Isolation and Identification of Two Antibacterial Agents from Chromolaena odorata L. Active against Four Diarrheal Strains


Chromolaena odorata L (Asteraceae) is a bad invasive plant, found in the humid tropics and sub-tropics worldwide. It is used against dysentery, diarrhea, malaria, wound healing, headache and toothache in traditional medicine. In the present study, we investigated the antibacterial activities of different leaves extracts of Chromolaena odorata L. (cyclohexane, dichloromethane, ethyl acetate and butanol) against four clinical diarrheal strains (Klebsiella oxytoca, Salmonella enterica, Shigella sonnei and Vibrio cholera). We demonstrated that C. odorata leaves extracts show an antibacterial activity between 0.156 and 1.25 mg/mL. Bioassay-guided chromatography by bioautography with iodonitrotetrazolium-based colorimetric assay allowed the isolation and identification of two active compounds. After the combination of RP-HPLC, mass spectrometry analysis, 1D and 2D-NMR spectroscopy, we isolated and characterized two active molecules corresponding to 3’,4’,5,6,7-Pentamethoxyflavone (Sinensetin) and4,5,6,7-Tetramethoxyflavone (Scutellareintetramethyl ether).

Share and Cite:

M. Atindehou, L. Lagnika, B. Guérold, J. Marc Strub, M. Zhao, A. Van Dorsselaer, E. Marchioni, G. Prévost, Y. Haikel, C. Taddéi, A. Sanni and M. Metz-Boutigue, "Isolation and Identification of Two Antibacterial Agents from Chromolaena odorata L. Active against Four Diarrheal Strains," Advances in Microbiology, Vol. 3 No. 1, 2013, pp. 115-121. doi: 10.4236/aim.2013.31018.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] World Health Organization (WHO), “La Diarrhea,” 2009. http://www.whoint/mediacentre/factsheets/fs330/fr/indexhtml.
[2] Institut de la Francophonie Pour la Médecine Tropicale (IFMT), “Maladies Diarrhé?ques: Vue D’ensemble,” 2005. http://www.ifmtauforg/IMG/pdf/maladies-diarrheiques-2pdf
[3] K. M. Hoffmann, A. Deutschmann, C. Weitzer, M. Joainig, E. Zechner, C. Hogenauer and A. C. Hauer, “Antibiotic-Associated Hemorrhagic Colitis Caused by Cyto-Toxin-Producing Klebsiellaoxytoca,” Pediatrics, Vol. 125, No. 4, 2010, pp. e960-e963. doi:10.1542/peds.2009-1751
[4] A. Plessier, J. Cosnes, J. P. Gendre and L. Beaugerie, “Inter-Current Klebsiellaoxytoca Colitis in a Patient with Crohn’s Disease,” Gastroenterologie Clinique et Biologique, Vol. 26, No. 8-9, 2002, pp. 799-800.
[5] G. Gorkiewicz, “Nosocomial and Antibiotic-Associated Diarrhoea Caused by Organisms Other than Clostridium difficile,” International Journal of Antimicrobial Agents, Vol. 33, No. S1, 2009, pp. S37-S41.
[6] World Health Organization (WHO), “Guidelines on Safety Monitoring of Herbal Medicines in Pharmacovigilance Systems,” 2004.
[7] A. Tuffs, “Three out of Four Germans Have Used Complementary or Natural Remedies,” British Medical Journal, Vol. 325, No. 7371, 2002, p. 990. doi:10.1136/bmj.325.7371.990/e
[8] C. Marwick, “US Report Calls for Tighter Controls on Complementary Medicine,” British Medical Journal, Vol. 324, No. 7342, 2002, p. 870. doi:10.1136/bmj.324.7342.870/a
[9] O. Sharma, R. Dawra, N. Kurade and P. Sharma, “A Review of the Toxicosis and Biological Properties of the Genus Eupatorium,” Natural Toxins, Vol. 6, No. 1, 1998, pp. 1-14.
