Comparative Spectrophotometric Studies of  Total Phenolics Content and Antioxidant Capacity Measured by DPPH and ABTS Methods in  Green Vegetables

Sofija Doganjić; Snežana Uskoković-Marković; Jelena Kukić Marković; Leposava Pavun; Aleksandra  Janošević Ležaić

doi:10.5937/arhfarm75-59496

Sofija Doganjić University of Belgrade - Faculty of Medicine, Belgrade, Serbia
Snežana Uskoković-Marković University of Belgrade – Faculty of Pharmacy, Department of Analytical Chemistry, Belgrade, Serbia
Jelena Kukić Marković University of Belgrade - Faculty of Pharmacy, Department of Pharmacognosy, Belgrade, Serbia
Leposava Pavun University of Belgrade – Faculty of Pharmacy, Department of Physical Chemistry and Instrumental Methods, Belgrade, Serbia
Aleksandra Janošević Ležaić University of Belgrade – Faculty of Pharmacy, Department of Physical Chemistry and Instrumental Methods, Belgrade, Serbia

DOI: https://doi.org/10.5937/arhfarm75-59496

Keywords: green vegetables, antioxidative activities, total phenol content (TPC), DPPH, ABTS

Abstract

Modern trends in nutrition imply the daily intake of fresh fruits and vegetables, with the statement that they are important sources of natural antioxidants that protect the body from oxidative stress and can contribute to the prevention of many chronic diseases. In these recommendations, green vegetables with high total phenolics content (TPC) are especially highlighted. However, one must ask whether the high total phenolics content of green vegetables, such as Brussels sprouts, cabbage, broccoli, parsley, borecole and spinach, is a reliable indicator of antioxidant activity for regulating free radicals in the body. The aim of this work is the spectrophotometric determination of TPC of hydromethanol extracts of selected green vegetables and its correlation with their antioxidant potential. For this purpose, DPPH and ABTS radical scavenging tests of extracts were performed. The obtained results indicate that Brussels sprouts and parsley exhibit the highest antioxidative activities, while cabbage extract shows lower values. The Pearson correlation used to test the correlation between the TPC obtained for green vegetable extracts and the results of the DPPH and ABTS tests shows a moderate correlation between TPC and ABTS values, whereas the correlation for TPC and DPPH is statistically significant.

References

Healthy lifestyle. Nutrition and healthy eating: Add antioxidants to your diet [Internet]. Mayo Clinic; 2025 [cited 2025 Jan 25]. Available from: https://www.mayoclinic.org/healthy-lifestyle/nutrition-and-healthy-eating/in-depth/add-antioxidants-to-your-diet/art-20546814?s=1.

Barsby JP, Cowley JM, Leemaqz SY, Grieger JA, McKeating DR, Perkins AV, et al. Nutritional properties of selected superfood extracts and their potential health benefits. PeerJ. 2021;9:e12525. doi: 10.7717/peerj.12525.

Soengas P, Velasco P, Fernández JC, Cartea ME. New Vegetable Brassica Foods: A Promising Source of Bioactive Compounds. Foods. 2021;10(12):2911. doi: 10.3390/foods10122911.

Ko S-H, Park J-H, Kim S-Y, Lee SW, Chun S-S, Park E. Antioxidant Effects of Spinach (Spinacia oleracea L.) Supplementation in Hyperlipidemic Rats. Prev Nutr Food Sci. 2014;19(1):19–26. doi: 10.3746/pnf.2014.19.1.019.

Waheeba EA, Ali AE-S, Ihssan MO. Antioxidant Activities of Parsley (Petroselinum crispum) on the Induced Biochemical and Histopathological Changes of Potassium Bromate-Fed Rats. PJN. 2020;19:80–5.

Shetty AA, Magadum S, Managanvi K. Vegetables as Sources of Antioxidants. J Food Nutr Disor. 2013;2:1. doi: 10.4172/2324-9323.1000104.

Rudrapal M, Khairnar SJ, Khan J, Dukhyil AB, Ansari MA. Dietary Polyphenols and Their Role in Oxidative Stress-Induced Human Diseases: Insights Into Protective Effects, Antioxidant Potentials and Mechanism(s) of Action. Front Pharmacol. 2022;13. doi: 10.3389/fphar.2022.806470.

