Antibacterial Potential of Dimocarpuslongan leaves extract

A Science Research Presented to the Faculty and Staff ofSaintMarys 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 ii iii iv v 1 1 2 2 2 2 3 4

Acknowledgement Dedication Abstract -

Table of Contents

CHAPTER I. BACKGROUND OF THE STUDY Rationale -

Statement of the Problem -

Statement of the Null Hypotheses Significance of the study Scope and Delimitation Definition of Terms -

CHAPTER II. REVIEW OF RELATED LITERATURE AND STUDIES CHAPTER III.MATERIALS AND METHODS Materials -

6 6

Methods

6 9 10 11 12 14

Research Environment The Project Set Up Experimental Design

CHAPTER IV. DATA PRESENTATION AND ANALYSIS

CHAPTER V. SUMMARY, CONCLUSIONS AND RECOMMENDATION BIBLIOGRAPHY 15 16 17 18

DOCUMENTATION COMMUNICATION CURRICULUM VITAE

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 Marys 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 Antibacterial 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 nations 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, dragons 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 antiinflammatory 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 Marys 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 Scissors Blender Beaker Digital weighing pan Aluminium foils ethyl alcohol filter paper funnel casserole stirring rod Erlenmeyer flask mortar and pestle rubber band masking tape inoculating loop petri dish ketokenozone

Alcohol lamp 15g agar 3g beef extract Tissue paper Methods

500 ml distilled water beaker/s filter paper disk streptomycine sulphate

bleach gulaman bar dishwashing liquid

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 its 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

E E C E E E

E C

Staphylococcus aureus

E. coli

Where: C-control E-extract of Dimocarpus longan leaves

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 Marys University College Biology and Chemistry Laboratories. SUMMARY OF FINDINGS

CONCLUSION AND RECOMMENDATIONS

REFERENCES

APPENDICES

COMMUNICATIONS

ACKNOWLEDGEMENT

DEDICATION

10 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) 1 Staphylococcus aureus E. Coli Control 2 3 4 5

11 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) 1 Extract Control 10.3 32.3 10 31.3 2 9.5 33.9 3 10.8 40.9 4 9.4 38 5

P<0.0005 If P<0.05 -There is significant difference on Staphylococcusaureus.

12 The table indicates that theres 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) 1 Extract 6 2 6 3 6 4 6 5 6

21.1 Control

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 didnt 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.

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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 Marys University Elementary: Saint Marys University Grade VI- with academic distinction Secondary: Saint Marys University

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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 Marys University Grade School Department Grade 1, 2, 3, 4, 5, 6: Top Ten Secondary: Saint Marys University Science High School

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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 Marys University Elementary: Saint Marys University Grade School Department With distinction Secondary: Saint Marys University Science High School

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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 Marys University Elementary: Saint Marys University Grade School Department Secondary: Saint Marys University Science High School

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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 Marys University Grade School Department Secondary: Saint Marys University Science High School

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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 Marys University Science High School

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