Extraction, Isolation, and Quantitative Determination of Flavonoids by HPLC

  • Ashim K. Sen
  • Dhanya B. Sen
  • Rajesh A. Maheshwari


Flavonoids are natural products, more precisely secondary metabolites with low molecular weight produced by plants. Unlike primary metabolites, they are non-essential for plant survival. These secondary metabolites are biologically active and more than 10,000 structural variants of flavonoids have been reported such as flavonols, flavones, flavanones, anthocyanidins, catechins, biflavins and many more. Majority of these compounds act as antioxidants and exist as sugar conjugates in food. Flavonoids are omnipresent in vegetables and fruits with broad range of properties. Plant-derived flavonoids are essential dietary parts of animal system and are known to have an expansive scope of properties including antimicrobial, antiviral, anticancer, antiatherosclerosis and antithrombotic activity. Hence, it is imperative to estimate flavonoid sources in food, which will indubitably lead to a new era of flavonoids in foods or pharmaceutical supplements. Different traditional methods like UV/Visible absorption spectroscopy and separation techniques like thin-layer chromatography (TLC), paper electrophoresis and polyamide chromatography were commonly used in earlier times for flavonoid separation. Evidently, TLC is still a preferable method for flavonoid analysis, because of its simplicity, rapidity and versatility. On the other hand, majority of recently published work refers to qualitative and quantitative determination of flavonoids by high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LC-MS), liquid chromatography-nuclear magnetic resonance (LC-NMR) and capillary electrophoresis (CE). The separation, identification and quantification of flavonoids can be performed in one operation by pairing HPLC to UV/Vis, MS or NMR detectors, especially if the study aims at determining individual flavonoids. Furthermore, HPLC enables the separation and quantification of the flavonoids simultaneously without requiring a preliminary derivatization. These factors have contributed to make HPLC analysis of flavonoids a commonly used technique.


Flavonoids Extraction Isolation Quantitative determination HPLC UV/Visible LC-MS/NMR 


  1. Agati G, Azzarello E, Pollastri S, Tattini M. Flavonoids as antioxidants in plants: location and functional significance. Plant Sci. 2012;196:67–76.PubMedCrossRefGoogle Scholar
  2. Aherne SA, Obrien NM. Dietary flavonols: chemistry, food content and metabolism. Nutrition. 2002;18:75–81.PubMedCrossRefGoogle Scholar
  3. Andlauer W, Martena MJ, Furst P. Determination of selected phytochemicals by reversed-phase high performance liquid chromatography combined with ultraviolet and mass spectrometric detection. J Chromatogr A. 1999;849:341–8.PubMedCrossRefGoogle Scholar
  4. Arts IC, Van de Putte B, Hollman PC. Catechin contents of foods commonly consumed in the Netherlands. 1. Fruits, vegetables, staple foods, and processed foods. J Agric Food Chem. 2000a;48:1746–51.PubMedCrossRefGoogle Scholar
  5. Arts ICW, Van de Putte B, Hollman PC. Catechin contents of foods commonly consumed in the Netherlands. 2. Tea, wine, fruit juices, and chocolate milk. J Agric Food Chem. 2000b;48:1752–7.PubMedCrossRefGoogle Scholar
  6. Bader AN, Pivovarenko VG, Demchenko AP, Ariese F, Gooijer C. Excited state and ground state proton transfer rates of 3-hydroxyflavone and its derivatives studied by shpol'skii spectroscopy: the influence of redistribution of electron density. J Phys Chem B. 2004;108:10589–95.CrossRefGoogle Scholar
  7. Bansal A, Chhabra V, Rawal RK, Sharma S. Chemometrics: a new scenario in herbal drug standardization. J Pharm Anal. 2014;4:223–33.PubMedPubMedCentralCrossRefGoogle Scholar
  8. Barbuch RJ, Coutant JE, Welsh MB, Setchell KDR. The use of thermospray liquid chromatography/tandem mass spectrometry for the class identification and structural verification of phytoestrogens in soy protein preparations. Biomed Environ Mass Spectrom. 1989;18:973–7.PubMedCrossRefGoogle Scholar
  9. Benavente-Garcia O, Castillo J, Marin FR, Ortuno A, Del-Rio JA. Uses and properties of citrus flavonoids. J Agric Food Chem. 1997;45:4505–15.CrossRefGoogle Scholar
  10. Benthin B, Danz H, Hamburger M. Pressurized liquid extraction of medicinal plants. J Chromatogr A. 1999;837:211–9.PubMedCrossRefGoogle Scholar
  11. Bevan CD, Marshall PS. The use of supercritical fluids in the isolation of natural products. Nat Prod Rep. 1994;11:451–66.PubMedCrossRefGoogle Scholar
  12. Block G, Patterson B, Subar A. Fruit, vegetables, and cancer prevention: a review of the epidemiological evidence. Nutr Cancer. 1992;18:1–29.PubMedCrossRefGoogle Scholar
  13. Borris RP. Natural products research: perspectives from a major pharmaceutical company. J Ethnopharmacol. 1996;51:29–38.PubMedCrossRefGoogle Scholar
