Separation and identification of diacylglycerols containing branched chain fatty acids by liquid chromatography-mass spectrometry

0540699 - MBÚ 2021 RIV NL eng J - Journal Article
Palyzová, Andrea - Řezanka, Tomáš
Separation and identification of diacylglycerols containing branched chain fatty acids by liquid chromatography-mass spectrometry.
Journal of Chromatography A. Roč. 1635, JAN 4 (2021), č. článku 461708. ISSN 0021-9673. E-ISSN 1873-3778
R&D Projects: GA MŠMT(CZ) LO1509
Institutional support: RVO:61388971
Keywords : Gram-positive bacteria * enzymatic hydrolysis * rp-hplc/ms-esi+ * branched fatty acids * phosphatidylglycerols * diacylglycerols
OECD category: Microbiology
Impact factor: 4.601, year: 2021
Method of publishing: Limited access
https://www.sciencedirect.com/science/article/abs/pii/S0021967320309821?via%3Dihub

A combination of two chromatographic and two enzymatic methods was used for the analysis of molecular species of lipids from Gram-positive bacteria of the genus Kocuria. Gram-positive bacteria contain a majority of branched fatty acids (FAs), especially isoand/or anteiso-FAs. Two strains K. rhizophila were cultivated at three different tem peratures (20, 28, and 37 degrees C) and the majority phospholipid, i.e., the mix ture of molecular species of phosphatidylglycerols (PGs) was separated by means of hydrophilic interaction liquid chromatography (HILIC). After enzymatic hydrolysis of PGs by phospholipase C and derivatization of the free OH group, the sn1,2-diacyl-3-acetyl triacylglycerols (AcTAGs) were separated by reversed phase HPLC. Molecular species such as i-15:0/i-15:0/2:0, ai-15:0/ai-15:0/2:0, and 15:0/15:0/2:0 (straight chains) were identified by liquid chromatography-positive electrospray ionization mass spectrometry. The tandem mass spectra of both standards and natural compounds containing iso, anteiso and straight chain FAs with the same carbons were identical. Therefore, for identification of the ratio of two regioisomers, i.e. i-15:0/ai-15:0/2:0 vs. ai-15:0/i-15:0/2:0, they were cleavage by pancreatic lipase. The mixture of free fatty acids (FFAs) and 2-monoacylglycerols (2-MAGs) was obtained. After their separation by TLC and esterification and/or transesterification, the fatty acid methyl esters were quantified by GC-MS and thus the ratio of regioisomers was determined.
Permanent Link: http://hdl.handle.net/11104/0318319