Tzimas Petros S, Petrakis Eleftherios A, Halabalaki Maria, Skaltsounis Leandros A
Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis Zografou, 15771, Athens, Greece.
Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis Zografou, 15771, Athens, Greece.
Anal Chim Acta. 2021 Mar 15;1150:338200. doi: 10.1016/j.aca.2021.338200. Epub 2021 Jan 18.
Cannabidiol (CBD) and cannabidiolic acid (CBDA) represent the most abundant non-psychoactive cannabinoids in fiber-type Cannabis sativa L. (hemp) and both have demonstrated high therapeutic potential. Hence, efficient extraction coupled with reliable determination of these compounds is crucial for informed utilization of hemp and is increasingly needed in the present state of harmonization efforts. In this context, a systematic approach for extraction optimization was followed, which initially involved comparison of three widely available extraction techniques, i.e. ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE), and dynamic maceration (DM). These were applied on samples of different hemp varieties (n = 3) using ethanol as a safe and efficient solvent. UAE showed the most promising results and was further optimized by means of response surface methodology (RSM), based on a circumscribed central composite design. The conditions maximizing CBD, CBDA, and total CBD content as well as extraction yield were determined with high desirability (0.97) and were experimentally confirmed. The optimized UAE method was also compared with a previously reported extraction procedure, demonstrating superior performance. For the quantitation of CBD and CBDA in hemp extracts, a reversed-phase UPLC-PDA method was developed and validated. Chromatographic separation was achieved in less than 10 min, while satisfactory results for linearity (R > 0.996), precision (RSD < 2.0%), and accuracy (recovery rates of 93.1-101.0%) were obtained for both analytes. Limits of detection were determined as 0.07 and 0.04 μg mL for CBD and CBDA, respectively, indicating sufficient sensitivity. The good performance of the method was verified by the evaluation of additional parameters (e.g. matrix effect, extraction recovery), which was largely enabled by the use of isolated standards. The whole analytical workflow, involving both optimized UAE extraction and UPLC-PDA determination, entails simplified manipulation and may offer a reliable and cost-effective approach for routine quality control of hemp regarding the principal cannabinoids.
大麻二酚（CBD）和大麻二酚酸（CBDA）是纤维型大麻（工业大麻）中含量最丰富的非精神活性大麻素，二者均已显示出很高的治疗潜力。因此，高效提取并可靠测定这些化合物对于合理利用大麻至关重要，并且在当前的协调工作状态下需求日益增加。在此背景下，采用了一种系统的提取优化方法，该方法首先涉及比较三种广泛使用的提取技术，即超声辅助提取（UAE）、微波辅助提取（MAE）和动态浸渍法（DM）。使用乙醇作为安全有效的溶剂，将这些技术应用于不同大麻品种的样品（n = 3）。UAE显示出最有前景的结果，并基于限定的中心复合设计，通过响应面法（RSM）进一步优化。确定了使CBD、CBDA和总CBD含量以及提取率最大化的条件，合意度高（0.97），并通过实验得到了证实。优化后的UAE方法还与先前报道的提取程序进行了比较，显示出优越的性能。为了定量分析大麻提取物中的CBD和CBDA，开发并验证了一种反相超高效液相色谱 - 光电二极管阵列检测（UPLC - PDA）方法。不到10分钟即可实现色谱分离，两种分析物的线性（R > 0.996）、精密度（RSD < 2.0%）和准确度（回收率93.1 - 101.0%）均获得了满意的结果。CBD和CBDA的检测限分别确定为0.07和0.04 μg/mL，表明灵敏度足够。通过评估其他参数（例如基质效应、提取回收率）验证了该方法的良好性能，这在很大程度上得益于使用了分离的标准品。整个分析流程，包括优化的UAE提取和UPLC - PDA测定，操作简便，可为大麻主要大麻素的常规质量控制提供可靠且经济高效的方法。