  14. Boue SM, Carter-Wientjes CH, Shih BY, Cleveland TE. Identification of flavone aglycones and glycosides in soybean pods by liquid chromatography–tandem mass spectrometry. J Chromatogr A. 2003;991:61–8.PubMedCrossRefGoogle Scholar
  15. Branco A, Pereira AD, Cardoso JN, De Aquino Neto FR, Pinto AC, Braz R. Further lipophilic flavonols in Vellozia graminifolia (Velloziaceae) by high temperature gas chromatography: quick detection of new compounds. Phytochem Anal. 2001;12:266–70.PubMedCrossRefGoogle Scholar
  16. Bravo L. Polyphenols: chemistry, dietary sources, metabolism, and nutritional significance. Nutr Rev. 1988;56:317–33.CrossRefGoogle Scholar
  17. Brown JE, Khodr H, Hider RC, Rice-Evans CA. Structural dependence of flavonoid interactions with Cu2+ ions: implications for their antioxidant properties. Biochem J. 1998;330:1173–8.PubMedPubMedCentralCrossRefGoogle Scholar
  18. Careri M, Mangia A, Musci M. Overview of the applications of liquid chromatography–mass spectrometry interfacing systems in food analysis: naturally occurring substances in food. J Chromatogr A. 1998;794:263–97.CrossRefGoogle Scholar
  19. Carlton RR, Gray AI, Lavaud C, Massiot G, Peter Waterman PG. Kaempferol-3-(2,3-diacetoxy-4-p-coumaroyl) rhamnoside from leaves of Myrica gale. Phytochemistry. 1990;29:2369–71.CrossRefGoogle Scholar
  20. Chu YH, Chang CL, Hsu HF. Flavonoid content of several vegetables and their antioxidant activity. J Sci Food Agric. 2000;80:561–6.CrossRefGoogle Scholar
  21. Clifford AH, Cuppett SL. Review: anthocyanins-nature, occurrence and dietary burden. J Sci Food Agric. 2000;80:1063–72.CrossRefGoogle Scholar
  22. Cook NC, Samman S. Review: flavonoids—chemistry, metabolism, cardioprotective effects, and dietary sources. J Nutr Biochem. 1996;7:66–76.CrossRefGoogle Scholar
  23. Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev. 1999;12:564–82.PubMedPubMedCentralCrossRefGoogle Scholar
  24. Cushnie TP, Lamb AJ. Antimicrobial activity of flavonoids. Int J Antimicrob Agents. 2005;26:343–56.PubMedPubMedCentralCrossRefGoogle Scholar
  25. Da Costa CT, Dalluge JJ, Welch MJ, Coxon B, Margolis SA, Horton D. Characterization of prenylated xanthones and flavanones by liquid chromatography/atmospheric pressure chemical ionization mass spectrometry. J Mass Spectrom. 2000;35:540–9.PubMedCrossRefGoogle Scholar
  26. Dawidowicz AL, Wianowska D, Gawdzik J, Danuta H, Smolarz DH. Optimization of ASE conditions for the HPLC determination of rutin and isoquercitrin in Sambucus nigra L. J Liq Chromatogr Relat Technol. 2003;26:2381–97.CrossRefGoogle Scholar
  27. De Rijke E, Zafra-Gomez A, Ariese F, Brinkman UA, Gooije C. Determination of isoflavone glucoside malonates in Trifolium pratense L. (red clover) extracts: quantification and stability studies. J Chromatogr A. 2001;932:55–64.PubMedCrossRefGoogle Scholar
  28. De Rijke E, Joshi HC, Sanderse HR, Ariese F, Brinkman UA, Gooijer C. Natively fluorescent isoflavones exhibiting anomalous Stokes' shifts. Anal Chim Acta. 2002;468:3–11.CrossRefGoogle Scholar
  29. De Rijke E, Zappey H, Ariese F, Gooijer C, Brinkman UA. Liquid chromatography with atmospheric pressure chemical ionization and electrospray ionization mass spectrometry of flavonoids with triple-quadrupole and ion-trap instruments. J Chromatogr A. 2003;984:45–58.PubMedCrossRefGoogle Scholar
  30. De Rijke E, Out P, Niessen WM, Ariese F, Gooijer C, Brinkman UA. Analytical separation and detection methods for flavonoids. J Chromatogr A. 2006;1112:31–63.PubMedCrossRefPubMedCentralGoogle Scholar
  31. Debnath PK, Banerjee S, Debnath P, Mitra A, Mukherjee PK. Ayurveda-opportunity for developing safe and effective treatment choice for the future. In: Mukherjee PK, editor. Evidence-based validation of herbal medicine. Amsterdam: Elsevier Science Publishing Co Inc; 2015. p. 427–54.CrossRefGoogle Scholar
  32. Deng F, Zito SW. Development and validation of a gas chromatographic-mass spectrometric method for simultaneous identification and quantification of marker compounds including bilobalide, ginkgolides and flavonoids in Ginkgo biloba L. extract and pharmaceutical preparations. J Chromatogr A. 2003;986:121–7.PubMedCrossRefGoogle Scholar
  33. Dey PM. Methods in plant biochemistry: plant phenolics. 1st ed. London: Academic Press Ltd; 1989.Google Scholar
  34. Dixon RA, Dey PM, Lamb CJ. Phytoalexins: enzymology and molecular biology. Adv Enzymol Relat Areas Mol Biol. 1983;55:1–136.PubMedGoogle Scholar
  35. Du F, Zhang F, Chen F, Wang A, Wang Q, Yin X, et al. Advances in microbial heterologous production of flavonoids. Afr J Microbiol Res. 2011;5:2566–74.CrossRefGoogle Scholar
  36. Duthie GG, Duthie SJ, Kyle JA. Plant polyphenols in cancer and heart disease: implications as nutritional antioxidants. Nutr Res Rev. 2000;13:79–106.PubMedCrossRefGoogle Scholar
