Main Article Content
Aims: To determine chemical constituents of the Leaf extracts of Centella asiatica using the LC-MS and GC-MS and their antimicrobial activities.
Study Design: Structural determination of compounds from the leaf extracts was done using GC-MS and LC-MS analysis. The antimicrobial properties of the extracts were done using disc diffusion method.
Place and Duration of Study: Pure and Applied Chemistry Department, Masinde Muliro University of Science and Technology, Kenya: Between 2016-2019.
Methodology: Plant materials of C. asiatica were sequentially extracted separately based on the polarity viz., hexane, ethyl acetate and methanol. Determination of chemical constituents was done using LC-MS and GC-MS analysis and phytochemical screening. The extracts were assayed against Staphylococcus aureus, Klebsiella pneumonia, Escherichia coli and Candida albicans. Standard antimicrobials viz. ampicillin and Nystatin were used as the control. Disc diffusion method was used and zones of inhibition, after respective incubation periods, were used to quantify antibacterial and antifungal activity.
Results: Phytochemical screening of the hexane, ethyl acetate and methanolic extracts showed that terpenoids, flavonoids, saponins, alkaloids, steroids, amino acids and proteins, reducing sugars and carbohydrates were present. LC-MS and GC-MS analyses of the methanolic extracts identified 22 and 33 compounds, respectively, by use of the national institute of standards and technology (NIST) library. The extracts showed appreciable activity against common microbes tested.
Conclusion: This study forms the basis for the biological characterization and significance of the compounds identified in the leaf and stem extracts of C. asiatica. These compounds are known to possess antibacterial and antifungal activities that could be established as potential candidates for future drug development. However, these extracts, need to be subjected to further chromatographic procedures to isolate the identified compounds and their bioactivities determined.
Evans CW. Trease and Evans Pharmacognosy (16 Ed.). London. Saunders Elservier Ltd; 2009
Mebberley D. The Plant Book-Apotrable Dictionary of Plants, their Classifications and Uses. New York. Cambridge University Press; 2008
Brinkhaus B, Lindner M, Schuppan D, Hahn EG. Chemical, pharmacological and clinical profile of the East Asian medical plant Centella asiatica. Phytomed. 2000; 7(5):427-448. DOI: 10.1016/s0944-7113(00)80065-3
Prakash V, Jaiswal N, Srivastava MA. Review on Medicinal Properties of Centella Asiatica. Asian J Pharm & Clin Res. 2017; 10(10): 69. DOI: 10.22159/ajpcr.2017.v10i10.20760
Mollik MAH, Hossain MF, Sen D, Hassan AI, Rahman MS; Traditional Asian medicine and leprosy in Bangladesh. European Journal of Integrative Medicine. 2009; 1(4):191–192. DOI: 10.1016/j.eujim.2009.08.096
Satake T, Kamiya K, An Y, Oishi(nee Taka) T, Yamamoto J. The Anti-thrombotic Active Constituents from Centella asiatica. Biological & Pharmaceutical Bulletin. 2007; 30(5):935–940. DOI: 10.1248/bpb.30.935
Brinkhaus B, Linda M. Chemical, Pharmacological and Clinical profile of the East Asian Medical Plant Centela asiatica, Review Article. Phytomed. 2000;7:427-448.
Hagemann RC, Burnham TH, Granick B, Neubauer D, St. Louis MO. Facts and Comparisons Division. J. B. Lippincott Co; Gotu kola, In, The Lawrence Review of Natural Products: facts and comparisons. 1996;41–2.
Gohil KJ, Patel J, Gajjar, AK. Pharmacological Review on Centella asiatica: A Potential Herbal Cure-all. Indian Journal of Pharmaceutical Sciences. 2010; 72(5):546–556. Available:https://doi.org/10.4103/0250-474X.78519
Orhan IE. Centella asiatica (L.) Urban: From traditional medicine to modern medicine with neuroprotective potential. Evidence-Based Complementary and Alternative Medicine. 2012;946259:1-8. DOI: 10.1155/2012/946259
George M and Joseph L. Anti-allergic, antipruritic, and anti-inflammatory activities of Centella asiatica extracts. Afr J Trad Comp & Alt Med. 2009;6(4):554–559.
