Department of Anatomy and Neurobiology, University of California, Irvine, California, USA.
Department of Neurobiology and Behavior, University of California, Irvine, California, USA.
Cannabis Cannabinoid Res. 2024 Feb;9(1):371-385. doi: 10.1089/can.2022.0216. Epub 2022 Nov 11.
Δ-tetrahydrocannabinol (THC) and cannabidiol (CBD) are major chemical constituents of cannabis, which may interact either directly or indirectly with the endocannabinoid and endocannabinoid-like ("paracannabinoid") systems, two lipid-based signaling complexes that play important roles in physiology. Legislative changes emphasize the need to understand how THC and CBD might impact endocannabinoid and paracannabinoid signaling, and to develop analytical approaches to study such impact. In this study, we describe a sensitive and accurate method for the simultaneous quantification of THC, its main oxidative metabolites [11-hydroxy-Δ-THC (11-OH-THC) and 11-nor-9-carboxy-Δ-THC (11-COOH-THC)], CBD, and a representative set of endocannabinoid [anandamide and 2-arachidonoyl--glycerol (2-AG)] and paracannabinoid [palmitoylethanolamide (PEA) and oleoylethanolamide (OEA)] compounds. Analyte separation relies on the temperature-dependent shape selectivity properties of polymerically bonded C18 stationary phases. Analytes are extracted from tissues using acetonitrile precipitation followed by phospholipid removal. The ultrahigh-performance liquid chromatography/tandem mass spectrometry protocol utilizes a commercially available C18 polymeric-bonded phase column and a simple gradient elution system. Ten-point calibration curves show excellent linearity (>0.99) over a wide range of analyte concentrations (0.02-500 ng/mL). Lowest limits of quantification are 0.05 ng/mL for anandamide, 0.1 ng/mL for 11-OH-THC and OEA, 0.2 ng/mL for THC and CBD, 0.5 ng/mL for 11-COOH-THC, 1.0 ng/mL for 2-AG, and 2.0 ng/mL for PEA. The lowest limits of detection are 0.02 ng/mL for anandamide, 0.05 ng/mL for 11-OH-THC and OEA, 0.1 ng/mL for THC and CBD, 0.2 ng/mL for 11-COOH-THC, 0.5 ng/mL for 2-AG, and 1.0 ng/mL for PEA. An application of the method is presented, which showed that phytocannabinoid administration elevates endocannabinoid levels in plasma and brain of adolescent male and female mice.
Δ-四氢大麻酚 (THC) 和大麻二酚 (CBD) 是大麻的主要化学成分，它们可能直接或间接与内源性大麻素和内源性大麻素样 ("拟大麻素") 系统相互作用，这两个脂质信号复合物在生理中发挥着重要作用。立法的变化强调了需要了解 THC 和 CBD 如何影响内源性大麻素和拟大麻素信号，并开发分析方法来研究这种影响。在这项研究中，我们描述了一种灵敏而准确的方法，用于同时定量 THC、其主要氧化代谢物 [11-羟基-Δ-THC (11-OH-THC) 和 11-去甲-9-羧酸-Δ-THC (11-COOH-THC)]、CBD 以及一组代表性的内源性大麻素 [花生四烯酸酰胺和 2-花生四烯酰基-甘油 (2-AG)] 和拟大麻素 [棕榈酰乙醇酰胺 (PEA) 和油酰乙醇酰胺 (OEA)] 化合物。分析物的分离依赖于聚合键合 C18 固定相的温度依赖性形状选择性特性。分析物使用乙腈沉淀从组织中提取，然后去除磷脂。超高效液相色谱/串联质谱法方案利用市售的 C18 聚合键合相柱和简单的梯度洗脱系统。十点校准曲线在宽浓度范围内（0.02-500ng/mL）表现出极好的线性 (>0.99)。花生四烯酸酰胺的最低定量下限为 0.05ng/mL，11-OH-THC 和 OEA 的最低定量下限为 0.1ng/mL，THC 和 CBD 的最低定量下限为 0.2ng/mL，11-COOH-THC 的最低定量下限为 0.5ng/mL，2-AG 的最低定量下限为 1.0ng/mL，PEA 的最低定量下限为 2.0ng/mL。花生四烯酸酰胺的最低检测限为 0.02ng/mL，11-OH-THC 和 OEA 的最低检测限为 0.05ng/mL，THC 和 CBD 的最低检测限为 0.1ng/mL，11-COOH-THC 的最低检测限为 0.2ng/mL，2-AG 的最低检测限为 0.5ng/mL，PEA 的最低检测限为 1.0ng/mL。该方法的应用表明，植物大麻素的给药会升高青春期雄性和雌性小鼠血浆和大脑中的内源性大麻素水平。