  37. Etievant P, Schlich P, Bertrand A, Symonds P, Bouvier JC. Varietal and geographic classification of French red wines in terms of pigments and flavonoid compounds. J Sci Food Agric. 1988;42:39–54.CrossRefGoogle Scholar
  38. Fabre N, Rustan I, De Hoffman E, Quetin-Leclercq J. Determination of flavone, flavonol, and flavanone aglycones by negative ion liquid chromatography electrospray ion trap mass spectrometry. J Am Soc Mass Spectrom. 2001;12:707–15.PubMedCrossRefPubMedCentralGoogle Scholar
  39. Fecka I, Kowalczyk A, Cisowski W. Optimization of the separation of flavonoid glycosides and rosmarinic acid from Mentha piperita on HPTLC plates. J Planar Chromatogr. 2004;17:22–5.CrossRefGoogle Scholar
  40. Ferry DR, Smith A, Malkhandi J, Fyfe DW, de Takats PG, Anderson D, et al. Phase I clinical trial of the flavonoid quercetin: pharmacokinetics and evidence for in vivo tyrosine kinase inhibition. Clin Cancer Res. 1996;2:659–68.PubMedGoogle Scholar
  41. Fiamegos YC, Konidari CN, Stalikas CD. Cyanuric acid trace analysis by extractive methylation via phase transfer catalysis and capillary gas chromatography coupled with flame thermoionic and mass-selective detection. Process parameter studies and kinetics. Anal Chem. 2003;75:4034–42.PubMedCrossRefGoogle Scholar
  42. Fiamegos YC, Nanos CG, Vervoort J, Stalikas CD. Analytical procedure for the in-vial derivatization--extraction of phenolic acids and flavonoids in methanolic and aqueous plant extracts followed by gas chromatography with mass-selective detection. J Chromatogr A. 2004;1041:11–8.PubMedCrossRefGoogle Scholar
  43. Franke AA, Custer LJ. High performance liquid chromatographic assay of isoflavonoids and coumestrol from human urine. J Chromatogr B Biomed Sci Appl. 1994;662:47–60.CrossRefGoogle Scholar
  44. Ganzler K, Szinai I, Salgo A. Effective sample preparation method for extracting biologically active compounds from different matrices by a microwave technique. J Chromatogr. 1990;520:257–62.PubMedCrossRefPubMedCentralGoogle Scholar
  45. Garcia-Viguera C, Bakker J, Bellworthy SJ, Reader HP, Watkins SJ, Bridle P. The effect of some processing variables on non-coloured phenolic compounds in port wines. Eur Food Res Technol. 1997;205:321–4.Google Scholar
  46. Gerdin B, Srensso E. Inhibitory effect of the flavonoid O-(beta-hydroxyethyl)-rutoside on increased microvascular permeability induced by various agents in rat skin. Int J Microcirc Clin Exp. 1983;2:39–46.PubMedPubMedCentralGoogle Scholar
  47. Gil-Izquierdo A, Gil MI, Ferreres F, Tomas-Barberan FA. In vitro availability of flavonoids and other phenolics in orange juice. J Agric Food Chem. 2001;49:1035–41.PubMedCrossRefPubMedCentralGoogle Scholar
  48. Grayer RJ, Kite GC, Abou-Zaid M, Archer LJ. The application of atmospheric pressure chemical ionization liquid chromatography–mass spectrometry in the chemotaxonomic study of flavonoids: characterization of flavonoids from Ocimum gratissimum var. gratissimum. Phytochem Anal. 2000;11:257–67.CrossRefGoogle Scholar
  49. Gutierrez AC, Gehlen MH. Time resolved fluorescence spectroscopy of quercetin and morin complexes with Al3+. Spectrochim Acta A Mol Biomol Spectrosc. 2002;58:83–9.PubMedCrossRefPubMedCentralGoogle Scholar
  50. Halliwell B, Gutteridge JMC. Free radicals in biology and medicine. 5th ed. Oxford: Oxford University Press; 1998.Google Scholar
  51. Hamburger M, Baumann D, Adler S. Supercritical carbon dioxide extraction of selected medicinal plants--effects of high pressure and added ethanol on yield of extracted substances. Phytochem Anal. 2004;15:46–54.PubMedCrossRefGoogle Scholar
  52. Hansen SH, Jensen AG, Cornett C, Bjornsdottir I, Taylor S, Wright B, et al. High-performance liquid chromatography on-line coupled to high-field NMR and mass spectrometry for structure elucidation of constituents of Hypericum perforatum L. Anal Chem. 1999;71:5235–41.CrossRefGoogle Scholar
  53. Haraguchi H, Tanimoto K, Tamura Y, Mizutani K, Kinoshita T. Mode of antibacterial action of retrochalcones from Glycyrrhiza inflata. Phytochemistry. 1998;48:125–9.PubMedCrossRefGoogle Scholar
  54. Harborne JB. Phytochemical methods: a guide to modern techniques of plant analysis. 2nd ed. New York: Chapman and Hall; 1988.Google Scholar
  55. Harborne JB, Turner BL. Plant chemosystematics. London: Academic Press; 1984.Google Scholar
  56. Hasler A, Sticher O, Meier B. Identification and determination of the flavonoids from Ginkgo biloba by high-performance liquid chromatography. J Chromatogr A. 1992;605:41–8.CrossRefGoogle Scholar
  57. Hawryl M, Hawryl A, Soczewinski E. Application of normal- and reversed-phase 2D TLC on a cyanopropyl-bonded polar stationary phase for separation of phenolic compounds from the flowers of Sambucus nigra L. J Planar Chromatogr. 2002;15:4–10.CrossRefGoogle Scholar
