Size exclusion chromatographic analysis of polyphenol-serum albumin complexes

Tsutomu Hatano, Mami Hori, Richard W. Hemingway, Takashi Yoshida

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Formation of water-soluble polyphenol-protein complexes was investigated by size-exclusion chromatography (SEC). The combination of (-)-epigallocatechin gallate (EGCG) and bovine serum albumin (BSA), which did not form a precipitate after the solutions were mixed, showed an SEC peak due to complex formation 2-24 h after mixing. Peak size of the complex varied with time, suggesting slow change of the conformation of the protein accompanied by complexation. Formation of the complex was substantiated by ultrafiltration of the mixture; the complex did not pass through a membrane with a 100,000 nominal molecular weight limit (NMWL). The SEC profile varied with the combination of compounds. The peaks due to the complexes showed that the apparent value of the number average molecular weight (Mn) of the EGCG-BSA complex was 2.8 × 105, while that of a pentagalloylglucose (PGG)-BSA complex was 9.5 × 105 under the conditions used. Dimeric hydrolyzable tannins, oenothein B and cornusiin A, also caused changes in the SEC profile of BSA, although the combinations did not show peaks attributable to formation of such large complexes observed for EGCG and PGG. Procyanidin B3 and (+)-catechin did not cause changes in the SEC profile of BSA. With cytochrome c, EGCG did not show any chromatographic changes.

Original languageEnglish
Pages (from-to)817-823
Number of pages7
JournalPhytochemistry
Volume63
Issue number7
DOIs
Publication statusPublished - Aug 2003

Keywords

  • Camellia sinensis
  • Complexation
  • Cytochrome c
  • Epigallocatechin gallate
  • Hydrolysable tannin
  • Pentagalloylglucose
  • Polyphenol
  • Proanthocyanidin
  • Serum albumin
  • Tea
  • Theaceae

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Plant Science
  • Horticulture

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