Antibacterial Potential of Dimocarpuslongan leaves extract

A Science Research Presented

to the Faculty and Staff ofSaintMary’s University High School and Science High

School

In Partial Fulfilment of the Course Requirement in

Science Research and Statistics 2 (SRAS II)

Student Investigators:

Zcyra Mae Billedo

Jeffrey Macadangdang

Dianne Cez Valeroso

Maria Paula Laureta

Rina Ananayo

Roselle Agulto

Research Adviser

Ms. Elsa Cajucom

SRAS2 Teacher

September 2012

TABLE OF CONTENTS

Page

Title Page - - - - - - - - i

Acknowledgement - - - - - - ii

Dedication - - - - - - - - iii

Abstract - - - - - - - - iv

Table of Contents - - - - - - - v

CHAPTER I. BACKGROUND OF THE STUDY - - 1

Rationale - - - - - - - 1

Statement of the Problem - - - - - 2

Statement of the Null Hypotheses- - - - 2

Significance of the study - - - - 2

Scope and Delimitation - - - - - 2

Definition of Terms - - - - - - 3

CHAPTER II. REVIEW OF RELATED LITERATURE AND - 4

STUDIES

CHAPTER III.MATERIALS AND METHODS - - - 6

Materials - - - - - - - 6

Methods - - - - - - - 6

Research Environment - - - - - 9

The Project Set Up - - - - - 10

Experimental Design - - - - - 11

CHAPTER IV. DATA PRESENTATION AND ANALYSIS - 12

CHAPTER V. SUMMARY, CONCLUSIONS AND RECOMMENDATION 14

BIBLIOGRAPHY - - - - - - - 15

DOCUMENTATION - - - - - - 16

COMMUNICATION - - - - - - 17

CURRICULUM VITAE - - - - - - 18

ABSTRACT

Title: The Antibacterial Potential of Dimocarpus Longan Leaves Extract

Student Researchers: Zcyra Mae C. Billedo, Jeffrey T. Macadangadang, Dianne Cez

Valeroso, Rina Ananayo, Maria Paula T. Laureta, Roselle M. Agulto

Research Adviser: Ms. Elsa Cajucom

Saint Mary’s University Science High School

The Objective of this research is to find an alternative plant that will be efficient

as anti-bacterial agent againstStaphylococcus aureus and Escheria coli. The researcher

used the extracted leaves of Dimocarpus longanin the Anti-bacterial experiment.

The study aimed to find if DimocarpusLongan in killing the bacteria. That can be

used as a product in killing disease-causing bacteria.

The result of the data of the longan leaves against Staphylococcus aureusis

partially active compared to the control while the result of the longan leaves against

Escheriacolli are known to have no significant difference between the controls.

Dimocarpus longan is proven effective against Staphylococcusaureus as Anti-

bacterial agent but not effective against Escherichia coli.

INTODUCTION

Background of the Study

Antibacterial is a compound known to kill bacteria or microbes. Staphylococcus

aureus is one of the most common bacteria which causes pus, known as abscess and

cellulitis. E. coli is also a common bacterium known to cause food and water

contamination.

Dimocarpuslongan, commonly known as longan, is a member of the

Sapindaceae family. It is a highly attractive fruit extensively distributed in China and

South East Asia including Thailand, Vietnam and the Philippines. Its leaves have a

description of dark green, shiny, leathery, up to 12 inches (30 cm) long with wavy

margins and blunt pointed tips. There are 6 to 9 pairs of leaflets per leaf. It is known to

improve the immunity system, heal wounds and replenish blood, and anti-ageing.

Our country today is facing problems especially in our economy. Disease is one

of the reasons. If the researchers prove the potential effect of Dimocarpuslongan

leaves, there is a possibility that the percentage of bacterial growth in our human body

can be lessened by the use natural sources.

Statement of the Problem

This project aimed to answer the following questions:

1. Would Dimocarpuslonganleaves extract be effective to kill

Staphylococcusaureus?

