• Md. Harun Al Rashid Pharmacognosy and Phytotherapy Research Laboratory, Division of Pharmacognosy, Department of Pharmaceutical Technology, Jadavpur University, Kolkata-700032, India
  • Deepak Bharadwaj P.v.p. Division of Molecular Medicine, Bose Institute, Kolkata-700054, India
  • Sayani Majumder Pharmacognosy and Phytotherapy Research Laboratory, Division of Pharmacognosy, Department of Pharmaceutical Technology, Jadavpur University, Kolkata-700032, India
  • Vivekananda Mandal Institute of Pharmacy, Guru Ghasidas Central University, Bilaspur-495009, India
  • Mahadeb Pal Division of Molecular Medicine, Bose Institute, Kolkata-700054, India
  • Subhash C Mandal Pharmacognosy and Phytotherapy Research Laboratory, Division of Pharmacognosy, Department of Pharmaceutical Technology, Jadavpur University, Kolkata-700032, India
  • Rajarajan Amirthalingam Thandavarayan Department of Cardiovascular Sciences, Center for Cardiovascular Regeneration, Houston Methodist Research Institute, TX-77030, USA.


Objective: The current study evaluates the antioxidant and cytotoxic prospective of the leaves from Diospyros melanoxylon Roxb. (D. melanoxylon).

Methods: Qualitative phytochemical analysis of the samples from D. melanoxylon was carried out for the detection of secondary metabolites. Total content of flavonoids, phenolics, triterpenoids, and tannins in D. melanoxylon was evaluated using colorimetric assay. Qualitative analysis of polyphenolic compounds was performed using HPLC method. The antioxidant activity was examined by assessing the various free radical scavenging assays. Five human cancerous cell lines (HeLa, MCF-7, HCT-116, PC-3, and HEK293), one rat colon cancer cell line (RCC 45) and two normal cell lines (NKE and WI-38) were used for evaluating the anticancer activity through methylene blue assay.

Results: The results indicated that ethyl acetate fraction (EAF) and aqueous fraction (AQF) exhibited the remarkable content of flavonoids, phenolics, triterpenoids, and tannins. Both EAF and AQF have cytotoxic activity with significant IC50 values but not showed significant antioxidant activity. However, the most powerful anticancer activity was recorded by EAF followed by AQF in a dose-dependent manner. It was also revealed that EAF and AQF were biocompatible with the normal cell lines.

Conclusion: Our principal finding exhibited a potent anticancer activity. Furthermore, it was also indicated that samples contain secondary metabolites like flavonoids, phenolics, triterpenoids, and tannins. We suggest that the further investigation on this herb is effective against various types of ailments including cancer.

Keywords: Diospyros melanoxylon Roxb., Scanning Electron Microscopy (SEM), High-Pressure Liquid Chromatography (HPLC), Polyphenols, Antioxidant, Anticancer

Keywords: Diospyros melanoxylon Roxb, Scanning Electron Microscopy (SEM), High-Pressure Liquid Chromatography (HPLC), Polyphenols, Antioxidant, Anticancer


