Department of Anesthesiology, University of Colorado, Anschutz Medical Campus, 12705 E Montview Blvd, Aurora, CO 80045, USA.
Clinical Research, Canopy Growth Corporation, 1 Hershey Drive, Smiths Falls, ON K7A 0A8, Canada.
J Anal Toxicol. 2024 Aug 21;48(7):499-506. doi: 10.1093/jat/bkae048.
In recent years, potential therapeutic applications of several different cannabinoids, such as Δ9-tetrahydrocannabinol (Δ9-THC), its isomer Δ8-THC and Δ9-tetrahydrocannabivarin (Δ9-THCV), have been investigated. Nevertheless, to establish dose-effect relationship and to gain knowledge of their pharmacokinetics and metabolism, sensitive and specific analytical assays are needed to measure these compounds in patients. For this reason, we developed and validated an online extraction high-performance liquid/liquid chromatography-tandem mass spectrometry (LC/LC-MS-MS) method for the simultaneous quantification of 13 cannabinoids and metabolites including the Δ8 and Δ9 isomers of THC, THCV and those of their major metabolites in human plasma. Plasma was fortified with cannabinoids at varying concentrations within the working range of the respective compound and 200 µL was extracted using a simple one-step protein precipitation procedure. The extracts were analyzed using online trapping LC/LC-atmospheric pressure chemical ionization-MS-MS running in the positive multiple reaction monitoring mode. The lower limit of quantification ranged from 0.5 to 2.5 ng/mL, and the upper limit of quantification was 400 ng/mL for all analytes. Inter-day analytical accuracy and imprecision ranged from 82.9% to 109% and 4.3% to 20.3% (coefficient of variance), respectively. Of 534 plasma samples following controlled oral administration of Δ8-THCV, 236 were positive for Δ8-THCV (median; interquartile ranges: 3.5 ng/mL; 1.8-11.9 ng/mL), 383 for the major metabolite (-)-11-nor-9-carboxy-Δ8-tetrahydrocannabivarin (Δ8-THCV-COOH) (95.4 ng/mL; 20.7-328 ng/mL), 260 for (-)-11-nor-9-carboxy-Δ9-tetrahydrocannabivarin (Δ9-THCV-COOH) (5.8 ng/mL; 2.5-16.1 ng/mL), 157 for (-)-11-hydroxy-Δ8-tetrahydrocannabivarin (11-OH-Δ8-THCV) (1.7 ng/mL; 1.0-3.7 ng/mL), 49 for Δ8-THC-COOH (1.7 ng/mL; 1.4-2.3 ng/mL) and 42 for Δ9-THCV (1.3 ng/mL; 0.8-1.6 ng/mL). We developed and validated the first LC/LC-MS-MS assay for the specific quantification of Δ8-THC, Δ9-THC and THCV isomers and their respective metabolites in human plasma. Δ8-THCV-COOH, 11-hydroxy-Δ8-THCV and Δ9-THCV-COOH were the major Δ8-THCV metabolites in human plasma after oral administration of 98.6% pure Δ8-THCV.
近年来，人们对几种不同大麻素的潜在治疗应用进行了研究，例如 Δ9-四氢大麻酚（Δ9-THC）、其异构体 Δ8-THC 和 Δ9-四氢大麻素（Δ9-THCV）。然而，为了建立剂量-效应关系，并了解它们的药代动力学和代谢情况，需要使用敏感和特异的分析方法来测量患者体内的这些化合物。出于这个原因，我们开发并验证了一种在线提取高效液相/液-液色谱-串联质谱（LC/LC-MS-MS）方法，用于同时定量分析 13 种大麻素及其代谢物，包括 Δ8 和 Δ9 异构体 THC、THCV 及其主要代谢物在人血浆中的含量。在各自化合物的工作范围内，用不同浓度的大麻素对血浆进行强化，并使用简单的一步蛋白沉淀程序提取 200 μL。使用在线捕获 LC/LC-大气压化学电离-MS-MS 以正多重反应监测模式进行分析。定量下限范围为 0.5 至 2.5ng/mL，所有分析物的定量上限均为 400ng/mL。在口服给予 98.6%纯 Δ8-THCV 后，534 个血浆样本的日内分析准确性和精密度范围分别为 82.9%至 109%和 4.3%至 20.3%（变异系数）。在这些样本中，236 个样本中检测到 Δ8-THCV（中位数；四分位间距：3.5ng/mL；1.8-11.9ng/mL），383 个样本中检测到主要代谢物（-）-11-去甲-9-羧基-Δ8-四氢大麻素（Δ8-THCV-COOH）（95.4ng/mL；20.7-328ng/mL），260 个样本中检测到（-）-11-去甲-9-羧基-Δ9-四氢大麻素（Δ9-THCV-COOH）（5.8ng/mL；2.5-16.1ng/mL），157 个样本中检测到（-）-11-羟基-Δ8-四氢大麻素（11-OH-Δ8-THCV）（1.7ng/mL；1.0-3.7ng/mL），49 个样本中检测到 Δ8-THC-COOH（1.7ng/mL；1.4-2.3ng/mL），42 个样本中检测到 Δ9-THCV（1.3ng/mL；0.8-1.6ng/mL）。我们开发并验证了首个用于在人血浆中特异性定量分析 Δ8-THC、Δ9-THC 和 THCV 异构体及其各自代谢物的 LC/LC-MS-MS 分析方法。在口服给予 98.6%纯 Δ8-THCV 后，Δ8-THCV 的主要代谢物为 Δ8-THCV-COOH、11-羟基-Δ8-THCV 和 Δ9-THCV-COOH。
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