Qamar Sadia, Manrique Yady J, Parekh Harendra S, Falconer James R
School of Pharmacy, Pharmacy Australia Centre of Excellence, The University of Queensland, Brisbane, QLD 4102, Australia.
Queensland University of Technology, Brisbane, QLD 4000, Australia.
Biology (Basel). 2021 May 28;10(6):481. doi: 10.3390/biology10060481.
In this study, the optimal setup of supercritical fluid extraction (SFE) was designed and developed, leading to the quantitation of 11 distinct cannabinoids (cannabidivann (CBDV), tetrahydrocannabivann (THCV), cannabidiol (CBD), cannabigerol (CBG) cannabidiolic acid (CBDA), cannabigerolic acid (CBGA), cannabinol (CBN), delta 9-tetrahydrocannabinol (Δ-THC), delta 8-tetrahydrocannabinol (Δ-THC), cannabichomere (CBC) and delta 9-tetrahydrocannabinol acid (THCA-A)) extracted from the flowers of medicinal cannabis (sp. ). Supercritical carbon dioxide (scCO) extraction was performed at 37 °C, a pressure of 250 bar with the maximum theoretical density of CO (893.7 kg/m), which generated the highest yield of cannabinoids from the flower-derived extract. Additionally, a cold separator (separating chamber) was used and positioned immediately after the sample containing chamber to maximize the yield. It was also found that successive washing of the extract with fresh scCO further increased yields. Ultra-high performance liquid chromatography coupled with DAD (uHPLC-DAD) was used to develop a method for the quantification of 11 cannabinoids. The C18 stationary phase was used in conjunction with a two solvent system gradient program resulting in the acquisition of the well-resolved chromatogram over a timespan of 32 min. The accuracy and precision of isolated cannabinoids across inter-and intra-day periods were within acceptable limits (<±15%). The assay was also fully validated and deemed sensitive from linearity, LOQ, and LOD perspective. The findings of this body of work are expected to facilitate improved conditions for the optimal extraction of select cannabinoids using scCO, which holds promise in the development of well-characterized medicinal cannabis formulations. As to our best knowledge, this is the first study to report the uHPLC quantification method for the analysis of 11 cannabinoids from scCO extract in a single run with more than 1 min peak separation.
在本研究中，设计并开发了超临界流体萃取（SFE）的最佳装置，用于定量从药用大麻（品种）花中提取的11种不同的大麻素（大麻二酚戊酸酯（CBDV）、四氢大麻酚戊酸酯（THCV）、大麻二酚（CBD）、大麻萜酚（CBG）、大麻二酚酸（CBDA）、大麻萜酚酸（CBGA）、大麻酚（CBN）、Δ9-四氢大麻酚（Δ-THC）、Δ8-四氢大麻酚（Δ-THC）、大麻环萜酚（CBC）和Δ9-四氢大麻酚酸（THCA-A））。超临界二氧化碳（scCO₂）萃取在37℃、250巴的压力下进行，此时CO₂的最大理论密度为893.7kg/m³，该条件从花提取物中产生了最高产量的大麻素。此外，使用了一个冷分离器（分离室），并将其紧邻含样品室放置以最大化产量。还发现用新鲜的scCO₂连续洗涤提取物可进一步提高产量。采用超高效液相色谱与二极管阵列检测器联用（uHPLC-DAD）开发了一种定量11种大麻素的方法。使用C18固定相结合双溶剂系统梯度程序，在32分钟的时间跨度内获得了分离良好的色谱图。跨日内和日间分离的大麻素的准确度和精密度在可接受范围内（<±15%）。该分析方法也经过了全面验证，从线性、定量限和检测限角度来看具有灵敏度。这项工作的研究结果有望为使用scCO₂优化提取特定大麻素提供更好的条件，这在开发特性明确的药用大麻制剂方面具有前景。据我们所知，这是第一项报道在单次运行中对scCO₂提取物中的11种大麻素进行uHPLC定量分析方法的研究，峰分离时间超过1分钟。
