Riassunto analitico
Phytoestrogens or estrogenic-like compounds from Cannabis sativa L. constitute an interesting and insufficiently explored field that could be extremely useful for their role in medicinal chemistry. In this context, the aim of this thesis was to identify this biological activity focusing on ten non-psychoactive (fiber-type) C. sativa (hemp) samples through effect-directed analysis (EDA). After extracting the samples, the detection of estrogenic activity was possible using high performance liquid chromatography (NP HPTLC-UV/Vis/FLD) hyphenated with EDA, via microchemical (Vanillin, AlCl3, Fast Blue Salt B) and enzymatic [pYES (planar yeast estrogen) and pYAS (planar yeast androgen) assays] derivatizations, and characterization with different types of mass spectrometric approached, which included DART-MS (direct analysis in real time mass spectrometry), ESI-MS (electrospray ionization mass spectrometry) and HRMS (high resolution mass spectrometry). Since the HPTLC plates were normal phase (NP) mass spectrometry (MS)-grade, trials at reducing the diffusion by using PEG (400, 8000), gelatine, GelRite and degalan were done to optimize the separation of the analytes. Hemp extracts are well-known for their complex chemical composition. So, enzymatic assays had to be done to test both the agonistic and antagonistic activity also using various pure compounds for comparison. This is also the reason why microchemical derivatizations were performed. Canniprene, cannflavins and cannabinoids were used to construct the IC50 curve as they showed a potential estrogenic-like activity by way of the pYES assay. In the end, only CPR showed estrogenic activity when compared to genistein, a natural phytoestrogen from soy.
This project was carried out in Professor Gertrud Morlock laboratory at the Food Science Department of the Justus Liebig University in Giessen (Germany) through the Erasmus+ in strict collaboration with Professor Federica Pellati.
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