title: High-throughput lipidomic analysis of fatty acid derived eicosanoids and N-acylethanolamines.
abstract: Fatty acid-derived eicosanoids and N-acylethanolamines (NAE) are important bioactive lipid mediators involved in numerous biological processes including cell signaling and disease progression. To facilitate research on these lipid mediators, we have developed a targeted high-throughput mass spectrometric based methodology to monitor and quantitate both eicosanoids and NAEs, and can be analyzed separately or together in series. Each methodology utilizes scheduled multiple reaction monitoring (sMRM) pairs in conjunction with a 25 min reverse-phase HPLC separation. The eicosanoid methodology monitors 141 unique metabolites and quantitative amounts can be determined for over 100 of these metabolites against standards. The analysis covers eicosanoids generated from cycloxygenase, lipoxygenase, cytochrome P450 enzymes, and those generated from non-enzymatic pathways. The NAE analysis monitors 36 metabolites and quantitative amounts can be determined for 33 of these metabolites against standards. The NAE method contains metabolites derived from saturated fatty acids, unsaturated fatty acids, and eicosanoids. The lower limit of detection for eicosanoids ranges from 0.1pg to 1pg, while NAEs ranges from 0.1pg to 1000pg. The rationale and design of the methodology is discussed.
authors.affiliation: Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, CA, USA.
authors.forename: Darren S
authors.initials: DS
authors.lastname: Dumlao
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Buczynski
authors.forename: Paul C
authors.initials: PC
authors.lastname: Norris
authors.forename: Richard
authors.initials: R
authors.lastname: Harkewicz
authors.forename: Edward A
authors.initials: EA
authors.lastname: Dennis
mesh_terms: D000818:Animals; D015232:Dinoprostone / Q000737:chemistry; D015777:Eicosanoids / Q000032:analysis* / Q000134:cerebrospinal fluid / Q000737:chemistry; D004983:Ethanolamines / Q000032:analysis* / Q000134:cerebrospinal fluid / Q000737:chemistry; D057166:High-Throughput Screening Assays / Q000379:methods*; D050356:Lipid Metabolism; D051381:Rats; D012015:Reference Standards; D012996:Solutions; D013997:Time Factors
chemical_list: D015777:Eicosanoids; D004983:Ethanolamines; C022203:N-acylethanolamines; D012996:Solutions; D015232:Dinoprostone