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(Decne.) CB Clarke Root Extracts and Their Anti-Bacterial Activity

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(Decne.) CB Clarke Root Extracts and Their Anti-Bacterial Activity Journal of Applied Pharmaceutical Science Vol. 9(05), pp 073-081, May, 2019 Available online at http://www.japsonline.com DOI: 10.7324/JAPS.2019.90509 ISSN 2231-3354 Gas chromatography/mass spectrometry profiling of the costus plant Saussurea lappa (Decne.) C.B. Clarke root extracts and their anti-bacterial activity Reem E. E. Omer1, Faisal Hammad Mekky Koua2,3*, Ibtihal M. Abdelhag4, Amar Mohamed Ismail2 1Faculty of Pharmacy, University of Medical Sciences & Technology-UMST, Khartoum, Sudan. 2Department of Biochemistry & Molecular Biology, Faculty of Science & Technology, Al Neelain University, Khartoum, Sudan. 3National University Research Institute, National University-Sudan, Khartoum, Sudan. 4Department of Histopathology, College of Medical Laboratory Sciences, Omdurman Islamic University, Omdurman, Sudan. ARTICLE INFO ABSTRACT Received on: 06/10/2018 The costus Saussurea lappa is a well-known medicinal plant that has been widely used in the traditional medicines Accepted on: 21/11/2018 in many Asian countries. Here, we report on the chemistry of the ethanol and water extracts of S. lappa roots using Available online: 08/05/2019 gas chromatography mass spectrometry (GC/MS) analysis. We further tested their antimicrobial activities against Staphylococcus aureus (Gram +ve) and Salmonella sp. (Gram –ve) isolates. The GC/MS analysis indicate that the ethanol extract has more compounds (37) than does the aqueous extract (18) with sesquiterpene lactones as the most Key words: abundant compounds in both extracts. The results reveal that the ethanol extract has significant antimicrobial activity Antimicrobial, medicinal against S. aureus isolate with 18–20 mm zone of inhibition, while no effect was observed against Salmonella sp. In plants, drug resistance, contrast, weak effects were found in the water extract against S. aureus and no effect against Salmonella sp. isolate. Costus, essential oil, GC/MS. Together with the GC/MS analyses, we conclude that the ethanol extract contains active secondary metabolites that may have a specific activity likely in a synergy effects with other metabolites against Gram +ve bacteria, but not Gram –ve isolate. Further experiments are needed to clarify the specific effects of S. lappa ethanol extracts against Gram +ve bacteria. INTRODUCTION pharmacology to intensively screen for the plants as promising The importance of plants as future sources of natural candidates for medicines to replace currently used synthetic antimicrobial agents holds great promise. Plants are inexpensive drugs (Abdallah, 2011; Cragg and Newman, 2013; Zulkipli and proved to be effective medicines to cure myriad of diseases et al., 2015). One of the big problems that is greatly challenging from mild microbial infections to complex diseases, including the public health is the rapid growing resistance developed cancers and devastating infectious diseases, such as tuberculosis, by infectious microorganisms against many synthetic drugs caused by Mycobacterium tuberculosis with much less side effects notably those frequently used against infectious diseases, such when compared to the synthetic drugs (Abuzeid et al., 2014; as food poisons, urinary tract, respiratory, and digestive systems Eklund et al., 2010; Koko et al., 2008). These advantages have (Cragg and Newman, 2013; Nascimento et al., 2000; Saga and encouraged scientists of different disciplines from chemistry to Yamaguchi, 2009). Some of the most threatening multi-drug resistant (MDR) microorganisms are Staphylococci, Streptococci, Salmonella, and M. tuberculosis (MDR-TB) owing to the high rates of horiztubeontal gene transfer that confers an MDR in these microorganisms even between evolutionarily distant species * Corresponding Author (Munita et al., 2017; Ventola, 2015). Hence, there is an urgent Faisal Hammad Mekky Koua, Department of Biochemistry & Molecular demand for antimicrobial agents, especially those with narrow Biology, Faculty of Science & Technology, Al Neelain University, Khartoum, Sudan. E-mail: amarqqqu @ yahoo.com spectra to minimize the rapid spread of resistance among infectious © 2019 Reem E. E. Omer et al. This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/). 