[10] K. Panyaphu, T. V. On, P. Sirisa-Ard, P. Srisa-Nga, S. Chansa Kaow and S. Nathakarnkitkul, “Medicinal Plants of the Mien (Yao) in Northern Thailand and Their Potential Value in the Primary Healthcare of Postpartum Women,” Journal of Ethnopharmacoly, Vol. 135, No. 2, 2011, pp. 226-237.
[11] Clinical and Laboratory Standards Institute, “Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically; Approved Standard-Seventh Edition,” Clinical and Laboratory Standards Institute, Vol. 26, No. 2, 2006, pp. 1-64.
[12] W. J. Begue and R. M. Kline, “The Use of Tetrazolium Salts in Bioautographic Procedure,” Journal of Chromatography, Vol. 64, No. 1, 1972, pp. 182-184.
[13] J. N. Eloff, “A Sensitive and Quick Microplate Method to Determine the Minimal Inhibitory Concentration of Plant Extracts for Bacteria,” Planta Medica, Vol. 64, No. 8, 1998, pp. 711-713. doi:10.1055/s-2006-957563
[14] A. Younes, A. Hamouda and S. G. Amyes, “First Report of a Novel Extended-Spectrum Beta-Lactamase KOXY-2 Producing Klebsiella oxytoca That Hydrolyses Cefotaxime and Ceftazidime,” Journal of Chemotherapy, Vol. 23, No. 3, 2011, pp. 127-130.
[15] F. Medalla, M. Sjolund-Karlsson, S. Shin, E. Harvey, K. Joyce, L. Theobald, B. N. Nygren, G. Pecic, K. Gay, J. Austin, A. Stuart, E. Blanton, E. D. Mintz, J. M. Whichard and E. J. Barzilay, “Ciprofloxacin-Resistant Salmonella enterica Serotype Typhi, United States, 1999-2008,” Emerging Infectious Diseases, Vol. 17, No. 6, 2011, pp. 1095-1098. doi:10.3201/eid1706.100594
[16] H. L. Alaoui, K. Oufdou and N. E. Mezrioui, “Determination of Several Potential Virulence Factors in Non-o1 Vibrio cholerae, Pseudomonas aeruginosa, Faecal Coliforms and Streptococci Isolated from Marrakesh Ground-Water,” Water Science and Technology, Vol. 61, No. 7, 2010, pp. 1895-1905. doi:10.2166/wst.2010.263
[17] O. Irobi, “Activities of Chromolaenaodorata (Compositae) Leaf Extract against Pseudomonas aeruginosa and Streptococcus faecalis,” Journal of Ethnopharmacology, Vol. 37, No. 1, 1992, pp. 8l-83.
[18] A. Cáceres, H. Menéndez, E. Méndez, E. Cohobón, B. Samayoa, E. Jauregui, E. Peralta and G. Carrillo, “Antigonorrhoeal Activity of Plants Used in Guatemala for the Treatment of Sexually Transmitted Diseases,” Journal of Ethnopharmacology, Vol. 48, No. 2, 1995, pp. 85-88. doi:10.1016/0378-8741(95)01288-O
[19] A. NgonoNgane, R. EbelleEtame, F. Ndifor, L. Biyiti, P. H. AmvamZollo and P. Bouchet, “Antifungal Activity of Chromolaena odorata (L.) King & Robinson (Asteraceae) of Cameroon,” Chemotherapy, Vol. 52, No. 2, 2006, pp. 103-106.
