A simple and reliable method was developed and validated to determine the endocannabinoids anandamide (AEA) and 2-arachidonoylglycerol (2-AG) in rat brain samples by micro salting-out assisted liquid-liquid extraction combined with ultra-high performance liquid chromatography tandem mass spectrometry (SALLLE/UHPLC-MS/MS). The SALLE parameters (brain homogenate volume, salting-out agent, salt concentration, salt solution volume, organic solvent, organic solvent volume, and centrifugation temperature) were optimized to improve sensitivity and selectivity of the method. The SALLE/UHPLC-MS/MS method presented linear ranges from 2.00 to 20.00 ng mL⁻¹ for AEA and from 0.300 to 10.00 μg mL⁻¹ for 2-AG, no significant matrix effect, and inter- and intra-assay precision and accuracy with CV and RSE values lower than 15%, respectively. This innovative method was successfully applied to determine AEA and 2-AG in brain hemispheres from a 6-OHDA animal model of Parkinson’s disease (PD).

建立了一种简单可靠的方法，并通过微盐析辅助液-液萃取与超高效液相色谱串联质谱法测定大鼠脑样品中的内源性大麻素（AEA）和2-花生四烯酸甘油酯（2-AG）光谱分析（SALLLE / UHPLC-MS / MS）。优化SALLE参数（脑匀浆体积，盐析剂，盐浓度，盐溶液体积，有机溶剂，有机溶剂体积和离心温度）以提高方法的灵敏度和选择性。SALLE / UHPLC-MS / MS方法的线性范围为2.00至20.00 ng mL ^-1（对于AEA）和0.300至10.00μgmL ^-1对于2-AG，CV和RSE值分别低于15％，无明显的基质效应，且批间和批内准确度均不高。此创新方法已成功应用于帕金森氏病（PD）的6-OHDA动物模型中测定大脑半球中的AEA和2-AG。