2. Would Dimocarpuslonganleaves extract be effective to kill E. coli?

Statement of the Null Hypotheses

1. Dimocarpuslongan leaves extract is not effective in killing Staphylococcusaureus.

2. Dimocarpuslongan leaves extract is not effective in killing E. coli.

OBJECTIVE OF THE STUDY

General Objective

The pbjective of this study is to help the growing nation’s knowledge about

Dimocarpus longan leaves extract for its widely use.

Specific Objective

The objective of this study is to find out whether or whether not Dimocarpus

longan leaves extract can annihilate Staphylococcus aureus and E. coli bacterias

SIGNIFICANCE OF THE STUDY

This studyproved that Dimocarpuslongan leaves extract has the efficiency in the

inhibitory growth of bacteria such asStaphylococcus aureus and E. coli. It can be used

as a product such as that may be used to avoid diseases.

SCOPE AND DELIMINATION

The research, with the availability of longanleaves can be done with the help of a

professional in a laboratory. It is intended to find out the antibacterial potential of longan

leaves that can be useful in making products like antiseptics, dishwashing liquid, etc.

REVIEW OF RELATED LITERATURE

Related Literature

Longan, Dimocarpuslongan, is a fruit tree closely related to the lychees. Longans

are also called ‘lungan’, ‘dragon’s eyes’, or ‘eyeball’. It is a large handsomeevergreen

tree and it is native of Southern western India occurs in the forest of Western ghats from

Kongan Southwards to Tinnevely hills.(Wikipedia)

An antibacterial is a compound or substance that kills bacteria. According to

study, most antibacterial chemically are synthetic modifications of various natural

compounds.(Wikipedia)

Longan leaves, according to a study in 2009, contains Hexadecanoic acid and

Octadecadienoic acid. Hexadecanoic acid or Palmitic acid is mainly use to produce

soaps, cosmetics and detergents. Octadecadienoic acid or Linoleic acid is use in anti-

inflammatory and antioxidants. Benzoic acid is also present which use for treating

fungal diseases.(African Journal of Biotechnology)

Longan leaves also contain 3.30% pentane, 8.44 % propane, 1.47% pyran,

1.47% methyl, 1.47% dihidro, 1.47% dihidroxyl, 0.40% benzoic acid, 1.20% amino acid,

1.20% benzoil fluoride, 5.39% Aziridinon, 5.39% tert,5.39% butyl, 5.39%

methylcyclohexy, 2.69% Hexadecane, 1.49% naphtalenone, 1.49% hexahydro and

5.46% Tetramethylchroman.(African Journal of Biotechnology)

Related Studies

A 2010 research was already conducted by Ducusin L. of Saint Mary’s University

High School that deal with the pythochemical analysis on longan leaves and gained

positive results. It was reviewed that longan leaves can be used as an antibacterial

agent.

The International Journal of Pharmaceutical Research and Development

conducted a research that goals to find the Gas Chromatography analysis on longan

leaves. Hexadexanoic acid and Octadecadienoic acid was found high in it.(African

Journal of Biotechnology)

RESEARCH METHODOLOGY

Conceptualization

MATERIALS AND METHODS

Materials

Plant extracts ethyl alcohol mortar and pestle

Scissors filter paper rubber band

Blender funnel masking tape

Beaker casserole inoculating loop

Digital weighing pan stirring rod petri dish

Aluminium foils Erlenmeyer flask ketokenozone

Alcohol lamp 500 ml distilled water bleach

15g agar beaker/s gulaman bar

3g beef extract filter paper disk dishwashing liquid

Tissue paper streptomycine sulphate

Methods

A. Gathering and Preparation of Longan Leaves

1. Gather longan leaves.

2. Wash the longan leaves.

3. Then dry it.

4. Cut into pieces.

5. Put into a container to avoid filth, close the container.

B. Air-Drying of Leaves

1. Select an area where the leaves are not exposed to the sun.

2. After air-dried, grind the leaves.

3. Keep the ground leaves in a sealed container.

C. Preparation of the Extracts

1. Transfer the pieces of longan leaves to the 150 mL beaker.

2. Treat the plant material with 95% of ethanol.

3. Stir it with aluminum foil and keep it for 48 hours.

4. After 48 hours, put the longan leaves on the water bath but not exceeding 40˚ to

remove the remaining alcohol to obtain pure essential.

