IN VITRO ANTIOXIDANT AND CYTOTOXIC ACTIVITY OF ISOLATED COMPOUNDS ON ROOTS OF CLERODENDRUM PHLOMIDIS LINN
Objective: The main aim of the study was to screen the isolated compounds of Clerodendrum phlomidis roots for its in vitro antioxidant and anticancer
activity and its efficacy against HeLa cell lines.
Methods: Pet ether, chloroform, ethyl acetate and ethanol extracts was prepared and assayed for the presence of phytochemicals. Three
compounds were isolated from ethanol extract of C. phlomidis by column chromatography such as ET1 (phenyl acetic acid), ET2 (ethyl-2-hydroxy-
4-methylbenzoate), ET3 (3,6,7-trihydroxy-2-(3-methoxyphenyl)-4H-chromen-4-one) characterized by IR (KBr), 1H-nuclear magnetic resonance
(NMR), 13C-NMR and Gas chromatographyâ€“mass spectrometry. The above isolated compounds were subjected to in vitro antioxidant activity against
2,2-diphenylpicrylhydrazyl (DPPH) radical, superoxide radical scavenging assay and iron chelating activity. The effect of isolated compounds on HeLa
cancer cell line was also evaluated by 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide colorimetric assay.
Results: ET2 has good in vitro antioxidant activity against DPPH radical, super oxide radical and iron chelating activity with an inhibitory concentration
50% value of 360 Î¼g/ml, 150 Î¼g/ml and 130 Î¼g/ml respectively. ET1 showed significant cytotoxic activity than the other two compounds on HeLa
cells with a percentage cell growth of 21.9% and a growth inhibition of 50% value of 180 Î¼g, respectively.
Conclusion: On the basis of obtained results, ET1 and ET2 isolated from the ethanolic extract of C. phlomidis represent a new group of cytotoxic
against HeLa cell line and antioxidant agents.
Keywords: Column chromatography, Clerodendrum phlomidis, In vitro antioxidant activity, 2,2-diphenylpicrylhydrazyl, Cytotoxicity,
3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide assay.
2. Larkin T. Herbs are often more toxic than magical. FDA Consum 1983;17:4-11.
3. Saxe TG. Toxicity of medicinal herbal preparations. Am Fam Physician 1987;35(5):135-42.
4. Shoeb M. Cytotoxic compounds from the Genus Centaurea. PhD Thesis. Aberdeen, UK: The Robert Gordon University; 2005.
5. Ferlay J, Shine HR, Bray F, Forman D, Mathers C, Parkin DM. Lyon, France: International Agency for Research in Cancer. [Last accessed on 2012 Oct 11].
6. Lozano R, Naghavi M, Foreman K, Lim S, Shibuya K, Aboyans V, et al. Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: A systematic analysis for the Global Burden of Disease Study 2010. Lancet 2012;380(9859):2095-128.
7. Merika E, Saif MW, Katz A, Syrigos K, Morse M. Review. Colon cancer vaccines: An update. In Vivo 2010;24(5):607-28.
8. Kirtikar KR, Basu BD. Indian Medicinal Plants. 2nd ed., Vol. II. Allahabad, India: Lalit Mohan Basu; 1935. p. 1492.
9. Grem JL, Allegra CJ. Toxicity of levamisole and 5-fluorouracil in human colon carcinoma cells. J Natl Cancer Inst 1989;81(18):1413-7.
10. Harborne JB. In: Phytochemical Methods. 11th ed. New York: Chapman & Hall; 1984. p. 4-5.
11. Kokate CK. Preliminary phytochemical screening. Practical Pharmacognosy. Ch. 6. New Delhi: Vallabh Prakashan; 1987. p. 106â€‘11.
12. Mensor LL, Menezes FS, LeitÃ£o GG, Reis AS, dos Santos TC, Coube CS, et al. Screening of Brazilian plant extracts for antioxidant activity by the use of DPPH free radical method. Phytother Res 2001;15(2):127â€‘30.
13. Winterbourn CC, Hawkins RE, Brian M, Carrell RW. The estimation of red cell superoxide dismutase activity. J Lab Clin Med 1975;85(2):337â€‘41.
14. Benzie IF, Strain JJ. The ferric reducing ability of plasma (FRAP) as a measure of â€œantioxidant powerâ€: The FRAP assay. Anal Biochem 1996;239(1):70-6.
15. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65(1-2):55-63.
16. Monks A, Scudiero D, Skehan P, Shoemaker R, Paull K, Vistica D, et al. Feasibility of a high-flux anticancer drug screen using a diverse panel of cultured human tumor cell lines. J Natl Cancer Inst 1991;83(11):757â€‘66.
17. Scudiero DA, Shoemaker RH, Paull KD, Monks A, Tierney S, Nofziger TH, et al. Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Res 1988;48(17):4827-33.
18. Sundaram S, Verma SK, Dwivedi P. In vitro cytotoxic activity of Indian medicinal plants used traditionally to treat cancer. Asian J Pharm Clin Res 2011;4(1):27-9.
19. Kemp W. Organic Spectroscopy. 3rd ed. United Kingdom: Palgrave Publishers; 2009. p. 127, 192.
20. Smith C, Halliwell B, Aruoma OI. Protection by albumin against the pro-oxidant actions of phenolic dietary components. Food Chem Toxicol 1992;30(6):483-9.
21. Duh PD, Tu YY, Yen GC. Antioxidant activity of water extract of Harng Jyur (Chrysanthemum morifolium Ramat). LMT Food Sci Technol 1999;32:269-77.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.