Measuring Procyanidin Concentration in Wines Using UHPLC

A recent study by the University of Bordeaux (Villenave d'Ornon, France) set out to develop and validate a method for rapid, reliable quantification of crown tannins in red and white wines through ultra-high performance liquid chromatography (UHPLC) coupled with a quadrupole time-of-flight (Q-TOF) mass spectrometry (MS). A paper based on this research was published in Methods and Protocols (1).

Widely distributed in plant-derived foods and beverages such as grapes, red wine, nuts, tea, apples, and chocolate, proanthocyanidins (also known as condensed tannins) are extensively studied for their ability to interact with other polyphenols or compounds (2). Proanthocyanidins contribute to various organoleptic properties in wine, such as its astringency and bitterness (3,4), and are also involved in the color stability and evolution of red wines (5,6). In 2015, a new subfamily of proanthocyanidins with an unusual structure was reported and named crown procyanidins (7); this subfamily appears to be more resistant to oxidation in wine than regular condensed tannins (8). The amount of crown procyanidins in grape skin is dependent on the variety of the grape; grape variety; to date, of the fifty different grape varieties (white and red varieties) analyzed to date, all exhibit these tannins (9,10).

The experimental assays for the authentication of the liquid chromatographic method were carried out using commercially available red and white wines (bag in box). The different red and white wines quantified were obtained from the microvinification performed at the Institute of Vine and Wine Sciences at the University of Bordeaux, from grapes collected at technical maturity. Prior to UHPLC-Q-TOF injection, red wines were diluted 20 times and white wines were diluted 10 times in water and were filtered through a 0.45 µm filter. The validation of the analytical method was carried out by assessing the following parameters: the matrix effect, linearity, accuracy, repeatability, intermediate reproducibility, limits of detection (LOD), and limits of quantification (LOQ) (1).

The authors report that their validation method, which followed international standards, demonstrated high sensitivity (LOQ = 0.033 mg/L), accuracy (recovery = 88.21% to 107.64%), repeatability (RSD = 1.99% to 11.03%), and intermediate reproducibility (RSD = 2.51% to 19.05%). Minimal matrix effects were observed, ensuring the reliable and precise quantification across both wine types. The applicability of the method was confirmed through the successful analysis of wine samples, leading to the first quantification of crown procyanidins in white wine. Concentrations ranged from 0.81 mg/L to 15.88 mg/L in the different analyzed wines (1).

This validated method provides a valuable tool for the study of crown procyanidin profiles in various wine matrices and establishes a foundation for future research into the role of crown procyanidins in wines and other food and beverage matrices where these compounds may be present. Furthermore, the authors wrote that future research could expand this methodology to other food and beverage matrices where crown procyanidins might be present, yielding further knowledge regarding their organoleptic properties. (1).

Red wine pouring into wine glass, close-up. © Africa Studio - stock.adobe.com

References

1. Ferreira, M.; Teissedre, P. L.; Jourdes, M. Development and Validation of a High-Throughput Quantification Method of Crown Procyanidins in Different Wines by UHPLC-Q-TOF. Methods Protoc. 2025, 8 (1), 7. DOI: 10.3390/mps8010007

2. Cheynier V. Polyphenols in Foods are More Complex Than Often Thought. Am. J. Clin. Nutr. 2005, 81 (1 Suppl), 223S-229S. DOI: 10.1093/ajcn/81.1.223S

3. Ma, W.; Guo, A.; Zhang, Y.; Wang, H.; Liu, Y.; Li, H. A Review on Astringency and Bitterness Perception of Tannins in Wine. Trends Food Sci. Technol. 2014,40(1), 6-19. DOI: 10.1016/j.tifs.2014.08.001

4. Cliff, M. A.; Stanich, K.; Edwards, J. E.; Saucier, C. T. Adding Grape Seed Extract to Wine Affects Astringency and Other Sensory Attributes. J. Food Qual.2012,35(4), 263-271. DOI: 10.1111/j.1745-4557.2012.00448.x

5. Zhang, B.; Liu, R.; He, F.; Zhou, P. P.; Duan, C. Q. Copigmentation of Malvidin-3-O-Glucoside with Five Hydroxybenzoic Acids in Red Wine Model Solutions: Experimental and Theoretical Investigations. Food Chem. 2015, 170, 226-233. DOI: 10.1016/j.foodchem.2014.08.026

6. Cheynier, V.; Duenas-Paton, M.; Salas, E.; Maury, C.; Souquet, J. M.; Sarni-Manchado, P.; Fulcrand, H. Structure and Properties of Wine Pigments and Tannins. Am.J. Enol. and Vitic. 2006,57(3), 298-305. DOI: 10.5344/ajev.2006.57.3.298

7. Zeng, L.; Pons-Mercadé, P.; Richard, T.; Krisa, S.; Teissèdre, P.L.; Jourdes, M. Crown Procyanidin Tetramer: A Procyanidin with an Unusual Cyclic Skeleton with a Potent Protective Effect against Amyloid-β-Induced Toxicity. Molecules 2019, 24, 1915. DOI: 10.3390/molecules24101915

8. Jouin, A.; Zeng, L.; Canosa, M.R.; Teissedre, P.L.; Jourdes, M. Evolution of the Crown Procyanidins’ Tetramer during Winemaking and Aging of Red Wine. Foods 2022, 11, 3194. DOI: 10.3390/foods11203194

9. Longo, E.; Rossetti, F.; Jouin, A.; Teissedre, P.L.; Jourdes, M.; Boselli, E. Distribution of Crown Hexameric Procyanidin and its Tetrameric and Pentameric Congeners in Red and White Wines. Food Chem. 2019, 299, 125125. DOI: 10.1016/j.foodchem.2019.125125

10. Jouin, A. Évolution et Caractérisation Chimique des Tanins Couronnes du Raisin au Vin [Internet]. Institute of Vine and Wine Sciences: University of Bordeaux. 2019. https://theses.hal.science/tel-03696540/document (accessed 2024-09-02).