Abstract
Flavonoids are a known group of organic compounds found in plants, where they act as dyes and natural fungicides. Together with the food of plant origin, they are delivered to the human body.
The chapter focuses on discussing the various modes of liquid chromatography used in the analysis of flavonoids in the food of plant origin. The most commonly used reversed-phase chromatography (RP-HPLC) has been compared with quite new hydrophilic interaction liquid chromatography (HILIC). Also, the potential of using two-dimensional chromatography in the analysis of polyphenolic compounds in the samples of plant origin is discussed. A separate subsection is devoted to chromatography combined with online antioxidant activity measurement. The whole chapter is completed by a short review of the detection methods used in the chromatographic determination of flavonoids.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Alpert, A.J. 1990. Hydrophilic interaction chromatography for the separation of peptides, nucleic acids and other polar compounds. Journal of Chromatography A 499: 177–196.
Beeiders, T., K.M. Kalili, E. Joubert, D. de Beer, and A. de Villiers. 2012. Comprehensive two dimensional liquid chromatographic analysis of rooibos (Aspalathus linearis) phenolics. Journal of Separation Science 35: 1808–1820.
Biesaga, M., A. Wach, M. Donten, J. Maik, and K. Pyrzyńska. 2006. Acidic hydrolysis and extraction of dyes present in plant and ancient textiles. Chemia Analityczna (Warsaw) 51: 251–265.
Bolarinwa, A., and J. Linseisen. 2005. Validated application of a new high-performance liquid chromatographic method for the determination of selected flavonoids and phenolic acids in human plasma using electrochemical detection. Journal of Chromatography B 823: 143–151.
Burnaz, N.A., M. Kucuk, and Z. Akar. 2017. An on-line HPLC system for detection of antioxidant compounds in some plant extracts by comparing three different methods. Journal of Chromatography B 1052: 66–72.
Cacciola, F., S. Farnetti, P. Dugo, P.J. Mariott, and L. Mondello. 2017. Comprehensive two-dimensional liquid chromatography for polyphenol analysis in food-stuffs. Journal of Separation Science 40: 7–24.
Cesla, P., T. Hajek, and P. Jandera. 2009. Optimization of two-dimensional gradient liquid chromatography separation. Journal of Chromatography A 1216: 3443–3457.
Chun, O.K., S.J. Chung, and W.O. Song. 2007. Estimated dietary flavonoid intake and major food sources of U.S. adults. Journal of Nutrition 127: 1244–1252.
Dai, J., and R.J. Mumper. 2010. Plant phenolics: Extraction, analysis and their antioxidant and anticancer properties. Molecules 15: 7313–7352.
De Beer, D., E. Joubert, C.J. Malherbe, and D.J. Brand. 2011. Use of countercurrent chromatography during isolation of 6-hydroxyluteolin-7-O-β-glucoside, a major antioxidant of Athrixia phylicoides. Journal of Chromatography A 1218: 6179–6186.
de Rijke, E., P. Out, W.M.A. Niessen, F. Ariese, C. Gooijer, and U.A. Brinkman. 2006. Analytical separation and detection methods for flavonoids. Journal of Chromatography A 1112: 31–63.
De Souza, L.M., T.R. Cipriani, C.F. Sant’Ana, M. Iacomini, P.A. Gorin, and G.L. Sasaki. 2009. Heart-cutting two dimensional (size exclusion x reversed phase) liquid chromatography-mass spectrometry analysis of flavonol glycosides from leaves of Maytenus ilicifolia. Journal of Chromatography A 1216: 99–105.
Donato, P., F. Rigano, F. Cacciola, M. Schure, S. Farnetti, M. Russo, P. Dago, and L. Mondelio. 2016. Comprehensive two-dimensional liquid chromatography- tandem mass spectrometry for the simultaneous determination of wine polyphenols and target contaminants. Journal of Chromatography A 145: 54–62.
Dugo, P., F. Cacciola, P. Donato, D. Airado-Rodriguez, M. Herrero, and L. Mondello. 2008a. Comprehensive two-dimensional liquid chromatography to quantify polyphenols in red wines. Journal of Chromatography A 1216: 7487–7493.
