Identification of the AntiListerial Constituents in Partially Purified Column Chromatography Fractions of Garcinia kola Seeds and Their Interactions with Standard Antibiotics
Partially purified fractions of the n-hexane extract of Garcinia kola seeds were obtained through column chromatography and their constituents were identified through the use of gas chromatography coupled to mass spectrometry (GC-MS). Three fractions were obtained by elution with benzene as the mobile phase and silica gel 60 as the stationery phase and these were named Benz1, Benz2, and Benz3 in the order of their elution. The antiListerial activities of these fractions were assessed through MIC determination and only Benz2 and Benz3 were found to be active with MIC’s ranging from 0.625 to 2.5？mg/mL. The results of the GC-MS analysis showed Benz2 to have 9 compounds whilst Benz3 had 7 compounds, with the major compounds in both fractions being 9,19-Cyclolanost-24-en-3-ol, (3.β.) and 9,19-Cyclolanostan-3-ol,24-methylene-, (3.β.). The Benz2 fraction was found to have mainly indifferent interactions with ampicillin and penicillin G whilst mainly additive interactions were observed with ciprofloxacin. The Benz3 fraction’s interactions were found to be 50% synergistic with penicillin G and 25% synergistic with ciprofloxacin and ampicillin. A commercially available 9,19-Cyclolanost-24-en-3-ol, (3.β.) was found not to exhibit any antiListerial activities at maximum test concentrations of 5？mg/mL, suggesting that the compound could be acting in synergy with the other compounds in the eluted fractions of Garcinia kola seeds. 1. Introduction Plants produce a vast diversity of secondary metabolites most of which are phytochemicals that have potential use in the pharmaceutical industry for new drug development purposes. Phytochemicals are naturally occurring bioactive plant compounds that act as a natural defence system for the host plant and also provide colour, aroma, and flavour [1]. Some phytochemicals have been shown to possess antimicrobial properties and these include terpenoids, essential oils, alkaloids, lectins, polypeptides, polyacetylenes, and phenolics, of which phenolics can be further divided into phenolic acids, flavonoids, quinones, tannins, coumarins, and simple phenols [2]. The terpenes are one of the largest and most diverse groups of plant secondary metabolites. They include complex compounds that are formed by the cyclization of 2,3-oxidosqualene [3]. They include terpenoids and sterols as well as essential oils which carry the fragrance of the plant. Terpenes possess antimicrobial properties and their mechanism of action is mainly through disruption of the bacterial membrane [2, 3]. Flavones, flavonoids, and flavonols are phenolic structures
References
[1]
Caribbean food and nutrition institute, “Phytochemicals,” Nyam News, vol. 1-2, pp. 1–4, 2005.
[2]
M. M. Cowan, “Plant products as antimicrobial agents,” Clinical Microbiology Reviews, vol. 12, no. 4, pp. 564–582, 1999.
[3]
N. S. Ncube, A. J. Afolayan, and A. I. Okoh, “Assessment techniques of antimicrobial properties of natural compounds of plant origin: current methods and future trends,” African Journal of Biotechnology, vol. 7, no. 12, pp. 1797–1806, 2008.
[4]
R. A. Dixon, P. M. Dey, and C. J. Lamb, “Phytoalexins: enzymology and molecular biology,” Advances in Enzymology and Related Areas of Molecular Biology, vol. 55, pp. 1–136, 1983.
[5]
K. Das, R. K. S. Tiwari, and D. K. Shrivastava, “Techniques for evaluation of medicinal plant products as antimicrobial agent: current methods and future trends,” Journal of Medicinal Plant Research, vol. 4, no. 2, pp. 104–111, 2010.
[6]
R. N. Bennett and R. M. Wallsgrove, “Secondary metabolites in plant defense mechanisms,” Phytotherapy Research, vol. 127, no. 4, pp. 617–633, 1994.
[7]
E. Haslam, “Natural polyphenols (vegetable tannins) as drugs: possible modes of action,” Journal of Natural Products, vol. 59, no. 2, pp. 205–215, 1996.
[8]
A. Sharma, V. K. Patel, and A. N. Chaturvedi, “Vibriocidal activity of certain medicinal plants used in Indian folklore medicine by tribals of Mahakoshal region of central India,” Indian Journal of Pharmacology, vol. 41, no. 3, pp. 129–133, 2009.
[9]
K. N. Varalakshmi, C. G. Sangeetha, A. N. Shabeena, S. R. Sunitha, and J. Vapika, “Antimicrobial and cytotoxic effects of Garcinia indica fruit rind extract,” American-Eurasian Journal of Agriculture and Environmental Science, vol. 7, no. 6, pp. 652–656, 2010.
[10]
M. M. Iwu, A. R. Duncan, and C. O. Okunji, “New antimicrobials of plant origin,” in Perspectives on New Crops and New Uses, J. Janick, Ed., pp. 457–462, ASHS Press, Alexandria, Va, USA, 1999.
