Optimized extraction of 2-arachidonyl glycerol and anandamide from aortic tissue and plasma for quantification by LC-MS/MS

卷:118

Garst, Christopher ; Fulmer, Makenzie ; Thewke, Doug ; Brown, Stacy

E Tennessee State Univ

关键词: Anandamide;    AEA;    2-Arachidonoylglycerol;    2-AG;    Endocannabinoids;    Liquid Chromatography;    Mass Spectrometry;    LC-MS;   

DOI  :  10.1002/ejlt.201500115

学科分类：食品科学和技术

【 摘 要 】

Atherosclerosis is a disease characterized by plaque formation due to an accumulation of fat, cholesterol, and immune cells in the walls of arteries. If a plaque ruptures, an occlusive thrombosis may form that causes either a heart attack or stroke. Macrophages express CB-2 receptors, and are one type of immune cell that plays a role in plaque destabilization and rupture. Endocannabinoids anandamide (AEA) and 2-arachidonyl glycerol (2-AG) have been found to have activity on CB-1 and CB-2 receptors throughout the body and immune system. In this study, we investigated several sample preparation options for the LC-MS quantification of AEA and 2-AG from plasma and aortic tissue. The extractions considered included liquid-liquid (LLE), solid-phase (SPE), and supported liquid (SLE). Some extraction protocols yielded high analyte recovery and prevention of 1-AG/2-AG isomerization. Our results indicate that a liquid-liquid extraction using toluene yields the highest recovery for both analytes, coupled with low ionization suppression in the mass spectrometer. This extraction and corresponding LC-MS/MS assay provides a simple, high throughput mechanism for the quantification of 2-AG and AEA in matrices relevant to the study of endocannabinoids' role in atherosclerosis. Practical applications : We developed an extraction method for AEA and 2-AG from plasma and aortic tissue samples and an LC-MS/MS assay for quantification of these compounds to help understand the role of endocannabinoids and CB-2 receptors in atherosclerosis. This assay can be applied toward the measurement of endocannabinoids, AEA and 2AG, in aortic tissue and plasma.