Skip to main content
SpringerLink
Log in

Characterization of oxidized tannins: comparison of depolymerization methods, asymmetric flow field-flow fractionation and small-angle X-ray scattering

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

Condensed tannins are a major class of plant polyphenols. They play an important part in the colour and taste of foods and beverages. Due to their chemical reactivity, tannins are not stable once extracted from plants. A number of chemical reactions can take place, leading to structural changes of the native structures to give so-called derived tannins and pigments. This paper compares results obtained on native and oxidized tannins with different techniques: depolymerization followed by high-performance liquid chromatography analysis, small-angle X-ray scattering (SAXS) and asymmetric flow field-flow fractionation (AF4). Upon oxidation, new macromolecules were formed. Thioglycolysis experiments showed no evidence of molecular weight increase, but thioglycolysis yields drastically decreased. When oxidation was performed at high concentration (e.g., 10 g L−1), the weight average degree of polymerization determined from SAXS increased, whereas it remained stable when oxidation was done at low concentration (0.1 g L−1), indicating that the reaction was intramolecular, yet the conformations were different. Differences in terms of solubility were observed; ethanol being a better solvent than water. We also separated soluble and non-water-soluble species of a much oxidized fraction. Thioglycolysis showed no big differences between the two fractions, whereas SAXS and AF4 showed that insoluble macromolecules have a weight average molecular weight ten times higher than the soluble ones.

Comparison of UV and MALLS fractograms obtained with water-soluble and -insoluble fractions of oxidized tannins in EtOH. Water-insoluble fractions exhibit a broader size distribution and has a weight average molecular weight about ten times larger than the -soluble one

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Cheynier V, Duenas-Paton M, Salas E, Maury C, Souquet JM, Sarni-Manchado P, Fulcrand H (2006) Structure and properties of wine pigments and tannins. Am J Enol Vitic 57:298–305

    CAS  Google Scholar 

  2. Fulcrand H, Duenas M, Salas E, Cheynier V (2006) Phenolic reactions during winemaking and aging. Am J Enol Vitic 57:289–297

    CAS  Google Scholar 

  3. Singleton VL, Rossi JA Jr (1965) Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. Am J Enol Vitic 16:144–158

    CAS  Google Scholar 

  4. Swain T, Hillis WE (1959) The phenolic constituents of Prunus domestica. I.—The quantitative analysis of phenolic constituents. J Sci Food Agric 10:63–68

    Article  CAS  Google Scholar 

  5. McMurrough I, McDowell J (1978) Chromatographic separation and automated analysis of flavanols. Anal Biochem 91:92–100

    Article  CAS  Google Scholar 

  6. Sun B, RicardodaSilva JM, Spranger I (1998) Critical factors of vanillin assay for catechins and proanthocyanidins. J Agr Food Chem 46:4267–4274

    Article  CAS  Google Scholar 

  7. Cooper KA, Campos-Gimenez E, JimenezAlvarez D, Nagy K, Donovan JL, Williamson G (2007) Rapid reversed phase ultra-performance liquid chromatography analysis of the major cocoa polyphenols and inter-relationships of their concentrations in chocolate. J Agr Food Chem 55:2841–2847

    Article  CAS  Google Scholar 

  8. Kelm MA, Johnson JC, Robbins RJ, Hammerstone JF, Schmitz HH (2006) High-performance liquid chromatography separation and purification of cacao (Theobroma cacao L.) procyanidins according to degree of polymerization using a diol stationary phase. J Agr Food Chem 54:1571–1576

    Article  CAS  Google Scholar 

  9. Taylor AW, Barofsky E, Kennedy JA, Deinzer ML (2003) Hop (Humulus lupulus L.) proanthocyanidins characterized by mass spectrometry, acid catalysis, and gel permeation chromatography. J Agr Food Chem 51:4101–4110

    Article  CAS  Google Scholar 

  10. Yanagida A, Kanda T, Shoji T, Ohnishi-Kameyama M, Nagata T (1999) Fractionation of apple procyanidins by size-exclusion chromatography. J Chromatogr A 855:181–190

    Article  CAS  Google Scholar 

  11. Kennedy JA, Taylor AW (2003) Analysis of proanthocyanidins by high-performance gel permeation chromatography. J Chromatogr A 995:99–107

    Article  CAS  Google Scholar 

  12. Matthews S, Mila I, Scalbert A, Pollet B, Lapierre C, duPenhoat C, Rolando C, Donnelly DMX (1997) Method for estimation of proanthocyanidins based on their acid depolymerization in the presence of nucleophiles. J Agr Food Chem 45:1195–1201

    Article  CAS  Google Scholar 

  13. Rigaud J, Perezilzarbe J, Ricardo-Da-Silva JM, Cheynier V (1991) Micro method for the identification of proanthocyanidin using thiolysis monitored by high-performance liquid-chromatography. J Chromatogr 540:401–405

    Article  CAS  Google Scholar 

  14. Souquet JM, Cheynier V, Brossaud F, Moutounet M (1996) Polymeric proanthocyanidins from grape skins. Phytochemistry 43:509–512

    Article  CAS  Google Scholar 

  15. Guyot S, Vercauteren J, Cheynier V (1996) Colourless and yellow dimers resulting from (+)-catechin oxidative coupling catalysed by grape polyphenoloxidase. Phytochemistry 42:1279–1288