Dugo, P., F. Cacciola, M. Herrero, P. Donato, and L. Mondello. 2008b. Use of partially porous column as second dimension in comprehensive two- dimensional system for analysis of polyphenolic antioxidants. Journal of Separation Science 31: 3297–3308.
Dugo, G., L. La Perla, L. Di Bella, V. Lo Turco, D. Pollicino, G.L. La Torre, and T.M. Pellicano. 2009. Sicilian virgin olive oils and red wines: a potentially rich source of antioxidant compounds in the Mediterraneam diet. Rivista Italiana Delle Sostanze Grasse 86: 165–171.
Epriliati, I., G. Kerven, B. D’Arcy, and M. Gidley. 2010. Chromatographic analysis of diverse fruit components using HPLC and UHPLC. Analytical Methods 2: 1606–1610.
Faller, A.L.K., and E. Fialho. 2010. Polyphenol content and antioxidant capacity in organic and conventional plant foods. Journal of Food Composition and Analysis 23: 561–568.
Fan, Y., Y. Fu, Q. Fu, J. Cai, H. Xin, M. Dai, and Y. Jin. 2016. Purification of flavonoids from licorice using an off-line preparative two-dimensional normal-phase liquid chromatography/reversed phase liquid chromatography method. Journal of Separation Science 39: 2710–2719.
Francois, I., A. de Villiers, B. Tienpont, F. David, and P. Sandra. 2008. Comprehensive two-dimensional liquid chromatography applying two parallel columns in the second dimension. Journal of Chromatography A 1178: 33–42.
Hajek, T., V. Skerikova, P. Cesla, K. Vynuchalova, and P. Jandera. 2008. Multidimensional LC x LC analysis of phenolic and flavone natural antioxidants with UV- electrochemical coulometric and MS detection. Journal of Separation Science 31: 3309–3328.
Harly, J.M., R.F. Doherty, G.R. Beecher, J.M. Holden, D.B. Haytowitz, S. Bhagwat, and S. Gerhard. 2006. Flavonoid content of U.S. fruits, vegetables and nuts. Journal of Agricultural and Food Chemistry 54: 9966–9977.
Hemstrom, P., and K. Irgum. 2006. Hydrophilic interaction chromatography. Journal of Separation Science 29: 1784–1821.
Hollands, W.J., J. Voorspoels, G. Jacobs, K. Aaby, A. Meisland, R. Garcia-Villalba, F.T. Barberan, M.K. Piskula, D. Mawson, I. Vovk, P.W. Needs, and P.A. Kroona. 2017. Development, validation and evaluation of an analytical method for the determination of monomeric and oligomeric procyanidins in apple extracts. Journal of Chromatography A 1495: 46–56.
Jafari, S., S. Saedinia, and M. Abdollah. 2014. Role of natural phenolic compounds in cancer chemoprevention via regulation of the cell cycle. Current Pharmaceutical Biotechnology 15: 409–421.
Jandera, P., and P. Janas. 2017. Recent advances in stationary phases and understanding of retention in hydrophilic interaction chromatography. A review. Analytica Chimica Acta 967: 12–32.
Jandera, P., J. Urban, V. Skerikova, P. Langmaier, R. Kubickova, and J. Planeta. 2010. Polimethacrylate monolithic and hybrid particle-monolithic columns for reversed phase and hydrophilic interaction capillary liquid chromatography. Journal of Chromatography A 1217: 22–33.
Jandera, P., T. Hajek, M. Stankova, K. Vynuchalova, and P. Cesla. 2012. Optimization of comprehensive two-dimensional gradient chromatography coupling in-line hydrophilic interaction and reserved phase liquid chromatography. Journal of Chromatography A 1268: 91–101.
Jing, J., H.S. Parekh, M. Wei, W.C. Ren, and S.B. Chen. 2013. Advances in analytical technologies to evaluate the quality of traditional Chinese medicines. Trends in Analytical Chemistry 44: 39–45.
Jovanovic, M., T. Rakic, and B. Jancic-Stojanovic. 2014. Theoretical and empirical models in hydrophilic interaction liquid chromatography. Instrumental Science and Technology 42: 230–266.
