4.7 Article

Effective isolation of cannabidiol and cannabidiolic acid free of psychotropic phytocannabinoids from hemp extract by fast centrifugal partition chromatography

Journal

ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 415, Issue 19, Pages 4827-4837

Publisher

SPRINGER HEIDELBERG
DOI: 10.1007/s00216-023-04782-9

Keywords

Hemp extract; Single step fractionation; Phytocannabinoids; Fast centrifugal partition chromatography; Pure cannabidiol; cannabidiolic acid; Removing ????????(9)-tetrahydrocannabinol; ?(9)-tetrahydrocannabinolic acid

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This study demonstrates the applicability of fast centrifugal partition chromatography (FCPC) as an effective method for the isolation of CBD and CBDA free of psychotropic compounds from Cannabis sativa L. plant extracts. The purity of isolated CBD and CBDA reached 98.9% and 95.1% (w/w), respectively, without the presence of non-psychotropic compounds, with only trace amounts of other biologically active compounds detected.
Cannabidiol (CBD), together with its precursor cannabidiolic acid (CBDA), is the major phytocannabinoid occurring in most hemp cultivars. To ensure the safe use of these compounds, their effective isolation from hemp extract is required, with special emphasis on the elimination of ?(9)-tetrahydrocannabinol (?(9)-THC) and ?(9)-tetrahydrocannabinolic acid (?(9)-THCA-A). In this study, we demonstrate the applicability of fast centrifugal partition chromatography (FCPC) as a challenging format of counter-current preparative chromatography for the isolation of CBD and CBDA free of psychotropic compounds that may occur in Cannabis sativa L. plant extracts. Thirty-eight solvent mixtures were tested to identify a suitable two-phase system for this purpose. Based on the measured partition coefficients (K-D) and separation factors (a), the two-phase system consisting of n-heptane:ethyl acetate:ethanol:water (1.5:0.5:1.5:0.5; v:v:v:v) was selected as an optimal solvent mixture. Employing UHPLC-HRMS/MS for target analysis of collected fractions, the elution profiles of 17 most common phytocannabinoids were determined. Under experimental conditions, the purity of isolated CBD and CBDA was 98.9 and 95.1% (w/w), respectively. Neither of ?(9)-THC nor of ?(9)-THCA-A were present; only trace amounts of other biologically active compounds contained in hemp extract were detected by screening against in-house spectral library using UHPLC-HRMS.

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