Cheng CL, Guo JS, Luk J, Koo MWL. The healing effects of Centella extract and asiaticoside on acetic acid induced gastric ulcers in rats. Life Sci. 2004;74(18):2237-2249.
Pingale SS. Evaluation of effect of Centella asiatica on CCl4 induced rat liver damage. Pharmacologyonline. 2008;3:537–543.
Wijeweera P, Arnason JT, Koszycki D, Merali Z. “Evaluation of anxiolytic properties of Gotukola—(Centella asiatica) extracts and asiaticoside in rat behavioral models. Phytomed. 2006;13(9-10):668–676.
Idris FN and Nadzir MM. Antimicrobial Activity of Centella asiatica on Aspergillus niger and Bacillus subtilis. Chem. Eng. trans. 2017;56:1381-185.
Raghavendra M, Maiti R, Kumar S, Trigunayat A, Mitra S, Acharya S. Role of Centella asiatica on cerebral postischemic reperfusion and long-term hypoperfusion in rats. Intl J. Green Pharm. 2009;3(2):88–96.
Babykutty S, Padikkala J, Sathiadeya, P, Vijayakurup V, Azis T, Siriniyas, P. Apoptosis Induction of Centella asiatica on Human Breast Cancer Cells. J Trad Comp & Alt Med. 2008;6:9-16.
Long HS, Stander MA, Van Wyk BE. Notes on the occurrence and significance of triterpenoids (asiaticoside and related compounds) and caffeoylquinic acids in Centella species. South Afr. J. Bot. 2012; 82: 53-59. DOI: 10.1016/j.sajb.2012.07.017
Antognoni F, Perellino NC, Crippa S, Dal Toso R, Danieli B, Minghetti A, Pressi G. Irbic acid, a dicaffeoylquinic acid derivative from Centella asiatica cell cultures. Fitoterapia. 2011;82(7):950–954. DOI: 10.1016/j.fitote.2011.05.008.
Alcazar MA, Wright K, Vaswani A, Caruso, M, Reed RL, Bailey CF, Maier, CS. Integration of mass spectral fingerprinting analysis with precursor ion (MS1) quantification for the characterization of botanical extracts: application to extracts of Centella asiatica (L.) Urban. Phytochem Anal. 2020;1-17. DOI: 10.1002/pca.2936
James JT, Dubery IA. Pentacyclic Triterpenoids from the Medicinal Herb, Centella asiatica (L.) Urban. Molecules. 2009;14:3922-3941. DOI: 10.3390/molecules14103922
James J, Dubery I. Identification and quantification of triterpenoid centelloids in Centella asiatica (L.) Urban by densitometric TLC. J. Plan Chromatography – Modern TLC. 2011; 24(1):82–87. DOI: 10.1556/jpc.24.2011.1.16
Subban R, Veerakumar A, Manimaran R. Two new flavonoids from Centella asiatica (Linn.). J Nat Med. 2008;62:369–373. DOI: 10.1007/s11418-008-0229-0
Bonfill M, Mangas S, Cusidó RM, Osuna L, Piñol MT, Palazón J. Identification of triterpenoid compounds of Centella asiatica by thin-layer chromatography and mass spectrometry. Biomed. Chromato-graphy. 2006; 20(2):151–153. DOI: 10.1002/bmc.564
Karting T. Clinical application of Centella asiatica (L) Urb. In herbs, spices and medicinal plants (Craker LE, Simon, JE Eds.). Oxyx press, phoenix; 1988.
Shukla A, Rasik A, Jain G, Shaikner R, Kulshrestha D, Dhawan B. Invitro and in vivo wound healing activity of asiaticoside isolated from Centella asiatica. J Ethnopharmacol. 1999;1:1-11.
Hanson JR. Natural products: the secondary metabolites Cambridge: Royal Society of Chemistry. ISBN 0–85404-490-6; 2003.
Inamdar P, Yeole R, Ghogare A, & Souza N. Determination of biologically active constituents in Centella asiatica. J Chromatograp. 1996;742:127-130.
Bonev B, Hooper J, Parisot J. Principals of assessing bacterial susceptibility to antibiotics using the agar diffusion method. J Antimicrob Chemotherapy. 2008;61(6): 1295-301.
Brown D, Kothari D. Comparison of antibiotic discs from different sources. J.Clin.Pathol. 1975;28(10):779-83.
