• Rajiniraja Muniyan Department of Biotechnology, School of Biosciences and Technology, VIT University, Vellore - 632 014, Tamil Nadu, India
  • Sabareesh Varatharajan Centre for Bio-Separation Technology, VIT University, Vellore - 632 014, Tamil Nadu, India
  • Saba Naz National Institute of Immunology, Aruna Asaf Ali Marg,New Delhi - 110 067, India
  • Vinay K Nandicoori National Institute of Immunology, Aruna Asaf Ali Marg,New Delhi - 110 067, India
  • Jayaraman Gurunathan Department of Biotechnology, School of Biosciences and Technology, VIT University, Vellore - 632 014, Tamil Nadu, India.



 Objectives: The purpose of the study is to characterize antimycobacterial phytoconstituent from ethyl acetate extract of dried bulbs of Allium sativum Linn. (Alliaceae) and elucidating the probable mode of action of the bioactive molecule.

Methods: Serial extraction, Mycobacterium tuberculosis assay by agar well diffusion method, minimal inhibitory concentration by microplate alamar blue assay, Fourier transform infrared (FT-IR), nuclear magnetic resonance (NMR) spectroscopy, liquid chromatography (LC)-electrospray ionization (ESI)-mass spectrometry (MS)/MS, cell leakage assay, scanning electron microscopy (SEM), inhibition property of linear alkylbenzene sulfonate (LAS) in the presence of rifampicin on M. tuberculosis were performed.

Results: Ethyl acetate extract displayed significant inhibition properties against M. tuberculosis H37Ra (MTCC 300). Subsequently, the bioactivity-guided fractionation was employed to purify the phytochemical. Analysis of FT-IR, LC-MS (ESI), 1H, and13C-NMR spectrum revealed that the bioactive phytochemicals are the variants of LAS, with C12-alkyl being predominant, and the minimum inhibitory concentration was found to be 5.56 μg/ml. Morphological examination by SEM and cell leakage assay indicated that these molecules change the membrane fluidity.

Conclusion: The results thus suggest the possibility of using low concentrations of LAS to effect changes in membrane fluidity, thereby enhancing the efficacy of antibiotic treatment.

Keywords: Bioactivity-guided fractionation, Microplate alamar blue assay, Liquid chromatography-mass spectrometry, Cell leakage assay, Scanning electron microscope.

Author Biography

Rajiniraja Muniyan, Department of Biotechnology, School of Biosciences and Technology, VIT University, Vellore - 632 014, Tamil Nadu, India

