De Palma Ryan, Matta Murali K, Patel Vikram, Florian Jeffry, Strauss David G, Rouse Rodney
Division of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug Administration, USA.
Division of Applied Regulatory Science, Office of Clinical Pharmacology, Center for Drug Evaluation and Research, US Food and Drug Administration, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2025 Sep 1;1263:124694. doi: 10.1016/j.jchromb.2025.124694. Epub 2025 Jun 3.
The removal of hemp from the definition of marijuana in the 2018 Agricultural Improvement Act has increased the number of cannabidiol-containing products available to consumers. Consequently, consumer use has also increased. Increased product availability and use drives the need for sensitive and specific analytical assays to measure the cannabidiol (CBD) and metabolites in patients, to establish dose-effect relationships and to gain knowledge of their pharmacokinetics. Here we describe the development and validation of a rapid high-throughput LC-MS/MS bioanalytical method for the quantification of cannabidiol and primary metabolites in human plasma to support an FDA-sponsored clinical study (NCT06192589). Sample preparation a single step protein precipitation followed by filtration through 96-well Phree™ Phospholipid removal plates. The method was validated over ranges of CBD: 1.95-500.00 ng mL; 7-hydroxy-cannabidiol (7-OH-CBD): 3.91-1000.00 ng mL; 7-carboxy-cannabidiol (7-COOH-CBD): 31.25-8000.00 ng mL. There was no cross-analyte interference, injection carryover, or matrix effect observed with this method. Analyte recoveries were consistent across three QC levels ranging from 83.90 to 90.85 %. Inter-day Accuracy across four QC level of all three analytes ranged from 93.87 to 107.31 % while precision ranged from 1.03 to 14.33 %. These results and other outlined in this manuscript met acceptance criteria as outline the current M10 Guidance for Bioanalytical Method Validation and Study Sample Analysis.
2018年《农业改进法案》将大麻从大麻的定义中移除，这增加了消费者可获得的含大麻二酚产品的数量。因此，消费者的使用量也有所增加。产品可得性和使用量的增加推动了对灵敏且特异的分析检测方法的需求，以测量患者体内的大麻二酚（CBD）及其代谢物，建立剂量效应关系并了解其药代动力学。在此，我们描述了一种快速高通量液相色谱-串联质谱（LC-MS/MS）生物分析方法的开发与验证，该方法用于定量人血浆中的大麻二酚及其主要代谢物，以支持美国食品药品监督管理局（FDA）赞助的一项临床研究（NCT06192589）。样品制备采用一步蛋白沉淀法，随后通过96孔Phree™磷脂去除板进行过滤。该方法在以下浓度范围内进行了验证：CBD：1.95 - 500.00 ng/mL；7-羟基大麻二酚（7-OH-CBD）：3.91 - 1000.00 ng/mL；7-羧基大麻二酚（7-COOH-CBD）：31.25 - 8000.00 ng/mL。该方法未观察到交叉分析物干扰、进样残留或基质效应。三种分析物在三个质量控制水平下的回收率一致，范围为83.90%至90.85%。所有三种分析物在四个质量控制水平下的日间准确度范围为93.87%至107.31%，精密度范围为1.03%至14.33%。本文所述的这些结果及其他内容符合现行生物分析方法验证和研究样品分析M10指南所述的验收标准。
