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Development of an Optimized LC-MS Method for the Detection of Specialized Pro-Resolving Mediators in Biological Samples
- Source :
- Frontiers in Pharmacology, Frontiers in Pharmacology, Frontiers, 2019, 10, ⟨10.3389/fphar.2019.00169⟩, Frontiers in Pharmacology, Vol 10 (2019)
- Publication Year :
- 2019
- Publisher :
- HAL CCSD, 2019.
-
Abstract
- International audience; The cardioprotective and anti-inflammatory effects of long chain omega-3 polyunsaturated fatty acids (n3 PUFA) are believed to be partly mediated by their oxygenated metabolites (oxylipins). In the last two decades interest in a novel group of autacoids termed specialized pro-resolving mediators (SPMs) increased. These are actively involved in the resolution of inflammation. SPMs are multiple hydroxylated fatty acids including resolvins, maresins, and protectins derived from the n3 PUFA eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) as well as lipoxins derived from arachidonic acid (ARA). In the present paper, we developed an LC-MS/MS method for a comprehensive set of 18 SPMs derived from ARA, EPA, and DHA and integrated it into our targeted metabolomics platform. Quantification was based on external calibration utilizing five deuterated internal standards in combination with a second internal standard for quality assessment of sample preparation in each sample. The tandem mass spectrometric parameters were carefully optimized for sensitive and specific detection. The influence of source parameters of the used AB Sciex 6500 QTRAP instrument as well as electronic parameters and the selection of transitions are discussed. The method was validated/characterized based on the criteria listed in the European Medicines Agency (EMA) guideline on bioanalytical method validation and method performance is demonstrated regarding recovery of internal standards (between 78 ± 4% and 87 ± 3% from 500 µL of human serum) as well as extraction efficacy of SPMs in spiked plasma (intra-day accuracy within ±20 and ±15% at 0.1 and 0.3 nM in plasma, respectively). Based on the lower limit of quantification of 0.02-0.2 nM, corresponding to 0.18-2.7 pg on column, SPMs were generally not detectable/quantifiable in plasma and serum supporting that circulating levels of SPMs are very low, i.e.
- Subjects :
- 0301 basic medicine
Bioanalysis
Resolution (mass spectrometry)
oxylipin
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Liquid chromatography–mass spectrometry
[CHIM.ANAL]Chemical Sciences/Analytical chemistry
Pharmacology (medical)
Sample preparation
[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology
Original Research
chemistry.chemical_classification
Pharmacology
Chromatography
lcsh:RM1-950
resolution
Eicosapentaenoic acid
3. Good health
LC-MS
030104 developmental biology
lcsh:Therapeutics. Pharmacology
chemistry
Docosahexaenoic acid
specialized pro-resolving mediators
inflammation
030220 oncology & carcinogenesis
Arachidonic acid
Polyunsaturated fatty acid
Subjects
Details
- Language :
- English
- ISSN :
- 16639812
- Database :
- OpenAIRE
- Journal :
- Frontiers in Pharmacology, Frontiers in Pharmacology, Frontiers, 2019, 10, ⟨10.3389/fphar.2019.00169⟩, Frontiers in Pharmacology, Vol 10 (2019)
- Accession number :
- edsair.doi.dedup.....7fa0e00597caa87737fd1765e748e445