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.

中文翻译：

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从主动脉组织和血浆中优化提取 2-花生四烯甘油和花生四烯酸，用于 LC-MS/MS 定量

动脉粥样硬化是一种以动脉壁中脂肪、胆固醇和免疫细胞积聚导致斑块形成为特征的疾病。如果斑块破裂，可能形成闭塞性血栓形成，导致心脏病发作或中风。巨噬细胞表达 CB-2 受体，是一种在斑块不稳定和破裂中起作用的免疫细胞。已发现内源性大麻素 anandamide (AEA) 和 2-花生四烯酸甘油 (2-AG) 对全身和免疫系统的 CB-1 和 CB-2 受体具有活性。在本研究中，我们研究了几种样品制备选项，用于对血浆和主动脉组织中的 AEA 和 2-AG 进行 LC-MS 定量。考虑的萃取包括液-液 (LLE)、固相 (SPE) 和支持液体 (SLE)。一些提取方案产生了高分析物回收率并防止了 1-AG/2-AG 异构化。我们的结果表明，使用甲苯进行液-液萃取时，两种分析物的回收率都最高，同时质谱仪中的电离抑制较低。这种提取和相应的 LC-MS/MS 分析为量化与内源性大麻素在动脉粥样硬化中的作用相关的基质中的 2-AG 和 AEA 提供了一种简单、高通量的机制。