[11]
J. N. Ogbulie, C. C. Ogueke, and F. C. Nwanebu, “Antibacterial properties of Uvaria chamae, Congronema latifolium, Garcinia kola, Vemonia amygdalina and Aframomium melegueta,” African Journal of Biotechnology, vol. 6, no. 13, pp. 1549–1553, 2007.
[12]
T. Sibanda, A. O. Olaniran, and A. I. Okoh, “In vitro antibacterial activities of crude extracts of Garcinia kola seeds against wound sepsis associated Staphylococcus strains,” Journal of Medicinal Plant Research, vol. 4, no. 8, pp. 710–716, 2010.
[13]
D. Penduka, O. O. Okoh, and A. I. Okoh, “In vitro antagonistic characteristics of crude aqueous and methanolic extracts of Garcinia kola (Heckel) seeds against some Vibrio bacteria,” Molecules, vol. 16, no. 4, pp. 2754–2765, 2011.
[14]
O. S. Adedeji, G. O. Farinu, S. A. Ameen, and T. B. Olayeni, “The effects of dietary Bitter kola (Garcinia kola) inclusion on body weight haematology and survival rate of pullet chicks,” Journal of Animal and Veterinary Advances, vol. 5, no. 3, pp. 184–187, 2006.
[15]
O. O. Adegbehingbe, S. A. Adesanya, T. O. Idowu, O. C. Okimi, O. A. Oyelami, and E. O. Iwalewa, “Clinical effects of Garcinia kola in knee osteoarthritis,” Journal of Orthopaedic Surgery and Research, vol. 3, no. 1, article 34, 2008.
[16]
A. O. Adesuyi, I. K. Elumm, F. B. Adaramola, and A. G. M. Nwokocha, “Nutritional and phytochemical screening of Garcinia kola,” Advance Journal of Food Science and Technology, vol. 4, no. 1, pp. 9–14, 2012.
[17]
R. T. Aplin, J. H. C. Blasdale, T. G. Halsall, and G. M. Hornby, “The isolation of cycloartenol and 24-methylenecycloartanol from false kola nuts (Garcinia kola Heckel),” Journal of the Chemical Society C, pp. 246–248, 1967.
[18]
I. I. Madubunyi, “Antimicrobial activities of the constituents of Garcinia kola seeds,” International Journal of Pharmacognosy, vol. 33, no. 3, pp. 232–237, 1995.
[19]
E. E. O. Odjadjare, L. C. Obi, and A. I. Okoh, “Municipal wastewater effluents as a source of listerial pathogens in the aquatic milieu of the eastern cape province of South Africa: a concern of public health importance,” International Journal of Environmental Research and Public Health, vol. 7, no. 5, pp. 2376–2394, 2010.
[20]
D. F. Basri and S. H. Fan, “The potential of aqueous and acetone extracts of galls of Quercus infectoria as antibacterial agents,” Indian Journal of Pharmacology, vol. 37, no. 1, pp. 26–29, 2005.
[21]
D. Penduka and A. I. Okoh, “In vitro anti-Listerial activities of crude n-hexane and aqueous extracts of Garcinia kola (heckel) seeds,” International Journal of Molecular Sciences, vol. 12, no. 10, pp. 6952–6965, 2011.
[22]
S. C. Selowa, L. J. Shai, P. Masoko, M. P. Mokgotho, and S. R. Magano, “Antibacterial activity of extracts of three Croton species collected in Mpumalanga region in South Africa,” African Journal of Traditional, Complementary and Alternative Medicines, vol. 7, no. 2, pp. 98–103, 2010.
[23]
J. N. Eloff, J. O. Famakin, and D. R. P. Katerere, “Combretum woodii (Combretaceae) leaf extracts have high activity against Gram-negative and Gram-positive bacteria,” African Journal of Biotechnology, vol. 4, no. 10, pp. 1161–1166, 2005.
[24]
EUCAST (European Committee for Antimicrobial Susceptibility Testing), “Determination of Minimum lnhibitory Concentration (MICs) of antimicrobial agents by broth dilution,” Clinical Microbiology and Infection, vol. 9, no. 8, pp. 1–7, 2003.
[25]
G. Miranda-Novales, B. E. Lea？os-Miranda, M. Vilchis-Pérez, and F. Solórzano-Santos, “In vitro activity effects of combinations of cephalothin, dicloxacillin, imipenem, vancomycin and amikacin against methicillin-resistant Staphylococcus spp. strains,” Annals of Clinical Microbiology and Antimicrobials, vol. 5, article 25, 2006.
[26]
K. P. Satish, R. C. Moellering, and G. M. Eliopoulos, “Antimicrobial combinations,” in Antibiotics in Laboratory Medicine, V. Lorian, Ed., pp. 365–440, Lippincott Williams & Wilkins, Philadelphia, Pa, USA, 2005.
[27]
O. D. Stefanovic, D. D. Stanojevic, and L. R. Comic, “Synergistic antibacterial activity of Salvia officinalis and Cichorium intybus extracts and antibiotics,” Drug Research, vol. 69, no. 3, pp. 457–463, 2012.
