Oliveira Igor Gustavo Carvalho, de Souza Israel Donizeti, de Crippa José Alexandre de Souza, Queiroz Maria Eugênia Costa
Department of Chemistry, Faculty of Philosophy, Sciences and Letters at Ribeirão Preto, University of São Paulo, São Paulo, Ribeirão Preto-SP, Brazil.
Department of Neuroscience and Behavior, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Ribeirão Preto-SP, Brazil.
J Sep Sci. 2025 Jan;48(1):e70068. doi: 10.1002/jssc.70068.
Cannabidiol (CBD) and Δ-tetrahydrocannabinol (THC), the main components of Cannabis sativa plants, can interact with specific cell receptors known as cannabinoid receptors (CBs). The endogenous compounds anandamide (AEA) and 2-arachidonoylglycerol (2-AG) are CB agonists, and, alongside enzymes, they constitute the endocannabinoid system (ECS) and take part in neuromodulation. Several LC-MS/MS methods have been developed to quantify these compounds in biological matrixes, but a fast and simple method that can determine these analytes in plasma samples simultaneously is not available. Here, we propose a disposable pipette extraction technique containing a zirconia-based sorbent (DPX(Zr)) combined with UHPLC-MS/MS analysis to determine CBD, THC, AEA, and 2-AG in plasma samples, simultaneously. The method combines simple protein precipitation (PPT) with a one-step DPX procedure to remove phospholipids, one of the most common endogenous interferents in biological samples. Optimization of the combined PPT-DPX sample preparation method reduced the matrix effect and improved the sensitivity of the analytical method. The validated DPX(Zr)-UHPLC-MS/MS method reported LLOQs of 0.1 ng mL for AEA and 2-AG and 1 ng mL for CBD and THC. The method demonstrated intra- and interassay accuracy and precision of less than 20% for the LLOQ, and less than 15% for the other calibration points. Additionally, no carryover or significant matrix effect was observed. We applied this method to determine AEA, 2-AG, and CBD in plasma samples obtained from obsessive-compulsive disorder patients treated with CBD.
大麻植物的主要成分大麻二酚（CBD）和Δ-四氢大麻酚（THC）可与称为大麻素受体（CB）的特定细胞受体相互作用。内源性化合物花生四烯乙醇胺（AEA）和2-花生四烯酸甘油酯（2-AG）是CB激动剂，它们与酶一起构成内源性大麻素系统（ECS）并参与神经调节。已经开发了几种液相色谱-串联质谱（LC-MS/MS）方法来定量生物基质中的这些化合物，但尚无一种能够同时测定血浆样品中这些分析物的快速简便方法。在此，我们提出一种包含基于氧化锆的吸附剂的一次性移液器萃取技术（DPX(Zr)），并结合超高效液相色谱-串联质谱（UHPLC-MS/MS）分析，以同时测定血浆样品中的CBD、THC、AEA和2-AG。该方法将简单的蛋白沉淀（PPT）与一步DPX程序相结合，以去除生物样品中最常见的内源性干扰物之一磷脂。对PPT-DPX组合样品制备方法的优化降低了基质效应并提高了分析方法的灵敏度。经过验证的DPX(Zr)-UHPLC-MS/MS方法报告的AEA和2-AG的定量下限（LLOQ）为0.1 ng/mL，CBD和THC的定量下限为1 ng/mL。该方法在LLOQ时的批内和批间准确度和精密度小于20%，在其他校准点时小于15%。此外，未观察到残留或显著的基质效应。我们应用该方法测定了接受CBD治疗的强迫症患者血浆样品中的AEA、2-AG和CBD。
