title: A quantitiative LC-MS/MS method for the measurement of arachidonic acid, prostanoids, endocannabinoids, N-acylethanolamines and steroids in human plasma.
abstract: Free arachidonic acid is functionally interlinked with different lipid signaling networks including those involving prostanoid pathways, the endocannabinoid system, N-acylethanolamines, as well as steroids. A sensitive and specific LC-MS/MS method for the quantification of arachidonic acid, prostaglandin E2, thromboxane B2, anandamide, 2-arachidonoylglycerol, noladin ether, lineoyl ethanolamide, oleoyl ethanolamide, palmitoyl ethanolamide, steroyl ethanolamide, aldosterone, cortisol, dehydroepiandrosterone, progesterone, and testosterone in human plasma was developed and validated. Analytes were extracted using acetonitrile precipitation followed by solid phase extraction. Separations were performed by UFLC using a C18 column and analyzed on a triple quadrupole MS with electron spray ionization. Analytes were run first in negative mode and, subsequently, in positive mode in two independent LC-MS/MS runs. For each analyte, two MRM transitions were collected in order to confirm identity. All analytes showed good linearity over the investigated concentration range (r>0.98). Validated LLOQs ranged from 0.1 to 190ng/mL and LODs ranged from 0.04 to 12.3ng/mL. Our data show that this LC-MS/MS method is suitable for the quantification of a diverse set of bioactive lipids in plasma from human donors (n=32). The determined plasma levels are in agreement with the literature, thus providing a versatile method to explore pathophysiological processes in which changes of these lipids are implicated.
authors.affiliation: Institute of Biochemistry and Molecular Medicine, NCCR TransCure, University of Bern, Bühlstrasse 28, 3012 Bern, Switzerland.
authors.forename: María Salomé
authors.initials: MS
authors.lastname: Gachet
authors.affiliation: Institute of Biochemistry and Molecular Medicine, NCCR TransCure, University of Bern, Bühlstrasse 28, 3012 Bern, Switzerland.
authors.forename: Peter
authors.initials: P
authors.lastname: Rhyn
authors.affiliation: Experimental and Clinical Pharmacopsychology, Department of Psychiatry, Psychotherapy and Psychosomatics, Psychiatric Hospital of the University of Zurich, Lenggstr. 31, CH-8032 Zürich, Switzerland.
authors.forename: Oliver G
authors.initials: OG
authors.lastname: Bosch
authors.affiliation: Experimental and Clinical Pharmacopsychology, Department of Psychiatry, Psychotherapy and Psychosomatics, Psychiatric Hospital of the University of Zurich, Lenggstr. 31, CH-8032 Zürich, Switzerland.
authors.forename: Boris B
authors.initials: BB
authors.lastname: Quednow
authors.affiliation: Institute of Biochemistry and Molecular Medicine, NCCR TransCure, University of Bern, Bühlstrasse 28, 3012 Bern, Switzerland. Electronic address: gertsch@ibmm.unibe.ch.
authors.forename: Jürg
authors.initials: J
authors.lastname: Gertsch
mesh_terms: D016718:Arachidonic Acid / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods*; D063388:Endocannabinoids / Q000097:blood*; D004983:Ethanolamines / Q000097:blood*; D006801:Humans; D011453:Prostaglandins / Q000097:blood*; D013256:Steroids / Q000097:blood*; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D063388:Endocannabinoids; D004983:Ethanolamines; C022203:N-acylethanolamines; D011453:Prostaglandins; D013256:Steroids; D016718:Arachidonic Acid
keywords: Arachidonic acid; Endocannabinoids; LC-MS/MS; Lipids; N-acylethanolamines; Noladin ether; Palmitoylethanolamide; Prostanoids; Steroids