Chandimali N, Bak SG, Park EH, Lim HJ, Won YS, Kim EK, et al. Free radicals and their impact on health and antioxidant defenses: a review. Cell Death Discov. 2025;11:19. doi: 10.1038/s41420-024-02278-8.

Lobo V, Patil A, Phatak A, Chandra N. Free radicals, antioxidants and functional foods: Impact on human health. Phcog Rev. 2010;4(8):118–26. doi: 10.4103/0973-7847.70902.

Langseth L. Oxidants, Antioxidants, and Disease Prevention. Washington: ILSI Press; 1995.

Halliwell B, Whitema M. Measuring reactive species and oxidative damage in vivo and in cell culture: how should you do it and what do the results mean? Br J Pharmacol. 2004;142(2):231–55.

Marčetić M, Arsenijević J. The Antioxidant activity of plant secondary metabolites. Arh farm. 2023;73(4):264–77. doi: 10.5937/arhfarm73-45560.

Kasote DM, Katyare SS, Hegde MV, Bae H. Significance of antioxidant potential of plants and its relevance to therapeutic applications. Int J Biol Sci. 2015;11(8):982–91.

Rappoport Z. The Chemistry of Phenols, Part 2. Chichester: John Wiley and Sons Ltd; 2003.

Dziki D, Różyło R, Gawlik-Dziki U, Świeca M. Current trends in the enhancement of antioxidant activity of wheat bread by the addition of plant materials rich in phenolic compounds. Trends Food Sci Technol. 2014;40(1):48–61. doi: 10.1016/j.tifs.2014.07.010.

Halliwell, B. The chemistry of free radicals. Toxicol Ind Health. 1993;9(1–2):1–21.

Pham-Huy LA, He H, Pham-Huy C. Free radicals, antioxidants in disease and health. IJBS. 2008;4(2):89–96.

Babović N, Djilas S, Jadranin M, Vajs V, Ivanović J, Petrović S, et al. Supercritical carbon dioxide extraction of antioxidant fractions from selected Lamiaceae herbs and their antioxidant capacity. IFSET. 2010;11(1):98–107.

Flieger J, Flieger W, Baj J, Maciejewski R. Antioxidants: Classification, Natural Sources, Activity/Capacity Measurements, and Usefulness for the Synthesis of Nanoparticles. Materials. 2021;14(15):4135. doi: 10.3390/ma14154135.

Khan NS, Ahmad A, Hadi S. Anti–oxidant, pro–oxidant properties of tannic acid and its binding to DNA. Chem Biol Interact. 2000;125(3):177–89.

Sule RO, Condon L, Gomes AV. A Common Feature of Pesticides: Oxidative Stress–The Role of Oxidative Stress in Pesticide-Induced Toxicity. Oxid Med Cell Longev. 2022;2022:5563759. doi: 10.1155/2022/5563759.

Sakanaka S, Tachibana Y, Okada Y. Preparation and antioxidant properties of extracts of Japanese persimmon leaf tea (kakinoha–cha). Food Chem. 2005;89(4):569–75.

Ainsworth EA, Gillespie KM. Estimation of total phenolic content and other oxidation substrates in plant tissues using Folin–Ciocalteu reagent. Nat Protoc. 2007;2(4):875–7.

Ford L, Theodoridou K, Sheldrake G, Walsh P. A critical review of analytical methods used for the chemical characterisation and quantification of phlorotannin compounds in brown seaweeds. Phytochem Anal. 2019;30(6):587–99.

Kumar S, Sandhir R, Ojha S. Evaluation of antioxidant activity and total phenol in different varieties of Lantana camara leaves. BMC Res Notes. 2014;7:560. doi: 10.1186/1756-0500-7-560.

Katanić J, Boroja T, Stanković N, Mihailović V, Mladenović M, Kreft S, et al. Bioactivity, stability and phenolic characterization of Filipendula ulmaria (L.) Maxim. Food Funct. 2015;6(4):1164–75.

Kedare SB. Genesis and development of DPPH method of antioxidant assay. J Food Technol. 2011;48(4):412–22.

Gledović A, Janošević Lezaić A, Krstonošić V, Djoković J, Nikolić I, Bajuk-Bogdanović D, et al. Low-energy nanoemulsions as carriers for red raspberry seed oil: Formulation approach based on Raman spectroscopy and textural analysis, physicochemical properties, stability and in vitro antioxidant/ biological activity. Plos one. 2020;15(4):e0230993. doi: 10.1371/journal.pone.0230993.

