Journal of Food Science and Technology

, Volume 52, Issue 9, pp 5790–5798 | Cite as

Reactivity of phenolic compounds towards free radicals under in vitro conditions

  • Sindhu MathewEmail author
  • T. Emilia Abraham
  • Zainul Akmar Zakaria
Original Article


The free radical scavenging activity and reducing power of 16 phenolic compounds including four hydroxycinnamic acid derivatives namely ferulic acid, caffeic acid, sinapic acid and p-coumaric acid, benzoic acid and its derivatives namely protocatechuic acid, gallic acid and vanillic acid, benzene derivatives namely vanillin, vanillyl alcohol, veratryl alcohol, veratraldehyde, pyrogallol, guaiacol and two synthetic antioxidants, butylated hydroxy anisole (BHA) and propyl gallate were evaluated using 1,1-Diphenyl-2-picrylhydrazyl radical (DPPH), 2,2′-Azinobis-3- ethylbenzothiazoline-6-sulfonic acid radical (ABTS+•), Hydroxyl radical (OH) and Superoxide radical (O2 •-) scavenging assays and reduction potential assay. By virtue of their hydrogen donating ability, phenolic compounds with multiple hydroxyl groups such as protocatechuic acid, pyrogallol, caffeic acid, gallic acid and propyl gallate exhibited higher free radical scavenging activity especially against DPPH and O2 •-. The hydroxylated cinnamates such as ferulic acid and caffeic acid were in general better scavengers than their benzoic acid counter parts such as vanillic acid and protocatechuic acid. All the phenolic compounds tested exhibited more than 85 % scavenging due to the high reactivity of the hydroxyl radical. Phenolic compounds with multiple hydroxyl groups also exhibited high redox potential. Exploring the radical scavenging and reducing properties of antioxidants especially those which are found naturally in plant sources are of great interest due to their protective roles in biological systems.


Free radical scavenging Antioxidant activity Phenolic compounds Reducing power 



Protocatechuic acid




Caffeic acid


Ferulic acid


p-Coumaric acid


Gallic acid


Vanillic acid


Sinapinic acid


Propyl gallate




Butylated hydroxy anisole


Veratryl alcohol


Vanillyl Alcohol




Benzoic acid





The authors are thankful to the Director, NIIST (CSIR), India for providing the facilities for carrying out this research work. The first author acknowledges CSIR, India for the Research Fellowship and Universiti Teknologi Malaysia (UTM) for the Postdoctoral Fellowship.