074 Omer et al. / Journal of Applied Pharmaceutical Science 9 (05); 2019: 073-081 microorganisms (Chandra et al., 2017; Ventola, 2015). To this end, To further our understanding on the phytochemistry researchers have turned eyes toward plants as a rich and ancient and biological activities of S. lappa root, we applied gas source of various active compounds of medicinal importance for chromatography/mass spectrometry (GC/MS) analyses on the producing next-generation drugs that could possibly overcome the essential oil composition of the aqueous and ethanol extracts and MDR challenges, especially for problematic infectious agent, such used these extracts to assess their antimicrobial activities against as MDR-TB (Chandra et al., 2017). two bacterial isolates the S. aureus and Salmonella sp. representing One of the most intriguing medicinal plants that has Gram +ve and Gram –ve strains, respectively. been used widely in the traditional medicine is the Costus Saussurea lappa (Decne.) C. B. Clarke., which is widely used, MATERIALS AND METHODS along with other costus species, in many developing countries in Plant materials Asia (Pandey et al., 2007; Waly, 2009). The S. lappa is a member Roots of S. lappa were purchased from general herbal of Asteraceae family, a family that is rich with well-known market in Khartoum, Sudan. Taxonomical identification and medicinal plants of highly economic importance. Many of which authentication were taken place at the Herbarium of Medicinal have been intensively studied for their phytochemical constituents and Aromatic Plants and Traditional Medicine Research Institute and medicinal applications, including S. lappa (Pandey et al., of the National Center for Research in Khartoum, Sudan. Voucher 2007). The S. lappa root in particular has been widely prescribed specimen was deposited to the same Herbarium of the National for various infectious and physiological diseases, including Center for Research in Khartoum, Sudan. jaundice, diarrhea, stomachache, respiratory tract infections, antispasmodic agents against spasms caused by asthma, Microorganisms rheumatism, leprosy, cholera, and chronic skin diseases among To assess the antibacterial properties of the S. lappa root others (Gwari et al., 2013; Kala and Manjerkar, 1999; Kapoor, extracts two bacterial isolates of Gram +ve S. aureus and Gram 2001). For example, many studies have also shown that different –ve Salmonella sp. were kindly provided by the Department of extracts of S. lappa possess various pharmacological activities, Microbiology and Molecular Biology, Faculty of Science and including hepatoprotective, anti-inflammatory, anti-cancer, and Technology at Al Neelain University, Khartoum, Sudan. These anti-ulcer properties (Chen et al., 1995; Cho et al., 2004; Li et al., bacterial strains were confirmed by applying conventional 2005). This wide range of biological activities can be confidently biochemical methods and prepared accordingly for further attributed to the richness of S. lappa in pharmacologically active antibacterial tests. ingredients (Pandey et al., 2007). Phytochemical constituents, such as diosgenin, prosapogenin B of dioscin, diosgenone, cycloartanol, Preparation of extracts 25-en-cycloartenol, and octacosanoic acid, were isolated from the Root specimens were grounded into a fine powder and costus rhizomes (Qiao et al., 2002). The major constituents of S. sample of 50 g was carefully loaded onto Soxhlet extractor (Pyrex, lappa were found to be sesquiterpene lactones, such as saussurea Germany) and extracted with 300 ml of 99.9% ethanol (248 Worli lactone (Rao and Verma, 1951) and isodeydrocostus lactone (Kalsi road, Mumbai-30) for 3 hours as described somewhere else et al., 1983). Active sesquiterpenes lactones, such as costunolide (Pandey and Tripathi, 2014). The solvent was separated from the and dehydrocostus lactone, were previously isolated from the crude extract using rotary evaporator at 60°C for 30 minutes and methanolic extracts of S. lappa (Shoji et al., 1986), and appeared then the extract was kept at the room temperature to allow drying to have an anti-inflammatory activity that causes an inhibition gently. For aqueous extraction the same steps were followed by of the production of the tumor necrosis factor alpha in murine replacing the ethanol with water. macrophage-like cells as well as possessing a strong anti-cancer activity when mixed together (Damre et al., 2003; Peng et al., Gas chromatography mass spectrometry analyses 2017; Sunkara et al., 2010). For qualitative and quantitative assessment of the Different extracts of S. lappa roots, including hexane, essential oil components of the ethanol and aqueous extracts chloroform, ethylacetate ether, methanol, and water extracts, have of the S. lappa roots, GC/MS analyses were performed using a been reported to have various antibacterial and antifungal activities Shimadzu GCMS-QP2010 Ultra (Shimadzu, Japan) equipped against several pathogenic bacteria, such as Staphylococcus with a capillary column Rtx-5MS of 30 m × 0.25 mm ID with aureus, Staphylococcus epidermidis, Salmonella typhi, a film thickness of 0.25 μm according to a standard protocol Salmonella typhimurium, Streptococcus mutans, Escherichia coli, (Pongpiachan
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