Department of Biotechnology

School of Bio-Sciences and Technology


1. World Health Organization. Global Tuberculosis Report. 20th ed. Geneva, Switzerland: World Health Organization (WHO); 2015.
2. Wells CD, Cegielski JP, Nelson LJ, Laserson KF, Holtz TH, Finlay A, et al. HIV infection and multidrug-resistant tuberculosis: The perfect storm. J Infect Dis 2007;196 Suppl 1:S86-107.
3. Ma Z, Lienhardt C, McIlleron H, Nunn AJ, Wang X. Global tuberculosis drug development pipeline: The need and the reality. Lancet 2010;375(9731):2100-9.
4. Chan ED, Iseman MD. Current medical treatment for tuberculosis. BMJ 2002;325(7375):1282-6.
5. Adhvaryu M, Vakharia B. Drug-resistant tuberculosis: Emerging treatment options. Clin Pharmacol 2011;3:51-67.
6. Newman DJ, Cragg GM, Snader KM. Natural products as sources of new drugs over the period 1981-2002. J Nat Prod 2003;66(7):1022-37.
7. Sukantha TA, Sripathi SK, Ravindran TN. Antibacterial activity of selected medicinal plant in traditional treatment of wound infection in Southeast India. Int J Pharm Pharm Sci 2014;6:511-3.
8. Douaouya L, Bouzerna N. Effect of garlic (Allium sativum L) on biochemical parameters and histopathology of pancreas of alloxan-induced diabetic rats. Int J Pharm Pharm Sci 2016;8:202-6.
9. Newton SM, Lau C, Wright CW. A review of antimycobacterial natural products. Phytother Res 2000;14(5):303-22.
10. Copp BR. Antimycobacterial natural products. Nat Prod Rep 2003;20(6):535-57.
11. Okunade AL, Elvin-Lewis MP, Lewis WH. Natural antimycobacterial metabolites: Current status. Phytochemistry 2004;65(8):1017-32.
12. Dini C, Fabbri A, Geraci A. The potential role of garlic (Allium sativum) against the multi-drug resistant tuberculosis pandemic: A review. Ann Ist Super Sanità 2011;47:465-73.
13. Gebreyohannes G, Gebreyohannes M. Medicinal values of garlic: A review. Int J Med Sci 2013;5:401-8.
14. Gupta R, Thakur B, Singh P, Singh HB, Sharma VD, Katoch VM, et al. Anti-tuberculosis activity of selected medicinal plants against multi-drug resistant Mycobacterium tuberculosis isolates. Indian J Med Res 2010;131:809-13.
15. Schauenberg P, Paris F. Guide to Medicinal Plants. 1st ed. Paris: Lutterworth Press; 1974.
16. Harborne JB. Phytochemical Methods. London: Chapman and Hall Ltd.; 1973.
17. Chaudhari RD. Herbal Drug Industry. 1st ed. New Delhi: Eastern Publishers; 1996.
18. Patra JK, Gouda S, Sahoo SK, Thatoi HN. Chromatography separation, 1H NMR analysis and bioautography screening of methanol extract of Excoecaria agallocha L. From Bhitarkanika, Orissa, India. Asian Pac J Trop Biomed 2012;2:S50-6.
19. Middlebrook G, Cohn ML. Bacteriology of tuberculosis: Laboratory methods. Am J Public Health Nations Health 1958;48(7):844-53.
20. Sánchez JG, Kouznetsov VV. Antimycobacterial susceptibility testing methods for natural products research. Braz J Microbiol 2010;41(2):270-7.
21. Rajiniraja M, Jayaraman G. Bioautography guided screening of selected Indian medicinal plants reveals potent antimycobacterial activity of Allium sativum extracts-implication of non-sulfur compounds in inhibition. Int J Pharm Pharm Sci 2014;6:671-6.
22. Collins L, Franzblau SG. Microplate alamar blue assay versus BACTEC 460 system for high-throughput screening of compounds against Mycobacterium tuberculosis and Mycobacterium avium. Antimicrob Agents Chemother 1997;41(5):1004-9.
23. Prescott ML, Harley J, Donald P. Microbiology. 2nd ed. USA: Brown Publisher, Antimicrob Chemother; 1999. p. 325.
24. Muniyan R, Gurunathan J. Lauric acid and myristic acid from Allium sativum inhibit the growth of Mycobacterium tuberculosis H37Ra: In silico analysis reveals possible binding to protein kinase B. Pharm Biol 2016;54(12):2814-21.
25. Llanos GG, Araujo LM, Jiménez IA, Moujir LM, Bazzocchi IL. Withaferin A-related steroids from Withania aristata exhibit potent antiproliferative activity by inducing apoptosis in human tumor cells. Eur J Med Chem 2012;54:499-511.
26. Oonmetta-Aree J, Suzuki T, Gasaluck P. Antimicrobial properties and action of galangal (Alpinia galanga Linn.) on Staphylococcus aureus. LWT 2006;39:1214-20.
27. Priester JH, Horst AM, Van de Werfhorst LC, Saleta JL, Mertes LA, Holden PA. Enhanced visualization of microbial biofilms by staining and environmental scanning electron microscopy. J Microbiol Methods 2007;68(3):577-87.
28. Agyare C, Koffuor GA, Boamah VE, Adu F, Mensah KB, Adu-Amoah L. Antimicrobial and anti-inflammatory activities of Pterygota macrocarpa and Cola gigantea (Sterculiaceae). Evid Based Complement Alternat Med 2012;2012:902394.
29. Berndtsson R. Transport and sedimentation of pollutants in a river reach: A chemical massbalance approach. Water Resour Res 1990;26(7):1549-58.
30. Denger K, Cook AM. LAS bioavailable to anaerobic bacteria as a source of sulphur. J Appl Microbiol 1999;86(1):165-8.
31. Tsuchiya H. Membrane interactions of phytochemicals as their molecular mechanism applicable to the discovery of drug leads from plants. Molecules 2015;20(10):18923-66.
32. Imelda F, Faridah DN, Kusumaningrum HD. Bacterial inhibition and cell leakage by extract of Polygonum minus Huds leaves. Int Food Res J 2014;21(2):553-60.
33. Takasaki A, Hashida T, Kato K, Moriyama T, Nishihara T. Action of a quaternary ammonium disinfectant on cell membrane of Staphylococcus aureus. Jpn J Toxicol Environ Health 1994;40:520-6.
34. Van Bambeke F, Mingeot-Leclercq MP, Schanck A, Brasseur R, Tulkens PM. Alterations in membrane permeability induced by aminoglycoside antibiotics: Studies on liposomes and cultured cells. Eur J Pharmacol 1993;247(2):155-68.
35. Maurin M, Raoult D. Use of aminoglycosides in treatment of infections due to intracellular bacteria. Antimicrob Agents Chemother 2001;45(11):2977-86.
36. Crowle AJ, May MH. Inhibition of tubercle bacilli in cultured human macrophages by chloroquine used alone and in combination with streptomycin, isoniazid, pyrazinamide, and two metabolites of vitamin D3. Antimicrob Agents Chemother 1990;34(11):2217-22.
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How to Cite
Muniyan, R., S. Varatharajan, S. Naz, V. K. Nandicoori, and J. Gurunathan. “ALLIUM SATIVUM LINN. CONTAINS LINEAR ALKYLBENZENE SULFONATES THAT ALTER MEMBRANE FLUIDITY FOR THE INHIBITION OF MYCOBACTERIUM TUBERCULOSIS H37RA”. Asian Journal of Pharmaceutical and Clinical Research, Vol. 10, no. 9, Sept. 2017, pp. 100-11, doi:10.22159/ajpcr.2017.v10i9.18693.
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