  1. Bilto YY, Suboh S, Aburjai T, Abdalla S (2012) Structure activity relationships regarding the antioxidant effects of the flavonoids on human erythrocytes. Natural Sci 4:740–747CrossRefGoogle Scholar
  2. Bountagkidou OG, Ordoudi SA, Tsimidou MZ (2010) Structure-antioxidant activity relationship study of natural hydroxybenzaldehydes using in vitro assays. Food Res Int 43:2014–2019CrossRefGoogle Scholar
  3. Brand-Williams W, Cuvelier ME, Berset C (1995) Use of free radical method to evaluate antioxidant activity. Lebensm Wiss Technol 28:25–30CrossRefGoogle Scholar
  4. Cai YZ, Mei S, Jie X, Luo Q, Corke H (2006) Structure-radical scavenging activity relationships of phenolic compounds from traditional Chinese medicinal plants. Life Sci 78:2872–2888CrossRefGoogle Scholar
  5. Crozier A, Jaganath IB, Clifford MN (2009) Dietary phenolics: chemistry, bioavailability and effects on health. Nat Prod Rep 26:1001–1043CrossRefGoogle Scholar
  6. Cuvelier ME, Richard H, Berset C (1992) Comparison of the antioxidant activity of some acid phenols: structure-activity relationship. Biosci Biotechnol Biochem 56:324–325CrossRefGoogle Scholar
  7. De Ancos B, Gonzàlez EM, Cano MP (2000) Ellagic acid, vitamin C and total phenolic contents and radical scavenging capacity affected by freezing and frozen storage in raspberry fruit. J Agric Food Chem 48:4565–4570CrossRefGoogle Scholar
  8. Dziedzic SZ, Hudson BJF (1983) Polyhydroxychalcones and flavanones as antioxidants for edible foods. Food Chem 12:205–212CrossRefGoogle Scholar
  9. Grootveld M, Halliwell B (1986) Aromatic hydroxylation as a potential measure of hydroxyl radical formation in vivo. Biochem J 237:499–504CrossRefGoogle Scholar
  10. Gul MZ, Bhakshu LM, Ahmad F, Kondapi AK, Qureshi IA, Ghazi IA (2011) Evaluation of Abelmoschus moschatus extracts for antioxidant, free radical scavenging, antimicrobial and antiproliferative activities using invitro assays. BMC Complement Altern Med 11:64–76CrossRefGoogle Scholar
  11. Halliwell B, Gutteridge JMC, Aruoma OI (1987) The deoxyribose method: a simple test tube assay for determination of rate constants for reactions of hydroxyl radicals. Anal Biochem 165:215–219CrossRefGoogle Scholar
  12. Haseloff RF, Blasig IE, Meffert H, Ebert B (1990) Hydroxyl radical scavenging and antipsoriatic activity of benzoic acid derivatives. Free Rad Biol Med 9:111–115CrossRefGoogle Scholar
  13. Heo BG, Park YJ, Park YS, Bae JH, Cho JY, Park K, Jastrzebski Z, Gorinstein S (2014) Anticancer and antioxidant effects of extracts from different parts of indigo plant. Ind Crop Prod 56:9–16CrossRefGoogle Scholar
  14. Hermann K (1989) Occurrence and content of hydroxycinnamic acid and hydroxybenzoic acid compounds in foods. Crit Rev Food Sci Nutr 28:315–347CrossRefGoogle Scholar
  15. Hsieh CL, Chang CH, Chiang SY, Li TC, Tang NY, Pon CZ, Hsieh CT, Lin JG (2000) Anticonvulsive and free radical scavenging activities of vanillyl alcohol in ferric chloride-induced epileptic seizures in Sprague–Dawley rats. Life Sci 67:1185–1195CrossRefGoogle Scholar
  16. Jing P, Zhao SJ, Jian WJ, Qian BJ, Dong Y, Pang J (2012) Quantitative studies on structure-DPPH scavenging activity relationships of food phenolic acids. Molecules 17:12910–12924CrossRefGoogle Scholar
  17. Kikuzaki H, Hisamoto M, Hirose K, Akiyama K, Taniguchi H (2002) Antioxidant properties of ferulic acid and its related compounds. J Agric Food Chem 50:2161–2168CrossRefGoogle Scholar
  18. Kumar SS, Priyadarsini KI, Sainis KB (2002) Free radical scavenging activity of vanillin and o-vanillin using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. Redox Rep 7:35–40CrossRefGoogle Scholar
  19. Kumbulainen JT, Salonen JT (1999) Natural antioxidants and anticarcinogens in nutrition: Health and diseases. Royal Society of Chemistry, Cambridge, p 12CrossRefGoogle Scholar
  20. Leonardis AD, Macciola V (2003) Effectiveness of caffeic acid as an antioxidant for cod liver oil. Inter J Food Sci Technol 38:475–480CrossRefGoogle Scholar
  21. Lin S, Lai TC, Chen L, Kwok HF, Lau CB, Cheung PCK (2014) Antioxidant and antiangiogenic properties of phenolic extract from Pleurotus tuber-regium. J Agric Food Chem 62:9488–9498CrossRefGoogle Scholar
  22. Lipinski B (2011) Hydroxyl radical and its scavengers in health and disease. Oxid Med Cell Longev 809696:1–9CrossRefGoogle Scholar
  23. Liu J, Mori A (1993) Antioxidant and pro-oxidant activities of p-hydroxybenzyl alcohol and vanillin: effects on free radicals, brain peroxidation and degradation of benzoate, deoxyribose, amino acids and DNA. Neuropharmacol 32:659–669CrossRefGoogle Scholar
  24. Lucini L, Pellizoni M, Pellegrino R, Molinari GP, Colla G (2015) Phytochemical constituents and in vitro radical scavenging activity of different Aloe species. Food Chem 170:501–507CrossRefGoogle Scholar
  25. Marinova EM, Yanishlieva NV (1992) Effect of temperature on the antioxidative action of inhibitors in lipid autoxidation. J Sci Food Agric 60:313–318CrossRefGoogle Scholar
