Coumarins content in wine: application of HPLC, fluorescence spectrometry, and chemometric approach

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In this work, high performance liquid chromatography (HPLC) and fluorescence spectrometry methods for determination of natural coumarins in Tokaj wine were developed and compared. Molecularly imprinted solid phase extraction procedure was applied for sample preparation. The separation of esculin, coumarin, herniarin, 4-methylumbelliferone, scoparone, scopoletin was performed on core–shell C18 type of stationary phase (100 × 4.6 mm, 5 µm) with a gradient elution of mobile phase containing 1% aqueous solution of acetic acid and methanol, UV–VIS (280 nm for coumarin) and fluorescence detection (Ex 320 nm, Em 450 nm for other coumarins). The HPLC method was validated in term of linearity, limit of detection, limit of quantification, precision and accuracy. Fluorescence detection offers high sensitivity with limit of detection in the ng mL−1 range. Scopoletine and 4-methylumbelliferone were identified and quantified in tested wines. Emission spectral data, synchronous fluorescence spectra of coumarins from fluorescence spectrometry and total concentrations of coumarins quantified by the HPLC method were used in the partial least squares regression. The linear regression between the concentrations predicted by the partial least squares model versus true values obtained by HPLC method confirmed good agreement between the two methods.

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  1. Aleixandre-Tudo JL, Buica A, Nieuwoudt H, Aleixandre JL, Du Toit W (2017) Spectrophotometric analysis of phenolic compounds in grapes and wines. J Agric Food Chem 65:4009–4026

  2. Barciela-Alonso MC, Otero-Lavandeira N, Bermejo-Barrera P (2017) Solid phase extraction using molecular imprinted polymers for phthalate determination in water and wine samples by HPLC-ESI-MS. Microchem J 132:233–237

  3. Bitar M, Cayot P, Bou-Maroun E (2014) Molecularly imprinted polymer solid phase extraction of fungicides from wine samples. Anal Methods 6:6467–6472

  4. Borges F, Roleira F, Milhazes N, Santana L, Uriarte E (2005) Simple coumarins and analogues in medicinal chemistry: occurrence, synthesis and biological activity. Curr Med Chem 12:887–916

  5. Büyüktuncel E, Porgali E, Özkara S (2018) Catechin-molecularly imprinted cryogel for determination of catechin in red wines by HPLC–DAD–fluorescence detector. Acta Chromatogr 30:54–61

  6. Canas S (2017) Phenolic compounds and related properties of aged wine spirits: influence of barrel characteristics. A review. Beverages 55:1–22

  7. Da Silva AA, Pereira do Nascimento ES, Cardoso DR, Franco DW (2009) Coumarins and phenolic fingerprints of oak and Brazilian woods extracted by sugarcane spirit. J Sep Sci 32:3681–3691

  8. De Rosso M, Panighel A, Dalla Vedova A, Stella L, Flamini R (2009) Changes in chemical composition of a red wine aged in acacia, cherry, chestnut, mulberry, and oak wood barrels. J Agric Food Chem 57:1915–1920

  9. De Simon BF, Sanz M, Cadahia E, Martinez J, Esteruelas E, Munoz AM (2014) Polyphenolic compounds as chemical markers of wine ageing in contact with cherry, chestnut, false acacia, ash and oak wood. Food Chem 143:66–76

  10. De Villiers A, Albertsa P, Tredoux AGJ, Nieuwoudt HH (2012) Analytical techniques for wine analysis: an African perspective; a review. Anal Chim Acta 730:2–23

  11. Garrido J, Borges F (2013) Wine and grape polyphenols: a chemical perspective. Food Res Int 54:1844–1858

  12. Giovannoli C, Passini C, Di Nardo F, Anfossi L, Baggiani C (2014) Determination of ochratoxin A in italian red wines by molecularly imprinted solid phase extraction and HPLC analysis. J Agric Food Chem 62:5220–5225

  13. Gomez FJ, Silva MF (2016) Microchip electrophoresis for wine analysis. Anal Bioanal Chem 408:8643–8653

  14. Huang J, Liang P, Xu J, Wu Y, Shen W, Xu B, Zhang D, Xia J, Zhuang S (2017) Qualitative and quantitative determination of coumarin using surface-enhanced Raman spectroscopy coupled with intelligent multivariate analysis. RSC Adv 7(77):49097–49101

  15. Izquierdo MEF, Granados JQ, Mir MV, Martinez MCL (2000) Comparison of methods for determining coumarins in distilled beverages. Food Chem 70:251–258

  16. Jandera P, Škeříková V, Řehová L, Hájek T, Baldriánová L, Škopová G, Kellner V, Horna A (2005) PR-HPLC analysis of phenolic compounds and flavonoids in beverages and plant extracts using a CoulArray detector. J Sep Sci 28:1005–1022

