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PHYTOCONSTITUENTS OF LEMONGRASS (Cymbopogon citratus) LEAVES

CRUDE EXTRACT POTENT AS AN ANTIOXIDANT A Science Investigatory
Project Individual Category for Pupils -Life Science Bue...

Research · January 2019

DOI: 10.13140/RG.2.2.33988.50569

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PHYTOCONSTITUENTS OF LEMONGRASS (Cymbopogon citratus)
LEAVES CRUDE EXTRACT POTENT AS AN ANTIOXIDANT

A Science Investigatory Project

Individual Category for Pupils – Life Science
Buenavista SPED Elementary School

An Official Entry to the

2019 National Children’s Science Congress
of the Science Club Advisers Association of the Philippines

ARIANNA TRISHA C. ABIVA – Research Presenter

FLOR E. CARINOSA – Project Adviser/Author

January, 2019
i
 ACKNOWLEDGMENT

The researcher would like to extend her thoughtful thanks to all those

who, in a way or another, contributed to the realization of this science

investigatory project;

To her science subject teacher, Mrs. Flor E. Cariñosa, for her

supervision and reinforcement, for approving the research plan, effort in

checking and improving this work, for guidance on what to do, and for ample

sharing of her knowledge and skills;

To the district science coordinator, Mrs. Rizalita C. Espera, for

extending her assistance in making this work a success;

To the resource speaker of SIP, Mrs. Analou O. Hermocilla for her

assessment, comment and suggestion to improve this study;

To the researcher’s parents, who offered their help and guidance for

the completion of this study for their understanding, prayers, and financial

provision;

To the researcher’s friends, for the good support to help us overcome

the challenges;

Most of all, thanks to Almighty God who gave us limitless guidance and

support to help us accomplish this research.

ii
 DEDICATION

This project is heartedly dedicated to Almighty God who

has been our initiator of life, the foundation of knowledge and

wisdom.

This modest determination is also committed to the

researcher’s parents, whose love, encouragement, and prayers

of day and night enable to complete such success.

Finally, it is our sincere gratitude and warmest care that

this effort is dedicated to the communities, students, advisers

and researchers who have been an endless source of

knowledge and inspiration.

ATCA

iii
 ABSTRACT

Plants have been a source of traditional medicines which exist for

thousands of years. Human has been depended to it and lives long because

of natural medicines. Over the years, the number of synthetic drugs has been

developed all over the world. And the number of diseases rises due to

commercialized synthetic drugs. This heightened the researcher’s ability to

investigate Lemongrass as a potent source of antioxidant. This study focused

on the determination of phytoconstituents found in Lemongrass (Cymbopogon

citratus) leaves crude extract. The result showed through phytochemical

screening that the occurrence of bioactive secondary metabolites of

Lemongrass (Cymbopogon citratus) leaves crude extract obtained higher

amount were alkaloids, flavonoids, steroids, and tannins, while the occurrence

of anthraquinones, cyanogenic-glycoside, and saponins was not observed.

The potential antioxidant activity of Lemongrass (Cymbopogon citratus) was

then observed through its occurrence of bioactive secondary metabolites.

Based on the findings, the researchers found out that the existence of the

higher amount of bioactive secondary metabolites in Lemongrass

(Cymbopogon citratus) leaves were the components potent as available

sources of natural antioxidant and its positivity in fatty acid which are involved

in the production of soap, detergents, and cosmetics can be used as an

antifungal, antibacterial as well as antimicrobial activities.

iv
 TABLE OF CONTENTS

TITLE PAGE i

ACKNOWLEGDMENT ii

DEDICATION iii

ABSTRACT iv

TABLE OF CONTENTS v - vi

LIST OF FIGURE vii

LIST OF TABLE viii

CHAPTER

1. Introduction

Background of the Study 1

Objectives of the Study 2

Significance of the Study 2

Scope and Limitations of the Study 2

Definition of Terms 3

2. Review of Related Literature 4

3. Methodology

Materials Used in the Study 8

Procedure 8

v
4. Results and Discussions

Phytochemical Screening of Lemongrass

(Cymbopogon citratus) Leaves Crude Extract 10

5. Summary, Findings, Conclusions,

and Recommendations

Summary 13

Findings 13

Conclusions 14

Recommendations 14

References 15

APPENDICES 17

vi
 LIST OF FIGURE

FIGURE TITLE

1 Flowchart of the Study 9

vii
 LIST OF TABLE

TABLE TITLE

1 Phytochemical Screening of Lemongrass

(Cymbopogon Citratus) Leaves Crude Extract 10

viii
 Chapter 1

Introduction

Background of the Study

Considering their usage in conventional medicine, natural resources

have long been a source of medicinal substances. An astounding number of

modern pharmaceuticals have been obtained from these sources. This is due

to control major diseases. The fundamental knowledge of modern medicine

comes from plants. Rich natural sources supply the fundamental chemical

and active structures needed to create synthetic fields (Preeti et al., 2010).

