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
Journal Title
Journal ISSN
Volume Title
Publisher
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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Keywords
Molecular Medicine