• Rizwana Abid Department of PG Studies and Research in Biotechnology and Bioinformatics, Jnanasahyadri, Kuvempu University, Shankaraghatta 577451, Shivamogga, Karnataka, India
  • Riaz Mahmood Department of PG Studies and Research in Biotechnology and Bioinformatics, Jnanasahyadri, Kuvempu University, Shankaraghatta 577451, Shivamogga, Karnataka, India
  • Rajesh K P Department of PG Studies and Research in Biotechnology and Bioinformatics, Jnanasahyadri, Kuvempu University, Shankaraghatta 577451, Shivamogga, Karnataka, India
  • Kumara Swamy B E Department of PG Studies and Research in Industrial Chemistry, Jnanasahyadri, Kuvempu University, Shankaraghatta 577451, Shivamogga, Karnataka, India


Objective: To evaluate the antioxidant potential and protective effect of Cassia fistula Linn. on hydrogen peroxide induced oxidative damage in erythrocytes.

Methods: The in vitro antioxidant potency was screened by various established chemical, biochemical and electrochemical techniques. The Chemical assays allowed an evaluation of their total phenolic and flavonoid contents, total antioxidant capacity, reducing power, radical scavenging activities and metal chelating activity. Both, CFE (C. fistula ethanolic extract) and CFA (C. fistula aqueous extract) were analyzed for phenolic acids and flavonoids by HPLC. The biochemical assays were employed to evaluate the lipid peroxidation and protective effect on induced oxidative damage in erythrocytes. Electrochemical measurement of CFE and CFA was determined by cyclic voltammetry.

Results: CFE showed strong antioxidant activity as well as >90% protection of erythrocytes. This might be due to the presence of high 464.3 ± 0.02 µg EGA mgˉ1total phenolics and 272.5 ± 0.5 µg EQ mgˉ1 total flavanoids while, CFA showed 75% antioxidant and protective activity possibly due to 250.84 ± 0.25 µg EGA mgˉ1 total phenolics and 195 ± 0.5 µg EQ mgˉ1 total flavonoid content. Moreover, HPLC of CFE and CFA exhibited widely known various good antioxidant molecules such as gallic acid, coumaric acid, ellagic acid, rutin, quercetin, myricetin and kaempferol. The cyclic voltammetric behavior shows the broad peak towards anodic potential represents the antioxidant property of CFE and CFA.

Conclusion: The present study clearly indicates that, C. fistula Linn. Fruit is an excellent source of natural antioxidant.


Keywords: Cassia fistula L, Antioxidant, Cyclic voltammetry, Oxidative damage inhibition, HPLC.


Download data is not yet available.