Lemongrass (Cymbopogon citratus) belongs to the family Poaceae, is

a tall perennial grass with slender sharp edge and green leaves that have a

pointed apex. Warm temperate and tropical climates are where it is native

(Bleasel et al., 2002). Used as a pesticide and preservative is lemon grass oil.

Used as a pesticide and preservative is lemon grass oil. Moreover, it functions

as an antifungal (Shadab et al., 2001). In soap, creams, candles, and

detergents, it serves as a fragrance. Fever, a cold, a cough, and upset

stomach can all be treated with lemon grass tea. It has diuretic properties and

water retention, making it helpful in an individual with high blood pressure. It

can also help to prevent typhoid fever, cancer, blurring of vision, lower

cholesterol levels, relieve menstrual problems and nausea. It can be used as

herbal medicine to treat nervous condition and inflammation, chest infections,

sores, muscle cramps and headache (Leite et al., 2000). Alkaloids, saponins,

tannins, anthraquinones, steroids, phenols, and flavonoids are all present in

1
lemongrass. These phytochemicals each have a variety of medicinal and

protective properties. (Nabiar and Matela, 2012).

Objectives of the Study

The main purpose of this study was to determine the phytoconstituents

found in Lemongrass (Cymbopogon citratus) leaves crude extract.

Specifically, this study aimed to:

1. Determine the bioactive secondary metabolites found

in Lemongrass (Cymbopogon citratus).

2. Determine if Lemongrass (Cymbopogon citratus) leaves

crude extract potent as an antioxidant.

Significance of the Study

The result of the study is significant to the communities to support them

in investigating medicinal plants as a possible source of active antioxidant. To

the future researchers, this study might be needed to determine its

phytoconstituents and its proper application to the human body.

Scope and Limitations

The study will be conducted at Buenavista SPED Elementary School,

located at Poblacion 5, Buenavista, Agusan del Norte on August 01, 2018 up

to September 12, 2018, with the supervision of the project adviser. The

analysis of phytoconstituents found in Lemongrass (Cymbopogon citratus)

leaves crude extract was referred from the Department of Chemistry, College

of Science and Mathematics, MSU-Iligan Institute of Technology, Iligan City.

2
This science investigatory project was limited only in using Lemongrass

(Cymbopogon citratus) leaves crude extract potent as an antioxidant.

Definition of Terms

To promote understanding and clarity on the concepts presented in this

study, the following terms were defined operationally.

Antioxidant. It is a substance that inhibits oxidation, especially one

used to counteract the deterioration of stored food products.

Bioactive compounds. These are distinguished from essential

nutrients.

Crude extract. It is a chemical, typically a physiologically active

ingredient, that is created by separating it from the original material using

solvents or evaporation.

Fatty acid. Particularly any of those appearing as esters in fats and

oils, it is a carboxylic acid made composed of a hydrocarbon chain and a

terminal carboxyl group.

Free radical. It describes a non-charged molecule with an unpaired

valence electron that is often extremely reactive and short-lived.

Metabolite. It is a substance formed in or necessary for metabolism.

Micronutrient. It is a chemical substance or element that is necessary

for the regular growth and development of living things in very little amounts.

Oxidative. It is relating to the process or result of oxidizing or being

oxidized.

Phytoconstituents. It refers to the synthetic intensifies that actually

happen in plants.

3
 Chapter 2

Review of Related Literature

This section provides the literature that has the relevance of the study.

Most of the biological effects ascribed top Cymbopogon citratus

extracts have been attributed to its primary bioactive constituents, derived

from its leaves, stem, and roots, and their secondary metabolites. Recently, a

large number of empirical studies have been carried out aiming to expand our

understanding of Cymbopogon citratus phytoconstituents including a number

of recently conducted studies. These investigations have shown that the

chemical composition of Cymbopogon citratus extracts varies according to the

geographical origin, genetic differences, part of the plant used, a method of

extraction, age/stage of maturity, and season of harvest. Despite these

differences, a number of classes of compounds are reproducibly found,

including tannins, saponins, flavonoids, alkaloids phenols, and

anthraquinones (Ekpenyong et al., 2014).

It shows that Lemongrass contains alkaloids, saponins, tannins,

anthraquinones, steroids, phenols, and flavonoids. These phytochemicals

each have a variety of medicinal and protective properties. For instance, it

was known that phenol modified erythrocyte membranes (Adesanya and

Sofowora, 1983).

