Center for Clinical Pharmacology, Children's Hospital of Philadelphia, Philadelphia, PA 19104, United States; Department of Anesthesiology and Critical Care Medicine, University of Pennsylvania, Philadelphia, PA 19104, United States.
Center for Clinical Pharmacology, Children's Hospital of Philadelphia, Philadelphia, PA 19104, United States.
J Chromatogr B Analyt Technol Biomed Life Sci. 2019 Nov 1;1130-1131:121828. doi: 10.1016/j.jchromb.2019.121828. Epub 2019 Oct 21.
Medical cannabis is increasingly used for the treatment of various ailments in children and adults. Three major cannabinoids in cannabis are delta-9-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN). There is a growing need to develop and utilize a patient-centric blood microsampling methodology to enable clinical trials and facilitate therapeutic drug monitoring. We have employed the volumetric absorptive microsampling (VAMS™) devices that enables accurate and precise collection of a fixed volume (20 µL) of blood, minimizing the impact of hematocriton accurate quantitation. We developed an ultra-performance liquid chromatographic method with tandem mass spectrometry detection for the quantification of three cannabinoids (THC, CBD, and CBN) employing deuterium labelled internal standards (THC-D, CBD-D, and CBN-D). Sample extraction of VAMS™ devices, followed by solid phase extraction, reverse phase chromatographic separation, and selective detection using tandem mass spectrometry with a 6-minute runtime per sample was developed. Standard curves were linear between 1 and 500 ng/mL for THC and 0.5-500 ng/mL for CBD and CBN. Intra-day accuracies were within 91.3-112% while inter-day accuracies were within 94.4-107% with both having precisions (CV (%)) of <13% based on quality control samples in a three day validation study for all three cannabinoids. Analytes were stable in human whole blood under assay conditions (60 h at room temperature and 24 h in autosampler post-extraction). Dried microsamples were stable for one week at 40 °C, two weeks (15 days) under different storage conditions (room temperature, 4, -20 and -78 °C), one month (29 days) at -20 and -78 °C and three months (68 days) at -78 °C. This assay provides an efficient quantitation of THC, CBD, and CBN in VAMS™ devices and is currently being implemented for pediatric clinical trials.
医用大麻越来越多地被用于治疗儿童和成人的各种疾病。大麻中的三种主要大麻素是 delta-9-四氢大麻酚（THC）、大麻二酚（CBD）和大麻酚（CBN）。人们越来越需要开发和利用以患者为中心的血液微采样方法，以进行临床试验并促进治疗药物监测。我们已经使用了容量吸收微采样（VAMS™）设备，该设备能够准确、精确地采集固定体积（20 µL）的血液，最大限度地减少了对血细胞比容准确定量的影响。我们开发了一种超高效液相色谱法与串联质谱检测法，用于定量分析三种大麻素（THC、CBD 和 CBN），采用氘标记内标物（THC-D、CBD-D 和 CBN-D）。采用 VAMS™设备进行样品提取，然后进行固相萃取、反相色谱分离，以及使用串联质谱进行选择性检测，每个样品的运行时间为 6 分钟。THC 的标准曲线在 1 到 500ng/mL 之间线性，CBD 和 CBN 的标准曲线在 0.5 到 500ng/mL 之间线性。在为期三天的验证研究中，所有三种大麻素的 QC 样品的日内准确度在 91.3-112%之间，日间准确度在 94.4-107%之间，均具有 <13%的精密度（CV（%））。在分析条件下（室温下 60 小时和提取后自动进样器中 24 小时），分析物在人全血中稳定。干燥微样本在 40°C 下稳定一周，在不同储存条件下（室温、4°C、-20°C 和-78°C）稳定两周（15 天），在-20°C 和-78°C 下稳定一个月（29 天），在-78°C 下稳定三个月（68 天）。该测定法可有效定量 VAMS™ 设备中的 THC、CBD 和 CBN，目前正在用于儿科临床试验。
