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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 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, PGB1, LTB4, t-LTB4, 20-OH-LTB4, PGE2, PGD2, PGF2α, TxB2) 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.
Author: | Louis Schmidt, Laura Sophie Burmeister, Andreas GreinacherORCiD, Stefanie König, Ulrike Garscha |
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URN: | urn:nbn:de:gbv:9-opus-77441 |
DOI: | https://doi.org/10.3390/metabo12121198 |
ISSN: | 2218-1989 |
Parent Title (English): | Metabolites |
Publisher: | MDPI |
Place of publication: | Basel |
Document Type: | Article |
Language: | English |
Date of first Publication: | 2022/11/30 |
Release Date: | 2024/04/23 |
Tag: | lipid mediators; monocytes; neutrophils; oxylipins; platelets; supercritical fluid chromatography; validation |
Volume: | 12 |
Issue: | 12 |
Article Number: | 1198 |
Page Number: | 15 |
Faculties: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Pharmazie |
Collections: | weitere DFG-förderfähige Artikel |
Licence (German): | Creative Commons - Namensnennung 4.0 International |