Neelamegam, R and Ezhilan, B; GC-MS analysis of phytocomponents in the ethanol extract of Polygonum chinense L. Pharmacognosy Research, 2012;4(1):11. DOI :10.4103/0974-8490.91028
Shen Y, Liu A, Ye M, Wang L, Chen J, Wang X, Han, C. Analysis of Biologically Active Constituents in Centella asiatica by Microwave-Assisted Extraction Combined with LC–MS. Chroma. 2009;70:431–438. Available:https://doi.org/10.1365/s10337-009-1152-6
Yadav RNS, Agarwala M. Phytochemical analysis of some medicinal plants. J Phytol. 2013;(12):10-14. [ISSN: 2075-6240]
Samy R, Ignacimuthu, Chow V. Antimicrobial and Phytochemical Analysisof Centella asiatica (L). Nature Procedings. 2011;6033.
Barne JS, Anderson LA, Philipson JD. Herbal Medicines (3rd Ed.).; London: Pharmaceutical Press; 2007.
Zakaria F, Ibrahim WNW, Ismail IS, Ahmad H, Manshoor N, Ismail N, Zainal Z, Shaari K. LCMS/MS Metabolite Profiling and Analysis of Acute Toxicity Effect of the Ethanolic Extract of Centella asiatica on Zebrafish Model. Pertanika J. Sci. & Technol. 2019;27 (2):971 -989.
Schoenroth LJ, Hart DA, Pollard KM, Fritzler MJ. The effect of the phytoestrogen coumestrol on the NZB/W F1 murine model of systemic lupus. J Autoimmun. 2004;23(4):323-332. DOI: 10.1016/j.jaut.2004.09.004
Yarnell E, Abascal K. Lupus Erythematosus and herbal medicine. Alt. & Comp. Ther. 2008;14(1):9–12. DOI: 10.1089/act.2008.14105
Jin X, Wang S, Zhao X, Jin Q, Fan C, Li J, Shan Z and Teng W. Coumestrol inhibits autoantibody production through modulating Th1 response in experimental autoimmune thyroiditis. Oncotarget. 2016; 7(33):52797–52809. DOI: 10.18632/oncotarget.10353
Siddiqui BS, Aslam H, Ali ST, Khan S, Begum S. Chemical constituents of Centella asiatica. J As Nat Prod Res. 2007;9(4):407–414. DOI: 10.1080/10286020600782454
Sugunabai J, Jeyaraj M, T Karpagam, Varalakshmi B, Rani SS, Shanmugapriya A, Kalaiyarasi G, Renuga R, Gomathi S. Outlining of Phytochemicals and GC-MS Profile of Centella asiatica. Intl J Pharmac & Dr. Anal. 2018;6(2):252–256. ISSN: 2348-8948 Available:http://ijpda.com
Ghosh K, Indra N. Phytochemistry, in vitro free radical scavenging, chelating and toxicity of Centella asiatica L. (apiaceae) ethanolic leaf extract. Intl J Pharmaceut Sci Rev & Res. 2014;29:328-334.
Santhi FC,Thomas MT. Essential oil profiling of Centella asiatica (L.) Urb.‐ a medicinally important herb. South Indian J Bio Sci. 2016;2(1):169‐173. ONLINE ISSN: 2454‐4787
Oyedeji OA, Afolayan AJ. Chemical composition and antibacterial activity of the essential oil of Centella asiatica Growing in South Africa. Pharmaceutical Bio. 2005; 43(3):249–252. DOI: 10.1080/13880200590928843
González-Rey C, Svenson SB, Bravo L, Siitonen A, Pasquale V, Dumontet S, Krovacek K. Serotypes and anti-microbial susceptibility of Plesiomonas shigelloides isolates from humans, animals and aquatic environments in different countries. Comp. Immunology, Microbiology and Infectious Diseases. 2004;27(2):129– 139. DOI: 10.1016/j.cimid.2003.08.001
Terry H, Ken M. Microbes and Diseases. London. Harper Collins; 2002.
Sakoulas G, Moellering Jr, RC. Increasing Antibiotic Resistance among Methicillin‐ Resistant Staphylococcus aureus Strains. Clinical Infectious Diseases. 2008; 46(S5): S360–S367. DOI: 10.1086/533592