D. Water Bath

1. This stage will enable the ethanol separated from extracted liquid.

2. The extract was in the water bath apparatus for 2 days.

7

E. Preparation of Culture Media

1. Mix 500 ml distilled water, 2.5 g peptone, 1.5g beef extract, 7.5 g agar using a stirring

rod in a casserole.

2. Stir while heating.

3. Place the liquid in Erlenmeyer flask. Cover with a stopper.

F. Preparation of Filter Paper Disks

1. Prepare the Filter Paper Disks.

2. Prepare the beakers of extract and positive control.

3. Soak disk in extract.

G. Dispensing Agar in Plates

1. Heat periphery of petri dish.

2. Distribute agar until it’s heavily covered at the bottom.

H. Inoculation

1. Heat periphery of Petri dishes. Wipe the moist with a paper towel.

2. The bacteria from the test tube will be poured to the first petri dish for S.A.

3. Swirl a couple of times.

4. After 5 minutes, the contents of Petri dish 1 will be transferred to Petri dish 2. Repeat

step 5 until the third Petri dish.

5. The contents will now be placed in a beaker.

6. Repeat step 2 to step 5 for E. coli.

7. Place the petri dishes in the container.

I. Data Gathering

1. Use a Digital VernierCaliper to measure the zone of inhibition.

J. Data Analysis

1. Use t-test to analyse data.

Flowchart

Experimental Design

EEC

C

Where:

C-control

E-extract of Dimocarpus longan leaves

C

EE

Staphylococcus aureus E. coli

E

E

EE

C

Statistical Treatment

RESULTS, ANALYSIS AND DISCUSSIONS

The longan leaves were collected at Batu Ferry, Bambang near the river. The

investigatory project is conducted at Saint Mary’s University College Biology and

Chemistry Laboratories.

SUMMARY OF FINDINGS

CONCLUSION AND RECOMMENDATIONS

REFERENCES

APPENDICES

COMMUNICATIONS

ACKNOWLEDGEMENT

DEDICATION

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Experimental Design

Title: Antibacterial Potential of Dimocarpus Longan Extract

Hypothesis: Dimocarpus longan is not effective in killing the bacteria, Staphylococcus

aureus and E. coli

IV: Longan Leaves extract

DV: Antibacterial potential

Constant: millimetre(mm)

Table 4.1 Zone of Inhibition (Staphylococcusaureus and E. coli)

Zone of Inhibition (mm)

Staphylococcus aureus1 2 3 4 5

E. ColiControl

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CHAPTER IV

DATA PRESENTATION AND ANALYSIS

In this chapter, the data are presented in the form of tables. The results were

computed by statistics. Below the table are the analysis and interpretation of the data.

Table 4.1Zone of inhibition (Staphylococcus aureus)

Zone of Inhibition (mm)

Extract1 2 3 4 5

10.3 10 9.5 10.8 9.4

Control32.3 31.3 33.9 40.9 38

P<0.0005

If P<0.05

-There is significant difference on Staphylococcusaureus.

12

The table indicates that there’s a significant difference between the data

gathered of the control variable and the extract on the Staphylococcusaureus, the data

also indicates that the longan extract is not that efficient as anti-bacterialagent than the

control.

Table 4.2 Zone of inhibition (E. coli)

Zone of Inhibition (mm)

Extract

1 2 3 4 5

6 6 6 6 6

Control

21.1 21.9 19.5 21.1 19.8

The table indicates that the Longan extract did not respond on the growth of E. coli

because the extract didn’t have a zone of inhibition. This indicates that longan is insufficient in

preventing E.coli bacteria.

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CHAPTER V

SUMMARY, CONCLUSIONS AND RECOMMENDATIONS

Summary

This research aimed to find an alternative herbal plant that can prevent

Staphylococcus aureus and E. coli bacteria. The researchers gathered fresh longan

leaves and they extract the needed essential nutrients by the use of ethyl alcohol, the

extracted product went to the process of water bath. After the process was done and

the extract was finished, the researchers went to the laboratory to test if the extract can

kill bacteria.