  17. Machyňáková A, Hroboňová K (2017a) Simultaneous determination of coumarin derivatives in natural samples by ultrahigh performance liquid chromatography. J Food Nutr Res 56:179–188

  18. Machyňáková A, Hroboňová K (2017b) Synthesis and evaluation of molecularly imprinted polymers as sorbents for selective extraction of coumarins. Chromatographia 80:1015–1024

  19. Machyňáková A, Lhotská I, Hroboňová K, Šatinsky D (2017) On-line coupling of molecularly imprinted solid phase extraction with liquid chromatography for the fast determination of coumarins from complex samples. J Pharm Biomed Anal 145:144–150

  20. Molinelli A, Weiss R, Mizaikoff B (2002) Advanced solid phase extraction using molecularly imprinted polymers for the determination of quercetin in red wine. J Agric Food Chem 50:1804–1808

  21. Poláček R, Májek P, Hroboňová K, Sádecká J (2015) Fluorescence spectroscopy as a tool for determination of coumarins by multivariate calibration. J Fluoresc 25:297–303

  22. Puech JL, Moutounet M (1988) Liquid chromatographic determination of scopoletin in hydroalcoholic extract of oak wood and in matured distilled alcoholic beverages. J Assoc Anal Chem 71:512–514

  23. Rentzsch M, Wilkens A, Winterhalter P (2009) In: Moreno-Arribas MV, Polo MC (eds) Wine chemistry and biochemistry. Springer, New York

  24. Ribéreau-Gayon P, Glories Y, Maujean A, Dubourdieu D (2006) Handbook of enology. The chemistry of wine: stabilization and treatment, vol 2, 2nd edn. Wiley, New York, pp 143–144

  25. Sádecká J, Jakubíková M, Májek P (2018) Fluorescence spectroscopy for discrimination of botrytized wines. Food Control 88:75–84

  26. Salagoity-Auguste MH, Tricard C, Sudral P (1987) Simultaneous determination of aromatic aldehydes and coumarins by high-preform liquid chromatography. Application to wines and brandies stored in oak barrels. J Chromatogr 392:379–387

  27. Sanagi MM, Salleh S, Ibrahim WAW, Naim AA (2011) Determination of organophosporus pesticides using molecular imprinted polymer solid phase extraction. Malaysian Journal of Analytical Sciences 15:175–183

  28. Sikorska E, Gliszczyńska-Świgło A, Insińska-Rak M, Khmelinskii I, De Keukeleire D, Sikorski M (2008) Simultaneous analysis of riboflavin and aromatic amino acids in beer using fluorescence and multivariate calibration methods. Anal Chim Acta 613:207–217

  29. Stauffer TM (2017) Ideas and applications toward sample preparation for food and beverages analysis. InTech, Rijeka

  30. Wang L, Wu HL, Yin XL, Hu Y, Gu HW, Yu RQ (2017) Simultaneous determination of umbelliferone and scopoletin in Tibetan medicine Saussurea laniceps and traditional Chinese medicine Radix angelicae pubescentis using excitation-emission matrix fluorescence coupled with second-order calibration method. Spectrochim Acta A Mol Biomol Spectrosc 170:104–110

  31. Wise BM, Gallagher NB, Bro R, Shaver JM, Windig W, Koch RS (2006) PLS toolboox 4.0 for use with Matlab™. Eigenvector Research Inc, Wenatchee

  32. Wold S, Sjostrom M, Eriksson L (2001) PLS-regression: a basic tool of chemometrics. Chemom Intell Lab Syst 58:109–130

  33. Yang M, Chen JL, Shi XF, Niu HJ (2011) Rapid determination of aesculin, aesculetin and fraxetin in cortex fraxini extract solutions based on ultraviolet spectroscopy. E J Chem 8(S1):S225–S236

  34. Zhang B, Cai J, Duan CQ, Reeves MJ, He F (2015) A review of phenolics in oak woods. Int J Mol Sci 16:6978–7014

  35. Žiak Ľ, Sádecká J, Májek P, Hroboňová K (2014) Simultaneous determination of phenolic acids and scopoletin in brandies using synchronous fluorescence spectrometry coupled with partial least squares. Food Anal Methods 7:563–570

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The authors thanks J & J Ostrožovič spol. s r.o. and Tokaj & CO, s.r.o. for cooperation and providing of wines. The authors thanks Eva Brokešová for cooperation in sample preparation.


This research was financially supported by the Slovak Research and Development Agency under the Contract No. APVV-15-0355.

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Correspondence to Katarína Hroboňová.

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Katarína Hroboňová declares that she has no conflict of interest. Jana Sádecká declares that she has no conflict of interest.

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Hroboňová, K., Sádecká, J. Coumarins content in wine: application of HPLC, fluorescence spectrometry, and chemometric approach. J Food Sci Technol 57, 200–209 (2020) doi:10.1007/s13197-019-04048-2

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  • Wine
  • Coumarins
  • High performance liquid chromatography
  • Fluorescence spectrometry