# New Methods for the Comprehensive Analysis of Bioactive Compounds in _Camabis sativa_ L. (hemp)
## 4 Conclusions  
A comprehensive analysis of the bioactive compounds in hemp infloresences was carried out in this research work for the first time. The chemical classes considered in this study encompassed non-psychoactive cannabinoids, flavonoids and related compounds, and volatile compounds.  
In detail, the qualitative and quantitative profiling of non-psychoactive cannabinoids was performed by applying an HPLC method with UV/DAD, ESI-MS and MS\({}^{2}\) detection. A new HPLC method with UV/DAD, ESI-MS and MS\({}^{2}\) detection was developed and validated for the analysis of hemp-specific flavonoids, combined with a selective and simple extraction procedure. Finally, the characterisation of the hemp volatile fraction was achieved by optimising a new HS-SPME-GC procedure with FID and MS detection.  
All the analytical methods developed in the present work clearly demonstrated the provision of a comprehensive and reliable analysis of the bioactive compounds present in hemp infloresences, which are known to be characterised by a very complex composition. Therefore, they can be applied for a detailed chemical characterisation of the plant material in order to monitor its composition and guarantee a better reproducibility of biological assays, since several compounds may act synergistically, and also in quality control to ensure the efficacy and safety of hemp-based products to be used in the pharmaceutical and nutraceutical fields.  
**Supplementary Materials:** The Supplementary Materials are available online. Table S1: linearity and sensitivity data for compounds used as hemp standards, Table S2: intra- and inter-day precision data for retention time (\(t_{\text{R}}\)) and peak area of the main flavonoids in hemp extracts (sample C6), Table S3: intra- and inter-day precision data for the extraction of the main flavonoids from hemp (sample C6).  
**Author Contributions:** Conceptualization, F.P.; Methodology, F.P. and V.B.; Software, V.B, L.M. and D.B.; Validation, F.P., V.B. and J.S.; Formal Analysis, F.P. and V.B.; Investigation, F.B., V.B. and J.S.; Resources, S.B.; Data Curation, F.P. and V.B.; Writing--Original Draft Preparation, F.P. and V.B.; Writing--Review and Editing, F.P.; Visualization, F.P.; Supervision, F.P. and S.B.; Funding Acquisition, S.B.  
**Funding:** This work was carried out thanks to: a research grant awarded to [PERSON] for her master thesis at the Department of Life Sciences of the University of Modena and Reggio Emilia (Italy) within the Erasmus+ program, under the scientific supervision of [PERSON]; a research grant (Fondo di Ateneo per la Ricerca, FAR 2017) awarded to [PERSON] from the Department of Life Sciences of the University of Modena and Reggio Emilia for the project entitled \"Active compounds from _Cannabis sativa_ L. for the prevention and complementary therapy of neurodegenerative diseases\".  
**Acknowledgments:** The authors are very grateful to [PERSON] of the Department of Drug Sciences of the University of Piemonte Orientale (Novara, Italy) for having kindly provided canniprene standard. [PERSON] from CREA-CIN centre (Rovigo, Italy) is also acknowledged for having provided fibre-type hemp samples.  
**Conflicts of Interest:** The authors declare no conflict of interest.