A column switching ultrahigh-performance liquid chromatography-tandem mass spectrometry method to determine anandamide and 2-arachidonoylglycerol in plasma samples.
This study reports a fast, sensitive, and selective column switching ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method to determine the endocannabinoids (eCBs), anandamide (AEA), and 2-arachidonoylglycerol (2-AG) in plasma samples. This bidimensional system used a restricted access media column (RP-8 ADS, 25 mm × 4 mm × 25 μM) in the first dimension and a core-shell Kinetex C18 (100 mm × 2, 1.7 mm × 1 μM) column in the second dimension, followed by detection in a mass spectrometer triple quadrupole (multiple reactions monitoring mode) operating in the positive mode. RP-8 ADS was used for trace enrichment of eCBs (reverse phase partitioning) and macromolecular matrix size exclusion; the core-shell column was used for the chromatographic separation. The column switching UHPLC-MS/MS method presented a linear range spanning from 0.1 ng mL-1(LOQ) to 6 ng mL-1for AEA and from 0.04 ng mL-1(LOQ) to 10 ng mL-1for 2-AG. Excluding the LLOQ values, the precision assays provided coefficients of variation lower than 8% and accuracy with relative standard error values lower than 14%. Neither carryover nor matrix effects were detected. This high-throughput column switching method compared to conventional methods is time saving as it involves fewer steps, consumes less solvent, and presents lower LLOQ. The column switching UHPLC-MS/MS method was successfully applied to determine AEA and 2-AG in plasma samples obtained from Alzheimer's disease patients. Graphical abstract A column switching ultra high-performance liquid chromatography-tandem mass spectrometry method using RP-8 ADS column and core shell column to determine endocannabinoids in plasma samples.
Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo (USP), Avenida Bandeirantes, 3900, São Paulo, Ribeirão Preto, CEP 14040-901, Brazil.
Camila
C
Marchioni
Departamento de Química, Universidade de São Paulo (USP), Avenida Bandeirantes, 3900, São Paulo, Ribeirão Preto, CEP 14040-901, Brazil.
Israel Donizeti
ID
de Souza
Departamento de Química, Universidade de São Paulo (USP), Avenida Bandeirantes, 3900, São Paulo, Ribeirão Preto, CEP 14040-901, Brazil.
Caroline Fernandes
CF
Grecco
Department of Neuroscience and Behavior, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.
José Alexandre
JA
Crippa
Department of Neuroscience and Behavior, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.
Vitor
V
Tumas
Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo (USP), Avenida Bandeirantes, 3900, São Paulo, Ribeirão Preto, CEP 14040-901, Brazil. mariaeqn@ffclrp.usp.br.|Departamento de Química, Universidade de São Paulo (USP), Avenida Bandeirantes, 3900, São Paulo, Ribeirão Preto, CEP 14040-901, Brazil. mariaeqn@ffclrp.usp.br.
Maria Eugênia Costa
MEC
Queiroz
D000544:Alzheimer Disease / Q000097:blood; D001095:Arachidonic Acids / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D063388:Endocannabinoids / Q000097:blood*; D004867:Equipment Design; D005989:Glycerides / Q000097:blood*; D006801:Humans; D057230:Limit of Detection; D053284:Polyunsaturated Alkamides / Q000097:blood*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods*
D001095:Arachidonic Acids; D063388:Endocannabinoids; D005989:Glycerides; D053284:Polyunsaturated Alkamides; C094503:glyceryl 2-arachidonate; C078814:anandamide
Column switching; Endocannabinoids; Liquid chromatography-tandem mass spectrometry; Plasma samples