title: Quantification of anandamide, oleoylethanolamide and palmitoylethanolamide in rodent brain tissue using high performance liquid chromatography-electrospray mass spectroscopy.
abstract: Reported concentrations for endocannabinoids and related lipids in biological tissues can vary greatly; therefore, methods used to quantify these compounds need to be validated. This report describes a method to quantify anandamide (AEA), oleoylethanolamide (OEA) and palmitoylethanolamide (PEA) from rodent brain tissue. Analytes were extracted using acetonitrile without further sample clean up, resolved on a C18 reverse-phase column using a gradient mobile phase and detected using electrospray ionization in positive selected ion monitoring mode on a single quadrupole mass spectrometer. The method produced high recovery rates for AEA, OEA and PEA, ranging from 98.1% to 106.2%, 98.5% to 102.2% and 85.4% to 89.5%, respectively. The method resulted in adequate sensitivity with a lower limit of quantification for AEA, OEA and PEA of 1.4 ng/mL, 0.6 ng/mL and 0.5 ng/mL, respectively. The method was reproducible as intraday and interday accuracies and precisions were under 15%. This method was suitable for quantifying AEA, OEA and PEA from rat brain following pharmacological inhibition of fatty acid amide hydrolase.
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY 40536, USA.
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Liput
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY 40536, USA.
authors.forename: Eleftheria
authors.initials: E
authors.lastname: Tsakalozou
authors.affiliation: AllTranz, Lexington, KY 40505, USA.
authors.forename: Dana C
authors.initials: DC
authors.lastname: Hammell
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, South College, Knoxville, TN 37922, USA.
authors.forename: Kalpana S
authors.initials: KS
authors.lastname: Paudel
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY 40536, USA.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Nixon
authors.affiliation: AllTranz, Lexington, KY 40505, USA ; Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, Baltimore, MD 21201, USA.
authors.forename: Audra L
authors.initials: AL
authors.lastname: Stinchcomb
keywords: Acylethanolamides; Anandamide; Endocannabinoids; LC–MS; OEA; PEA