Bidimensional heart-cut achiral-chiral liquid chromatography coupled to high-resolution mass spectrometry for the separation of the main chiral phytocannabinoids and enantiomerization studies of cannabichromene and cannabichromenic acid.

In this work, a heart-cut bidimensional achiral-chiral liquid chromatography method coupled to high-resolution mass spectrometry was developed for the separation of the main carboxylated phytocannabinoids, namely cannabidiolic acid (CBDA), tetrahydrocannabinolic acid (THCA), cannabichromenic acid (CBCA), and cannabicyclolic acid (CBLA), and decarboxylated derivatives, namely cannabidiol (CBD), Δ9-tetrahydrocannabinol (Δ9-THC), cannabichromene (CBC), and cannabicyclol (CBL), and the evaluation of their enantiomeric composition in extracts of different Cannabis sativa L. varieties. Optimal conditions for the chiral analysis of CBC- and CBL-type compounds were found with methanol and water (95:5, v/v , with 0.1% formic acid, 1.5 mL/min) on an amylose-based chiral stationary phase. These settings also allowed to evaluate the parameters responsible for CBC and CBCA racemization. [Display omitted] • Phytocannabinoids were analyzed by a heart-cut 2D achiral-chiral RP-HPLC-HRMS method. • CBCA enantiomers were separated for the first time in different cannabis varieties. • Enantiomerization studies were performed on isolated enantiomers of CBCA and CBC. • Temperature and UV light were investigated as responsible for erosion of enantiopurity. [ABSTRACT FROM AUTHOR]