  26. Mathew S, Abraham TE (2006) Invitro antioxidant activity and scavenging effects of Cinnamomum verum leaf extract assayed by different methodologies. Food Chem Toxicol 44:198–206CrossRefGoogle Scholar
  27. Matthaus B (2002) Antioxidant activity of extracts obtained from residues of different oilseeds. J Agric Food Chem 50:3444–3452CrossRefGoogle Scholar
  28. Moure A, Cruz JM, Franco D (2001) Natural antioxidants from residual sources. Food Chem 72:145–171CrossRefGoogle Scholar
  29. Natella F, Nardini M, di Felice M, Scaccini C (1999) Benzoic and cinnamic acid derivatives as antioxidants: Structure-activity relation. J Agric Food Chem 47:1453–1459CrossRefGoogle Scholar
  30. Nenadis N, Wang LF, Tsimidou M, Zhang HY (2004) Estimation of scavenging activity of phenolic compounds using the ABTS assay. J Agric Food Chem 52:4669–4674CrossRefGoogle Scholar
  31. Nicodemus KK, Jacobs DR, Folsom AR (2001) Whole and refined grain intake and risk of incident postmenopausal breast cancer. Cancer Cause Control 12:917–925CrossRefGoogle Scholar
  32. Pacifico S, Gallicchio M, Lorenz P, Duckstein SM, Potenza N, Galasso S, Marciano S, Fiorentino A, Stintzing FC, Monaco P (2014) Neuroprotective potential of Laurus nobilis antioxidant polyphenol enriched leaf extracts. Chem Res Toxicol 27:611–626CrossRefGoogle Scholar
  33. Pino E, Campos AM, Lopez-Alarcon C, Aspee A, Lissi E (2006) Free radical scavenging capacity of hydroxycinnamic acids and related compounds. J Phys Org Chem 19:759–764CrossRefGoogle Scholar
  34. Pokorny J (1987) Major factors affecting the autooxidation of lipids. In: Chan HWS (ed) Autooxidation of unsaturated lipids. Academic, London, pp 141–206Google Scholar
  35. Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice-Evans C (1999) Antioxidant activity applying an improved ABTS radical cation decolorisation assay. Free Radical Biol Med 26:1231–1237CrossRefGoogle Scholar
  36. Rice Evans CA, Miller NJ, Paganga G (1996) Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Rad Biol Med 20:933–956CrossRefGoogle Scholar
  37. Robbins RJ (2003) Phenolic acids in foods: An overview of analytical methodology. J Agric Food Chem 51:2866–2887CrossRefGoogle Scholar
  38. Shahidi F, Wanasundara PKJ (1992) Phenolic antioxidants. Crit Rev Food Sci Nutr 32:67–103CrossRefGoogle Scholar
  39. Siquet C, Paiva-Martins F, Lima JL, Reis S, Borges F (2006) Antioxidant profile of dihydroxy- and trihydroxyphenolic acids–a structure activity relationship study. Free Radical Res 40:433–442CrossRefGoogle Scholar
  40. Sivakumar PM, Prabhakar PA, Doble M (2011) Synthesis, antioxidant evaluation and quantitative structure-activity relationship studies of chalcones. Med Chem Res 20:482–492CrossRefGoogle Scholar
  41. Teixeira J, Gaspar A, Garrido EM, Borges F (2013) Hydroxycinnamic acid antioxidants: An electrochemical overview. Biomed Res Int 2013, Article ID 251754, 11 pagesGoogle Scholar
  42. Torres de Pinedo A, Penalver P, Morales JC (2007) Synthesis and evaluation of new phenolic-based antioxidants: structure-activity relationship. Food Chem 103:55–61CrossRefGoogle Scholar
  43. Vafiadis AP, Bakalbassis EG (2003) A computational study of the structure–activity relationships of some p-hydroxybenzoic acid antioxidants. J Am Oil Chem Soc 80:1217–1223CrossRefGoogle Scholar
  44. Velkov ZA, Kolev MK, Tadjer AV (2007) Modeling and statistical analysis of DPPH scavenging activity of phenolics. Collec Czech Chem Commun 72:1461–1471CrossRefGoogle Scholar
  45. von Gadow A, Joubert E, Hansmann CF (1997) Comparison of the antioxidant activity of aspalathin with that of other plant phenols of Rooibos tea (Aspalathus linearis), alpha-Tocopherol, BHT, and BHA. J Agric Food Chem 45:632–638CrossRefGoogle Scholar
  46. Wardman P (1989) Reduction potentials of one-electron couples involving free radicals in aqueous solutions. J Phys Chem Ref Data 18:1637–1755CrossRefGoogle Scholar
  47. Yamagami C, Akamatsu M, Motohashi N, Hamada S, Tanahashi T (2005) Quantitative structure-activity relationship studies for antioxidant hydroxybenzalacetones by quantum chemical-and 3-D-QSAR(CoMFA) analyses. Bioorg Med Chem Lett 15:2845–2850CrossRefGoogle Scholar
  48. Yanishlieva N, Marinova EM (1995) Effects of antioxidants on the stability of triacylglycerols and methyl esters of fatty acids of sunflower oil. Food Chem 54:377–382CrossRefGoogle Scholar
  49. Zhang J, Stanley RA, Adaim A, Melton LD, Skinner MA (2006) Free radical scavenging and cytoprotective activities of phenolic antioxidants. Mol Nutr Food Res 50:996–1005CrossRefGoogle Scholar

Copyright information

© Association of Food Scientists & Technologists (India) 2015

Authors and Affiliations

  • Sindhu Mathew
    • 1
    • 3
    Email author
  • T. Emilia Abraham
    • 1
  • Zainul Akmar Zakaria
    • 2
  1. 1.Chemical Science and Technology DivisionNational Institute of Interdisciplinary Science and Technology (NIIST, CSIR)TrivandrumIndia
  2. 2.Institute of Bioproduct DevelopmentUniversiti Teknologi MalaysiaJohor BahruMalaysia
  3. 3.Institute of Bioproduct DevelopmentUniversiti Teknologi MalaysiaJohor BahruMalaysia

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