Wongumpornpinit Vorawut, Temkitthawon Prapapan, Khumpirapang Nattakanwadee, Paenkaew Sujittra, Saesong Tongchai, Boonnoun Panatpong, Wongwad Eakkaluk, Waranuch Neti, Ingkaninan Kornkanok
Center of Excellence for Natural Health Product Innovation, Faculty of Pharmaceutical Sciences, Naresuan University, Phitsanulok, Thailand.
Department of Pharmaceutical Chemistry and Pharmacognosy, Faculty of Pharmaceutical Sciences and Center of Excellence for Innovation in Chemistry, Naresuan University, Phitsanulok, Thailand.
Med Cannabis Cannabinoids. 2025 May 14;8(1):65-79. doi: 10.1159/000546263. eCollection 2025 Jan-Dec.
Cannabinoids, a class of compounds found in L., possess a wide range of pharmacological properties. While Δ-tetrahydrocannabinol (Δ-THC) is strictly regulated owing to its psychoactive effects, cannabidiol (CBD), a nonpsychoactive compound, is permitted in certain countries. This study aimed to optimize the preparation of ethanolic cannabis extracts using response surface methodology (RSM) and develop an effective system for removing Δ-THC through centrifugal partition chromatography (CPC) to produce broad-spectrum CBD (hemp extract containing CBD and other compounds with minimal or no Δ-THC).
Three variables and six responses were assessed to optimize extraction conditions. Predictions were made using Design-Expert software, and the experimental conditions were identified using the Box-Behnken design (BBD). The extracts were analyzed using high-performance liquid chromatography and a chromameter. Optimal conditions were used for pilot-scale extraction, and the CPC process was optimized by determining the partition coefficient of the target cannabinoids in various solvent systems and maximum sample load.
The optimal extraction conditions were -31°C for 33 min and a sample-to-solvent ratio of 1:8% w/v, with a desirability value of 0.576. Temperature was the most influential factor. Although the total yield decreased, this condition provided the highest concentration of light-colored cannabinoids and was successfully scaled up for the three other cannabis samples. The optimal CPC solvent system, consisting of hexane/0.1% FA in ACN/20 mm ammonium formate at a ratio of 10/6.5/3.5 v/v/v, demonstrated a yield recovery of 89.3 ± 0.21% w/w with a maximum load of 5 g of sample per run. The resulting broad-spectrum CBD extract had a high CBD content (73.3 ± 0.37% w/w) and minimal Δ-THC content (0.2 ± 0.00% w/w).
BBD-RSM optimization of ethanolic cannabis extraction provided the highest cannabinoid concentration with a short extraction time and desirable appearance. The CPC process successfully separated Δ-THC, yielding a high-purity broad-spectrum CBD extract.
大麻素是一类存在于大麻中的化合物，具有广泛的药理特性。虽然Δ-四氢大麻酚（Δ-THC）因其精神活性作用受到严格管制，但非精神活性化合物大麻二酚（CBD）在某些国家是允许使用的。本研究旨在使用响应面法（RSM）优化乙醇大麻提取物的制备，并开发一种通过离心分配色谱法（CPC）去除Δ-THC的有效系统，以生产广谱CBD（含有CBD和其他化合物且Δ-THC含量极低或无Δ-THC的大麻提取物）。
评估了三个变量和六个响应以优化提取条件。使用Design-Expert软件进行预测，并使用Box-Behnken设计（BBD）确定实验条件。使用高效液相色谱法和色差仪对提取物进行分析。将最佳条件用于中试规模提取，并通过确定目标大麻素在各种溶剂系统中的分配系数和最大样品负载量来优化CPC过程。
最佳提取条件为-31°C下33分钟，样品与溶剂比例为1:8% w/v，可取性值为0.576。温度是最具影响力的因素。虽然总产率下降，但该条件提供了最高浓度的浅色大麻素，并成功扩大到其他三种大麻样品。最佳CPC溶剂系统由己烷/0.1% 甲酸乙腈溶液/20 mM甲酸铵以10/6.5/3.5 v/v/v的比例组成，产率回收率为89.3 ± 0.21% w/w，每次运行最大样品负载量为5 g。所得的广谱CBD提取物具有高CBD含量（73.3 ± 0.37% w/w）和极低的Δ-THC含量（0.2 ± 0.00% w/w）。
乙醇大麻提取物的BBD-RSM优化提供了最高的大麻素浓度，提取时间短且外观理想。CPC过程成功分离了Δ-THC，得到了高纯度的广谱CBD提取物。