1. Zima TS, Fialova L, Mestek O, Janebova M, Crkovska J, Malbohan I, et al. Oxidative stress, metabolism of ethanol and alcohol-related diseases. J Biomed Sci 2001;8:59-70.
2. Singh U, Jialal I. Oxidative stress and atherosclerosis. Pathophysiology 2006;13:129-42.
3. Kinnula VL, Crapo JD. Superoxide dismutases in malignant cells and human tumors. Free Radic Biol Med 2004;36:718-44.
4. Sas K, Robotka H, Toldi J, Vecsei L. Mitochondrial, metabolic disturbances, oxidative stress and kynurenine system, with focus on neurodegenerative disorders. J Neurol Sci 2007;257:221-39.
5. Guidi I, Galimberti D, Lonati S, Novembrino C, Bamonti F, Tiriticco M, et al. Oxidative imbalance in patients with mild cognitive impairment and Alzheimer’s disease. Neurobiol Aging 2006;27:262-9.
6. Bolton JL, Trush MA, Penning TM, Dryhurst G, Monks TJ. Role of quinones in toxicology. Chem Res Toxicol 2000;13:135-60.
7. Arteel GE. Oxidants and antioxidants in alcohol induced liver disease. Gastroenterol 2003;124:778-90.
8. Ramakrishna BS, Varghese R, Jayakumar S, Mathan M, Balasubramanian KA. Circulating antioxidants in ulcerative colitis and their relationship to disease severity and activity. J Gastroenterol Hepatol 1997;12:490-4.
9. Hyun DH, Hernandez JO, Mattson MP, De Cabo R. The plasma membrane redox system in aging. Aging Res Rev 2006;5:209-20.
10. Upston JM, Kritharides L, Stocker R. The role of vitamin E in atherosclerosis. Prog Lipid Res 2003;42:405-22.
11. Anderson D. Antioxidant defences against reactive oxygen spe-cies causing genetic and other damage. Mut Res 1999;350:103-8.
12. Halliwell B, Guttridge JMC. Free Radicals in Biology and Medicine. 2nd ed. Japan: Scientific Societies Press; 1989.
13. Sadrazadeh SMH, Graf E, Panter SS, Hallaway PE, Eaten JW. Hemoglobin, a biological Fenton reagent. J Biol Chem 1984;259:14354-6.
14. Sukalski KA, LaBerge TP, Johnson WT. In vivo oxidative modification of erythrocyte membrane proteins in copper deficiency. Free Radic Biol Med 1997;22:835-42.
15. Rice-Evans CA, Miller NJ, Bolwell PG, Bramley PM, Pridham JB. The relative antioxidant activities of plant-derived polyphenolic flavonoids. Free Radical Res 1995;23:375-83.
16. Samia M, El-Sawad, Samir A, Ross. A phytochemical study of some Cassia species cultivated in Egypt. J Nat Prod 1982;46:431-2.
17. Bahorun T, Vidushi S, Neergheen P, Okezie I, Aruoma OI. Phytochemical constituents of Cassia fistula. Afr J Biotech 2005;4:1530-40.
18. Khanna VG, Kannabiran K. Larvicidal effect of Hemidesmus indicus, Gymnema sylvestre and Eclipta prostrata against Culex qinquifaciatus mosquito larvae. Afr J Biotechnol 2006;6:307-11.
19. Chang C, Yang M, Wen H, Chern J. Estimation of total flavonoid content in propolis by two complementary colorimetric methods. J Food Drug Anal 2002;10:178–82.
20. Zhishen J, Mengcheng T, Jianming W. The determination of flavonoid content in mulberry and their scavenging effects on superoxide radicals. Food Chem 1999;64:555-9.
21. Prieto P, Pineda M, Aguilar M. Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Anal Biochem 1999;269:337-41.
22. Oyaizu M. Studies on products of browning reaction: Antioxidative activities of products of browning reaction prepared from glucosamine. Jpn J Nutr 1986;44:307-15.
23. Braca A, Tommasi ND, Bari LD, Pizza C, Politi M, Morelli I. Antioxidant principles from Bauhinia terapotensis. J Nat Prod 2001;64:892-5.
24. Arnao MB, Cano A, Acosta M. The hydrophilic and lipophilic contribution to total antioxidant activity. Food Chem 2001;73:239-44.
25. Nishimiki M, Rao NA, Yagi K. The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem Biophys Res Commun 1972;46;849-53.
26. Halliwell B, Gutteridge JMC, Aruoma OI. The deoxyribose method: A simple ‘test tube’ assay for determination of rate constants for reactions of hydroxyl radicals. Anal Biochem 1987;165:215-9.
27. Marcocci L, Packer L, Sckaki A, Albert GM. Antioxidant action of Ginko biloba extracts EGb 761. Methods Enzymol 1994;234:462-75.
28. Dinis TCP, Madeira VMC, Almeida LM. Action of phenolic derivatives (acetaminophen, salicylate and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation as peroxyl radical scavenging effects. Chem Pharm Bull 1994;36:2090-7.
29. Yang HL, Chen SC, Chang NW, Chang JM, Lee ML, Tasi PE, et al. Protection from oxidative damage using Bidens pilosa extracts in normal human erythrocytes. Food Chem Toxicol 2006;44:1513-21.
