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DOI: 10.1055/s-0036-1596294
Purification of bioactive compounds from Centaurium erythraea by conventional and new generation designed Centrifugal Partition Chromatography column
Publication History
Publication Date:
14 December 2016 (online)
Centaurium erythraea Rafn. (Gentianaceae) is a widespread herbaceous plant used in a traditional medicine in more than twenty countries. Hepatoprotective, antibacterial or hypoglycemic activities are reported in the literature. Seco-iridoid glycosides, xanthones, phenolic derivatives and triterpenes were identified as major compounds. In order to recover efficiently the polar constituents, the raw material was macerated in MeOH, EtOH, or EtOH/H2O 1/1, or infused in boiling H2O. UHPLC-MS-UV profiles of each extract enabled to establish the yield in swertiamarin (ST) and sweroside (SR), two structurally closely-related target natural products.
Centrifugal Partition Chromatography (CPC), also called hydrostatic countercurrent chromatography (HSCCC), a support-free liquid-liquid chromatography based on the partitioning of analytes between two non-miscible liquid phases, was used to isolate ST and SR from the extract. Experimental conditions were optimized on isocratic and gradient mode using a mixture EtOAc/EtOH/H2O on a conventional CPC 250 mL column. These data were then transposed to a new generation 250 mL column. These latter allowed us to perform a five times faster separation compared with the conventional 250 mL column. By gradient run mode, we obtained enriched fraction of each mainly presented seco-iridoids whereof swertiamarin major. The structures of the recovered compounds were unambiguously ascertained by UPLC-ESI-MS/MS and extensive NMR analyses.
This study has proven the ability of new generation CPC column to purify higher amount of a polar extract in a timeless duration run. The choice of an environment-friendly solvent mixture including the recycling of EtOAc was also noticeable and may contribute to upgrade CPC as a potential industrial production tool.
Keywords: Seco-iridoids, Centaurium erythraea, centrifugal partition chromatography, counter current chromatography.
References:
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