Conclusions

1. Longan extract is effective in killing the bacteria, Staphylococcus aureus.

2. Longan extract is not effective in killing the bacteria, E. coli

Recommendations

1. The future researchers should try experimenting on the antifungal effects of

longan extract.

2. A product out of the extract could be tried like antiseptic, dishwashing liquid, etc.

14

BIBLIOGRAPHY

Ducusin, L. (2009) The Phytochemical Analysis of Dimocarpus

Longan leaves. SMU Unpolished

Research.

Longan Leaves. http://en.wikipedia.org.

Date accessed: September 04, 2012

Longan Leaves. http://www.ijrpd.com

Date accesed: September 05, 2012

Yuge L., Liqin L., Yiwei M., Changbin W., Lingling L. and Ping L.(2012) African Journal

of Biotechnology

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DOCUMENTATION

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CURRICULUM VITAE

I. PERSONAL INFORMATION

Name: Jeffrey TottocMacadangdang

Age: 14

Birthday: October 25, 1997

Birthplace: Bayombong, Nueva Vizcaya

Father: Jessie Miguel Macadangdang

Mother: EstrelitaTottocMacadangdang

II. EDUCATIONAL ATTAINMENT AND ACHIEVEMENT

Preschool: Kiddie College, Saint Mary’s University

Elementary: Saint Mary’s University

Grade VI- with academic distinction

Secondary: Saint Mary’s University

18

CURRICULUM VITAE

I. PERSONAL INFORMATION

Name: Zcyra Mae C. Billedo

Age: 14

Birthday; February 20, 1997

Birthplace: Bayombong, Nueva Vizcaya

Father: Ericson TiongsonBilledo

Mother: Amelia Castro Billedo

II. EDUCATIONAL ATTAINMENT AND ACHEIVEMENT

Preschool: Kids Workshop Learning Center

Nursery: with distinction

Kinder: Top 4

Elementary: Saint Mary’s University Grade School Department

Grade 1, 2, 3, 4, 5, 6: Top Ten

Secondary: Saint Mary’s University Science High School

19

CURRICULUM VITAE

I. PERSONAL INFORMATION

Name: Roselle M. Agulto

Age: 16

Birthday: October 6, 1996

Birthplace:

Father: RoniAgulto

Mother: Carol ManaloAgulto

II. EDUCATIONAL ATTAINMENT AND ACHIEVEMENT

Preschool: Saint Mary’s University

Elementary: Saint Mary’s University Grade School Department

With distinction

Secondary: Saint Mary’s University Science High School

20

CURRICULUM VITAE

I. PERSONAL INFORMATION

Name: Dianne CezValeroso

Age: 14

Birthday: November 11, 1997

Birthplace: Bayombong, Nueva Vizcaya

Father: Rowel Damaso Valeroso

Mother: Mary Ann Ramos Valeroso

II. EDUCATIONAL ATTAINMENT AND ACHIEVEMENT

Preschool: Saint Mary’s University

Elementary: Saint Mary’s University Grade School Department

Secondary: Saint Mary’s University Science High School

21

CURRICULUM VITAE

I. PERSONAL INFORMATION

Name: Maria Paula T. Laureta

Age: 15

Birthday: June 18, 1997

Birthplace: Bayombong, Nueva Vizcaya

Mother: Maria Dolores Laureta

II. EDUCATIONAL ATTAINMENT AND ACHIEVEMENT

Preschool: Day Care Center

Elementary: Bayombong South Elementary School

Saint Mary’s University Grade School Department

Secondary: Saint Mary’s University Science High School

22

CURRICULUM VITAE

I. PERSONAL INFORMATION

Name: Rina Guimpayan Ananayo

Age: 15

Birthday: June 10, 1997

Bithplace: Lagawe, Ifugao

Father: Richard ButicAnanayo

Mother: DonalynGuimpayanAnanayo

II. EDUCATIONAL ATTAINMENT AND ACHIEVEMENT

Preschool: Kingsway Christian Academy

Preschool: Top Ten

Elementary: Kingsway Christian Academy (Grade 1, Grade 3-6)

United Methodist Christian School (Grade 2)

Grade 1, 2,3,5, and 6: Top ten

Secondary: Saint Mary’s University Science High School

23