In Vitro Evaluation Of Antifungal, Antioxidant And Cytotoxicity Activity Of Couroupita Guianensis With Identification Of Antifungal Active Fraction

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Date
2015-12
Authors
Sivapragasam, Gothai
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Universiti Sains Malaysia
Abstract
The current study was undertaken to evaluate the antifungal, antioxidant and cytotoxicity activities of Couroupita guianensis flower extract. The Couroupita guianensis flower extract was selected from 125 local medicinal plant by disc diffusion method against Streptococcus aureus, E. coli and C. albicans. It was found that the C. guianensis flower extract possessed a broad spectrum of antimicrobial properties. With respect to the emergence of susceptibility of C. albicans to antifungal agent, this study was conducted to determine the anti-Candida activity of C. guianensis flower extract. Anti-Candida test was carried out using disc diffusion assay, broth dilution method, and time killing profile. The effect of C. guianensis flower extracts upon the morphological and structure of C. albicans were examined by scanning electron microscope (SEM). Anti-Candida activity indicated that the mean zone of inhibition was 18.2 ± 0.3 mm. The MIC and MFC value for C. albicans found to be 12.5 mg/mL and 25 mg/mL respectively. Time kill experiments indicated an intense concentration-dependent fungicidal effect against C. albicans. Imaging of SEM revealed a gradual alteration over time on the outer microstructure of C. albicans. A complete destruction of C. albicans cells were observed after 36 hours of exposure to flower extracts C. guianensis. Qualitative phytochemical screening C. guianensis flower extract showed the presence of alkaloids, flavonoids, tannins, steroids, glycosides and saponin. The Total phenolic content of the C. guianensis flower extract was determined and exhibited moderate value with 32.2 ± 0.22 mg of GAE/100 gram of extract. The free radical scavenging activity of C. guianensis flower extract was assessed by 2, 2-diphenyl-1-picryl hydrazyl (DPPH) radical scavenging assay and hydrogen peroxide radical scavenging activity. The IC50 values for C. guianensis flower extract for DPPH and hydrogen peroxide free radicals scavenging activities was 93.2 μg/mL ± 0.011 and 46.48 ± 0.13 μg/mL respectively. The cytotoxic effect of the C. guianensis flower extract was evaluated using brine shrimp assay and MTT assay. The compounds in C. guianensis flower extract exhibited non-toxic results with LC50= 1210.65 μg/mL and IC50= 513.22 μg/mL. Genotoxicity study on C. guianensis flower extract was assessed by the single cell gel electrophoresis (comet assay). Genotoxicity study results showed that treatment with C. guianensis flower extract did not cause obvious DNA damage in vero cells. The flower exract of C. guianensis was serially extracted with solvents of varying polarities, with hexane, ethyl acetate, butanol and water and tested for anticandida activity by disk diffusion method. Ethyl acetate fraction was regarded as active fraction on the basis of highest zone of inhibition (18.99 mm). The active ethyl acetate fraction were analysed with Gas Chromatography-Mass Spectrometry (GC-MS). The GC-MS analysis showed the presence of docosanoic acid, eicosanoic acid, n-hexadecanoic acid, benzamide, 2, 3, 4, 5-tetrafluoro-N(-3-methylthio-1, 2, 4- triazol-5-yl), 9, 12-octodecadienoic acid (Z, Z), octadecadienoic acid, pentadecanoin acid, 14-methyl-, squalene and tetradecanoic acid in the ethyl acetate fraction which may responsible for observed antimicrobial and antifungal activities in this study. The results of this study suggested that C. guianensis flower extract possess good antifungal and antioxidant activity. Further study is required to identify the chemical compounds responsible for their biological activity
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Molecular Medicine
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