[20] P. Wafo, R. S. Kamdem, Z. Ali, S. Anjum, A. Begum, O. O. Oluyemisi, S. N. Khan, B. T. Ngadjui, X. F. Etoa and M. I. Choudhary, “Kaurane-Type Diterpenoids from Chromoleanaodorata, Their X-Ray Diffraction Studies and Potent Alpha-Glucosidase Inhibition of 16-Kauren-19-Oic Acid,” Fitoterapia, Vol. 82, No. 4, 2011, pp. 642-646. doi:10.1016/j.fitote.2011.02.003
[21] V. B. Owoyele, J. O. Adediji and A. O. Soladoye, “Anti-Inflammatory Activity of Aqueous Leaf Extract of Chromolaenaodorata,” Inflammopharmacology, Vol. 13, No. 5-6, 2005, pp. 479-484. doi:10.1163/156856005774649386
[22] T. P. Thang, A. H. Margaret, W. C. George, T. T. Le and H. M. Pham, “An Aqueous Extract of the Leaves of Chromolaena odorata (Formerly Eupatorium odoratum) (Eupolin) Inhibits Hydrated Collagen Lattice Contraction by Normal Human Dermal Fibroblasts,” The Journal of Alternative and Complementary Medicine, Vol. 2, No. 3, 1996, pp. 335-343.
[23] K. SrinivasaRao, P. K. Chaudhury and A. Pradhan, “Evaluation of Anti-Oxidant Activities and Total Phenolic Content of Chromolaena odorata,” Food and Chemical Toxicology, Vol. 48, No. 2, 2009, pp. 729-732.
[24] A. Suksamrarn, A. Chotipong, T. Suavansri, S. Boongird, P. Timsuksai, S. Vimuttipong and A. Chuaynugul, “Antimycobacterial Activity and Cytotoxicity of Flavonoids from the Flowers of Chromolaena odorata,” Archives of Pharmacal Research, Vol. 27, No. 5, 2004, pp. 507-511. doi:10.1007/BF02980123
[25] T. M. Hung, T. D. Cuong, N. H. Dang, S. Zhu, P. Q. Long, K. Komatsu and B. S. Min, “Flavonoid Glycosides from Chromolaena odorata Leaves and Their in Vitro Cytotoxic Activity,” Chemical and Pharmaceutical Bulletin (Tokyo), Vol. 59, No. 1, 2011, pp. 129-131. doi:10.1248/cpb.59.129
[26] P. B. Kouame, C. Jacques, G. Bedi, V. Silvestre, D. Loquet, S. Barille-Nion, R. J. Robins and I. Tea, “Phytochemicals Isolated from Leaves of Chromolaena odorata: Impact on Viability and Clonogenicity of Cancer Cell Lines,” Phytotherapy Research, 2012.
[27] P. Taylor, M. Arsenak, M. J. Abad, A. Fernandez, B. Milano, R. Gonto, M. C. Ruiz, S. Fraile, S. Taylor, O. Estrada and F. Michelangeli, “Screening of Venezuelan Medicinal Plant Extracts for Cytostatic and Cytotoxic Activity against Tumor Cell Lines,” Phytotherapy Research, 2012.
[28] R. Gonzalez, I. Ballester, R. Lopez-Posadas, M. D. Suarez, A. Zarzuelo, O. Martinez-Augustin and F. Sanchez de Medina, “Effects of Flavonoids and Other Polyphenols on Inflammation,” Critical Reviews in Food Science and Nutrition, Vol. 51, No. 4, 2011, pp. 331-362. doi:10.1080/10408390903584094
[29] R. N. Barua, R. P. Sharma, G. Thyagarajan and W. Hertz, “Flavonoids of Chromolaenaodorata,” Phytochemistry, Vol. 17, No. 10, 1978, pp. 1807-1808.
[30] S. I. Kang, H. S. Shin, H. C. Ko and S. J. Kim, “Effects of Sinensetin on Lipid Metabolism in Mature 3T3-L1 Adipocytes,” Phytotherapy Research, Vol. 27, No. 1, 2013, pp. 131-134.
[31] A. Barra, “Factors Affecting Chemical Variability of Essential Oils: A Review of Recent Developments,” Natural Product Communications, Vol. 4, No. 8, 2009, pp. 1147-1154.
[32] R. Bruni and G. Sacchetti, “Factors Affecting Polyphenol Biosynthesis in Wild and FIELD GROWN St. John’s Wort (Hypericumperforatum L. Hypericaceae/Guttiferae),” Molecules, Vol. 14, No. 2, 2009, pp. 682-725.

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.