Bioassay Guided Fractionation of Allium austroiranicum by Cytotoxic Effects against Ovary and Cervical Cancer Cell Lines

Document Type: Original paper


1 Department of Pharmacognosy, Faculty of Pharmacy, Isfahan University of Medical Sciences,Isfahan, Iran

2 Department of Pharmacognosy, Isfahan University of Medical Sciences, Isfahan, Iran.

3 Department of Pharmaceutical Biotechnology, Isfahan University of Medical Sciences, Isfahan, Iran.



Background and objectives: Cancer is a major health problem in the world. The aim of this study was to extract the flowers of Allium austroiranicum used by Iranian people as a condiment or for its medicinal effects followed by bioassay guided fractionation of the extracts and fractions, using anti-proliferative effects against ovarian and cervical cancer cells. Methods: The air-dried flowers of Allium austroiranicum were extracted in a four-step extraction method, resulting hexan, chloroform, chloroform: methanol (9: 1), butanol and aqueous extracts. Anti-proliferative effects of the extracts were evaluated by MTT assay against OVCAR-3, HeLa, and HUVEC cell lines. The most potent cytotoxic extract was then subjected to fractionation by MPLC method on a RP-18 silicagel column. Finally, the cytotoxic effects of resulted fractions were analyzed again and the most potent cytotoxic fraction and its IC50 were determined. Results: Statistical analysis showed thatbutanol extract of A. austroiranicum showed the most potent cytotoxic effects against OVCAR-3, HeLa and HUVEC cell lines with IC50 values of 38±2, 56±1.4, and 60±3.5 µg/mL, respectively. On the other hand, for 7 fractions resulting from fractionation of the butanol extract, MTT assay results showed that 6th fraction (F) was the most cytotoxic fraction with IC50 of 2.7±0.26 and 7.5±0.5 µg/mL for OVCAR-3 and HeLa cancer cell lines, respectively. Primary evaluation of the fraction by TLC and NMR analysis suggested the steroidal saponins as the main constituents. Conclusion: Allium austroiranicum showed significant cytotoxic effects against ovarian cancer cell line especiallyfractions assumed to contain steroidal saponins. The fraction constituents have the potential of being strong cytotoxic agents and the isolation and identification of compounds are suggested.


[1] Segal R, Miller K, Jemal A. Cancer statistics, 2018. Ca Cancer J Clin. 2018; 68(1): 7-30.

[2] Torre LA, Trabert B, DeSantis CE, Miller KD, Samimi G, Runowicz CD, Gaudet MM, Jemal A, Siegel RL. Ovarian cancer statistics, 2018. Ca Cancer J Clin. 2018; 68(4): 284-296.

[3] Reid BM, Permuth JB, Sellers TA. Epidemiology of ovarian cancer: a review. Cancer Biol Med. 2017; 14(1): 9-32.

[4] Desai AG, Qazi GN, Ganju RK, El-Tamer M, Singh J, Saxena AK, Bedi YS, Taneja SC, Hari KB. Medicinal plants and cancer chemoprevention. Cur Drug Met. 2008; 9(7): 581-591.

[5] Richard TS, Kamdje AHN, Mukhtar F. Medicinal plants in breast cancer therapy. J Dis Med Plants. 2015; 1(1): 19-23.

[6] Chehri Z, Zolfaghari B, Dinani MS. Isolation of cinnamic acid derivatives from the bulbs of Allium tripedale. Adv Biomed Res. 2018; 7(1): 1-5.

[7] Fenwick GR, Hanley AB, Whitaker JR. The genus Allium-part 1. Crit Rev Food Sci Nutr. 1985; 22(3): 199-271.

[8] Thomson M, Ali M. Garlic [Allium sativum]: a review of its potential use as an anti-cancer agent. Curr Cancer Drug Targets. 2003; 3(1): 67-81.

[9] Azzini E, Durazzo A, Foddai MS, Temperini O, Venneria E, Valentini S, Giuseppe M. Phytochemicals content in Italian garlic bulb (Allium sativum L.) varieties. J Food Res. 2014; 3(4): 26-32.

