, Volume 76, Issue 17–18, pp 1107–1115 | Cite as

Preparative Purification and Isolation of Pyranoanthocyanins from Red Wine and Evaluation of Their Antioxidant Activity

  • Petr Marhol
  • Petr BednářEmail author
  • Eva Tománková
  • Barbora Papoušková
  • Jan Stávek
  • Pavel Híc
  • Petr Barták
  • Vladimír Křen
  • Josef Balík


Untreated silica gel was used as the stationary phase for a simple and cost-effective prepurification of 5-carboxypyranoanthocyanins from red wine enriched by the addition of pyruvic acid. The composition of the fractions obtained was analyzed by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC/MS2) and the results enabled the changes in 5-carboxypyranoanthocyanin profile during preparative separation to be described and explained. Repeated purification on silica gel followed by reversed-phase separation enabled the isolation of 5-carboxypyranomalvidin-3-glucoside in high purity and represents a cheap and robust alternative to the commonly used procedures. Reversed-phase preparative chromatography with an isocratic elution [15 % acetonitrile in water (v/v)] provided sufficient resolution, showing that the addition of an acid is not necessary in anthocyanin separation. The antioxidant capacity of 5-carboxypyranomalvidin-3-glucoside was compared with malvidin-3-glucoside and a group of common polyphenols (e.g., gallic acid, quercetin, rutin, and catechin).


Flash chromatography Silica gel Pyranoanthocyanins Wine Antioxidant capacity 



The financial support by projects no. P206/10/0625 and P301/11/0662 from the Czech Science Foundation, project no. CZ.1.05/2.1.00/03.0058 of the Ministry of Education, Youth and Sports of the Czech Republic, Operational Program Research Development for Innovations—European Regional Development Fund, and grant from Palacky University no. PrF_2013_020 is gratefully acknowledged.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Petr Marhol
    • 1
  • Petr Bednář
    • 2
    Email author
  • Eva Tománková
    • 3
  • Barbora Papoušková
    • 2
  • Jan Stávek
    • 3
  • Pavel Híc
    • 3
  • Petr Barták
    • 2
  • Vladimír Křen
    • 1
  • Josef Balík
    • 3
  1. 1.Laboratory of Biotransformation, Institute of MicrobiologyAcademy of Sciences of the Czech RepublicPrague 4Czech Republic
  2. 2.Department of Analytical Chemistry, Faculty of Science, Regional Centre of Advanced Technologies and MaterialsPalacký UniversityOlomoucCzech Republic
  3. 3.Department of Post-Harvest Technology of Horticultural Products, Faculty of Horticulture in LedniceMendel University in Brno, Czech RepublicLedniceCzech Republic

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