Method for the isolation of diterpene pinusolide in Thuja occidentalis Linn (Cupressaceae)

摘要

Isolation of the diterpene pinusolide in Thuja occidentalis Linn. (Cupressaceae). This patent relates to the description of the methodological steps necessary for the isolation and identification of the diterpene pinusolide, belonging to the class of labdanes.The pinusolide, according to several reports in the literature, is assigned as the main property the potent and specific receptor antagonist ligand action of platelet activating factor, in addition to the anti-inflammatory activity and anti leukaemic.For the purpose of isolation, the drug plant was extracted for two days with dichloromethane and the plant material obtained was again re extracted to exhaustion of drug plant. After the extract was filtered and exposed to dryness.Then, through the analysis on the chromatographic column wet filter with various systems of organic solvents was the fraction F3, eluted with hexane / ethyl acetate (50: 50). The fraction obtained was taken to dryness and placed in isolation for fractionating column.The chromatographic profile of fraction F3 was visualized by high performance liquid chromatography (HPLC), and this fraction previously treated by solid phase extraction (SPE). However, to optimize the method of isolation of the substance of interest, it is proposed to the system isocru00e1tico with H ~ 2 ~ and ACN, with separation factor k = 20 containing the system H ~ 2 ~ / ACN (38, 62).Despite the good resolution of F3, the organic solvent is further weakened and, using the nomu00f3grafo reverse phase to replace the ACN by MeOH, keeping constant the strength of the mobile phase, it is the system H ~ 2 ~: MeOH (25: 75), which was further weakened when - if the H ~ 2 ~ / MeOH (30: 70), which provided sufficient conditions for isolation of the isolated substance (S1).The substance S1 has been collected in HPLC and identified by means of various techniques of nuclear magnetic resonance (NMR).