title: In vivo solid-phase microextraction liquid chromatography-tandem mass spectrometry for monitoring blood eicosanoids time profile after lipopolysaccharide-induced inflammation in Sprague-Dawley rats.
abstract: A fast and non-lethal in vivo solid-phase microextraction (SPME) sampling method for rat blood coupled to liquid chromatography and tandem mass spectrometry (LC-MS/MS) was developed for monitoring rapid changes in concentrations of eicosanoids - lipid mediators involved in the development of inflammatory conditions - using diffusion-based calibration. Sampling rates of target eicosanoids were pre-determined under laboratory conditions with a precision of ≤10%, and directly used for quantification of analyte concentrations in blood after lipopolysaccharide-induced inflammation in Sprague-Dawley rats. Results showed significant changes in unbound plasma concentrations of arachidonic acid (AA) and 12-hydroxyeicosatetraenoic acid (12-HETE) in response to the treatment. Next, performance of the proposed method was compared with protein precipitation (PP) of plasma, a conventional sample preparation technique. Finally, percentages of plasma protein binding (PPB) of specific eicosanoids were determined. PPB of target eicosanoids was in agreement with literature values, ranging from 99.3 to 99.9% for 12-HETE and DHA, respectively. We envision that the proposed method is a particularly suitable alternative to lethal sampling and current methods based on sample depletion in animal studies for accurate monitoring of rapid changes in blood concentrations of small molecules.
authors.affiliation: Department of Chemistry, University of Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Vincent
authors.initials: V
authors.lastname: Bessonneau
authors.affiliation: Department of Chemistry, University of Waterloo, Ontario N2L 3G1, Canada.
authors.forename: Yanwei
authors.initials: Y
authors.lastname: Zhan
authors.affiliation: InterVivo Solutions Inc., 120 Carlton Street, Suite 203, Toronto, Ontario M5A 4K2, Canada.
authors.forename: Inés A M
authors.initials: IA
authors.lastname: De Lannoy
authors.affiliation: Department of Chemistry, University of Waterloo, Ontario N2L 3G1, Canada; InterVivo Solutions Inc., 120 Carlton Street, Suite 203, Toronto, Ontario M5A 4K2, Canada.
authors.forename: Victor
authors.initials: V
authors.lastname: Saldivia
authors.affiliation: Department of Chemistry, University of Waterloo, Ontario N2L 3G1, Canada. Electronic address: janusz@uwaterloo.ca.
authors.forename: Janusz
authors.initials: J
authors.lastname: Pawliszyn
mesh_terms: D000818:Animals; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods; D015777:Eicosanoids / Q000097:blood*; D006801:Humans; D007249:Inflammation / Q000097:blood; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D017207:Rats, Sprague-Dawley; D052617:Solid Phase Microextraction / Q000379:methods; D013048:Specimen Handling; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D015777:Eicosanoids; D008070:Lipopolysaccharides
keywords: Eicosanoids; In vivo sampling; Inflammation; Liquid chromatography; Mass spectrometry; Solid-phase microextraction