Se requiere una suscripción a JoVE para ver este contenido. Inicie sesión o comience su prueba gratuita.
Capítulos
- 00:05Título
- 01:26Collection of Media and Cell Samples
- 02:00Extraction of Lipids from Media
- 03:02Extraction of Lipids from Cell Lysate
- 05:16Pentafluorylbenzylbromide (PFB) Derivatization
- 05:40SID-LC-ECAPCI-MS Analysis
- 07:46Targeted Lipidomics Profile
- 09:03Conclusion
This protocol will demonstrate the extraction and analysis of free and esterified bioactive fatty acids from cells. Fatty acids are accurately quantified using stable isotope dilution, chiral liquid chromatography, electron capture atmospheric chemical ionization multiple reaction monitoring mass spectrometry (SID-LC-ECAPCI-MRM/MS).
Tags
Cellular Lipid ExtractionTargeted Stable Isotope DilutionLiquid Chromatography-mass Spectrometry AnalysisFatty AcidsArachidonic AcidLinoleic AcidOxidized Bioactive LipidsStereoisomersEnzymatic MetabolismReactive Oxygen SpeciesCyclooxygenaseLipoxygenaseCytochromes P450Cell LysateLiquid-liquid ExtractionStable Isotope Internal StandardsIon SuppressionMass Spectrometry Analysis
