Analysis of Free and Esterified Sterol Content and Composition in Seeds Using GC and ESI-MS/MS.
Chromatography, Gas
/ methods
Esterification
Esters
/ analysis
Fatty Acids
/ metabolism
Glycosphingolipids
/ metabolism
Lipid Metabolism
/ physiology
Lipids
/ chemistry
Phytosterols
/ analysis
Plants
/ metabolism
Seeds
/ chemistry
Spectrometry, Mass, Electrospray Ionization
/ methods
Sterols
/ analysis
Tandem Mass Spectrometry
/ methods
ESI-MS/MS
Free sterol
GC-FID
HPLC
MRM
Saponification
Silylation
Steryl ester
Undecanoyl chloride
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2021
2021
Historique:
entrez:
28
5
2021
pubmed:
29
5
2021
medline:
23
6
2021
Statut:
ppublish
Résumé
Total sterol content and composition in plant tissues can be easily determined by gas chromatography (GC) after saponification of the total lipid extract. However, in oleogenic tissues a significant proportion of the sterol is esterified to fatty acids, with GC methodologies unable to provide information about the proportion and the molecular species composition of intact steryl esters (SEs). Here we describe an electrospray ionization-tandem mass spectrometry (ESI-MS/MS) and Multiple Reaction Monitoring (MRM) method which, in parallel with GC analysis, allows for the accurate determination of both free and esterified sterol content and composition in seeds. After extraction of seed oil with hexane, free sterols are derivatized with undecanoyl chloride, total steryl esters are then purified from triacylglycerol (TAG) by liquid chromatography, infused and ionized as ammonium adducts, with molecular species identified and quantified by fragmentation in the presence of internal standards.
Identifiants
pubmed: 34047978
doi: 10.1007/978-1-0716-1362-7_11
doi:
Substances chimiques
Esters
0
Fatty Acids
0
Glycosphingolipids
0
Lipids
0
Phytosterols
0
Sterols
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
179-201Subventions
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/L002124/1
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/P012663/1
Pays : United Kingdom
Références
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