“Antibacterial Property of Pandan Leaves Extract (Pandanus amaryllifolius)

on Staphylococcus aureus”

A research presented to the faculty and staff of the

Virgen Milagrosa University Foundation

Special Science High School

M.P Posadas Ave. San Carlos City Pangasinan

In partial fulfilment of the

Requirement in Science Research I

A thesis proposed by:

Alexandra Marie M. Aquino

Arvie Joyce G. Cayabyab

Ernest John G. Gonzales

Princess Marie F. Marquez

Kathleen Claire V. Ursua

THE PROBLEM

This chapter presents the problem of the study and its subtopics. It further

includes background of the study, conceptual framework, conceptual paradigm, statement

of the problem, null hypothesis, and significance of the study, scope and delimitation, and

definition of terms.

Background of the study

Pandan is a tropical plant in the Pandanus (screwpine) genus which is commonly

known as pandan leaves and is usually found in Southeast Asia

(http:eb.m.wikipedia.org/wiki/Pandanus amaryllifolius).It is an upright green plant with

fan-shaped sprays of long, harrow, blade like leaves and woody aerial roots. The leaves

and other parts of screw pine contain alkaloids and glycosides, organic compounds that

can provide health benefits to humans and animals.

Pandan leaves are the leaves of the plant Pandanus amaryllifolius, which is also

called pandan plant. Pandan plant is native to Asia and even tropical parts of Australia.

Most predominantly used the Southeast Asian cooking, this upright green plant has fan

shaped sprays like structure of leaves that are narrow and blade-like. These leaves are

attached to the woody aerial roots of the plant. Pandan plant is known to be sterile and it

is often propagated by cutting. The leaves are dark green in color, with strong nutty

aroma.
 Pandan flowers rarely. It prefers to be in filtered light to partial sun and likes

average moisture. It can reach a height of 6 to 8 feet tall. It is a tropical plant so avoid

freezing temperatures.

Bacteria are one-celled organisms visible only through a microscope.

Biologists classify them as prokaryotes. Prokaryotes are simple, single-celled organisms

that lack a defined nucleus. Bacteria inhabited Earth long before human beings or other

living things appeared. The earliest bacteria that scientists have discovered, in fossil

remains in rocks, probably lived about 3.5 billion years ago. (Microsoft Student 2009,

DVD)

Unlike in multicellular organisms, increases in cell size (cell growth and

reproduction by cell division) are tightly linked in unicellular organisms. Bacteria grow

to a fixed size and then reproduce through binary fission, a form of asexual

reproduction. Under optimal conditions, bacteria can grow and divide extremely rapidly,

and bacterial populations can double as quickly as every 9.8 minutes. In cell division, two

identical clone daughter cells are produced. Some bacteria, while still reproducing

asexually, form more complex reproductive structures that help disperse the newly

formed daughter cells.(http://en.wikipedia.org/wiki/Bacteria#Growth_and_reproduction)

Bacteria, like any other organisms in nature have good and bad effects on the

environment and in the human world. Beneficial roles of bacteria includes the enhancing

of soil fertility, helps in the decay and decomposition of organic matter, helps in the

industry and medicine and an important tool in biotechnology.

(http://www.preservearticles.com/2011122118779/five-important-beneficial-effects-of-

bacteria.html)
 But if bacteria form a parasitic association with other organisms, they are classed

as pathogens. Pathogenic bacteria are a major cause of human death and disease and

cause infections such as tetanus, typhoid fever, diphtheria, syphilis, cholera, foodborne

illness, leprosy and tuberculosis.

Each species of pathogen has a characteristic spectrum of interactions with its

human hosts. Some organisms, such as Staphylococcus or Streptococcus, can cause skin

infections, pneumonia, meningitis and even overwhelming sepsis, a

systemic inflammatory response producing shock, massive vasodilation and death.

(http://en.wikipedia.org/wiki/Bacteria)

Due to the earlier accounts and statements which provided information about the

antibacterial properties of pandan, the researchers came up with an idea of conducting a

research about the antibacterial property of Pandan (Pandanus amaryllifolius) to

determine its effectiveness in fighting the bacteria Staphylococcus aureus.

The study will be focusing on the antibacterial property of pandan leaves extract

on Staphylococcus aureus. The independent variable or the input of the study is pandan

leaves extract which will be tested and undergo several procedures in order to get the

desired result. The methods or the processes that will be used in the study are gathering

procedures, extraction, disc-diffusion method and phytochemical analysis. Through this

study, results will be drawn and with these results, the researchers will be able to identify

the antibacterial property of pandan leaves extract on Staphylococcus aureus.

