Extensive Screening for Edible Herbal Extracts with Potent Scavenging Activity against Superoxide Anions


To search for edible herbal extracts with potent antioxidant activity, we conducted a large scale screening based on the superoxide scavenging activity. That is, scavenging activity against superoxide anions were extensively screened from ethanol extracts of approximately 1,000 kinds of herbs by applying an electron spin resonance (ESR)-spin trapping method. Among them we chose four edible herbal extracts with prominently potent ability to reduce the signal intensity of 5,5-dimethyl-1-pyrroline-N-oxide (DMPO)–OOH, a spin adduct formed by DMPO and superoxide anion. They are the extracts from Punica granatum (Peel), Syzygium aromaticum (Bud), Mangifera indica (Kernel), and Phyllanthus emblica (Fruit), and are allowed to be used as foodstuffs according to the Japanese legal regulation. The ESR-spin trapping method coupled with steady state kinetic analysis showed that all of the four extracts directly scavenge superoxide anions, and that the superoxide scavenging potential of any of the extracts was comparable to that of L-ascorbic acid. Furthermore, polyphenol determination indicates that the activity is at least in part attributable to polyphenols. These results with such large scale screening might give useful information when choosing a potent antioxidant as a foodstuff.

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superoxide dismutase


electron spin resonance




xanthine oxidase




  1. 1.

    Skulachev VP (2005) Biogenetic aspects of apoptosis, necrosis and mitoptosis. Apoptosis 4:473–485

    Google Scholar 

  2. 2.

    Wallace DC (2005) A mitochondrial paradigm of metabolic and degenerative diseases, aging, and cancer: a dawn for evolutionary medicine. Annu Rev Genet 39:359–407

    Article  CAS  Google Scholar 

  3. 3.

    Jezak P, Hlavata L (2005) Mitochondria in homeostasis of reactive oxygen species in cell, tissues, and organism. Int J Biochem Cell Biol 37:2478–2503

    Article  CAS  Google Scholar 

  4. 4.

    Keele BB, McCord JM, Fridovoch I (1970) Superoxide dismutase from Escherichia coli B. J Biol Chem 245:6176–6181

    CAS  Google Scholar 

  5. 5.

    Goldstein S, Meyerstein D, Czapski G (1993) The Fenton reagents. Free Radic Biol Med 15:435–445

    Article  CAS  Google Scholar 

  6. 6.

    Haber F, Weiss J (1934) The catalytic decomposition of hydrogen peroxide by iron salts. Proc R Soc London Ser A 147:332–351

    CAS  Google Scholar 

  7. 7.

    Buettner GR (1987) Spin trapping: ESR parameters of spin adducts. Free Rad Biol Med 3:259–303

    Article  CAS  Google Scholar 

  8. 8.

    Rosen H, Klebanoff SJ (1979) Hydroxyl radical generation by polymorphonuclear leukocytes measures by electron spin resonance spectroscopy. J Clin Invest 64:1725–1729

    Article  CAS  Google Scholar 

  9. 9.

    Tanabe K, Masuda A, Hirayama S, Nagase S, Kono I, Kuno S (2006) Effect of spontaneous exercise on antioxidant capacity in rat muscles determined by electron spin resonance. Acta Physiol 186:119–125

    Article  CAS  Google Scholar 

  10. 10.

    Opie LH, Lecour S (2007) The red wine hypothesis: from concepts to protective signaling molecules. Eur Heart J 28:1683–1693

    Article  CAS  Google Scholar 

  11. 11.

    Karatzi KN, Papamichael CM, Karatzis EN, Papaioannou TG, Aznaouridis KA, Katsichti PP, Stamatelopoulos KS, Zampelas A, Lekakis JP, Mavrikakis ME (2005) Red wine acutely induces favorable effects wave reflections and central pressures in coronary artery disease patients. Am J Hypertens 18:1161–1167

    Article  CAS  Google Scholar 

  12. 12.

    Papamichael C, Karatzi K, Karatzis E, Papaioannou TG, Katsichti P, Zampelas A, Lekakis J (2006) Combined acute effects of red wine consumption and cigarette smoking on haemodynamics of young smokers. J Hypertens 24:1287–1292

    Article  CAS  Google Scholar 

  13. 13.

    Sato M, Maulik N, Das DK (2002) Cardioprotection with alcohol: role of both alcohol and polyphenolic antioxidants. Ann NY Acad Sci 957:122–135

    CAS  Article  Google Scholar 

  14. 14.

    Martinez J, Moreno JJ (2000) Effect of resveratrol, a natural polyphenolic compound, on reactive oxygen species and prostaglandin production. Biochem Pharmacol 59:865–870

    Article  CAS  Google Scholar 

  15. 15.

