Brain and Mind Centre, Translational Research Collective, Faculty of Medicine and Health, The University of Sydney, Sydney, Australia.
Department of Psychiatry and Psychotherapy, Central Institute of Mental Health, Medical Faculty Mannheim, Heidelberg University, Mannheim, Germany.
Cannabis Cannabinoid Res. 2023 Aug;8(4):657-669. doi: 10.1089/can.2021.0181. Epub 2022 May 17.
The primary compounds of , delta-9-tetrahydrocannabinol (Δ-THC) and cannabidiol (CBD), inflict a direct influence on the endocannabinoid system-a complex lipid signaling network with a central role in neurotransmission and control of inhibitory and excitatory synapses. These phytocannabinoids often interact with endogenously produced endocannabinoids (eCBs), as well as their structurally related -acylethanolamines (NAEs), to drive neurobiological, nociceptive, and inflammatory responses. Identifying and quantifying changes in these lipid neuromodulators can be challenging owing to their low abundance in complex matrices. This article describes a robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the extraction and quantification of the eCBs anandamide and 2-arachidonoylglycerol, along with their congener NAEs oleoylethanolamine and palmitoylethanolamine, and phytocannabinoids CBD, Δ-THC, and 11-Nor-9-carboxy-Δ-tetrahydrocannabinol, a major metabolite of Δ-THC. Our method was applied to explore pharmacokinetic and pharmacodynamic effects from intraperitoneal injections of Δ-THC and CBD on circulating levels of eCBs and NAEs in rodent serum. Detection limits ranged from low nanomolar to picomolar in concentration for eCBs (0.012-0.24 pmol/mL), NAEs (0.059 pmol/mL), and phytocannabinoids (0.24-0.73 pmol/mL). Our method displayed good linearity for calibration curves of all analytes (>0.99) as well as acceptable accuracy and precision, with quality controls not deviating >15% from their nominal value. Our LC-MS/MS method reliably identified changes to these endogenous lipid mediators that followed a causal relationship, which was dependent on both the type of phytocannabinoid administered and its pharmaceutical preparation. We present a rapid and reliable method for the simultaneous quantification of phytocannabinoids, eCBs, and NAEs in serum using LC-MS/MS. The accuracy and sensitivity of our assay infer it can routinely monitor endogenous levels of these lipid neuromodulators in serum and their response to external stimuli, including cannabimimetic agents.
大麻素的主要成分，即 δ-9-四氢大麻酚（Δ-THC）和大麻二酚（CBD），对内源性大麻素系统产生直接影响，该系统是一个复杂的脂质信号网络，在神经传递和抑制性及兴奋性突触的控制中发挥核心作用。这些植物大麻素经常与内源性产生的内源性大麻素（eCBs）以及它们结构相关的酰基乙醇胺（NAEs）相互作用，从而驱动神经生物学、痛觉和炎症反应。由于这些脂质神经调节剂在复杂基质中的含量较低，因此识别和量化它们的变化具有挑战性。本文描述了一种强大的液相色谱-串联质谱（LC-MS/MS）方法，用于提取和定量分析 eCB 大麻酰胺和 2-花生四烯酰甘油，以及它们的同系物 NAEs 油酰乙醇胺和棕榈酰乙醇胺，以及植物大麻素 CBD、Δ-THC 和 11-去甲-9-羧基-Δ-四氢大麻酚，这是 Δ-THC 的主要代谢物。我们的方法用于探索腹腔注射 Δ-THC 和 CBD 对啮齿动物血清中环流 eCB 和 NAE 水平的药代动力学和药效学影响。检测限在 eCB（0.012-0.24 pmol/mL）、NAE（0.059 pmol/mL）和植物大麻素（0.24-0.73 pmol/mL）的浓度中从纳摩尔到皮摩尔不等。我们的方法对所有分析物的校准曲线均表现出良好的线性（>0.99），并且具有可接受的准确性和精密度，质量控制品的偏差不超过其标称值的 15%。我们的 LC-MS/MS 方法可靠地识别了这些内源性脂质介质的变化，这些变化遵循因果关系，这取决于给予的植物大麻素的类型及其药物制剂。我们提出了一种快速可靠的方法，用于使用 LC-MS/MS 同时定量血清中的植物大麻素、eCB 和 NAE。我们的测定方法的准确性和灵敏度推断，它可以常规监测血清中这些脂质神经调节剂的内源性水平及其对包括大麻类似物在内的外部刺激的反应。