Conceptual Paradigm

Input Process Output

Gathering
Procedures,
Leaf Extract of Antibacterial
Pandan Extraction
Property on
Null hypothesis
(Pandanus Disc-Diffusion, Staphylococcus
Amaryllifolius) aureus
Phytochemical
Analysis

Figure1. The paradigm of the study

This study aims to determine the Antibacterial property of the leaf extract of Pandan

(Pandanus amaryllifolius) on Staphylococcus aureus.

Generally, the study attempts to answer the following questions:

1. What are the active constituents present in the leaf extract of pandan?

2. What is the level of effectiveness of antibacterial property of pandan?

3. Is there a significant difference between the antibacterial property of pandan and the

positive control?

Hypothesis

Using the 0.01 level of significance, there is no significant difference between the

antibacterial property of the leaf extract of pandan and the positive control on

Staphylococcus aureus.

Significance of the study

This study will be of benefit, to the following and users:

To the Researchers. This study will enable them to realize the use of plants as an

alternative medicine in lowering blood-cholesterol level.

To Future Researchers. This study will serve as a guide to enable them to further study

on the other beneficial uses of Neem.

To Health Care Providers. This will serve as an eye opener and give additional

knowledge about an ideal alternative medicine in lowering blood-cholesterol

level.
To Pharmacists. This will give them an idea to produce more effective products that

would lower blood-cholesterol level.

To Pharmaceutical Companies. This study will be able to provide them knowledge that

they can use in creating new and effective drugs in lowering blood-cholesterol

level.

To the community. This will contribute to their wellness without spending much money

because these leaves are readily available in the community.

Scope and Delimitation

This study will be focusing on the antibacterial property of pandan leaves extract

on Staphylococcus aureus. In this study, the researchers will use pandan leaves that are to

be gathered at San Carlos City, Pangasinan. To determine the active constituents of

pandan, leaves extraction and phytochemical screening will be conducted at the College

of Pharmacy of Virgen Milagrosa University Foundation. The researchers will also

conduct disc diffusion method in the Microbiology Department of Virgen Milagrosa

University Foundation to determine the level of effectiveness of extracted pandan leaves

on Staphylococcus aureus. This study will be conducted in the year of 2013-2015.

Definition of Terms

Antibacterial. Any drug that destroys bacteria or inhibits their growth.

Disc Diffusion. Method of antibiotic susceptibility testing is the most practical method for

determining antibiotic resistance of microorganisms to different antimicrobial

pandan against staphylococcus aureus.

Extraction. The process of extracting something or of being extracted. The process of

obtaining the leaf extracts of Pandan for the purpose of examining the plant’s

antibacterial property against staphylococcus aureus.

Pandan. Any of the various Old World tropical palm like trees having huge prop roots

and edible cone like fruits and leaves like pineapple leaves. In the study, pandan

leaves extract drawn will be tested to determine its antibacterial effectiveness.

Phytochemical Analysis. Use to determine those chemical that may have biological

significance, for example antioxidants, but are not established as essential

nutrients. In the study, it will be used as a procedure to determine the antibacterial

property of pandan.

Staphylococcus aureus. Bacterium that is a member of the Firmicutes and is frequently

found in the human respiratory tract and on the skin. In this study, the bacterium

is used as a variable in order to determine the effectiveness of antibacterial

property of pandan.
 Chapter II

REVIEW OF RELATED LITERATURE

This chapter presents facts and information from different sources that will give

the readers additional knowledge about Pandan (Pandanus amaryllifolius).

Related Literature

Local

Pandan has been found to have medicinal benefits containing tannins, glycosides

and alkaloids, which said to be the reason for the effectiveness of such various health

concerns.

As a matter of fact, Pandan leaves is extremely useful for healing various wounds

and diseases such as smallpox, and is said to be a great pain reliever in such headache,

chest pain, arthritis, earache and a fever reducer as well. It is also believed to freshen the

breath and reduce gum and mouth pain by chewing its leaf. In addition, each leaf of

pandan is also effective in reducing stomach cramps and stomach spasm. It the same

manner, many people discovered and proved that it is an effective remedy for cough as

well. (http://www.drinkbenefits.com/pandan-leaves-health-benefits.htm)

Pandan also functions as a pain reliever, mostly for headaches and pain caused by

arthritis, and even hangover. It can also be used as antiseptic and antibacterial, which

makes it ideal for healing wounds. (http://lmc.lorma.org/pdf/Pandan(October).pdf)