    Leonard SS, Xia C, Jiang BH, Stinefelt B, Klandorf H, Harris GK, Shi X (2003) Resveratrol scavenges reactive oxygen species and effects radical-induced cellular responses. Biochem Biophys Res Commun 309:1017–1026

    Article  CAS  Google Scholar 

  16. 16.

    Frankel EN, Kanner J, German JB, Parks E, Kinsella JE (1993) Inhibition of human low-density lipoprotein by phenolic substances in red wine. Lancet 341:454–457

    Article  CAS  Google Scholar 

  17. 17.

    Kanner J, Frankel EN, Granit R, German B, Kinsella JE (1994) Natural antioxidant in grapes and wines. J Agric Food Chem 42:64–69

    Article  CAS  Google Scholar 

  18. 18.

    Kinsella JE, Frankel EN, German JB, Kanner J (1993) Possible mechanisms for the protective role of antioxidants in wine and plant foods. Food Technol 47(4):85–89

    CAS  Google Scholar 

  19. 19.

    Sato E, Kohno M, Hamano H, Niwano Y (2006) Increased anti-oxidative potency of garlic by spontaneous short-term fermentation. Plant Foods Hum Nutr 61:157–160

    Article  CAS  Google Scholar 

  20. 20.

    Sato E, Niwano Y, Matsuyama Y, Kim D, Nakashima T, Oda T, Kohno M (2007) Some dinophycean red tide planktons generate a superoxide scavenging substance. Biosci Biotechnol Biochem 71:704–710

    Article  CAS  Google Scholar 

  21. 21.

    Niwano Y, Sato E, Kohno M, Matsuyama Y, Kim D, Oda T (2007) Antioxidant properties of aqueous extracts from red tide plankton cultures. Biosci Biotechnol Biochem 71:1145–1153

    Article  CAS  Google Scholar 

  22. 22.

    Kohno M, Mizuta Y, Kusai M, Masumizu T, Makino K (1994) Measurements of superoxide anion radical and superoxide anion scavenging activity by electron spin resonance spectroscopy coupled with DMPO spin trapping. Bull Chem Soc Jpn 67:1085–1090

    Article  CAS  Google Scholar 

  23. 23.

    Schanderl SH (1970) Tannins and related phenolics. In: Joslyn MA (ed) Methods in Food Analysis. Academic, New York, pp 701–724

    Google Scholar 

  24. 24.

    Watts RW, Wats JA, Seegmiller LE (1965) Xanthine oxidase activity in human tissues and its inhibition by allopurinol (4-hydroxypyarazolo[3,4-d] pyrimidine). J Lab Clin Med 66:688–697

    CAS  Google Scholar 

  25. 25.

    Nicoué EE, Savard S, Belkacemi K (2007) Anthocyanins in Wild Blueberries of Quebec: Extraction and Identification. J Agric Food Chem 55:5626–5635

    Article  CAS  Google Scholar 

  26. 26.

    Cui T, Nakamura K, Tian S, Kayahara H, Yian YL (2006) Polyphenolic content and physiological activities of Chinese hawthorn extracts. Biosci.Biotechnol Biochem 50:2948–2956

    Article  CAS  Google Scholar 

  27. 27.

    Cerdá B, Llorach R, Cerón JJ, Espín JC, Tomás-Barberán FA (2003) Evaluation of the bioavailability and metabolism in the rat of punicalagin, an antioxidant polyphenols from pomegranate juice. Eur J Nutr 42:18–28

    Article  CAS  Google Scholar 

  28. 28.

    Prabhu S, Jainu M, Sabitha KE, Devi CS (2006) Role of mangiferin on biochemical alterations and antioxidant status in isoproterenol-induced myocardial infarction in rats. J Ethnopharmacol 107:126–133

    Article  CAS  Google Scholar 

  29. 29.

    Habib-ur-Rehman, Yasin KA, Choudhary MA, Khaliq N, Atta-ur-Rahman, Choudhary MI, Malik S (2007) Studies on the chemical constituents of Phyllanthus emblica. Nat Prod Res 21:775–781

    Article  CAS  Google Scholar 

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This work was supported by a grant of Kampou Science Foundation.

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Correspondence to Yoshimi Niwano.

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Saito, K., Kohno, M., Yoshizaki, F. et al. Extensive Screening for Edible Herbal Extracts with Potent Scavenging Activity against Superoxide Anions. Plant Foods Hum Nutr 63, 65–70 (2008). https://doi.org/10.1007/s11130-008-0071-2

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  • Edible herbal extracts
  • Superoxide scavenging activity