[28]
A. J. Cummins, A. K. Fielding, and J. McLauchlin, “Listeria ivanovii infection in a patient with AIDS,” Journal of Infection, vol. 28, no. 1, pp. 89–91, 1994.
[29]
C. Guillet, O. Join-Lambert, A. Le Monnier et al., “Human listeriosis caused by Listeria ivanovii,” Emerging Infectious Diseases, vol. 16, no. 1, pp. 136–138, 2010.
[30]
G. Todeschini, S. Friso, S. Lombardi, M. Casaril, R. Fontana, and R. Corrocher, “A case of Listeria murray/grayi bacteremia in a patient with advanced Hodgkin's disease,” European Journal of Clinical Microbiology and Infectious Diseases, vol. 17, no. 11, pp. 808–810, 1998.
[31]
A. Rapose, S. D. Lick, and N. Ismail, “Listeria grayi bacteremia in a heart transplant recipient,” Transplant Infectious Disease, vol. 10, no. 6, pp. 434–436, 2008.
[32]
H. Salimnia, D. Patel, P. R. Lephart, M. R. Fairfax, and P. H. Chandrasekar, “Listeria grayi: vancomycin-resistant, gram-positive rod causing bacteremia in a stem cell transplant recipient,” Transplant Infectious Disease, vol. 12, no. 6, pp. 526–528, 2010.
[33]
C. T. Seanego and R. N. Ndip, “Identification and antibacterial evaluation of bioactive compounds from Garcinia kola (Heckel) seeds,” Molecules, vol. 17, pp. 6569–6584, 2012.
[34]
A. E. O. Floriani, J. Ferreira, A. R. S. Santos, F. Monache, R. A. Yunes, and V. Cechinel-Filho, “Analgesic compounds from Epidendrum mosenii stems,” Pharmazie, vol. 53, no. 6, pp. 426–427, 1998.
[35]
I. Abe, M. Rohmer, and G. D. Prestwich, “Enzymatic cyclization of squalene and oxidosqualene to sterols and triterpenes,” Chemical Reviews, vol. 93, no. 6, pp. 2189–2206, 1993.
[36]
M.-H. Lee, J.-H. Jeong, J.-W. Seo et al., “Enhanced triterpene and phytosterol biosynthesis in Panax ginseng overexpressing squalene synthase gene,” Plant and Cell Physiology, vol. 45, no. 8, pp. 976–984, 2004.
[37]
K. Ohyama, M. Suzuki, J. Kikuchi, K. Saito, and T. Muranaka, “Dual biosynthetic pathways to phytosterol via cycloartenol and lanosterol in Arabidopsis,” Proceedings of the National Academy of Sciences of the United States of America, vol. 106, no. 3, pp. 725–730, 2009.
[38]
M. F. Adegboye, D. A. Akinpelu, and A. I. Okoh, “The bioactive and phytochemical properties of Garcinia kola (Heckel) seed extract on some pathogens,” African Journal of Biotechnology, vol. 7, no. 21, pp. 3934–3938, 2008.
[39]
C. Y. Ragasa, K. Jorvina, and J. A. Rideout, “Antimicrobial compounds from Artocarpus heterophyllus,” Philippine Journal of Science, vol. 133, no. 2, pp. 97–101, 2004.
[40]
G. Tegos, F. R. Stermitz, O. Lomovskaya, and K. Lewis, “Multidrug pump inhibitors uncover remarkable activity of plant antimicrobials,” Antimicrobial Agents and Chemotherapy, vol. 46, no. 10, pp. 3133–3141, 2002.
[41]
L. O. Regasini, G. M. Vieira-Júnior, D. C. Fernandes, V. D. S. Bolzani, A. J. Cavalheiro, and D. H. S. Silva, “Identification of triterpenes and sterols from Pterogyne nitens (fabaceae-caesalpinioideae) using high-resolution gas chromatography,” Journal of the Chilean Chemical Society, vol. 54, no. 3, pp. 218–221, 2009.
[42]
R. J. Owen and E. A. Palombo, “Anti-listerial activity of ethanolic extracts of medicinal plants, Eremophila alternifolia and Eremophila duttonii, in food homogenates and milk,” Food Control, vol. 18, no. 5, pp. 387–390, 2007.
[43]
F. C. Tenover, “Mechanisms of antimicrobial resistance in bacteria,” American Journal of Medicine, vol. 119, no. 6, pp. S3–S10, 2006.
[44]
J. Sikkema, J. A. M. de Bont, and B. Poolman, “Mechanisms of membrane toxicity of hydrocarbons,” Microbiological Reviews, vol. 59, no. 2, pp. 201–222, 1995.
[45]
D. D. Glover, D. Lalka, and G. R. G. Monif, “Ampicillin Vs. Penicillin for in utero therapy,” Infectious Diseases Obstetrics Gynecology, vol. 4, no. 1, pp. 43–46, 1996.