[10] Cook N, Samman S. Flavonoids-chemistry, metabolism, cardioprotective effects, and dietary sources. J Nutri Biochem. 1996; 7(2): 66-76.

[11] Lacaille-Dubois MA. Bioactive saponins with cancer related and immunomodulatory activity: recent developments.  In: Atta-Ur-Rahman, Ed. Studies in natural products chemistry. Amesterdam: Elsevier, 2005.

[12] Bianchini F, Vainio H. Allium vegetables and organosulfur compounds: do they help prevent cancer? Environ Health Perspect. 2001; 109(9): 893-902.

[13] Sautour M, Mitaine-Offer AC, Lacaille-Dubois MA. The Dioscorea genus: a review of bioactive steroid saponins. J Nat Med. 2007; 61(2): 91-101.

[14] Fattorusso E, Lanzotti V, Taglialatela-Scafati O, Di Rosa M, Ianaro A. Cytotoxic saponins from bulbs of Allium porrum L. J Agric Food Chem. 2000; 48(8): 3455-3462.

[15] Tong QY, He Y, Zhao QB, Qing Y, Huang W, Wu XH. Cytotoxicity and apoptosis-inducing effect of steroidal saponins from Dioscorea zingiberensis Wright against cancer cells. Steroids. 2012; 77(12): 1219-1227.

[16] Fritsch RM, Abbasi M. New taxa and other contributions to the taxonomy of Allium L. (Alliaceae) in Iran. Rostaniha. 2009; 9(2): 1-77.

[17] Rezaee F, Zolfaghari B, Dinani MS. Isolation of dioscin-related steroidal saponin from the bulbs of Allium paradoxum L. with leishmanicidal activity. Res Pharm Sci. 2018; 13(5): 469-475.

[18] Kazemi M, Zolfaghari B, Keivanloo Shahrestanaki M, Sadeghi Dinani M. Cytotoxic effects of Allium affine Ledeb butanolic fraction on breast and ovary cancer cell lines. J Med Plants. 2018; 4(64): 83-90.

[19] Mahmoudvand H, Ezzatkhah F, Sharififar F, Sharifi I, Dezaki ES. Antileishmanial and cytotoxic effects of essential oil and methanolic extract of Myrtus communis L. Korean J Parasitol. 2015; 53(1): 21-27.

[20] Bakht J, Muhammad T, Ali H, Islam A, Shafi M. Effect of different solvent extracted sample of Allium sativum (Linn) on bacteria and fungi. Afr J Biotechnol. 2011; 10(31): 5910-5915.

[21] Hadjzadeh M, Tavakol Afshari J, Ghorbani A, Shakeri M. The effects of aqueous extract of Garlic (Allium sativum L.) on laryngeal cancer cells (Hep-2) and L929 cells in vitro. J Med Plants. 2006; 2(18): 41-48.

[22] Ivanova A, Mikhova B, Najdenski H, Tsvetkova I, Kostova I. Chemical composition and antimicrobial activity of wild garlic Allium ursinum of Bulgarian origin. Nat Prod Commun. 2009; 4(8): 1059-1062.

[23] Zolfaghari B, Sadeghi M, Troiano R, Lanzotti V. Vavilosides A1/A2–B1/B2, new furostane glycosides from the bulbs of Allium vavilovii with cytotoxic activity. Bioorg Med Chem. 2013; 21(7): 1905-1910.

[24] Zolfaghari B, Sadeghi M, Troiano R, Lanzotti V. 3, 4-Seco-spirostane sapogenins with cytotoxic activity from Allium umbilicatum Boiss. Phytochem Lett. 2015; 12: 291-295.

[25] Oliveira Filho CC, Kampke EH, Vargas TS, Salustriano NA, Scherer R, Fronza M, Campagnaro BP. In vitro cytotoxic activity of five commercial samples of Tribulus terrestris Linn in Espírito Santo (Brazil). Braz J Pharm Sci. 2017; 53(4): 1-8.

[26] Chan PK. Acylation with diangeloyl groups at C21-22 positions in triterpenoid saponins is essential for cytotoxcity towards tumor cells. Biochem Pharmacol. 2007; 73(3): 341-350.