  58. He Q, Kim J, Sharma RP. Silymarin protects against liver damage in BALB/c mice exposed to fumonisin B1 despite increasing accumulation of free sphingoid bases. Toxicol Sci. 2004;80:335–42.PubMedCrossRefGoogle Scholar
  59. Heim KE, Tagliaferro AR, Bobilya DJ. Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. J Nutr Biochem. 2002;13:572–84.PubMedCrossRefGoogle Scholar
  60. Hertog MGL, Hollman PCH, Katan MB. Content of potentially anticarcinogenic flavonoids of 28 vegetables and 9 fruits commonly consumed in the Netherlands. J Agric Food Chem. 1992;40:2379–83.CrossRefGoogle Scholar
  61. Hertog MG, Hollman PC, Katan MB. Intake of potentially anticarcinogenic flavonoids and their determinants in adults in the Netherlands. Nutr Cancer. 1993a;20:21–9.PubMedCrossRefGoogle Scholar
  62. Hertog MGL, Hollman PCH, Van de Putte B. Content of potentially anticarcinogenic flavonoids of tea infusions, wine sand fruit juices. J Agric Food Chem. 1993b;41:1242–6.CrossRefGoogle Scholar
  63. Ho CT, Lee CY, Huang MT. Phenolic compounds in food and their effects on health I: analysis. Occurrence & Chemistry: American Chemical Society; 1992.CrossRefGoogle Scholar
  64. Ho CT, Osawa T, Huang MT, Rosen RT. Food phytochemicals for cancer prevention II: teas, spices, and herbs. Oxford University Press: American Chemical Society; 1994.CrossRefGoogle Scholar
  65. Hollman PC, De Vries JH, Van Leeuwen SD, Mengelers MJ, Katan MB. Absorption of dietary quercetin in healthy ileostomy volunteers. Am J Clin Nutr. 1995;62:1276–82.PubMedCrossRefGoogle Scholar
  66. Hollman PCH, Van Trijp JMP, Buysman MNCP. Fluorescence detection of flavonols in HPLC by post column chelation with aluminium. Anal Chem. 1996;68:3511–5.PubMedCrossRefGoogle Scholar
  67. Hollman PC, Van Trijp JM, Buysman MN, Van Der Gaag MS, Mengelers MJ, De Vries JH, et al. Relative bioavailability of the antioxidant flavonoid quercetin from various foods in man. FEBS Lett. 1997;418:152–6.PubMedCrossRefGoogle Scholar
  68. Hostettmann K, Marston A, Hostettmann M. Preparative chromatography techniques: applications in natural product isolation. 2nd ed. Berlin: Springer-Verlag; 1998.CrossRefGoogle Scholar
  69. Huang MT, Osawa T, Ho CT, Rosen RT. Food phytochemicals for cancer prevention I: fruits and vegetables. 1st ed. Washington DC: Am Chem Society; 1994.Google Scholar
  70. Huck CW, Bonn GK. Evaluation of detection methods for the reversed-phase HPLC determination of 3′,4′,5′-trimethoxyflavone in different phytopharmaceutical products and in human serum. Phytochem Anal. 2001;12:104–9.PubMedCrossRefGoogle Scholar
  71. Hunter T. Protein kinases and phosphatases: the yin and yang of protein phosphorylation and signalling. Cell. 1995;80:225–36.PubMedCrossRefGoogle Scholar
  72. Hussein L, Fattah MA, Salem E. Characterization of pure proanthocyanidins isolated from the hulls of faba beans. J Agric Food Chem. 1990;38:95–8.CrossRefGoogle Scholar
  73. Issaq HJ. Capillary electrophoresis of natural products-II. Electrophoresis. 1999;20:3190–202.PubMedCrossRefGoogle Scholar
  74. Jamshidi A, Adjvadi M, Husain SW. Determination of Kaempferol and Quercetin in the extract of Ginkgo biloba by high-performance thin layer chromatography (HPTLC). J Planar Chromatogr. 2000;13:57–9.Google Scholar
  75. Janeczko Z, Krzek J, Pisulewska E, Sobolewska D, Dabrowska-Tylka M, Hubicka U, et al. Densitometric determination of genistin and daidzin in different cultivars of soy (Glycine max). J Planar Chromatogr. 2004;17:32–5.CrossRefGoogle Scholar
  76. Jarvis AP, Morgan ED. Isolation of plant products by supercritical fluid extraction. Phytochem Anal. 1997;8:217–22.CrossRefGoogle Scholar
  77. Jasprica I, Smolcic-Bubalo A, Mornar A, Medic-Saric M. Investigation of the flavonoids in Croatian propolis by thin-layer chromatography. J Planar Chromatogr. 2004;17:95–101.CrossRefGoogle Scholar
  78. Juan AD, Tauler R. Chemometrics applied to unravel multicomponent processes and mixtures: revisiting latest trends in multivariate resolution. Anal Chim Acta. 2003;500:195–210.CrossRefGoogle Scholar
  79. Justesen U, Knuthsen P, Leth T. Quantitative analysis of flavonols, flavones, and flavanones in fruits, vegetables and beverages by high-performance liquid chromatography with photo-diode array and mass spectrometric detection. J Chromatogr A. 1998;799:101–10.PubMedCrossRefGoogle Scholar
  80. Kaufmann B, Christen P. Recent extraction techniques for natural products: microwave assisted extraction and pressurized solvent extraction. Phytochem Anal. 2002;13:105–13.PubMedCrossRefGoogle Scholar
  81. Kim SM, Kang K, Jho EH, Jung YJ, Nho CW, Um BH, et al. Hepatoprotective effect of flavonoid glycosides from Lespedeza cuneata against oxidative stress induced by tert-butyl hyperoxide. Phytother Res. 2011;25:1011–7.PubMedCrossRefGoogle Scholar
