Madre de Cacao (Gliricidia Sepium) Leaves Extract As An Antibacterial Hand Sanitizer

Madre De Cacao (Gliricidia Sepium) Leaves
Extract as an antibacterial hand sanitizer

OBJECTIVE
S
1. GENERAL OBJECTIVES:
The goal of this research is to study the activity of Antibacterial

in Madre De Cacao and use it to make sanitizer. Researchers will
cultivate bacteria to test the effectiveness of the product. Researches
propose a solution to help people what we are currently facing the Corona
Virus pandemic, using with what we see around us like the leaves
of Madre de cacao that will be beneficial to the community.
2. SPECIFIC OBJECTIVES:
2.1. To determine if there is an Antibacterial properties in Madre de

Cacao.
2.2. To determine the effectiveness of Antibacterial properties of

Madre de Cacao to formulate Hand sanitizers
2.3. To identify how long the effectiveness of the Antibacterial

properties of Madre De cacao is.
2.4. To Formulate the Hand sanitizer using Madre Cacao leaves

extract.
METHODOLOGY
CONCEPTUAL FRAMEWORK
INPUT PROCESS OUTPUT
Experimental
Antibacterial Effective
Different
agent in formulation of
hand
Madre de hand sanitizer sanitizer
Cacao Time kill test for E.
coli and S. aureus
II. METHODOLOGY
The research method used in this study is center on the growth of

Staphylococcus aureus and Streptococcus using Madre de Cacao extract formulated
as compared with different types of commercial brand hand sanitizers contain with
ethyl and isopropyl alcohol. The treatment is managed as it is with gram positive
controls and experimental bunch since regulating negative control probably will not
appear any significant antibacterial activity.
Material and Method
Plant Sample
 Gliricidia Sepium Leaves (Madre de Cacao)
Medium
 Nutrient broth and agar
 Mueller Hinton agar
Sample
 Volunteer sample: Swab from the hand skin of the volunteer without any clinical
signs of disease is included in this study.
 Bacteria sample: Staphylococcus aureus and Streptococcus

Preparation of culture
Collecting of Plant
media and Inoculation
Material
of bacteria.
Preparation of Procurement of pure

sample and extract Preparation for the culture
Experimental Procedures
Preparation of
commercial hand Formulation of Madre
sanitizer de Cacao hand sanitizer
Collecting of Plant Material
The fresh leaves of Gliricidia Sepuim (Madre de Cacao) plant are collect at Brgy. San Jose Antipo
city. The fresh leaves of kakawate are collect; wash altogether to expel undesirable contaminants and
other particles which may influence in the microbial activity of the plant sample. It is finely chop or cu
into smaller pieces using knife and wash once more to maintain contamination free.
Preparation of sample and extract

250 grams of finely cut G.sepium leaves are soaked with 95% ethyl for 72 hours then the mixture is filter
and the marc is discarded. The Filtrate is dissipated until it comes to a syrup consistency. An excess solvent
is evacuated by setting the remaining volume in hot air oven (at 40 oC). The residue will soaked again, all
filter is will collect and vanish. The compacted extract is liquefied by 20% of surfactant and distilled water.
All extract is set in a firmly cover holder and keep it refrigerated at 28 oC until advance uses. 1% of solution
of G.sepium extract will undergo Phytochemical screening for the detection of different classes constituents.
The extract will be used to formulate the Madre de Cacao hand sanitizer.
Preparation of commercial hand sanitizer
Ethanol and isopropanol are the common solutions found in most hand sanitizers. They kill organisms
without causing any hurt in the skin tissues, hence making them effective hand sanitizers ingredient and
disinfectant. The hand sanitizers that utilize in this study are 2 varying brand names with different
manufactures. The brand name are lifebuoy hand sanitizers and alcogel hand sanitizers.
Formulation of Madre de Cacao hand sanitizer

Gelatin powder is set in the beaker and dissolves with a portion of glycerin with the help of warm.
This is previously pre-dissolves in alcohol, the remaining alcohol and glycerin is also added. With the
use of spatula it is enthusiastically mix until it is fully mix together. The Madre de cacao leaf extract is
include to the blend which is then transfer to a container.
Procurement of pure culture
The microbial culture required for testing are procure at (tentative laboratory). Upon securing there
pure culture are sealed in a fix holder to avoid contaminations while being transfer. It should also be
labeled appropriately for the security purposes. It will be preserve at refrigeration and use paraffin method
before being held in the organic testing. Staphylococcus aureus and Streptococcus are gotten through
segregation of unadulterated culture by the utilize of quadrant streak, isolation procedures and streaking
method.
Preparation of culture media and Inoculation of bacteria.

Before getting culture S. aureus and Streptococcus arrangement of the culture medium is need to be done. 20g of
nutrient agar dissolve in water and boil to complete dissolution. The agar solution is autoclave in 220 degree
centigrade for 15 minutes. The sterile solution is pour with 20ml in each petri dish cover with foil. The agar will
cool at room temperature and set aside for refrigeration after the preparation. the pure culture is obtain by
inoculation.(Biwa’s &Rogers,2013).

Antibacterial Measurement of the
susceptibility Zone inhibition
testing
Test and Analytical

Screening of Antibacterial
Methods Microbial Test
activity by Agar Well
Diffusion Assay
Determination of Minimum
Inhibitory concentration (MIC)
of the extract
After the preparation proceeds to the Method that will be used in this study these are the following:
Methods:
Antibacterial susceptibility testing
The formulated Madre de Cacao hand sanitizers and commercial brand hand sanitizer are going to
test using standard Mueller Hinton II plate to detect the antibacterial activity of these solutions.
Screening of Antibacterial activity by Agar Well Diffusion Assay

The agar well diffusion method (Perez et al., 1990) is employing. Mueller Hinton agar is the
medium inoculate with the microorganism which is Staphylococcus aureus and Streptococcus.
Determination of Minimum Inhibitory concentration (MIC) of the extract
MIC is the defined lowest concentration that completely inhibits the solution of microorganisms for
24hours. The plate’s agar will incubate at 37oC for 24 hour. The lower concentration of extract showing a
clear zone of inhibition is considered as MIC.
Measurement of the Zone inhibition

A larger zone bacteria-free zone that encompasses a hand sanitizer disk implies that the microscopic
organisms are more sensitive to the sanitizers that the disk contains. To measure the distance across of the
surrounding area is will be measure in millimeters (mm) of the zone, the zone that
is repressed whereas keeping the plate’s cover input.
Microbial Test
Microbial test is analyses before applying sanitizer and after applying sanitizer
REFERENCES
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Madre De Cacao (Gliricidia Sepium) Leaves

Extract as an antibacterial hand sanitizer

The goal of this research is to study the activity of Antibacterial

2.1. To determine if there is an Antibacterial properties in Madre de

2.2. To determine the effectiveness of Antibacterial properties of

2.3. To identify how long the effectiveness of the Antibacterial

2.4. To Formulate the Hand sanitizer using Madre Cacao leaves

INPUT PROCESS OUTPUT

The research method used in this study is center on the growth of

 Bacteria sample: Staphylococcus aureus and Streptococcus

Preparation of Procurement of pure

Preparation of sample and extract

Formulation of Madre de Cacao hand sanitizer

Preparation of culture media and Inoculation of bacteria.

Test and Analytical

Screening of Antibacterial activity by Agar Well Diffusion Assay

Measurement of the Zone inhibition

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