Foreign
 The Pandan Leaf comes from the Screwpine Tree, which can be found in tropical

areas of Asia and Europe. The plant has many uses. It has medicinal properties. The

medicinal uses for this tree are as follows: The entire plant is used as a diuretic, the

roots have anti-diabetic properties, and the leaves are used for treating diseases of the

skin..(http://www.essortment.com/many-uses-pandan-leaf-53654.html)

Related Studies

Local

In the study conducted by Dumaoalet.al., (2010). entitled “In Vitro Activity of

Pandan (Pandanus amaryllifolius) Leaves Crude Extract Against Selected Bacterial

Isolates”, Pandan (Pandanus amaryllifolius), crude extract can inhibit the growth of S.

aureus resulted from the biological screening and can kill the said bacterial isolate. The

study implied that the use of pure concentration may result in inhibition and in killing of

the infecting organism.

(http://philair.ph/publication/index.php/jpair/article/view/103/326)

Foreign

(Gazali, 2007) states in his study, Extraction of Pandanus Amaryllifolius

Essential Oil from Pandan Leaves thatPandan leaves are very beneficial for various

health conditions. Pandan leaves consist of essential oils, traces of tannin, glycosides and

alkaloids as well. As a matter of fact, wholepandan plant is considered to be diuretic and

is extremely useful for healing various wounds and diseases like smallpox. Pandan leaves

are said to be pain relievers and used that way to cure chest pain, headache, reduce fever,
arthritis, earache, etc.Pandan leaves are also used as a healthy laxative for children. They

are also found to be effective in recovery of women with weakness after childbirth. It has

a cooling effect and is excellent for the treatment of internal inflammations, urinary

infections, colds, coughs, measles, bleeding gums and skin diseases.

Aside from that, Pandan leaves are also anti-carcinogenic, while benefits

ofpandan leaves for diabetes are also very significant. Pandan leaves are also useful for

treating several skin disordersincluding leprosy. Pandan leaves are also used for

preparation of various herbal teas, with other herbs like lemongrass, mulberry leaves,

safflower, green tea and other such herbs.Bathing with water having boiled pandan

leaves, is useful for treating skin diseases and sunburns.

(http://umpir.ump.edu.my/2709/1/NUR_SYAZWANI_BINTI_GHAZALI.PDF)

Chapter III
 RESEARCH METHODOLOGY

This chapter denotes to exhibit and demonstrate the methods and procedures that

will be utilized in the study. This chapter includes the research design, research subject,

reagents, materials and statistical treatment used in the study.

Research Design

The research design that will be implied by the researchers in the study is the

two -group design. Penicillin will be used as the control group while pandan will be

wielded as experimental group used in the study.

Research Subject

The bacterium Staphylococcus aureus that will be cultured from the Microbiology

Department of Virgen Milagrosa University Foundation will be used as the research

subject of the study.

Locale of the Study

Pandan leaves will be collected by the researchers at Brgy. Manzon, San Carlos

City, Pangasinan. The phytochemical screening that will be used in the study will be

conducted at the Department of Pharmacy in Virgen Milagrosa University Foundation

while the disc-diffusion method will be conducted at the Microbiology Department of

Virgen Milagrosa University Foundation.

Research Materials/Reagents
 For extraction of leaf extracts: Pandan (Pandanus amaryfollus) leaves, tap

water, casserole, stove and sanitized vial or small glass bottle.

For antibacterial analysis: Extract product of Pandan (Pandanus amaryfollus)

leaves, pure culture of Staphylococcus aureus, test tube, petri dish, inoculating loop,

cotton swab, thumb forcep, alcohol lamp, beaker, caliper or ruler, incubator , autoclave

and paper disc soaked with the extract product.

Data Gathering Procedure

The researchers will gather pandan (Pandanus amaryfollus) leaves from Brgy.

Manzon, San Carlos City, Pangasinan. The leaves of the said plant will be washed first

with tap water to ensure its cleanliness. Then the leaves will be steamed in a casserole to

produce the extracts needed. The extricated extracts will then be transferred into a clean

vial.

The obtained extracts from pandan will be tested using the disc diffusion,

screening methods, and phytochemical analysis to determine its antibacterial property

against the bacteria Staphylococcus aureus.

Statistical treatment

The statistical treatment that will be used in the study of antibacterial property of

the leaf extract of pandan on Staphylococcus aureus will be the t-test method. The

assumptions are that they are samples from normal distribution.

t= −¿ ¿

√( )(
Ss 1+ Ss 2 1 1
+
n1 +n2−2 n1 n2 )
Where:

is the mean for Group 1.

is the mean for Group 2.

is the number of people in Group 1.

is the number of people in Group 2.

Ss1is the variance for Group 1.

Ss2is the variance for Group 2.