RESUMEN
INTRODUCTION: Plant extracts are usually complex mixtures of various polarity compounds and their study often includes a purification step, such as solid-phase extraction (SPE), to isolate interest compounds prior analytical investigations. Molecularly imprinted polymers (MIPs) are a new promising type of SPE material which offer tailor-made selectivity for the extraction of trace active components in complex matrices. Numerous specific cavities that are sterically and chemically complementary of the target molecules, are formed in imprinted polymers. A molecularly imprinted polymer (MIP) was synthesised in order to trap a specific class of triterpene, including betulin and betulinic acid from a methanolic extract of plane bark. METHODOLOGY: Imprinted polymers were synthesised by thermal polymerisation of betulin as template, methacrylic acid (MAA) or acrylamide (AA) as functional monomer, ethylene glycol dimethacrylate as crosslinking agent and chloroform as porogen. Afterwards, MAA- and AA-MIPs were compared with their non-imprinted polymers (NIPs) in order to assess the selectivity vs betulin and its derivatives. Recovered triterpenes were analysed by HPLC during MIP-SPE protocol. RESULTS: After SPE optimisation, the MAA-imprinted polymer exhibited highest selectivity and recovery (better than 70%) for betulin and best affinity for its structural analogues. Thus, a selective washing step (chloroform, acetonitrile) removed unwanted matrix compounds (fatty acids) from the SPE cartridge. The elution solvent was methanol. Finally, the MAA-MIP was applied to fractionate a plane bark methanolic extract containing betulin and betulinic acid. CONCLUSION: This study demonstrated the possibility of direct extraction of betulin and its structural analogues from plant extracts by MIP technology.
Asunto(s)
Betulaceae/química , Impresión Molecular/métodos , Polímeros/química , Extracción en Fase Sólida/métodos , Triterpenos/aislamiento & purificación , Triterpenos Pentacíclicos , Corteza de la Planta/química , Polímeros/síntesis química , Extracción en Fase Sólida/instrumentación , Ácido BetulínicoRESUMEN
The study of the oligosaccharides extracted from Tephrosia purpurea seeds was undertaken using the instant controlled pressure drop (DIC) as a pre-treatment prior to conventional solvent extraction. This DIC procedure provided structural modification in terms of expansion, higher porosity and improvement of specific surface area; diffusion of solvent inside such seeds and availability of oligosaccharides increase notably. In this paper, we investigated and quantified the impact of the different DIC operative parameters on the yields of ciceritol and stachyose extracted from T. purpurea seeds. The treatment could be optimized with a steam pressure (P) (P=0.2 MPa), initial water content (W) (W=30% dry basis (DB)) and thermal treatment time (t) (t=30s). By applying DIC treatment in these conditions, the classic process of extraction was intensified in both aspects of yields (145% of ciceritol and 185% of stachyose), and kinetics (1h of extraction time instead of 4h for conventional process). The scanning electron microscopy micrographs provided evident modifications of structure of seeds due to the DIC treatment.
Asunto(s)
Oligosacáridos/aislamiento & purificación , Semillas/química , Tephrosia/química , Microscopía Electrónica de Rastreo , Extractos Vegetales/química , Presión , Semillas/ultraestructuraRESUMEN
Reverse pharmacognosy aims at finding biological targets for natural compounds by virtual or real screening and identifying natural resources that contain the active molecules. We report herein a study focused on the identification of biological properties of meranzin, a major component isolated from Limnocitrus littoralis (Miq.) Swingle. Selnergy, an IN SILICO biological profiling software, was used to identify putative binding targets of meranzin. Among the 400 screened proteins, 3 targets were selected: COX1, COX2 and PPARgamma. Binding tests were realised for these 3 protein candidates, as well as two negative controls. The predictions made by Selnergy were consistent with the experimental results, meaning that these 3 targets can be modulated by an extract containing this compound in a suitable concentration. These results demonstrate that reverse pharmacognosy and its inverse docking component is a powerful tool to identify biological properties for natural molecules and hence for plants containing these compounds.