Profilo fitochimico, DNA fingerprinting e valutazione delle proprietà funzionali di diverse varietà di Perilla frutescens

Perilla frutescens (L.) Britton is an aromatic, annual, short-day herbal plant, belonging to the family Lamiaceae/Labiatae, that is widely cultivated in all Asian countries, especially in the South-east, however, China is probably considered a primary genetic center for this species. The medical history of Perilla begins in antiquity, indeed all parts of the plant find wide uses in traditional Chinese medicine. Being a plant of Asian origin, over the centuries it has become relevant from a socio-cultural and economic point of view as a functional food throughout Asian communities. However, in western countries Perilla is not very well known for these aspects. Precisely because of the historical popularity and importance of this plant in traditional Chinese medicine and its rich use in Asian cuisine, the world of scientific research has also shown interest in Perilla in recent years, using it as an object of study in various fields, among European countries as well, and the interest in bringing this plant here to Europe is growing. The first part of the experimental work consists of a bibliometric analysis to highlight the increasing interest that world scientific research is demonstrating towards Perilla frutescens, using the last 15 years (2008-2022) as a time frame. Bibliometrics is a science that uses mathematical and statistical techniques to analyze information distribution patterns, dealing in particular with the measurement of scientific publications and their impact within the scientific community. The second part of the work consists of a phytochemical analysis of the bioactive metabolites. Phytochemistry is a scientific discipline that has as its object the isolation, study, analysis, purification and characterization of the chemical structure and biological activity of the active principles of plants. The starting samples were dry leaves of eight varieties of Perilla frutescens, and quantification assays for phenolic compounds, High Performance Liquid Chromatography (HPLC) analysis, and Antioxidant activity (AOA) assays were performed. To quantify phenolic compounds, the total phenolic content (TPC) by Folin-Ciocâlteu assay, the total flavonoid content (TFC) by the aluminum chloride method, the total proanthocyanidin content (TPAC) by the DMAC assay, and finally the total anthocyanidins content (TAC) by the pH jump method were performed. To measure the AOA, assays for radical scavenging activity (ABTS and DPPH), and for ferric reducing antioxidant power (FRAP) were performed. Furthermore, gas chromatography (GC) analyzes coupled to flame ionization detector (FID) or mass spectrometry (MS) were conducted to obtain the fatty acid profile of the plants. Finally, high performance liquid chromatography (HPLC) coupled to diode array detector (DAD) or MS/MS techniques were used to identify and quantify the main phytoconstituents present in the extracts of the different Perilla varieties. In the third part of the thesis, a DNA fingerprinting was carried out on the eight samples of Perilla. DNA fingerprinting is a molecular method developed to identify biological entities and based on the analysis of the variability of one or more molecular markers, i.e. standard regions of DNA, which provides rapid and accurate identification of species. The molecular markers chosen for the analysis were: ● ITS, Internal Transcribed Spacer (ITS-p5 - ITS-u4) ● TrnL-F, Intergenic spacer of the chloroplast DNA