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1. Rajesh R, Chitra K, Paarakh PM, Chidambaranathan N. Anticancer activity of aerial parts of aerva lanata linn juss ex schult against dalton's ascitic lymphoma. Eur J Integr Med 2011 3:e245-e50.
2. Das M, Mohanty C, Sahoo SK. Ligand-based targeted therapy for cancer tissue. Expert Opin Drug Delivery 2009;6:285-304.
3. Sahoo S, Parveen S, Panda J. The present and future of nanotechnology in human health care. Nanomed Nanotechnol Biol Med 2007;3:20-31.
4. Mihailovic V, Misic D, Matic S, Mihailovic M, Stanic S, Vrvic MM, et al. Comparative phytochemical analysis of gentiana cruciata l. Roots and aerial parts, and their biological activities. Ind Crops Prod 2015;73:49-62.
5. Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Telser J. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 2007;39:44-84.
6. Wiseman H, Halliwell B. Damage to DNA by reactive oxygen and nitrogen species: role in inflammatory disease and progression to cancer. Biochem J 1996;313:17.
7. Al Rashid MH, Mandal V, Mandal SRa SC. Herbal nanotechnology: an emerging tool in cancer therapy. Biology, Biotechnology, and Sustainable Development; 2015. p. 86.
8. Nurhanan M, Asiah O, Ilham MM, Syarifah MS, Norhayati I, Sahira HL. Anti-proliferative activities of 32 malaysian plant species in breast cancer cell lines. J Trop For Sci 2008;77-81.
9. Lo PC, Skeath JB, Gajewski K, Schulz RA, Frasch M. Homeotic genes autonomously specify the anteroposterior subdivision of the drosophila dorsal vessel into aorta and heart. Dev Biol 2002;251:307-19.
10. Raza H, John A. In vitro effects of tea polyphenols on redox metabolism, oxidative stress, and apoptosis in pc12 cells. Ann N Y Acad Sci 2008;1138:358-65.
11. Brewer M. Natural antioxidants: sources, compounds, mechanisms of action, and potential applications. Compr Rev Food Sci F 2011;10:221-47.
12. Dewick PM. Medicinal natural products: a biosynthetic approach. John Wiley and Sons; 2002.
13. Halliwell B, Gutteridge JM. Free radicals in biology and medicine. Oxford University Press, USA; 2015.
14. Aiyegoro OA, Okoh AI. Preliminary phytochemical screening and in vitro antioxidant activities of the aqueous extract of helichrysum longifolium dc. BMC Complement Altern Med 2010;10:21.
15. Mallavadhani U, Panda AK, Rao Y. Review article number 134 pharmacology and chemotaxonomy of diospyros. Phytochemistry 1998;49:901-51.
16. Chopea R. Poisonous plants of india. Poisonous plants of India; 1949.
17. Sastri B. The wealth of india. A dictionary of indian raw materials and industrial products. Raw materials. The Wealth of India A Dictionary of Indian Raw Materials and Industrial Products Raw Materials; 1950.
18. Banik R, Pandey D. Optimizing conditions for oleanolic acid extraction from lantana camara roots using response surface methodology. Ind Crops Prod 2008;27:241-8.
19. Asaduzzaman M, Uddin MJ, Kader M, Alam A, Rahman AA, Rashid M, et al.: In vitro acetylcholinesterase, inhibitory activity and the antioxidant properties of aegle marmelos leaf extract: implications for the treatment of Alzheimer's disease. Psychogeriatrics 2014;14:1-10.
20. Ordonez A, Gomez J, Vattuone M. Antioxidant activities of sechium edule (jacq.) swartz extracts. Food Chem 2006; 97:452-8.
21. Fatiha B, Didier H, Naima G, Khodir M, Martin K, Léocadie K, et al. Phenolic composition, in vitro antioxidant effects and tyrosinase inhibitory activity of three algerian mentha species: M. Spicata (l.), m. Pulegium (l.) and m. Rotundifolia (l.) huds (lamiaceae). Ind Crops Prod 2015;74:722-30.
22. Koncic MZ, Kremer D, Gruz J, Strnad M, Bisevac G, Kosalec I, et al. Antioxidant and antimicrobial properties of moltkia petraea (tratt.) griseb. Flower, leaf and stem infusions. Food Chem Toxicol 2010;48:1537-42.
23. Fan JP, He CH. Simultaneous quantification of three major bioactive triterpene acids in the leaves of diospyros kaki by high-performance liquid chromatography method. J Pharm Biomed Anal 2006;41:950-6.
24. Russo D, Valentao P, Andrade PB, Fernandez EC, Milella L. Evaluation of antioxidant, antidiabetic and anticholinesterase activities of smallanthus sonchifolius landraces and correlation with their phytochemical profiles. Int J Mol Sci 2015;16:17696-718.
25. Singh R, Kumari N. Comparative determination of phytochemicals and antioxidant activity from leaf and fruit of sapindus mukorrossi gaertn.–a valuable medicinal tree. Ind Crops Prod 2015;73:1-8.
26. Sharma OP, Bhat TK. Dpph antioxidant assay revisited. Food Chem 2009;113:1202-5.
27. Ashraf A, Sarfraz RA, Mahmood A, ud Din M. Chemical composition and in vitro antioxidant and antitumor activities of eucalyptus camaldulensis dehn. Leaves Ind Crops Prod 2015;74:241-8.