Kalili, K.M., and A. de Villiers. 2009. Off-line comprehensive 2-dimensional hydrophilic interaction x reserved phase liquid chromatography analysis of procyanidins. Journal of Chromatography A 1187: 18–24.
Kalili, K.M., and A. de Villiers. 2010. Off-line comprehensive two-dimensional hydrophilic interaction x reversed phase liquid chromatographic analysis of green tea phenolics. Journal of Separation Science 33: 853–863.
Kalili, K.M., and A. de Villiers. 2013. Systematic optimisation and evaluation of on-line, off-line and stop-flow comprehensive hydrophilic interaction chromatography x reversed phase chromatographic analysis of procyanidins. Part II: Application to cocoa procyanidins. Journal of Chromatography A 1289: 69–70.
Klejdus, B., J. Vacek, L. Benesova, J. Kopecky, O. Lapcik, and V. Kuban. 2007. Rapid resolution HPLC with spectrometric detection for the determination and identification of isoflavones in soy preparation and plant extracts. Analytical and Bioanalytical Chemistry 389: 2277–2285.
Klejdus, B., J. Vacek, L. Lojkova, L. Beneskova, and V. Kuban. 2009. Ultra-high pressure liquid chromatography of isoflavonoids and phenolic acids on different stationary phases. Journal of Chromatography A 1195: 52–59.
Kusznierewicz, B., A. Piasek, A. Bartoszek, and J. Namieśnik. 2011. The optimisation of analytical parameters for routine profiling of antioxidants in complex mixtures by HPLC coupled post-column derivatisation. Phytochemical Analysis 22: 392–402.
Lee, J.E., G.S. Kim, S. Park, H.Y. Kim, B.Y. Kim, W.S. Lee, W. Jeong, S.J. Lee, S.J. Jin, and S.C. Shin. 2014. Determination of chokeberry (Aronia melanocarpa) polyphenol components using liquid chromatography-tandem mass spectrometry: Overall contribution to antioxidant activity. Food Chemistry 146: 1–5.
Lei, Z., B.W. Sumner, A. Bhatia, S.J. Sarma, and L.W. Sumner. 2019. UHPLC-MS analyses of plant flavonoids. Current Protocols in Plant Biology 4: e20085.
Magiera, S., I. Baranowska, and A. Lautenszleger. 2015. UHPLC-UV method for the determination of flavonoids in dietary supplements and foe evaluation of their antioxidant activities. Journal of Pharmaceutical and Biomedical Analysis 102: 468–475.
Malherbe, C.J., D. De Beer, and E. Joubert. 2012. Development of on-line high performance liquid chromatography (HPLC)-biochemical detection methods as tools in the identification of bioactives. International Journal of Molecular Science 13: 3101–3133.
McCalley, D.V. 2017. Understanding and manipulating the separation in hydrophilic interaction liquid chromatography- a review. Journal of Chromatography A 1523: 49–71.
Meyer, R., G. Stecher, R. Wuerzner, R.C. Silva, T. Sultana, L. Trojer, I. Feuerstein, C. Krieg, G. Abel, M. Popp, O. Bobleter, and G.K. Bonn. 2008. Procyanidins: Target compounds as antibacterial agents. Journal of Agricultural and Food Chemistry 56: 6959–6966.
Michalkiewicz, A., M. Biesaga, and K. Pyrzyńska. 2008. Solid-phase extraction procedure for determination of phenolic acids and some flavonols in honey. Journal of Chromatography A 1187: 18–24.
Mojzer, E.B., M. Hrncic, Z. Knez, and U. Bren. 2016. Polyphenols: Extraction methods, antioxidative action, bioavailability and anticarcinogenic effect. Molecules 21: 901–939.
Motilva, M.J., A. Serra, and A. Macia. 2013. Analysis of food polyphenols by ultra-high performance liquid chromatography coupled to mass spectrometry: An overview. Journal of Chromatography A 1292: 66–82.
Oszmianski, J., J. Kolniak-Ostek, and A. Wojdyła. 2013. Application of ultra performance liquid chromatography photodiode detector quadrupole/time of flight mass spectrometry (UHPLC-PDA-Q/TOF-MS) method for the characterization of phenolic compounds of Lepidium sativum L. sprouts. European Journal of Food Research and Technology 236: 699–706.