  82. Klejdus B, Vacek J, Adam V, Zehnalek J, Kizek R, Trnkova L, et al. Determination of isoflavones in soybean food and human urine using liquid chromatography with electrochemical detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2004;806:101–11.PubMedCrossRefGoogle Scholar
  83. Koen B, Ruth V, Guido V, Johannes VS. Induction of cancer cell apoptosis by flavonoids is associated with their ability to inhibit fatty acid synthase activity. J Biol Chem. 2005;280:5636–45.CrossRefGoogle Scholar
  84. Kumar S, Pandey AK. Antioxidant, lipo-protective and antibacterial activities of phytoconstituents present in Solanum xanthocarpum root. Int Review Biophys Chem. 2012;3:42–7.Google Scholar
  85. Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: an overview. Sci World J. 2013a;162750:16. Scholar
  86. Kumar S, Pandey AK. Phenolic content, reducing power and membrane protective activities of Solanum xanthocarpum root extracts. Vegetos. 2013b;26:301–7.Google Scholar
  87. Kumar S, Gupta A, Pandey AK. Calotropis procera root extract has the capability to combat free radical mediated damage. ISRN Pharmacol. 2013a;691372:8. Scholar
  88. Kumar S, Mishra A, Pandey AK. Antioxidant mediated protective effect of Parthenium hysterophorus against oxidative damage using in vitro models. BMC Complement Altern Med. 2013b;13:120. Scholar
  89. Lamson DW, Brignall MS. Antioxidants and cancer, part 3: quercetin. Altern Med Rev. 2000;5:196–208.PubMedGoogle Scholar
  90. Lee MJ, Wang ZY, Li H, Chen L, Sun Y, Gobbo S, et al. Analysis of plasma and urinary tea polyphenols in human subjects. Cancer Epidemiol Biomark Prev. 1995;4:393–9.Google Scholar
  91. Leopoldini M, Russo N, Chiodo S, Toscano M. Iron chelation by the powerful antioxidant flavonoid quercetin. J Agric Food Chem. 2006;54:6343–51.PubMedCrossRefGoogle Scholar
  92. Leutert T, Von Arx E. Praparative Mitteldruck-flussigkeitschromatographie (preparative medium-pressure liquid chromatography). J Chromatogr A. 1984;292:333–44.CrossRefGoogle Scholar
  93. Liang YZ, Xie P, Chan K. Quality control of herbal medicines. J Chromatogr B Analyt Technol Biomed Life Sci. 2004;812:53–70.PubMedCrossRefGoogle Scholar
  94. Liggins J, Bluck LJ, Runswick S, Atkinson C, Coward WA, Bingham SA. Daidzein and genistein content of fruits and nuts. J Nutr Biochem. 2000;11:326–31.PubMedCrossRefGoogle Scholar
  95. Loewus FA, Ryan CA. Recent advances in phytochemistry. 1st ed. New York: Springer US; 1981.Google Scholar
  96. Ma YL, Vedernikova I, Heuvel HVD, Claeys M. Internal glucose residue loss in protonated O-diglycosyl flavonoids upon low-energy collision-induced dissociation. J Am Soc Mass Spectrom. 2000;11:136–44.PubMedCrossRefGoogle Scholar
  97. Mabry T, Markham KR, Thomas MB, editors. The systematic identification of flavonoids. 1st ed. New York: Springer-Verlag; 1970.Google Scholar
  98. Mahomoodally MF, Gurib-Fakim A, Subratty AH. Antimicrobial activities and phytochemical profiles of endemic medicinal plants of Mauritius. Pharm Biol. 2005;43:237–42.CrossRefGoogle Scholar
  99. Males Z, Medic-Saric M. Optimization of TLC analysis of flavonoids and phenolic acids of Helleborus atrorubens Waldst. Et kit. J Pharm Biomed Anal. 2001;24:353–9.PubMedCrossRefGoogle Scholar
  100. Marchart E, Krenn L, Kopp B. Quantification of the flavonoid glycosides in Passiflora incarnata by capillary electrophoresis. Planta Med. 2003;69:452–6.PubMedCrossRefGoogle Scholar
  101. Markaverich BM, Roberts RR, Alejandro MA, Johnson GA, Middleditch BS, Clark JH. Bioflavonoid interaction with rat uterine type II binding sites and cell growth inhibition. J Steroid Biochem. 1988;30:71–8.PubMedCrossRefGoogle Scholar
  102. Mattila P, Astola J, Kumpulainen J. Determination of flavonoids in plant material by HPLC with diode-array and electro-array detections. J Agric Food Chem. 2000;48:5834–41.PubMedCrossRefGoogle Scholar
  103. McKee TC, Bokesch HR, McCormick JL, Rashid MA, Spielvogel D, Gustafson KR, et al. Isolation and characterization of new anti-HIV and cytotoxic leads from plants, marine, and microbial organisms. J Nat Prod. 1997;60:431–8.PubMedCrossRefGoogle Scholar
  104. Medic-Saric M, Stanic G, Bosnjak I. The use of information theory and numerical taxonomy methods for evaluating the quality of thin-layer chromatographic separations of flavonoid constituents of Matricariae flos. Pharmazie. 2001;56:156–9.PubMedGoogle Scholar
  105. Medic-Saric M, Jasprica I, Mornar A, Smolcic-Bubalo A, Golja P. Quantitative analysis of flavonoids and phenolic acids in propolis by two-dimensional thin layer chromatography 2004.Google Scholar
  106. Merken HM, Beecher GR. Measurement of food flavonoids by high-performance liquid chromatography: a review. J Agric Food Chem. 2000;48:577–99.PubMedCrossRefGoogle Scholar
  107. Middleton E. The flavonoids. Trends Pharmacol Sci. 1984;5:335–8.Google Scholar
  108. Middleton EJ. Effect of plant flavonoids on immune and inflammatory cell function. Adv Exp Med Biol. 1998;439:175–82.PubMedCrossRefGoogle Scholar