28. Rao M. Nitric oxide scavenging by curcuminoids. J Pharm Pharmacol 1997;49:105-7.
29. Luqman S, Kumar R, Kaushik S, Srivastava S, Darokar MP, Khanuja SP. Antioxidant potential of the root of vetiveria zizanioides (l.) nash; 2009.
30. Irshad M, Zafaryab M, Singh M, Rizvi M. Comparative analysis of the antioxidant activity of cassia fistula extracts. Int J Med Chem 2012.
31. Khan RA, Khan MR, Sahreen S, Shah NA, Khan AM, Khan YM, et al. Effect of various fractions of launaea procumbens on antioxidant enzymes in rats liver: oxidative stress induced by potassium bromate (kbro3). Afr J Pharm Pharmacol 2012;6:512-5.
32. Rathore K, Singh VK, Jain P, Rao SP, Ahmed Z, Singh VD. In vitro and in vivo antiadipogenic, the hypolipidemic and antidiabetic activity of diospyros melanoxylon (roxb). J Ethnopharmacol 2014;155:1171-6.
33. Kedar P, Chakrabarti C. Effects of bittergourd (momordica charantia) seed and glibenclamide in streptozotocin induced diabetes mellitus. Indian J Exp Biol 1982;20:232.
34. Mandal V, Dewanjee S, Mandal SC. Microwave‐assisted extraction of total bioactive saponin fraction from gymnema sylvestre with reference to gymnemagenin: a potential biomarker. Phytochem Anal: Int J Plant Chem Biochem Techniques 2009;20:491-7.
35. Yıldırım A, Mavi A, Kara AA. Determination of antioxidant and antimicrobial activities of rumex crispus l. Extracts. J Agric Food Chem 2001;49:4083-9.
36. Mandal V, Mandal SC. Design and performance evaluation of a microwave-based low carbon yielding extraction technique for naturally occurring bioactive triterpenoid: oleanolic acid. Biochem Eng J 2010;50:63-70.
37. Blois MS. Antioxidant determinations by the use of a stable free radical. Nature 1958;181:1199.
38. Bala A, Kar B, Haldar PK, Mazumder UK, Bera S. Evaluation of anticancer activity of cleome gynandra on ehrlich's ascites carcinoma treated mice. J Ethnopharmacol 2010;129:131-4.
39. Fotsis T, Pepper MS, Aktas E, Breit S, Rasku S, Adlercreutz H, et al. Flavonoids, dietary-derived inhibitors of cell proliferation and in vitro angiogenesis. Cancer Res 1997;57:2916-21.
40. Lee WJ, Zhu BT. Inhibition of DNA methylation by caffeic acid and chlorogenic acid, two common catechol-containing coffee polyphenols. Carcinogenesis 2005;27:269-77.
41. Lee WJ, Shim JY, Zhu BT. Mechanisms for the inhibition of DNA methyltransferases by tea catechins and bioflavonoids. Mol Pharmacol; 2005.
42. Yoshioka K, Kataoka T, Hayashi T, Hasegawa M, Ishi Y, Hibasami H. Induction of apoptosis by gallic acid in human stomach cancer kato iii and colon adenocarcinoma colo 205 cell lines. Oncol Rep 2000;7:1221-4.
43. Aires V, Limagne E, Cotte AK, Latruffe N, Ghiringhelli F, Delmas D. Resveratrol metabolites inhibit human metastatic colon cancer cells progression and synergize with chemotherapeutic drugs to induce cell death. Mol Nutr Food Res 2013;57:1170-81.
44. Roleira FM, Tavares-da-silva EJ, Varela CL, Costa SC, Silva T, Garrido J, et al. Plant-derived and dietary phenolic antioxidants: anticancer properties. Food Chem 2015;183:235-58.
45. Taylor J, Rabe T, McGaw L, Jäger A, Van Staden J. Towards the scientific validation of traditional medicinal plants. Plant Growth Regul 2001;34:23-37.
46. Moon JK, Shibamoto T. Antioxidant assays for plant and food components. J Agric Food Chem 2009;57:1655-66.
47. Rice Evans CA, Miller NJ, Paganga G. Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Radical Biol Med 1996;20:933-56.
48. Ghasemi K, Ghasemi Y, Ebrahimzadeh MA. Antioxidant activity, phenol and flavonoid contents of 13 citrus species peels and tissues. Pak J Pharm Sci 2009;22:277-81.
49. Sengul M, Yildiz H, Gungor N, Cetin B, Eser Z, Ercisli S. Total phenolic content, antioxidant and antimicrobial activities of some medicinal plants. Pak J Pharm Sci 2009;22:102-6.
50. Al Rashid H. Preparation and characterization of plga loaded nanoparticles obtained from d. Melanoxylon roxb. Leaves for their antiproliferative and antidiabetic activity. Int J Green Pharm 2017;11:S438-S447.
51. Pemmaraju D, Appidi T, Minhas G, Singh SP, Khan N, Pal M, et al. Chlorophyll rich biomolecular fraction of a. Cadamba loaded into polymeric nanosystem coupled with photothermal therapy: a synergistic approach for cancer theranostics. Int J Biol Macromol 2018;110:383-91.
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How to Cite
Al Rashid, M. H., D. B. P.v.p., S. Majumder, V. Mandal, M. Pal, S. C. Mandal, and R. A. Thandavarayan. “ANTIOXIDANT AND ANTICANCER ACTIVITY OF EXTRACT AND FRACTIONS OBTAINED FROM DIOSPYROS MELANOXYLON ROXB. LEAVES AND CORRELATION WITH THEIR POLYPHENOLIC PROFILES”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 10, no. 11, Nov. 2018, pp. 6-16, doi:10.22159/ijpps.2018v10i11.21111.
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