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Pilot-Scale Preparation of Broad-Spectrum CBD: Extraction Optimization and Purification using Centrifugal Partition
Vorawut Wongumpornpinit1,2, Prapapan Temkitthawon1,2, Nattakanwadee Khumpirapang2
1Center of Excellence for Natural Health Product Innovation, Faculty of Pharmaceutical Sciences, Naresuan University, Phitsanulok, Thailand.
Introduction:
Cannabinoids, a class of compounds found in Cannabis sativa L., possess a wide range of pharmacological properties. While Δ9-tetrahydrocannabinol (Δ9-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 Δ9-THC through centrifugal partition chromatography (CPC) to produce broad-spectrum CBD (hemp extract containing CBD and other compounds with minimal or no Δ9-THC).
Methods:
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.
Results:
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 Δ9-THC content (0.2 ± 0.00% w/w).
Conclusion:
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 Δ9-THC, yielding a high-purity broad-spectrum CBD extract.