30. Tedesco I, Russo M, Russo P, Lacomino GG, Russo GL, Carrasturo A, Faruolo C, et al. Antioxidant effect of red wine polyphenols on red blood cells. J Nutr Biochem 2000;11:114-9.
31. Niranjana E, Kumara Swamy BE, Raghavendra RN, Sherigara BS, Jayadevappa H. Electrochemical investigations of potassium ferricyanide doamine by sodium dodecyl sulfate modified carbon paste electrode: a cyclic voltammetrc study. J Electroanal Chem 2009;631:1-9.
32. Megala J, Geetha A. Free radical-scavenging and H+, K+-ATPase inhibition activities of Pithecellobium dulce. Food Chem 2010;121:1120-8.
33. Chang WT, Shao ZH, Vanden Hoek TL, McEntee E, Mehendale SR, Li J, et al. Cardioprotective effects of grape seed proanthocyanidins, baicalin and wogonin: comparison between acute and chronic treatments. Am J Chin Med 2006;34:363-5.
34. Huang D, Ou B, Prior RL. The chemistry behind antioxidant capacity assays. J Agric Food Chem 2005;53:1841-56.
35. Tosun M, Ercisli S, Sengul M, Ozer H, Polat T. Antioxidant properties and total phenolic content of eight Salvia species from Turkey. Biol Res 2009;41:175-81.
36. Sirappuselvi S, Chitra M. In vitro antioxidant activity of Cassia tora Lin. Int Res J Biological Sci 2012;1:57-61.
37. Irshad M, Zafaryab M, Man Singh, Moshahid MAR. Comparative analysis of the antioxidant activity of Cassia fistula extracts. Int J Med Chem 2012;doi: 10.1155/2012/157125.
38. Bhalodia NR, Nariya PB, Acharya RN, Shukla VJ. Evaluation of in vitro antioxidant activity of flowers of Cassia fistula Linn. Int J Pharm Tech Res 2011;3:589-99.
39. Siddhuraju P, Mohan PS, Becker K. Studies on the antioxidant activity of Indian Laburnum (Cassia fistula L.), a preliminary assessment of crude extracts from stem bark, leaves, flowers and fruit pulp. J Food Chem 2002;70:61-7.
40. Sumathy R, Sankaranarayanan S, Bama P, Ramachandran J, Vijayalakshmi M, Deecaraman M. Antibacterial and antioxidant activity of flavanoid rich fraction from the petals of Cassia auriculata – an in-vitro study. Int J Pharm Pharm Sci 2013;5:492-7.
41. Winterbourne CC, Kettle AJ. Radical–radical reactions of superoxide; a potential route to toxicity. Biochem Biophys Res Commun 2003;305:729-36.
42. Lui SL, Chan LY, Zhang XH, Zhu W, Chan TM, Fung PC, Lai KN. Effect of mycophenolate mofetil on nitric oxide production and inducible nitric oxide synthase gene expression during renal ischaemia-reperfusion injury. Nephrol Dial Transplant 2001;16:1577-82.
43. Middleton E, Kandaswami C, Theoharides TC. The effects of plant flavonoids on mammalian cells: Implications for inflammation, heart disease, and cancer. Pharmacol Rev 2000;52:673-751.
44. Liu ZQ, Luo XY, Sun YX, Chen YP, Wang ZC. Can ginsenosides protect human erythrocytes against free radical induced haemolysis? Biochimica Biophysica Acta 2002;1572:58-66.
45. Magalhaes AS, Silva BM, Pereira JA, Andrade PB, Valentao P, Carvalho M. Protective effect of quince (Cydonia oblonga Miller) fruit against oxidative haemolysis of human erytrhocytes. Food Chem Toxicol 2009;47:1372-7.
46. Singh N, Rajini PS. Antioxidant-mediated protective effects of potato peel extract in erythrocytes against oxidative damage. Chem-Biol Interact 2008;173:97-104.
47. Linderkamp O, Kiau U, Ruef P. Cellular and membrane deformability of red blood cells in protein infants with and without growth retardation. Clinical Hemorheology and Microcirculation 1997;17:279-83.
48. Ajila CM, Prasada Rao UJS. Protection against hydrogen peroxide induced oxidative damage in rat erythrocytes by Mangifera indica L. peel extract. Food Chem Toxicol 2008;4:303-9.
49. Sun-Waterhouse D, Smith BG, O’Connor CJ, Melton LD. Effect of raw and cooked onion dietary fibre on the antioxidant activity of ascorbic acid and quercetin. Food Chem 2008;111:580-5.
50. Slusarczyk S, Hajnos M, Skalicka WK, Matkowski A. Antioxidant activity of polyphenols from Lycopus lucidus Turcz. Food Chem 2009;113:134-8.
452 Views | 744 Downloads
How to Cite
Abid, R., R. Mahmood, R. K. P, and K. S. B. E. “POTENTIAL IN VITRO ANTIOXIDANT AND PROTECTIVE EFFECT OF CASSIA FISTULA LINN. FRUIT EXTRACTS AGAINST INDUCED OXIDATIVE DAMAGE IN HUMAN ERYTHROCYTES”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 6, no. 9, 1, pp. 497-05, https://innovareacademics.in/journals/index.php/ijpps/article/view/2572.
Original Article(s)