• Agrawal Jyoti
  • Kar Anand
  • Panda Sunanda


Objective: The present study was aimed to find out the comparative in vitro antioxidative activity of three commercially available polyherbal formulations (PHFs) and Gymnema sylvestre, the major constituent of all the test formulations.

Methods: Different in-vitro free radical scavenging assays such as 1, 1-Diphenyl-2-picrylhydrazyl (DPPH), Nitric oxide (NO), reducing power assay and ferrous sulphate (FeSO4) induced lipid peroxidation (LPO) in liver and kidney homogenates were performed in four parallel sets for each drug. Total polyphenols and flavonoid contents were also estimated.

Results: Although all the test drugs exhibited significant free radical scavenging activity, Madhuhari was found to be the most effective one against DPPH, NO and FeSO4 induced free radical systems. However, their higher concentrations were peroxidative in nature.

Conclusion: Poly herbal formulations were more antiperoxidative as compared to mono herb preparation. As all the test drugs showed decreased efficacy and/or peroxidative effects at their higher concentrations, their higher dose should be avoided. Significant amount of polyphenols and flavonoids present in the test drugs supported their antioxidative property. This study appears to be the first one to demonstrate a comparative protective as well as harmful effect(s) at higher concentration of three herbal formulations.

Keywords: Antioxidants, DPPH, harmful effects, herbal drugs, lipid peroxidation


1. Kekuda TRP, Swarnalatha SP, Preethi HR, Surabhi KS, Sudharshan SJ, Vinayaka KS. Antioxidant and antihelminthic activity of two polyherbal antidiabetic formulations used in Ayureda. Drug Inven Today 2009; 1: 50-54.
2. Cohen PA, Ernst E. Safety of Herbal Supplements: A Guide for Cardiologists. Cardiovasc Ther 2010; 28: 246–53.
3. Halliwell B. Lipid peroxidation, antioxidants and cardiovascular disease: how should we move forward? Cardiovasc Res 2000; 47: 410–18.
4. Samuel V, Johncy S. Evaluation of Lipid Peroxidation and Antioxidant Status in Diabetes with and without Complications. J Biomed Sci Res 2010; 2: 162-66.
5. Rachh PR, Patel SR, Hirpara HV, Rupareliya MT, Rachh MR, Bhargava AS et al. In vitro evaluation of antioxidant activity of Gymnema sylvestre r. br. Leaf extract. Rom J Biol Plant Biol 2009; 54: 141–48.
6. Kajaria DK, Gangwar M, Sharma AK, Nath G, Tripathi YB, Tripathi JS et al. Comparative Evaluation of Phenol and Flavonoid Content of Polyherbal Drugs. Pharmacologyonline 2011; 3: 1365-73.
7. Ji HF, Li XJ, Zhang HY. Natural products and drug discovery. Eur Mol Biol Org 2009; 10: 194-200.
8. Parmar HS, Kar A. Antiperoxidative, Antithyroidal, Antihyperglycemic and Cardioprotective Role of Citrus sinensis Peel Extract in Male Mice. Phytotherapy Res 2008; 22: 791–95.
9. Khanna N, Arora D, Halder S, Mehta AK, Garg GR, Sharma SB et al. Comparative effects of Ocimum sanctum, Commiphora mukul, folic acid ramipril on lipid peroxidation in experimentally-induced hyperlipidemia. Ind j Exp Biol 2010;48: 299-305.
10. Chatterjee K, Ali KM, De D, Panda DK, Ghosh D. Antidiabetic and Antioxidative activity of Ethyl acetate Fraction of Hydromethanolic Extract of Seed of Eugenia jambolana Linn Through In-Vivo and In-Vitro Study and its Chromatographic Purification. Free Rad and Antioxidants 2012;2: 21-30.
11. Ernst E. How the public is being misled about complementary/alternative medicine. R S Med 2008;101: 528–30.
12. Ai G, Huang Z, Wang D, Liu Z. Study on toxicity of hyperoside in rat embryo- fetal development. China Chinese materia medica 2012;37: 2452-55.
13. Yama OE, Bassey RB, Amah CI, Oyebadejo SA, Oremosu A. An Antagonistic salubrious effect of Tahitian Noni in Momordica charantia-induced cytoarchitectural alterations in rat testes: Parallel light microscopic findings. Int J Nutr Met 2012;4: 40-44.
14. Panda S, Kar A. How safe is neem extract with respect to thyroid function in male mice. Pharmacol res 2000;41: 419-22.
15. Kiyohara H, Matsumoto T, Yamada H. Combination Effects of Herbs in a Multi-herbal Formula: Expression of Juzen-taiho-to’s Immuno-modulatory Activity on the Intestinal Immune System. eCAM 2004;1: 83–91.
16. Chan E, Tan M, Xin J, Sudarsanam S, Johnson DE. Interactions between traditional Chinese medicines and Western therapeutics. Curr Opin Drug Disc Dev 2010;13: 50-65.
17. Posadzki P, Watson L, Ernst E. Contamination and adulteration of herbal medicinal products (HMPs): an overview of systematic reviews. Eur J Clin Pharmaco 2013; 69: 295–307.
18. Olesen C, Harbig P, Barat I, Damsgaard EM. Absence of 'over-the-counter' medicinal products in on-line prescription records: a risk factor of overlooking interactions in the elderly. Pharmaco epidemiol Drug Saf 2013;22: 145-50.
19. Panda S, Kar A. Piperine lowers the serum concentrations of thyroid hormones, glucose and hepatic 5’D activity in adult male mice. Horm Met Res 2003; 35: 523-26.