Pace, B., I. Capotorto, M. Cefola, P. Minasi, N. Montemurro, and V. Carbone. 2020. Evaluation of quality, phenolic and carotenoid composition of fresh-cut purple Polignano carrots stored in modified atmosphere. Journal of Food Composition and Analysis 86: 10336–10344.
Pedan, V., R. Fischer, and S. Rohn. 2016. An online NP-HPLC-DPPH method for the determination of the antioxidant activity of condensed polyphenols in cocoa. Food Research International 89: 890–900.
Peterson, J., and J. Dwyer. 1998. Flavonoids: Dietary occurrence and biochemical activity. Nutrition Research 18: 1995–2008.
Rice-Evans, C.A., M. Miller, and G. Paganga. 1997. Antioxidant properties of phenolic compounds. Trends in Plant Science 2: 152–159.
Robbins, R.J., J. Leonczak, J.C. Johnson, J. Li, C. Kwik-Uribe, R.L. Prior, and L. Gu. 2009. Method performance and multi-laboratory assessment of normal phase high pressure liquid chromatography-fluorescence detection method for the quantification of flavonols and procyanidins in cocoa and chocolate containing samples. Journal of Chromatography A 1216: 4830–4840.
Sakakibara, H., Y. Honda, S. Nakagawa, H. Ashida, and K. Kanazawa. 2003. Simultaneous determination of all polyphenols in vegetables, fruits and teas. Journal of Agricultural and Food Chemistry 51: 571–581.
Schwarz, M., M.C. RodrÃguez, D.A. Guillén, and C.G. Barosso. 2009. Development and validation of UPLC for the determination of phenolic compounds and furanic derivatives in Brandy de Jerez. Journal of Separation Science 32: 1782–1790.
Sentkowska, A., M. Biesaga, and K. Pyrzyńska. 2013. Effect of the operation parameters on HILIC separation of flavonoids on zwitterionic column. Talanta 115: 284–290.
Sentkowska, A., M. Biesaga, and K. Pyrzyńska. 2015a. Retention study of flavonoids under different chromatographic modes. Journal of Chromatographic Sciences 54: 516–522.
Sentkowska, A., M. Biesaga, and K. Pyrzyńska. 2015b. Polyphenolic composition and antioxidative properties of lemon balm (Melissa officinalis L.) extract affected by different brewing processes. International Journal of Food Properties 18: 2009–2014.
Sentkowska, A., M. Biesaga, and K. Pyrzyńska. 2016a. Hydrophilic interaction chromatographic analysis of quercetin and its glycosides. Current Analytical Chemistry 12: 60–64.
Sentkowska, A., M. Biesaga, and K. Pyrzyńska. 2016b. Application of hydrophilic interaction liquid chromatography for the quantification of flavonoids in Genista tinctoria extract. Journal of Analytical Methods in Chemistry 2016: 3789348.
Serrut, M., G. Cretier, and S. Heinisch. 2014. Theoretical and practical interest in UHPLC technology for 2D-LC. Trends in Analytical Chemistry 63: 104–112.
Shahidi, F., and P. Ambigaipalan. 2015. Phenolics and polyphenolics in foods, beverages and spices: Antioxidant activity and health effects – A review. Journal of Functional Foods 15: 820–897.
Shahidi, F., and Y. Zhong. 2015. Measurement of antioxidant activity. Journal of Functional Foods 18: 757–781.
Shi, S.Y., Y.P. Zhang, X.Y. Jiang, X.Q. Chen, K.L. Huang, H.H. Zhou, and X.Y. Jiang. 2009. Coupling HPLC to on-line, post-column (bio)chemical assays for high-resolution screening of bioactive compounds from complex mixtures. Trends in Analytical Chemistry 28: 865–877.
Silva, C.L., J. Pereira, V.G. Wouter, C. Giro, and J.S. Camara. 2011. Fast method using a new hydrophilic lipophilic balanced sorbent in combination with ultra-high performance liquid chromatography for quantification of significant bioactive metabolites in wines. Talanta 86: 82–90.
Skerikova, V., and P. Jandera. 2010. Effects of the operation parameters on hydrophilic interaction liquid chromatography separation of phenolic acids on zwitterionic monolithic capillary columns. Journal of Chromatography A 1217: 7891–7989.