Hsu CF, Peng H, Basle C, Travas-Sejdic J, Kilmartin PA. ABTS•+ scavenging activity of polypyrrole, polyaniline and poly(3,4-ethylenedioxythiophene). Polym Int. 2011;60(1):69–77.

Ilyasov IR. ABTS/PP Decolorization Assay of Antioxidant Capacity Reaction Pathways. Int J Mol Sci. 2020;21(3):1131. doi: 10.3390/ijms21031131.

Kukić-Marković J. Methods for the determination of antioxidant activity of plant extracts in vitro. Arch Pharm. 2023;73:318–35.

Apak R, Gorinstein S, Böhm V, Schaich KM, Özyürek M, Güçlü K. Methods of measurement and evaluation of natural antioxidant capacity/activity (IUPAC Technical Report). Pure Appl Chem. 2013;85(5):957–98. doi: 10.1351/pac-rep-12-07-15.

Li Y, Zhang J-J, Xu D-P, Zhou T, Zhou Y, Li S, et al. Bioactivities and health benefits of wild fruits. Int J Mol Sci. 2016;17(8):1258. doi: 10.3390/ijms17081258.

Podsędek A. Natural antioxidants and antioxidant capacity of Brassica vegetables: A review. LWT – Food Sci Technol. 2007;40(1):1–11.

Doniec J, Florkiewicz A, Dziadek K, Filipiak-Florkiewicz A. Hydrothermal Treatment Effect on Antioxidant Activity and Polyphenols Concentration and Profile of Brussels sprouts (Brassica oleracea var. gemmifera) in an In Vitro Simulated Gastrointestinal Digestion Model. Antioxidants. 2022;11(3):446. doi: 10.3390/antiox11030446.

Kamboj A, Sharma S, Singh VP, Sinha A, Yadav KS, Lal B, et al. Phytochemical and therapeutic potential of broccoli (Brassica oleracea): A review. Pharma Innovation. 2023;12(6):633–8.

Statilko O, Tsiaka T, Sinanoglou VJ, Strati IF. Overview of Phytochemical Composition of Brassica oleraceae var. capitata Cultivars. Foods. 2024;13(21):3395. doi: 10.3390/foods13213395.

Cartea ME, Francisco M, Soengas P, Velasco P. Phenolic Compounds in Brassica Vegetables. Molecules. 2011;16(1):251–80. doi: 10.3390/molecules16010251.

Vallejo F, Tomas-Barberan F, Garcia–Viguera C. Health–promoting compounds in broccoli as influenced by refrigerated transport and retail sale period. J Agric Food Chem. 2003;51(10):3029–34.

Pérez M, Dominguez–López I, Lamuela–Raventós RM. The Chemistry Behind the Folin–Ciocalteu Method for the Estimation of (Poly)phenol Content in Food: Total Phenolic Intake in a Mediterranean Dietary Pattern. J Agric Food Chem. 2023;71(46):17543–53.

Torres P, Osaki S, Silveira E, dos Santos DYAC, Chow F. Comprehensive evaluation of Folin-Ciocalteu assay for total phenolic quantification in algae (Chlorophyta, Phaeophyceae, and Rhodophyta). Algal Res. 2024;80:103503. doi: 10.1016/j.algal.2024.103503.

Halvorsen BL, Holte K, Myhrstad MCW, Barikmo I, Hvattum E, Remberg SF, et al. A systematic screening of total antioxidants in dietary plants. J Nutr. 2002;132(3):461–71. doi: 10.1093/jn/132.3.461.

Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, Rice–Evans C. Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med. 1999;26(9–10):1231–7.

Prior RL, Wu X, Schaich K. Standardized methods for the determination of antioxidant capacity and phenolics in foods and dietary supplements. J Agric Food Chem. 2005;53(10):4290–302.

Thaipong K, Boonprakob U, Crosby K, Cisneros-Zevallos L, Hawkins Byrne D. Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimating antioxidant activity from guava fruit extracts. J Food Compost Anal. 2006;19(6-7):669–75.

George J, Edwards D, Pun S, Williams D. Evaluation of Antioxidant Capacity (ABTS and CUPRAC) and Total Phenolic Content (Folin–Ciocalteu) Assays of Selected Fruit, Vegetables, and Spices. Int J Food Sci. 2022;2022:2581470. doi: 10.1155/2022/2581470.