1H NMR-based metabolomics differentiation and real time PCR analysis of medicinal Cannabis organs

Cannabis sativa L. (Cannabaceae), is an annual plant that has been cultivated and used as a source of medicine since prehistoric time. In this report, H-1 NMR-based metabolomics along with real time PCR (RT-PCR) analysis was performed to study metabolite profiles and cannabinoids biosynthesis in different organs of Cannabis. Two medicinal cultivars, 'Bedrocan' and 'Bedica', were analyzed. Principal component analysis (PCA) models have been successfully applied to classify metabolite profiles of Cannabis samples. Among those, tetrahydrocannabinolic acid (THCA) was the most important discriminant compound. 13 compounds in the water extracts were identified from the water and chloroform extract contributing in the classification of metabolite profiles of these samples. The absence of asparagine signals in the H-1 NMR spectra indicated this compound as the leading differentiating compound in the water extracts. RT-PCR analysis in the medicinal plants revealed that the highest expression level of THCA synthase (THCAS) was in the leaves of the plants.