Slimestad, R., T. Fossen, and C. Brede. 2020. Flavonoids and other phenolics in herbs commonly used in Norwegian commercial kitchens. Food Chemistry 309: 125678.
Soto-Vaca, A., A. Gutierrez, J.N. Losso, Z. Xu, and J.W. Finley. 2012. Evaluation of phenolic compounds from color and flavor problems to health benefits. Journal of Agricultural Food Chemistry 60: 6658–6677.
Stander, M.A., W. Kulan, and N.F. Hiten. 2013. Survey of South Africa fruit juices using a fast screening HILIC-MS method. Food Additives and Contaminants Part A 30: 1473–1484.
Steevensz, A., S.L. MacKinnon, R. Hankinson, C. Craft, S. Connan, D.B. Stengel, and J.E. Melanson. 2012. Profiling phlorotannins brown macroalgae by liquid chromatography-high resolution mass spectrometry. Phytochemical Analysis 23: 547–553.
Stevenson, D., and R. Hurst. 2007. Polyphenolic phytochemicals- just antioxidant or much more? Cellular and Molecular Life Science 64: 2900–2916.
Surowiec, I., J. Orska-Gawryś, M. Biesaga, M. Trojanowicz, M. Hutta, R. Halko, and K. Urbaniak-Walczak. 2003. Identification of natural dyestuff in archeological Coptic textiles by HPLC with fluorescence detection. Analytical Letters 36: 1211–1229.
Wach, A., K. Pyrzyńska, and M. Biesaga. 2007. Quercetin content in some food and herbal samples. Food Chemistry 100: 699–704.
Wang, J., L. Chen, L. Qu, K. Li, K. Zhao, Z. Wang, Y. Li, X. Zhang, Y. Jin, and X. Liang. 2019. Isolation and bioactive evaluation of flavonoid glycosides from Lobelia chinesis Lour using two-dimensional liquid chromatography combined with label-free cell phenotypic assays. Journal of Chromatography A 1601: 224–231.
Wawer, I., P. Eggert, and B. Hołub. 2012. Aronia super owoc (Aronia super fruit). Warszawa: Wektor (in Polish).
West, C., and E. Lesseltier. 2006. Characterisation of stationary phases in subcritical fluid chromatography with the solvation model: III, polar stationary phase. Journal of Chromatography A 1110: 200–213.
Willemse, C.M., M.A. Stander, A.G.J. Tredoux, and A. de Villiers. 2014. Comprehensive two-dimensional liquid chromatographic analysis of anthocyanins. Journal of Chromatography A 1359: 189–201.
Yanagida, A., H. Murao, M. Ohnishi-Kameyama, Y. Yamakawa, A. Shoji, M. Tagashira, T. Kanda, H. Shindo, and Y. Shibusawa. 2007. Retention behaviour of oligomeric proanthocyanidins in hydrophilic interaction chromatography. Journal of Chromatography A 1143: 153–162.
Zhao, Y., Y. Wang, Z.T. Jiang, and R. Li. 2017. Screening and evaluation of active compounds in polyphenol mixtures by HPLC coupled with chemical methodology and its application. Food Chemistry 227: 187–193.
Zheng, J., X. Zhang, Z. Guo, J. Feng, J. Zheng, X. Xue, and L. Liang. 2012. Separation and identification of flavonoids from complex samples using off-line two-dimensional liquid chromatography tandem mass spectrometry. Journal of Chromatography A 1220: 50–56.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Nature Switzerland AG
About this chapter
Cite this chapter
Sentkowska, A. (2021). Application of Liquid Chromatography for the Analysis of Flavonoids in Food: An Overview. In: Jeszka-Skowron, M., Zgoła-Grześkowiak, A., Grześkowiak, T., Ramakrishna, A. (eds) Analytical Methods in the Determination of Bioactive Compounds and Elements in Food. Food Bioactive Ingredients. Springer, Cham. https://doi.org/10.1007/978-3-030-61879-7_2
Download citation
DOI: https://doi.org/10.1007/978-3-030-61879-7_2
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-030-61878-0
Online ISBN: 978-3-030-61879-7
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)