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Centrifugal partition chromatography enables selective enrichment of trimeric and tetrameric proanthocyanidins for biomaterial development
Journal article   Open access   Peer reviewed

Centrifugal partition chromatography enables selective enrichment of trimeric and tetrameric proanthocyanidins for biomaterial development

Rasika S. Phansalkar, Joo-Won Nam, Shao-Nong Chen, James B. McAlpine, Ariene A. Leme, Berdan Aydin, Ana-Karina Bedran-Russo and Guido F. Pauli
Journal of Chromatography A, Vol.1535, pp.55-62
02/02/2018
DOI: 10.1016/j.chroma.2017.12.050
PMCID: PMC5801063
PMID: 29331224
url
https://doi.org/10.1016/j.chroma.2017.12.050View
Published (Version of record) Open Access

Abstract

•Development of proanthocyanidins (PACs) as restorative dental biomaterials.•Separation of PAC trimers/tetramers by centrifugal partition chromatography (CPC).•Advantage of methyl acetate replacing methanol in HEMWat solvent systems for PACs.•Evaluation of selectively enriched PAC oligomers in dentin biomechanical assay.•Demonstration of DESIGNER extract concept for PAC-rich plant extracts. Proanthocyanidins (PACs) find wide applications for human use including food, cosmetics, dietary supplements, and pharmaceuticals. The chemical complexity associated with PACs has triggered the development of various chromatographic techniques, with countercurrent separation (CCS) gaining in popularity. This study applied the recently developed DESIGNER (Depletion and Enrichment of Select Ingredients Generating Normalized Extract Resources) approach for the selective enrichment of trimeric and tetrameric PACs using centrifugal partition chromatography (CPC). This CPC method aims at developing PAC based biomaterials, particularly for their application in restoring and repairing dental hard tissue. A general separation scheme beginning with the depletion of polymeric PACs, followed by the removal of monomeric flavan-3-ols and a final enrichment step produced PAC trimer and tetramer enriched fractions. A successful application of this separation scheme is demonstrated for four polyphenol rich plant sources: grape seeds, pine bark, cinnamon bark, and cocoa seeds. Minor modifications to the generic DESIGNER CCS method were sufficient to accommodate the varying chemical complexities of the individual source materials. The step-wise enrichment of PAC trimers and tetramers was monitored using normal phase TLC and Diol-HPLC-UV analyses. CPC proved to be a reliable tool for the selective enrichment of medium size oligomeric PACs (OPACs). This method plays a key role in the development of dental biomaterials considering its reliability and reproducibility, as well as its scale-up capabilities for possible larger-scale manufacturing.
Centrifugal partition chromatography Degree of polymerization Dental biomaterials Diol-HPLC Enrichment Oligomeric proanthocyanidins

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