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Separation and identification of diacylglycerols containing branched chain fatty acids by liquid chromatography-mass spectrometry
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SYSNO ASEP 0540699 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Separation and identification of diacylglycerols containing branched chain fatty acids by liquid chromatography-mass spectrometry Author(s) Palyzová, Andrea (MBU-M) RID, ORCID
Řezanka, Tomáš (MBU-M) ORCIDArticle number 461708 Source Title Journal of Chromatography A. - : Elsevier - ISSN 0021-9673
Roč. 1635, JAN 4 (2021)Number of pages 10 s. Language eng - English Country NL - Netherlands Keywords Gram-positive bacteria ; enzymatic hydrolysis ; rp-hplc/ms-esi+ ; branched fatty acids ; phosphatidylglycerols ; diacylglycerols Subject RIV EE - Microbiology, Virology OECD category Microbiology R&D Projects LO1509 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Limited access Institutional support MBU-M - RVO:61388971 UT WOS 000604226500012 EID SCOPUS 85096475930 DOI 10.1016/j.chroma.2020.461708 Annotation 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. Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2021 Electronic address https://www.sciencedirect.com/science/article/abs/pii/S0021967320309821?via%3Dihub
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