  109. Middleton EJ, Kandaswami C. Effects of flavonoids on immune and inflammatory cell functions. Biochem Pharmacol. 1992;43:1167–79.PubMedCrossRefGoogle Scholar
  110. Middleton E, Kandaswami C. In: Harborne JB, editor. The flavonoids - advances in research since 1986. London: Chapman and Hall; 1993. p. 619.Google Scholar
  111. Miller LP. Phytochemistry. California: Van Nostrand Reinhold; 1973.Google Scholar
  112. Mishra AK, Mishra A, Kehri HK, Sharma B, Pandey AK. Inhibitory activity of Indian spice plant Cinnamomum zeylanicum extracts against Alternaria solani and Curvularia lunata, the pathogenic dematiaceous moulds. Ann Clin Microbiol Antimicrob. 2009;8:1–7. Scholar
  113. Mishra A, Kumar S, Bhargava A, Sharma B, Pandey AK. Studies on in vitro antioxidant and antistaphylococcal activities of some important medicinal plants. Cell Mol Biol. 2011;57:16–25.PubMedGoogle Scholar
  114. Mishra A, Kumar S, Pandey AK. Scientific validation of the medicinal efficacy of Tinospora cordifolia. Sci World J. 2013a;292934:8. Scholar
  115. Mishra A, Sharma AK, Kumar S, Saxena AK, Pandey AK. Bauhinia variegata leaf extracts exhibit considerable antibacterial, antioxidant and anticancer activities. Biomed Res Int. 2013b;915436:10. Scholar
  116. Moerman DE. An analysis of the food plants and drug plants of native North America. J Ethnopharmacol. 1996;52:1–22.PubMedCrossRefGoogle Scholar
  117. Mori A, Nishino C, Enoki N, Tawata S. Antibacterial activity and mode of action of plant flavonoids against Proteus vulgaris and Staphylococcus aureus. Phytochemistry. 1987;26:2231–4.CrossRefGoogle Scholar
  118. Morton M, Arisaka O, Miyake A, Evans B. Analysis of phyto-oestrogens by gas chromatography-mass spectrometry. Environ Toxicol Pharmacol. 1999;7:221–5.PubMedCrossRefGoogle Scholar
  119. Narasimhachari N, Rudloff EV. Gas–liquid chromatography of some flavonoid compounds and hydroxy diphenyls. Can J Chem. 1962;40:1123–9.CrossRefGoogle Scholar
  120. Narayana KR, Reddy MS, Chaluvadi MR, Krishna DR. Bioflavonoids classification, pharmacological, biochemical effects and therapeutic potential. Indian J Pharmacol. 2001;33:2–16.Google Scholar
  121. Nishizuka Y. The molecular heterogeneity of protein kinase C and its implications for cellular regulation. Nature. 1988;334:661–5.PubMedCrossRefGoogle Scholar
  122. Ohemeng KA, Schwender CF, Fu KP, Barrett JF. DNA gyrase inhibitory and antibacterial activity of some flavones. Bioorg Med Chem Lett. 1993;3:225–30.CrossRefGoogle Scholar
  123. Pan MH, Lai CS, Ho CT. Anti-inflammatory activity of natural dietary flavonoids. Food Funct. 2010;1:15–31.PubMedCrossRefGoogle Scholar
  124. Pan S, Zhou S, Gao S, Yu Z, Zhang S, Tang M, et al. New perspectives on how to discover drugs from herbal medicines: CAM's outstanding contribution to modern therapeutics. Evid Based Complement Altern Med. 2013:25.Google Scholar
  125. Pan S, Litscher G, Gao S, Zhou S, Yu Z, Chen H, et al. Historical perspective of traditional indigenous medical practices: the current renaissance and conservation of herbal resources. Evid Based Complement Alternat Med. 2014;525340:20. Scholar
  126. Pandey AK. Anti-staphylococcal activity of a pan-tropical aggressive and obnoxious weed Parihenium histerophorus: an in vitro study. Natl Acad Sci Lett. 2007;30:383–6.Google Scholar
  127. Pandey AK, Mishra AK, Mishra A, Kumar S, Chandra A. Therapeutic potential of C. zeylanicum extracts: an antifungal and antioxidant perspective. Int J Biol Med Res. 2010;1:228–33.Google Scholar
  128. Pandey AK, Mishra AK, Mishra A. Antifungal and antioxidative potential of oil and extracts derived from leaves of Indian spice plant Cinnamomum tamala. Cell Mol Biol. 2012;58:142–7.PubMedGoogle Scholar
  129. Pandey MM, Rastogi S, AKS R. Indian traditional ayurvedic system of medicine and nutritional supplementation. Evid Based Complement Alternat Med. 2013:12. Scholar
  130. Pereira ADS, Padilha MC, Neto FRDA. Two decades of high temperature gas chromatography (1983-2003): what’s next? Microchem J. 2004;77:141–9.CrossRefGoogle Scholar
  131. Peterson J, Dwyer J. Flavonoids: dietary occurrence and biochemical activity. Nutr Res. 1998;18:1995–2018.CrossRefGoogle Scholar
  132. Peyrat-Maillard MN, Bonnely S, Berset C. Determination of the antioxidant activity of phenolic compounds by coulometric detection. Talanta. 2000;51:709–16.PubMedCrossRefGoogle Scholar
  133. Pineiro Z, Palma M, Barroso CG. Determination of catechins by means of extraction with pressurized liquids. J Chromatogr A. 2004;1026:19–23.PubMedCrossRefGoogle Scholar
  134. Rahman A. Studies in natural products chemistry. 1st ed. Amsterdam: Elsevier Science Ltd; 1989.Google Scholar
  135. Rastija V, Mornar A, Jasprica I, Srecnik G, Medic-Saric M. Analysis of phenolic components in Croatian red wines by thin-layer chromatography. J Planar Chromatogr. 2004;17:26–31.CrossRefGoogle Scholar