    Article  CAS  Google Scholar 

  16. Kusano R, Tanaka T, Matsuo Y, Kouno I (2007) Structures of epicatechin gallate trimer and tetramer produced by enzymatic oxidation. Chem & Pharml Bull 55:1768–1772

    Article  CAS  Google Scholar 

  17. Tanaka T, Matsuo Y, Kouno I (2005) A novel black tea pigment and two new oxidation products of epigallocatechin-3-O-gallate. J Agr Food Chem 53:7571–7578

    Article  CAS  Google Scholar 

  18. Schimpf ME, Caldwell K, Giddings JC (2000) Field flow fractionation handbook. Wiley-Interscience, New York

    Google Scholar 

  19. Purawatt S, Siripinyanond A, Shiowatana J (2007) Flow field-flow fractionation-inductively coupled optical emission spectrometric investigation of the size-based distribution of iron complexed to phytic and tannic acids in a food suspension: implications for iron availability. Anal Bioanal Chem 389:733–742

    Article  CAS  Google Scholar 

  20. Pedersen JS (1997) Analysis of small-angle scattering data from colloids and polymer solutions: modeling and least-squares fitting. Adv Colloid Interface Sci 70:171–210

    Article  CAS  Google Scholar 

  21. Poncet-Legrand C, Cabane B, Bautista-Ortin AB, Carrillo S, Fulcrand H, Perez J, Vernhet A (2010) Tannin oxidation: intra-versus intermolecular reactions. Biomacromolecules 11:2376–2386

    Article  CAS  Google Scholar 

  22. Poncet-Legrand C, Cartalade D, Putaux J-L, Cheynier V, Vernhet A (2003) Flavan-3-ol aggregation in model ethanolic solutions: incidence of polyphenol structure, concentration, ethanol content and ionic strength. Langmuir 19:10563–10572

    Article  CAS  Google Scholar 

  23. Burchard W (1974) Statistics of star-shaped molecules. I. Stars with polydisperse side chains. Macromolecules 7:835–841

    Article  CAS  Google Scholar 

  24. Debye P (1947) Molecular weight determination by light scattering. J Phys Colloid Chem 51:18–32

    Article  CAS  Google Scholar 

  25. Fisher ME, Burford RJ (1967) Theory of critical-point scattering and correlations. I. The Ising model. Phys Rev 156:583

    Article  CAS  Google Scholar 

  26. Beaucage G (1996) Small-angle scattering from polymeric mass fractals of arbitrary mass-fractal dimension. J Appl Crystallogr 29:134–146

    Article  CAS  Google Scholar 

  27. Takeshita M, Ishida Y, Akamatsu E, Ohmori Y, Sudoh M, Uto H, Tsubouchi H, Kataoka H (2009) Proanthocyanidin from blueberry leaves suppresses expression of subgenomic hepatitis C virus RNA. J Biol Chem 284:21165–21176

    Article  CAS  Google Scholar 

  28. de Gennes PG (1979) Scaling concepts in polymer physics. Cornell University Press, Ithaca, New York

    Google Scholar 

Download references

Acknowledgements

We thank Javier Pérez for helping us on the beamline Swing and Christine Char-Raluy for her kind assistance in measuring the dn/dc. AF4 experiments were realized on the PlantLipPol-Green platform with the financial support of Agropolis foundation. Ana-Belén Bautista-Ortín and Stéphanie Carrillo's help in thioglycolysis is greatly appreciated.

Author information

Authors and Affiliations

  1. INRA, UMR1083 SPO, 2 place Viala, 34060, Montpellier cedex 1, France

    Aude Vernhet, Hélène Fulcrand & Céline Poncet-Legrand

  2. Montpellier SupAgro, UMR1083 SPO, 2 place Viala, 34060, Montpellier cedex 1, France

    Aude Vernhet, Hélène Fulcrand & Céline Poncet-Legrand

  3. Université Montpellier 1, UMR1083 SPO, 2 place Viala, 34060, Montpellier cedex 1, France

    Aude Vernhet, Hélène Fulcrand & Céline Poncet-Legrand

  4. Montpellier SupAgro, UMR1208 IATE, PlantLipPol-Green Platform, 34060, Montpellier, France

    Stéphane Dubascoux & Eric Dubreucq

  5. PMMH, UMR7636 (CNRS-ESPCI-P6-P7), 10 rue Vauquelin, 75231, Paris cedex 05, France

    Bernard Cabane

Authors
  1. Aude Vernhet
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stéphane Dubascoux
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bernard Cabane
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hélène Fulcrand
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Eric Dubreucq
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Céline Poncet-Legrand
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Céline Poncet-Legrand.

Additional information

Published in the special issue Wine Analysis with Guest Editor Isabelle Pianet.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 25 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vernhet, A., Dubascoux, S., Cabane, B. et al. Characterization of oxidized tannins: comparison of depolymerization methods, asymmetric flow field-flow fractionation and small-angle X-ray scattering. Anal Bioanal Chem 401, 1559–1569 (2011). https://doi.org/10.1007/s00216-011-5076-2

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-011-5076-2

Keywords

Search

Navigation