Phytochemical study of Stachys sylvatica L. (Lamiaceae)

The subject of this thesis is the phytochemical study of the aerial parts of the Stachys sylvatica L., perennial herbaceous stolonifera, belonging to the family of the Lamiaceae and mostly present in the Mediterranean region and south-west Asiatic regions. It is generally used in popular medicine for genital cancer treatments, spleen diseases, inflammatory processes, cough and ulcers. The aerial parts were picked in the vicinity of the Pisan mountainside, dried, grinded, underwent extraction by maceration, at room temperature, with polarity increasing solvents: n-hexane, chloroform, chloroform-methanol (9:1) and methanol. The extracts that were considered for the analysis are the chloroform extract (RC), chloroform-methanol extract (RCM) and methanol extract (RM) The methanol extract (RM) was subjected to repartition between n-BuOH/H20. The n-butanol extract was separated with molecular exclusion chromatography Sephadex® LH-20, resulting in 18 main fractions (1-18). Based on the TLC (Thin Layer Chromatography) chromatography profile the fractions 5th, 7th, 8th, 12th, 14th, and 16th considered were then subsequently subjected to RP-HPCPC (High Performance Centrifugal Partition Chromatography). The Chloroform-methanol extract (RCM) was separated through molecular exclusion chromatography Sephadex® LH-20, resulting in 16 main fractions (1-16). The 7th, 9th and 13th fractions were subjected to RP-HPCPC (High Performance Centrifugal Partition Chromatography), the 6th extraction was subjected to silica gel column chromatography using the flash purification Biotage®/Isolera™ Spektra. The chloroform extract, lastly, was subjected to silica gel column chromatography using the flash purification Biotage®/Isolera™ Spektra, resulting in 16 main fractions (1-16). Subsequently, the considered fractions obtained from this extract were separated with reverse phasal high pressure chromatography RP-HPCPC (High Performance Centrifugal Partition Chromatography). In conclusion, the phytochemical study on the aerial parts of the Stachys sylvatica L. enabled the isolation of a total of 16 compounds members of different secondary metabolites classes. The structural determination of the isolated compounds was possible with the use of mono and bidimensional NMR spectoscopic techniques (1H-NMR, 13C-NMR, HSQC, HMBC, 1D-TOCSY, DQF-COSY) and mass spectoscopic ESI-MS tests