Influence of alternative liquid chromatography techniques on the chemical complexity and bioactivity of isolated proanthocyanidin mixtures

Jessica L. Alwerdt, David S. Seigler, Elvira Gonzalez De Mejia, Gad G. Yousef, Mary Ann Lila

Research output: Contribution to journalArticlepeer-review

Abstract

Extracts rich in proanthocyanidins, which are implicated in multiple human health benefits, were comparatively separated using alternative separation methods [vacuum or open column liquid chromatography], separation matrices [Toyopearl, Sephadex, or silica gel], and degrees of subfractionation [8 or 12 subfraction series], to evaluate the influence of separation technique on the resolution of the chemical composition and the biological activity of separated proanthocyanidin mixtures in individual subfractions. Bioactivity was assessed using a DNA human topoisomerase II bioassay and structural composition by acid thiolysis (average degree of polymerization, DP) and HPLC-ESI/MS. The amount of parent fraction needed to inhibit 50% of topoisomerase II was 3.38 ng/mL with an average DP of 25.5. A 23 factorial analysis revealed that the vacuum and open column strategies for separation, when individually considered, did not yield significantly different results in terms of mass recovery, DP, or bioactivity; however, interactions with other factors such as matrix or subfraction series resulted in distinctive shifts in fraction profiles and biological activity. In general, Sephadex as a matrix permitted elution and separation of discrete, polymerized subfractions with potent inhibition against human topoisomerase II. Sephadex vacuum chromatography, Toyopearl open column chromatography, and Toyopearl vacuum chromatography separation techniques eluted highly polymerized proanthocyanidin mixtures, but the inhibitory bioactivity was attenuated as compared to the parent fraction, whereas Sephadex open column chromatography eluted highly polymerized subfraction mixtures that retained bioactive potential.

Original languageEnglish (US)
Pages (from-to)1896-1906
Number of pages11
JournalJournal of Agricultural and Food Chemistry
Volume56
Issue number6
DOIs
StatePublished - Mar 26 2008

Keywords

  • Proanthocyanidins
  • Sephadex LH-20
  • Silica gel type G
  • Topoisomerase II bioassay
  • Toyopearl HW-40
  • Vaccinium angustifolium Ait.
  • Wild blueberry

ASJC Scopus subject areas

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

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