Chen Jing, Paudel Kalpana S, Derbenev Andrei V, Smith Bret N, Stinchcomb Audra L
Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, KY 40536-0082, USA.
Chromatographia. 2009 Jan 1;69(1-2):1-7. doi: 10.1365/s10337-008-0841-x.
A quantitative method has been developed and validated for the simultaneous determination of anandamide (AEA), docosatetraenylethanolamide (DEA) and N-arachidonyldopamine (NADA) in dorsal vagal complex (DVC) of rat brainstem by liquid chromatographic-electrospray ionization mass spectrometry. The analytes were extracted from the tissue samples of rat brainstem by a single step liquid extraction technique using acetonitrile. The chromatographic separation was conducted on a C18 column using a gradient mobile phase consisting of methanol and water at a flow rate of 0.3 mL min(-1). The analytes were quantified by positive electrospray ionization mass spectrometry with selected ion monitoring (SIM) mode. The limits of detection (LOD) for AEA, DEA and NADA were 0.5, 1 and 0.5 ng mL(-1), respectively. This method required only simple processing of the samples and could be applied to monitor the change in the level of these compounds in DVC of the rat brain tissue. Time dependent (10-70 min) accumulation of the endocannabinoids (AEA, DEA, and NADA) in brain tissue was also studied, which included a novel examination of the accumulation of DEA as a function of time in rat brain tissue after decapitation.
已开发并验证了一种定量方法，用于通过液相色谱-电喷雾电离质谱法同时测定大鼠脑干背侧迷走神经复合体（DVC）中的花生四烯酸乙醇胺（AEA）、二十二碳四烯酸乙醇胺（DEA）和N-花生四烯酰多巴胺（NADA）。使用乙腈通过单步液体萃取技术从大鼠脑干组织样品中提取分析物。在C18柱上进行色谱分离，使用由甲醇和水组成的梯度流动相，流速为0.3 mL min(-1)。通过正电喷雾电离质谱法在选择离子监测（SIM）模式下对分析物进行定量。AEA、DEA和NADA的检测限（LOD）分别为0.5、1和0.5 ng mL(-1) 。该方法仅需对样品进行简单处理，可用于监测大鼠脑组织DVC中这些化合物水平 的变化。还研究了内源性大麻素（AEA、DEA和NADA）在脑组织中的时间依赖性（10-70分钟）积累情况，并包括一项对大鼠脑组织断头后DEA积累随时间变化情况 的全新研究 。