20. Parmar HS, Kar A. Comparative analysis of free radical scavenging potential of several fruit peel extracts by in vitro methods. Drug Disc Therap 2009; 3: 49-55.
21. Kundu PK, Chatterjee PS. Meta-analysis of Diabecon Tablets: Efficacy and Safety Outcomes from 15 Clinical Trials in Diabetes Mellitus. Ind J Clin Prac 2010;20: 653-59.
22. Alam N, Bristi NJ, Zaman R. Review on in vivo and in vitro methods evaluation of antioxidant activity. R Saudi Pharma 2013;21: 143–52.
23. Leontowicz M, Gorinstein S, Leontowicz H, Krzeminski R, Lojek A, Katrich E et al. Apple and pear peel and pulp and their influence on plasma lipids an antioxidant potentials in rats fed cholesterol-containing diets. J Agri Food Chem 2003;51: 5780-85.
24. Dixit Y, Kar A. Antioxidative activity of some vegetable peels determined in vitro by inducing liver lipid peroxidation. Food Res Int 2009;42: 1351–54.
25. Wang CC, Li L, Tang LY, Leung PC. Safety evaluation of commonly used Chinese herbal medicines during pregnancy in mice. Hum Reprod 2012;27: 2448-56.
26. Lim YY, Lim TT, Tee JJ. Antioxidant properties of several tropical fruits: A comparative study. Food Chem 2007;103: 1003-08.
27. Ohkawa H, Ohishi N, Yagi K. Assays of lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Bioche 1979;95: 351-58.
28. Nain P, Saini V, Nain J. Antidiabetic and antioxidant potential of Emblica officinalis Gaertn. leaves extract in streptozotocin-induced type-2 diabetes mellitus (T2DM) rats. J Ethanopharmacol 2012;26: 65-71.
29. Zhou W, Chai H, Lin PH, Lumsden AB, Yao Q, Chen C. Molecular mechanisms and clinical applications of ginseng root for cardiovascular disease. Med Sci Monit 2004;10: 187-92.
30. Gutierrez RMP, Guzman MD. Meliacinolin: A Potent α-Glucosidase and α-Amylase Inhibitor Isolated from Azadirachta indica Leaves and in Vivo Antidiabetic Property in Streptozotocin-Nicotinamide-Induced Type 2 Diabetes in Mice. Biol Pharma Bul 2012;35: 1516–24.
31. Sole SS, Srinivasan BP, Akarte AS. Anti-inflammatory action of Tamarind seeds reduces hyperglycemic excursion by repressing pancreatic β-cell damage and normalizing SREBP-1c concentration. Pharma Biol 2013;51: 350-60.
32. Naowaboota J, Pannangpetcha P, Kukongviriyapana V, Kukongviriyapanb U, Nakmareongb S, Itharatc A. Mulberry leaf extract restores arterial pressure in streptozotocin-induced chronic diabetic rats. Nutr Res 2009;29: 602–08.
33. Baliga MS, Jagetia GC, Ulloor JN, Baliga MP, Venkatesh P, Reddy R et al. The evaluation of the acute toxicity and long term safety of hydroalcoholic extract of Sapthaparna (Alstonia scholaris) in mice and rats. Toxicol. Lett 2004;15: 317-26.
34. Hussain SA, Shafiuddin M, Suresh DK, Parmar J. Hepatoprotective and antioxidant activity of polyherbal formulation. Int J Drug form res 2011;2: 192-210.
35. Parasuram S, Kumar EP, Kumar A, Emerson SF. Free radicals scavenging property and diuretic effects of triglize, a poly herbal formulation in the experimental model. J pharma pharmac ther 2010;1: 38-41.
36. Wu SJ, Ng LT. Antioxidant and free radical scavenging activities of wild bitter melon (Momordica charantia Linn. var. abbreviata Ser.) in Taiwan. Food Sci Tech 2008;41: 323–30.
37. Ramadas D, Srinivas L. Antioxidant effects of 28kda antioxidant protein from turmeric (Curcuma longa). Asian J Pharm Clin Res 2011; 4: 119-23.
38. Niki E, Yoshida Y, Saito Y, Noguchi N. Lipid peroxidation: Mechanisms, inhibition, and biological effects. Biochem Biophys Res Commun 2005;338: 668–76.
39. Marlesa RJ, Farnsworth NR. Antidiabetic plants and their active constituents. Phytomed 1995;2: 137-89.
40. Mansoory N, Jarald E, Edwin S, Agarwal N, Silawat N. Antidiabetic Activity of Hydroalcholic Extract of Herbal Marketed Product Madhuhari Churna in Alloxan - Induced Diabetic Rats. Int J Pharma Biol Arc 2010;1: 164-68.
41. Purwantiningsih, Hussin AH, Chan KM. Herb­drug interaction studies of Eurycoma longifolia extract taf­273 on the Metabolism of rosiglitazone, an antidiabetic drug. Asian J Pharm Clin Res 2011; 4: 88-92.
42. Mitra R, Mazumder PM, Sasmal D. Comparative evaluation of the efficacy of some Ayurvedic formulations in attenuating the progression of diabetic nephropathy. Pharmacologyonline 2010;1: 709-17.
43. Chen CW, Ho CT. Antioxidant properties of polyphenols extracted from green and black tea. Food Lipids 1995;1: 35-46.
44. Parmar HS, Kar A. Protective role of Citrus sinensis, Musa paradisiaca, and Punica granatum peels against diet-induced atherosclerosis and thyroid dysfunctions in rats. Nutr Res 2007; 27: 710–18.
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How to Cite
Jyoti, A., K. Anand, and P. Sunanda. “SYNERGISTIC ACTION OF PHYTOCHEMICALS AUGMENTS THEIR ANTIOXIDATIVE EFFICACY: AN IN VITRO COMPARATIVE STUDY”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 6, no. 8, Sept. 2013, pp. 121-6,