Brighenti Virginia, Pellati Federica, Steinbach Marleen, Maran Davide, Benvenuti Stefania
Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 103, Modena 41125, Italy.
Department of Life Sciences, University of Modena and Reggio Emilia, Via G. Campi 103, Modena 41125, Italy.
J Pharm Biomed Anal. 2017 Sep 5;143:228-236. doi: 10.1016/j.jpba.2017.05.049. Epub 2017 Jun 4.
The present work was aimed at the development and validation of a new, efficient and reliable technique for the analysis of the main non-psychoactive cannabinoids in fibre-type Cannabis sativa L. (hemp) inflorescences belonging to different varieties. This study was designed to identify samples with a high content of bioactive compounds, with a view to underscoring the importance of quality control in derived products as well. Different extraction methods, including dynamic maceration (DM), ultrasound-assisted extraction (UAE), microwave-assisted extraction (MAE) and supercritical-fluid extraction (SFE) were applied and compared in order to obtain a high yield of the target analytes from hemp. Dynamic maceration for 45min with ethanol (EtOH) at room temperature proved to be the most suitable technique for the extraction of cannabinoids in hemp samples. The analysis of the target analytes in hemp extracts was carried out by developing a new reversed-phase high-performance liquid chromatography (HPLC) method coupled with diode array (UV/DAD) and electrospray ionization-mass spectrometry (ESI-MS) detection, by using an ion trap mass analyser. An Ascentis Express C column (150mm×3.0mm I.D., 2.7μm) was selected for the HPLC analysis, with a mobile phase composed of 0.1% formic acid in both water and acetonitrile, under gradient elution. The application of the fused-core technology allowed us to obtain a significant improvement of the HPLC performance compared with that of conventional particulate stationary phases, with a shorter analysis time and a remarkable reduction of solvent usage. The analytical method optimized in this study was fully validated to show compliance with international requirements. Furthermore, it was applied to the characterization of nine hemp samples and six hemp-based pharmaceutical products. As such, it was demonstrated to be a very useful tool for the analysis of cannabinoids in both the plant material and its derivatives for pharmaceutical and nutraceutical applications.
本研究旨在开发并验证一种新的、高效且可靠的技术，用于分析不同品种纤维型大麻（Cannabis sativa L.）花序中的主要非精神活性大麻素。本研究旨在鉴定生物活性化合物含量高的样品，以强调衍生产品质量控制的重要性。为了从大麻中获得高产量的目标分析物，应用并比较了不同的提取方法，包括动态浸渍法（DM）、超声辅助提取法（UAE）、微波辅助提取法（MAE）和超临界流体萃取法（SFE）。室温下用乙醇（EtOH）动态浸渍45分钟被证明是提取大麻样品中大麻素的最合适技术。通过开发一种新的反相高效液相色谱（HPLC）方法，并结合二极管阵列（UV/DAD）和电喷雾电离质谱（ESI-MS）检测，使用离子阱质量分析仪，对大麻提取物中的目标分析物进行了分析。选择Ascentis Express C柱（150mm×3.0mm内径，2.7μm）进行HPLC分析，流动相由水和乙腈中均含有0.1%甲酸组成，采用梯度洗脱。与传统颗粒固定相相比，熔融核技术的应用使我们能够显著提高HPLC性能，缩短分析时间并显著减少溶剂用量。本研究中优化的分析方法经过充分验证，以表明符合国际要求。此外，它还应用于九个大麻样品和六种大麻基药品的表征。因此，它被证明是分析植物材料及其用于制药和营养保健品应用的衍生物中大麻素的非常有用的工具。