  136. Ratty AK, Das NP. Effects of flavonoids on nonenzymatic lipid peroxidation: structure-activity relationship. Biochem Med Metab Biol. 1988;39:69–79.PubMedCrossRefGoogle Scholar
  137. Rauha JP, Vuorela H, Kostiainen R. Effect of eluent on the ionization efficiency of flavonoids by ion spray, atmospheric pressure chemical ionization, and atmospheric pressure photoionization mass spectrometry. J Mass Spectrom. 2001;36:1269–80.PubMedCrossRefGoogle Scholar
  138. Rechner AR, Smith MA, Kuhnle G, Gibson GR, Debnam ES, Srai SK, et al. Colonic metabolism of dietary polyphenols: influence of structure on microbial fermentation products. Free Radic Biol Med. 2004;36:212–25.PubMedCrossRefGoogle Scholar
  139. Reinhold L, Harborne JB, Swain T. Progress in Phytochemlstry. 1st ed. Oxford: Pergamon Press; 1977.Google Scholar
  140. Rice-Evans CA, Miller NJ, Bolwell PG, Broamley PM, Pridham JB. The relative antioxidant activities of plant-derived polyphenolic flavonoids. Free Radic Res. 1995;22:375–83.PubMedCrossRefGoogle Scholar
  141. Rice-Evans CA, Miller NJ, Paganga G. Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Radic Biol Med. 1996;20:933–56.PubMedCrossRefPubMedCentralGoogle Scholar
  142. Rodriguez-Delgado MA, Malovana S, Perez JP, Borges T, Garcia Montelongo FJ. Separation of phenolic compounds by high-performance liquid chromatography with absorbance and fluorimetric detection. J Chromatogr A. 2001;912:249–57.PubMedCrossRefGoogle Scholar
  143. Rosenberg ZRS, Jenkins DJ, Diamandis EP. Flavonoids and steroid hormone-dependent cancers. J Chromatogr B Analyt Technol Biomed Life Sci. 2002;777:219–32.CrossRefGoogle Scholar
  144. Sakakibara H, Honda Y, Nakagawa S, Ashida H, Kanazawa K. Simultaneous determination of all polyphenols in vegetables, fruits, and teas. J Agric Food Chem. 2003;51:571–81.PubMedCrossRefGoogle Scholar
  145. Saleh MM, Hashem FAEM, Glombitza KW. Study of Citrus taitensis and radical scavenger activity of the flavonoids isolated. Food Chem. 1998;63:397–400.CrossRefGoogle Scholar
  146. Saller R, Meier R, Brignoli R. The use of silymarin in the treatment of liver diseases. Drugs. 2001;61:2035–63.PubMedCrossRefGoogle Scholar
  147. Sargenti SR, Vichnewski W. Sonication and liquid chromatography as a rapid technique for extraction and fractionation of plant material. Phytochem Anal. 2000;11:69–73.CrossRefGoogle Scholar
  148. Schaufelberger D, Hostettmann K. Analytical and preparative reversed-phase liquid chromatography of secoiridoid glycosides. J Chromatogr A. 1985;346:396–400.CrossRefGoogle Scholar
  149. Seal T. Quantitative HPLC analysis of phenolic acids, flavonoids and ascorbic acid in four different solvent extracts of two wild edible leaves, Sonchus arvensis and Oenanthe linearis of north-eastern region in India. J App Pharm Sci. 2016;6:157–66.CrossRefGoogle Scholar
  150. Sen S, Chakraborty R. Revival, modernization and integration of Indian traditional herbal medicine in clinical practice: importance, challenges and future. J Tradit Complement Med. 2017;7:234–44.PubMedCrossRefGoogle Scholar
  151. Sengupta PK, Kasha M. Excited state proton-transfer spectroscopy of 3-hydroxyflavone and quercetin. Chem Phys Lett. 1979;68:382–5.CrossRefGoogle Scholar
  152. Setchell KD, Brown NM, Desai P, Zimmer-Nechemias L, Wolfe BE, Brashear WT, et al. Bioavailability of pure isoflavones in healthy humans and analysis of commercial soy isoflavone supplements. J Nutr. 2001;131:1362S–75S.PubMedCrossRefPubMedCentralGoogle Scholar
  153. Shankar D, Majumdar B. Beyond the biodiversity convention: the challenges facing the biocultural heritage of India's medicinal plants. In: Bodeker G, KKS B, Burley J, Vantomme P, editors. Medicinal plants for Forest conservation and health care. Rome: Food and Agriculture Organization (FAO); 1997. p. 87–99.Google Scholar
  154. Singleton VL, Noble AC. Wine flavour and phenolic substances. In: Charalambous G, Katz I, editors. Phenolic, sulfur, and nitrogen compounds in food flavors. Chicago: American Chemical Society; 1976. p. 47–70.CrossRefGoogle Scholar
  155. Soczewinski E, Wojciak-Kosior M, Matysik G. Analysis of glycosides and aglycones of flavonoid compounds by double-development thin-layer chromatography. J Planar Chromatogr. 2004;17:261–3.CrossRefGoogle Scholar
  156. Sonnenbichler J, Zetl I. Biochemical effects of the flavonolignan silibinin on RNA, protein and DNA synthesis in rat livers. Prog Clin Biol Res. 1986;213:319–31.PubMedPubMedCentralGoogle Scholar
  157. Spencer JP, Vauzour D, Rendeiro C. Flavonoids and cognition: the molecular mechanisms underlying their behavioural effects. Arch Biochem Biophys. 2009;492:1–9.PubMedCrossRefPubMedCentralGoogle Scholar
