Rapid reversed phase ultra-performance liquid chromatography analysis of the major cocoa polyphenols and inter-relationships of their concentrations in chocolate

Karen A. Cooper, Esther Campos-Giménez, Diego Jiménez Alvarez, Kornél Nagy, Jennifer L. Donovan, Gary Williamson

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133 Citations (Scopus)


Chocolate and other cocoa-containing products are a rich source of polyphenols. This paper describes an ultra-performance liquid chromatography (UPLC) method that can separate and quantify in 3 min six of the major chocolate polyphenols: catechin; epicatechin; B2 (epicatechin-4β-8-epicatechin); B5 (epicatechin-4β-6-epicatechin); C1 (epicatechin-4β-8-epicatechin- 4β-8-epicatechin); and tetramer D (epicatechin-4β-8-epicatechin- 4β-8-epicatechin-4β-8-epicatechin). A survey of 68 chocolate samples indicated that there was a strongly predictive relationship between epicatechin and the other individual polyphenols, especially procyanidin B2 (R2 = 0.989), even though the chocolates came from varied sources and manufacturers. The relationship was less strong with catechin, and so further work to explore the reasons for this difference was performed. Chiral analysis on a subset of 23 chocolates showed that (-)-epicatechin had a predictive relationship with (+)-catechin in line with the other polyphenols, but not with (-)-catechin (the predominant form). This indicates that (-)-catechin is the most affected by manufacturing conditions, possibly formed through epimerization from (-)-epicatechin during processing. The results show that epicatechin concentrations can be used to predict the content of other polyphenols, especially B2 and C1, and total polyphenols content. Finally, the (-)-catechin content is not predictable from the epicatechin content, and it is concluded that this is the main form of polyphenol that varies according to manufacturing conditions and cocoa origin.

Original languageEnglish
Pages (from-to)2841-2847
Number of pages7
JournalJournal of Agricultural and Food Chemistry
Issue number8
Publication statusPublished - 18 Apr 2007
Externally publishedYes


  • Antioxidant
  • Chocolate
  • Cocoa
  • Epicatechin
  • Polyphenols
  • Procyanidins
  • UPLC

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