University of Liège, CIRM, Laboratory of Pharmaceutical Analytical Chemistry, B36 Tower 4 Avenue Hippocrate 15, 4000 Liège, Belgium.
Azienda USL Toscana centro, Laboratory of Clinical Toxicology and Antidoping, Via di San Salvi 12, 50135 Florence, Italy.
J Pharm Biomed Anal. 2019 Mar 20;166:326-335. doi: 10.1016/j.jpba.2019.01.032. Epub 2019 Jan 21.
Cannabidiol (CBD) and Δ-tetrahydrocannabinol (Δ-THC) are considered as the most interesting cannabinoids in Cannabis sativa L. for the clinical practice. Since 2013, the Italian law allows pharmacists to prepare and dispense cannabis extracts to patients under medical prescription, and requires the evaluation of CBD and Δ-THC content in cannabis extracts before sale. Cannabis olive oil extracts are prepared from dried female cannabis inflorescences, but a standard protocol is still missing. In this study, a fast RP-HPLC/UV method has been developed to quantify CBD and Δ-THC in cannabis olive oil extracts. The analytical quality by design strategy has been applied to the method development, setting critical resolution and total analysis time as critical method attributes (CMAs), and selecting column temperature, buffer pH and flow rate as critical method parameters. Information from Doehlert Design in response surface methodology combined to Monte-Carlo simulations led to draw the risk of failure maps and to identify the method operable design region. The method was validated according to the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines and then implemented in routine analysis. A control strategy based on system control charts was planned to monitor the developed method performances. Evaluation data were recorded over a period of one year of routine use, and both the CMAs showed values within the specifications in every analysis performed. Hence, a new risk evaluation for the future performances of the method was achieved by using a Bayesian approach based on the routine use data, computing the future distribution of the two CMAs. Finally, a study focusing on the monitoring of CBD and Δ-THC concentrations in cannabis olive oil extracts was carried out. The developed method was applied to 459 extracts. The statistical analysis of the obtained results highlighted a wide variability in terms of concentrations among different samples from the same starting typology of cannabis, underlining the compelling need of a standardised procedure to harmonise the preparation of the extracts.
大麻二酚（CBD）和Δ-四氢大麻酚（Δ-THC）被认为是大麻属植物中最具临床应用价值的两种大麻素。自 2013 年以来，意大利法律允许药剂师根据医疗处方为患者配制和分发大麻提取物，并要求在销售前评估大麻提取物中 CBD 和 Δ-THC 的含量。大麻橄榄油提取物是由干燥的雌性大麻花序制备而成，但目前仍缺乏标准的制备方案。在本研究中，建立了一种快速反相高效液相色谱/紫外检测法（RP-HPLC/UV）用于定量分析大麻橄榄油提取物中的 CBD 和 Δ-THC。采用分析设计策略（Doehlert 设计和蒙特卡罗模拟）来开发该方法，将关键分离度和总分析时间（TAT）作为关键方法属性（CMAs），并选择柱温、缓冲液 pH 值和流速作为关键方法参数。响应曲面法（Doehlert 设计）和蒙特卡罗模拟相结合的信息用于绘制风险失败图，以确定方法可操作设计区域。该方法按照国际人用药品注册技术协调会（ICH）指南进行验证，并随后应用于常规分析。根据控制策略，计划使用系统控制图来监测开发方法的性能。在常规使用的一年中记录评估数据，并且在每次分析中，两个 CMA 均显示在规格范围内。因此，通过使用基于常规使用数据的贝叶斯方法对方法未来的性能进行了新的风险评估，计算了两个 CMA 的未来分布。最后，进行了一项关于大麻橄榄油提取物中 CBD 和 Δ-THC 浓度监测的研究。该方法应用于 459 个提取物。对获得的结果进行统计学分析，结果表明，即使是来自同一起始大麻类型的不同样本，其浓度也存在很大差异，这突出表明需要标准化程序来统一提取物的制备。
