Development of SFC-MS Method for Quantification of Eicosanoids Biosynthesized in Primary Human Blood Cells

Development of SFC-MS Method for Quantification of Eicosanoids Biosynthesized in Primary Human Blood Cells

Eicosanoids are lipid mediators generated from arachidonic acid with pro- and anti-inflammatory properties. Despite these lipid mediators being known for decades, quantitative determination in biological samples is still challenging due to low abundance, instability, the existence of regio- and ster...

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Bibliographic Details
Published in:Metabolites Vol. 12; no. 12; p. 1198
Main Authors: Schmidt, Louis, Burmeister, Laura Sophie, Greinacher, Andreas, König, Stefanie, Garscha, Ulrike
Format: Journal Article
Language:English
Published: Switzerland MDPI AG 30-11-2022
MDPI
Subjects:
Acetonitrile
Amylose
Analysis
Anti-inflammatory agents
Arachidonic acid
Biosynthesis
Blood cells
Calibration
Chromatography
Eicosanoic acid
Eicosanoids
Evaluation
Inflammation
Mass spectrometry
Mass spectroscopy
monocytes
Neutrophils
oxylipins
Physiology
platelets
Propanol
Properties
Prostaglandin E2
Scientific imaging
Stereoisomers
supercritical fluid chromatography
validation
Zileuton
Germany
United States > US
lipid mediators
monocytes
supercritical fluid chromatography
oxylipins
neutrophils
platelets
validation
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Summary:Eicosanoids are lipid mediators generated from arachidonic acid with pro- and anti-inflammatory properties. Despite these lipid mediators being known for decades, quantitative determination in biological samples is still challenging due to low abundance, instability, the existence of regio- and stereoisomers, and a wide polarity range that hampers chromatographic separation. In this study, we developed a supercritical fluid chromatography mass spectrometry (SFC-MS) platform for the quantification of relevant eicosanoids. Application of a chiral amylose-based column and modifier combination of 2-propanol/acetonitrile offered separation and sufficient resolution of 11 eicosanoids (5-, 12-, 15-HETE, PGB , LTB , t-LTB , 20-OH-LTB , PGE , PGD , PGF , TxB ) with baseline separation of isobaric analytes within 12 min. The method was validated in terms of range (78-2500 ng/mL), linearity, accuracy, precision, and recovery according to EMA guidelines. Finally, we confirmed the method's applicability by quantifying eicosanoid levels in human primary blood cells. In conclusion, we present a validated SFC-MS method for the determination of relevant eicosanoids in biological samples with a wide range of polarity while maintaining baseline separation of isobars, which allows coupling to a single quadrupole mass detector.
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ISSN:2218-1989
2218-1989
DOI:10.3390/metabo12121198