PHYTOCHEMICAL SCREENING
AND ANTIBACTERIAL ACTIVITIES
OF Cucumis metuliferus
Purity Cheptoo
(DSLT/306J/2021)
Supervisor. Dr. Ali Shee
Introduction
Free radicals are oxidizing agents because they have reactive
oxygen species.
They have a surplus of one or more free floating electron
rather than having paired electron therefore they are very
reactive and unstable.
Often deprived from normal body metabolism or external
source like X-ray irradiation.
Failure to inactive this free radicals their chemical reactivity
can damage cellular substrate like lipids which can cause
chronic infection like diabetes and ulcers.
Therefore in humans, antioxidants help solve this potential
hazards of free radicals by scavenging them
Introduction…
Chemical antioxidants are bioactive compounds that protect
against oxidation of various substates.
They block propagations stage in oxidative chain reactions
by preventing the formation of these radicals.
Antibiotics have been used to solve this.
However the clinical efficiency of many existing antibiotics is
being threatened by the emergency of multidrug resistance
pathogens.
The resistance problem demands that renewed effort be
made to screen various medicinal plants for the potential
antimicrobials traits
Problem statement
Free radicles are healthy hazards because they cause
oxidative and chronic disease in humans.
The emergence and spread of drug resistance pathogens
continues to threaten our ability to treat common infections
with existing antimicrobials drugs.
Home made melon have been used in Kenya traditional to
treat variety of chronic condition such as ulcers, stomach
aches and diarrhea.
But there lacks adequate information of home made
melon in terms of antioxidant and antibacterial potential as
well as phytochemical composition
Objectives
General objective
To determine phytochemical constituents, antibacterial and
antioxidant potential of Cucumis metuliferus extracts
Specific objectives
1.To determine the phytochemicals of Cucumis metuliferus
2.To evaluate antioxidant activities of the extracts of
Cucumis metuliferus
3.To determine antimicrobials activities of the extracts of
Cucumis metuliferus.
Methodology
The samples of Cucumis metuliferus will be air-dried for at
least four weeks in a shade away from sunlight, grounded
using pestle and mortar to homogenous powder to
increase surface area for extraction.
Sample will be extracted using methanol in 1:10 ratio, in a
shaker for 12 h.
The extracted sample will be filtered and centrifuged at
1000 rpm for 10 min.
The supernatant collected will be filtered using Whatman
filter paper No. 1 and concentrated at 70 ⁰C evaporated
using vacuum rotary evaporator to obtain the extract.
The extract will be stored in the refrigerator at 4 ⁰C until
use.
Methodology…
Phytochemical tests were will be carried out to determine the classes of
natural molecules contained in the extracts of Cucumis metuliferus as
described by Gul et al., (2017); Singh et al., (2022).
Test for alkaloids (Mayers test). 0.5 g of crude extract will be put in a
clean test tube and few drops of Mayer’s reagent added. The appearances
of a creamy or white precipitate indicate presence of alkaloids.
Test for flavonoids (Alkaline reagent test). 0.5 g of crude extract will be
placed in a test tube and few drops of dilute NaOH solution added.
Appearances of intense yellow colour indicate presence of flavonoids.
Test for tannin (Gelatin test). 2 g of crude extract was weighed and
dissolved in 5 mL of distilled water in a test tube, filtered and few drops of
ferric chloride reagent added. The appearances of yellow to deep green
precipitate indicate presence of tannin.
Test for Terpenoids (Salkowski test). 0.5 g of crude extract will be
weighed, dissolved in 2 mL of chloroform in a test tube and 5 ml of
concentrated sulphuric acid added to form a layer. The appearance of foam
indicate presence of terpenoids.
Methodology…
Test for saponins (Frothing test). 2 g of crude extract will be weighed,
dissolved in 5 mL of distilled water in a test tube, shaken vigorously to
form froth then few drops of olive oil added and shaken vigorously.
Appearances of foam indicate presence of saponins.
Test for steroids (Salkowski test). 0.5 g of crude extract will be
weighed, placed in a test tube. 2 mL of chloroform and concentrated
sulphuric acid will be added. Appearances of red color on the lower
chloroform layer indicate presence of steroids.
Test for phenols (Ferric Chloride Test). 2 mL of crude extract will be
measured, placed in a test tube and 3-4 drops of FeCl 3 added.
Appearances of bluish black colour indicate presence of phenol.
Test for glycosides (Killer killiani test). 0.5 g was weighed, mixed
with 2 mL of glacial acetic acid in a test tube then 1-2 drops of 5% FeCl 3
and concentrated H2SO4 added
Methodology…
In vitro antimicrobial susceptibility test
The agar disk-dilution method of European Committee on
Antimicrobial Susceptibility Testing (EUCAST, 2013) will be
employed to determine MIC using MHA against S. aureus
as described by (Bauer et al., 1966).
Serial dilution will be employed to yield a series of different
concentrations (5, 2, 0.8, 0.4, 0.1, 0.05, 0.03 and 0.01
mg/mL) of both plant fractions in test tubes labelled T1 toT8.
Disc of 0.7 cm will be soaked into these tubes for 15 min
and later these impregnated discs will be spotted on
inoculated agar plate and then incubated at 37°C for 24 h.
The same will be repeated, now with the extract mixed. After
24 h, zones of inhibition will be measured in millimeters to
ascertain bioactivity of the extracts.
For negative controls sterile water will be used instead of
extract and amoxicillin for positive control.
Work plan
Activity Month
October,2023 November,2023 December,2023 January,2024 February,2024 March,2024 April,
2024
Concept
development
Proposal
writing
Proposal
presentation
Data
collection and
analysis
Report writing
Report
presentation
Budget
ITEM COST (Ksh)
Equipment cost 1,600
Infrastructure costs 1,200
Raw material cost 8,000
Labor cost 1,500
Research and Development 1,500
Miscellaneous cost 1000
TOTAL COST 17500
THANK YOU!
