CHempion - High revenue-generating molecules from industrial hemp: cost-effective, chemical classification-driven solutions for their production

The overriding goal of the present research is the valorization of major and minor molecules present in industrial hemp inflorescence, leaves, and roots. Cannabis (Cannabis sativa L.) is one of the most versatile plant species historically used as a source of food, forage, fiber, oil, and narcotics. The interest in this herbaceous plant has recently seen a resurgence because of its multipurpose applications in the pharmaceutical, cosmetic, and nutraceutical industries, with appliances ever-expanding in the latest years. The global market of industrial hemp, i.e., the varieties of C. sativa whose tetrahydrocannabinol (THC) content is lower than 0.2%, is expected to annually grow by 34% from 4,6 billion dollars in 2019 to 26,6 billion in 2025. However, the range of bioactive compounds present in industrial hemp is still scarcely investigated and valorized: even now, cannabidiol (CBD) extraction represents the main industrial application.

To expand the profitability of hemp agribusiness, this project will develop a wide array of analytical tools for the characterization, isolation, and purification of major and minor bioactive components of industrial hemp inflorescence, together with the by-products arising from industrial CBD extraction and hemp leaves, and roots. Besides phytocannabinoids also phenolic compounds, terpenes and terpenoids, cannflavins, cannabisins, triterpenes, triterpenoids and alkaloids are largely present in industrial hemp flowers and their by-products and represent valuable supplies for industrial applications (nutraceutical, pharmaceutical, and essential oil industry). To this purpose, the influence of different industrial hemp genotypes characterized by various sexual reproduction,

flowering time, and chemotype will be investigated using high-throughput analytical platforms based on hyphenated chromatographic techniques coupled to MS analyzers. The large raw dataset will be  processed by dedicated bio-informatics software programs and multivariate chemometric tools to build validated classification models of hemp accessions. At a later stage, specific attention will be paid to the development of cutting-edge, cost-effective analytical solutions for the purification of selected bioactive compounds from the extracts. These strategies will be based on an innovative multi-column, highly automated technology, in which separation is accomplished under continuous countercurrent conditions to maximize purification yield and recovery. Purification of chiral compounds with chiral columns to obtain separated bioactive enantiomers will be verified. The purified fractions will be tested with novel biological assays on cell lines, ensuring more reliable results than traditional bioassays, and results will be interpreted by multivariate regression methods.