Institute of Microbiology Czech Academy of Sciences Vídeňská 1083 142 20 Prague 4 Czech Republic

Institute of Medical Biochemistry and Laboratory Diagnostics 1st Faculty of Medicine Charles University and General University Hospital Prague U Nemocnice 2 128 08 Prague 2 Czech Republic; Centre for Experimental Medicine Institute for Clinical and Experimental Medicine Vídeňská 1958 9 140 21 Prague 4 Czech Republic

Institute of Microbiology Czech Academy of Sciences Vídeňská 1083 142 20 Prague 4 Czech Republic

Institute of Microbiology Czech Academy of Sciences Vídeňská 1083 142 20 Prague 4 Czech Republic; 4th Medical Department 1st Faculty of Medicine Charles University and General Faculty Hospital Prague U Nemocnice 2 128 08 Praha 2 Czech Republic

Institute of Microbiology Czech Academy of Sciences Vídeňská 1083 142 20 Prague 4 Czech Republic; Faculty of Chemical Technology University of Chemistry and Technology Prague Technická 5 166 28 Prague 6 Czech Republic

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