  158. Stoggl WM, Huck CW, Bonn GK. Structural elucidation of catechin and epicatechin in sorrel leaf extracts using liquid-chromatography coupled to diode array-, fluorescence-, and mass spectrometric detection. J Sep Sci. 2004;27:524–8.PubMedCrossRefGoogle Scholar
  159. Suntornsuk L. Capillary electrophoresis of phytochemical substances. J Pharm Biomed Anal. 2002;27:679–98.PubMedCrossRefPubMedCentralGoogle Scholar
  160. Tapas AR, Sakarkar DM, Kakde RB. Flavonoids as nutraceuticals: a review. Trop J Pharm Res. 2008;7:1089–99.CrossRefGoogle Scholar
  161. Tomas-Barberan FA. Capillary electrophoresis: a new technique in the analysis of plant secondary metabolites. Phytochem Anal. 1995;6:177–92.CrossRefGoogle Scholar
  162. Tomas-Barberan FA, Clifford MN. Flavanones, chalcones and dihydrochalcones–nature, occurrence and dietary burden. J Sci Food Agric. 2000;80:1073–80.CrossRefGoogle Scholar
  163. Tsao R, Deng Z. Separation procedures for naturally occurring antioxidant phytochemicals. J Chromatogr B. 2004;812:85–99.CrossRefGoogle Scholar
  164. Tsuchiya H, Iinuma M. Reduction of membrane fluidity by antibacterial sophoraflavanone G isolated from Sophora exigua. Phytomedicine. 2000;7:161–5.PubMedCrossRefGoogle Scholar
  165. Tunon MJ, Garcia-Mediavilla MV, Sanchez-Campos S, Gonzalez-Gallego J. Potential of flavonoids as anti-inflammatory agents: modulation of pro-inflammatory gene expression and signal transduction pathways. Curr Drug Metab. 2009;10:256–71.PubMedCrossRefGoogle Scholar
  166. Urbanek M, Blechtova L, Pospisilova M, Polasek M. On-line coupling of capillary isotachophoresis and capillary zone electrophoresis for the determination of flavonoids in methanolic extracts of Hypericum perforatum leaves or flowers. J Chromatogr A. 2002;958:261–71.PubMedCrossRefGoogle Scholar
  167. Van Beek TA, Breteler H. Phytochemistry and agriculture (proceedings of phytochemical Society of Europe). Oxford: Clarendon Press; 1993.Google Scholar
  168. Wachtel-Galor S, Benzie IFF. Herbal medicine: an introduction to its history, usage, regulation, current trends, and research needs. In: Benzie IFF, Wachtel-Galor S, editors. Herbal medicine: biomolecular and clinical aspects. 2nd ed. Boca Raton (FL): CRC Press/Taylor & Francis; 2011.Google Scholar
  169. Wang J, Sporns P. Analysis of anthocyanins in red wine and fruit juice using MALDI-MS. J Agric Food Chem. 1999;47:2009–15.PubMedCrossRefPubMedCentralGoogle Scholar
  170. Wang J, Sporns P. MALDI-TOF MS analysis of food flavonol glycosides. J Agric Food Chem. 2000a;48:1657–62.PubMedCrossRefPubMedCentralGoogle Scholar
  171. Wang J, Sporns P. MALDI-TOF MS analysis of isoflavones in soy products. J Agric Food Chem. 2000b;48:5887–92.PubMedCrossRefPubMedCentralGoogle Scholar
  172. Watson DG, Atsriku C, Oliveira EJ. Review role of liquid chromatography–mass spectrometry in the analysis of oxidation products and antioxidants in biological systems. Anal Chim Acta. 2003;492:17–47.CrossRefGoogle Scholar
  173. Weston A, Brown PR. HPLC and CE: principles and practice. Sun Diego: Academic Press; 1997.Google Scholar
  174. WHO, General Guidelines for Methodologies on Research and Evaluation of Traditional Medicines, 2000, p. 1–74.Google Scholar
  175. Wojciak-Kosior M, Matysik G, Skalska A. Densitometric determination of kinetics of hydrolysis of flavonoid glycosides. J Planar Chromatogr. 2004;17:286–9.CrossRefGoogle Scholar
  176. Wolfbeis OS, Knierzinger A, Schipfer R. pH-dependent fluorescence spectroscopy XVII: first excited singlet state dissociation constants, phototautomerism and dual fluorescence of flavonol. J Photochem. 1983;21:67–79.CrossRefGoogle Scholar
  177. Yao LH, Jiang YM, Shi J, Tomas-Barberan FA, Datta N, Singanusong R, et al. Flavonoids in food and their health benefits. Plant Foods Hum Nutr. 2004;59:113–22.PubMedCrossRefGoogle Scholar
  178. Zhong D, Yang B, Chen X, Li K, Xu J. Determination of scutellarin in rat plasma by high-performance liquid chromatography with ultraviolet detection. J Chromatogr B Analyt Technol Biomed Life Sci. 2003;796:439–44.PubMedCrossRefGoogle Scholar
  179. Zhu W, Jia Q, Wang Y, Zhang Y, Xia M. The anthocyanin cyanidin-3-O-𝛽-glucoside, a flavonoid, increases hepatic glutathione synthesis and protects hepatocytes against reactive oxygen species during hyperglycemia: involvement of a cAMP-PKA-dependent signaling pathway. Free Radic Biol Med. 2012;52:314–27.PubMedCrossRefGoogle Scholar
  180. Zogg GC, Nyiredy S, Sticher O. Operating conditions in preparative medium pressure liquid chromatography (MPLC).II. Influence of solvent strength and flow rate of the mobile phase, capacity and dimensions of the column. J Liq Chromatogr. 1989;12:2049–65.Google Scholar

Copyright information

© Springer Nature Singapore Pte Ltd. 2020

Authors and Affiliations

  • Ashim K. Sen
    • 1
  • Dhanya B. Sen
    • 1
  • Rajesh A. Maheshwari
    • 1
  1. 1.Department of PharmacySumandeep Vidyapeeth Deemed to be UniversityVadodaraIndia

Personalised recommendations