CHROMATOGRAPHIC ISOLATION OF ALLIUM CEPA (SSP. RED ONION) AND ITS CYTOTOXIC ACTIVITY AGAINST HUMAN LIVER CARCINOMA CELL LINES (HEPG2)
Keywords:
Allium cepa, Cytotoxic, Human liver carcinoma cell lines HepG2, Chromatographic isolationAbstract
Objective: The present study aimed to find out cytotoxic agent from Allium cepa L. (ssp. red onion) family Alliaceae as well as chromatographic isolation of its active constituents.
Methods: The defatted methanol extract of Allium cepa (ssp. red onion) was fractionated with chloroform, ethyl acetate and n-butanol. Each of ethyl acetate and butanolic fraction was separately submitted to chromatographic isolation using different chromatographic techniques such as column chromatography (CC), thin layer chromatography (TLC) and paper chromatography (PC). The structures of the isolated compounds were elucidated through spectroscopic analysis.
Results: Four compounds were isolated from ethyl acetate fraction and identified as kaempferol (1), quercetin (2) isorhamnetin (3) and isorhamentin-4`-O-β- D-glucopyranoside (4) whereas two compounds were isolated and identified from the butanolic fraction as α -D-glucopyranose (5), β-D-glucopyranosyl- α- D-glucopyranoside (6). The cytotoxic activity of defatted methanolic extract of A. cepa (ssp red onion), ethyl acetate and butanolic fractions against human liver carcinoma cell line (HepG2) was IC50 = 10.9, 6.08 and 9.95 µg/ ml respectively. Also, compounds 4 isolated from ethyl acetate fraction and compound 6 isolated from butanolic fraction showed cytotoxic activity (IC50 = 11.90 and 22.0 µg/ ml) respectively.
Conclusion: It was appeared that the ethyl acetate fraction was the most active due to it contain high amount of phenolic compounds.
Downloads
References
Seyfried NT, Laura MS. Cancer as a metabolic disease. J Nutr Metabol 2010;7 (7):1-22.
Parikh S, David H. Hepatocellular Cancer: A Guide for the Internist. Am J Med 2007;120:194-202.
Gao J, Li X, Wan-Shui Y, Wei Z, Shan G, Jing W, Yong-Bing X. Risk Factors of Hepatocellular Carcinoma-Current Status and Perspectives. Asian Pac J Cancer Prev 2012;13:743-52.
Kaneria M, Baravalia Y, Vaghasiya Y, Chanda S. Determination of antibacterial and antioxidant potential of some medicinal plants from Saurashtra region, India. Indian J Pharma Sci 2009;71-406.
Dai J and Russell JM. Plant Phenolics:Extraction, Analysis and Their Antioxidant and Anticancer Properties. J Molecules 2010;15:7313-52.
Saxena M, Jyoti S, Rajeev N, Dharmendra S, Abhishek G. Phytochemistry of Medicinal Plants. J Pharm Phytochem 2013;1 (6):168-82.
Peteros PN, Mylene MU. Antioxidant and cytotoxic activities and phytochemical screening of four Philippine medicinal plants. J Med Plants Res 2010;4(5):407-14.
Benkeblia N. Free-radical scavenging capacity and antioxidant properties of some selected onions (Allium cepa L.) and garlic (Allium sativum L.) extracts. J Brazilian Arch Bio Tech 2005;48:753-9.
Lanzotti V. The analysis of onion and garlic. J Chromatogr A 2006;1112:3-22.
Skehan PR, Storeng D, Scudiero. New colorimetric cytotoxicity assay for anticancer drug screening. J Nat Cancer Inst 1990;82 (13):1107-12.
Mabry TJ, Markham KR, Thomas, MB. The systematic identification of flavonoids. Springer-Verlag. New York:Heidlberg Berlin;1970,p. 354.
Abdel-Moaty HI. Evaluation of the primary and secondary products of Nepeta septemcrenata Erenb. J Nat Prod 2009;2:81-8.
Mohamed GA, Ibrahim SRM and Sayed HM. Phenolic constituents of Cucurbita pepo L. cv Eskandrani` (Summer Squash) Flowers. J Bull Pharm Sci Assiut Univ 2009;32(2):311-9.
El-Sayed MM, Maher MA, Mahfouz MA, El-Sayed SA, Wafaa SA, Ezzat EA. Chemical constituents and cytotoxic activity of Cassia glauca Linn. leaves. Life Sci J 2013;10(3):1617-25.
Rajvaidhya S, Gajendra KS, Badri PN, Saroj KP. Extraction, isolation and chemical structure elucidation of isorhamnetin from leaves and stems of Argemone mexicana Linn of Bhopal, Madhya Pradesh, India. Indian J Appl Res 2014;4(2):10-13.
Sikorska M and Irena M. Kaempferol and isorhamnetin and their glycosides in the flowers of Asclepias syriaca L. J Acta Pol Pharm Drug Res 2001;58(4):269-72.
Park YK and Chang YL. Identification of isorhamnetin 4`-glucoside in onions. J Agric Food Chem 1996;44:34-6.
Guvenalp ZL, Omur D. Flavonol glycosides from Asperula arvensis L. Turk J Chem 2005;29:163-9.
Roslund, MU, Petri T, Matthias N, Rainer S. Complete assignments of the 1H and 13C chemical shifts and JH,H coupling constants in NMR spectra of D-glucopyranose and all D-glucopyranosyl-D-glucopyranosides. J Carbohydrate Res 2008;343:101-12.
Leeuwen SS, Bas RL, Gerrit JG, Johannis PK. Development of a 1H-NMR structural-reporter-group concept for the primary structural characterization of α-D-glucans. J Carbohydrate Res 2008;343:1114-19.
Corchado JC, Sa´nchez ML, Aguilar MA. Theoretical study of the relative stability of rotational conformers of alpha and beta-D-glucopyranose in gas phase and aqueous solution. J Am Chem Soc 2004;126, 7311-19.
Nijveldt JR, Els N, Danny ECH, Petra GB, Klaske N, Paul AML. Flavonoids:a review of probable mechanisms of action and potential applications. Am J Clin Nutr 2001;74:418-25.
Bulbul IJ, Laizuman N and Mahmuda H. Antibacterial, cytotoxic and antioxidant activity of chloroform, n-hexane and ethyl acetate extract of plant Coccinia cordifolia. Agric Bio J North Am 2001;2(4):713-9.
Boik J. Natural compounds in cancer therapy. Oregon Medical Press, Princeton, Minn, USA 2001.
Abdel-Hameed ES, Salih AB, Mahmood SS. Characterization of the phytochemical constituents of Taif rose and its antioxidant and anticancer activities. J Biomed Res Int 2013;1:1-13.
Kathrin P, Gabriele K, Eleni K, Elke R, Peter R, Axel R, Peter S, Carsten S. Structure-activity relationships of flavonoid-induced cytotoxicity on human leukemia cells. J Arch Biochem Biophys 2007; 460:1-9.