The cited literature focused on the phytoconstituents of Cymbopogon

citratus.

4
 Hot Water Extract had significantly higher DPPH radical scavenging

ability, Fe2+ chelating ability and OH* scavenging ability than Cold Water

Extract. Conclusively, heat treatment may affect antioxidant properties due to

the release of phenolic phytochemicals, hence contributing to the health

promoting and disease preventing abilities of Cymbopogon citratus (Oboh et

al., 2010).

Patel and Mehta 2006, evaluated the compounds having antioxidant

property in lemon Grass and found that dry lemon grass contains more phenol

and flavonoids than a fresh one. Hence we can infer that the dried lemon

grass has greater antioxidant potential than the fresh lemon grass.

The cited literature focused on the antioxidant activities of

Cymbopogon citratus.

Cymbopogon citratus is used as traditional medicine in the treatment of

the nervous condition, gastrointestinal disturbances, fever, and hypertension.

Coughs, elephantiasis, the flu, gingivitis, headaches, leprosy, malaria,

ophthalmia, pneumonia, and vascular problems can all be treated with lemon

grass. It is principally taken in the form of tea as a remedy for digestive

problems, diarrhea and stomach ache. Lemon grass has historically been

used as a carminative and an insect repellent (Dama et al., 2011). Studies on

extract leaves have demonstrated anti-inflammatory, vasorelaxing, a diuretic

and valuable remedy in treating ringworm as a local application (Gore et al.,

2010).

5
 Cymbopogon citratus is an Angolan medicinal plant use by local people

for treating high fever, stomach, gut problems and headache. Due to the

unpleasant aroma of its infusion, this plane is widely known and used in

tropical and sub-tropical countries. Moreover, it has antidepressant and mood-

enhancing properties. Additional studies have revealed antioxidant, anxiolytic,

sedative, hypolipidemic, and hypoglycemic effects (Blanco et al., 2009).

Numerous plant species have been discovered to have medicinal uses (Ali-

Shtayeh and Abu, 1999).

Numerous plant species have been discovered to have medicinal uses

(Ali-Shtayeh and Abu, 1999). Several essential oils display antibacterial

(Ozcan et al., 2006), antifungal, and other therapeutic properties. Essential

oils are fragrant compounds that are produced from various plant parts by

steam distillation (Chee and Lee, 2007) and antiviral (Khan et al., 2005)

activity. Lemon grass (Cymbopogon citratus is widely used in traditional

medicine in many countries around the world. Among its attributable

properties are those related to antibacterial and antifungal activities (Inouye et

al., 2001, Inoue et al., 2006; Bansod and Rai, 2008; Revathi et al., 2012),

along with having painkilling and anti-inflammatory qualities (Negrelle and

Gomes, 2007).

The cited literature focused on the herbal and traditional medicinal

uses of Cymbopogon citratus.

Cymbopogon citratus has been used over many years to make

caffeine-free tea and as an herbal drink, suggesting that it may be a healthier

alternative to caffeine-containing tea products. In comparison to other tea

6
brands consumed among Nigerians (Lipton tea, Nescafe, Green tea, and Top

tea); Cymbopogon citratus tea was a good source of antioxidants such as

flavonoids, and therefore a nutritionally acceptable and medicinally valuable

beverage. While some research has indicated that it contains tannins,

coumarins, saponins, and anthraquinones, which have been linked to modest

harmful effects, the limited bioavailability of these phytochemicals in people

makes these effects less of a concern may confer a measure of protection

against toxicity. The average concentrations of anti-nutrients such as phylate

are not significantly higher in Cymbopogon citratus while cardiac glycosides,

cyanates, phlobatannins, and heavy metals like lead and mercury are absent.

Therefore, Cymbopogon citratus is considered to be safe for human

consumption on the basis of its phytochemical constitution, both nutritional

and non-nutritional (Akande et al., 2014).

The cited literature focused on the use of Cymbopogon citratus as a

tea for protection against toxicity and diseases.

In the United States of America and some regions of Asia, where they

are grown for commercial purposes as industrial raw materials to cosmetic

and insecticide manufacturers, Cymbopogon citratus is a plant of enormous

economic importance. It is used as a scent in many product including soap,

perfume candle, mosquito and other insect repellents (Praditvarn and

Samhandharaksa, 1990).

The cited literature focused on the industrial use of Cymbopogon

citratus raw materials.

7
 Chapter 3

Methodology

This chapter presents the materials used in the study, procedure, and

utilization of Lemongrass (Cymbopogon citratus) leaves for phytochemical

screening.

Materials Used in the Study

Lemongrass (Cymbopogon citratus) leaves, basket, basin, knife,

chopping board, strainer, and sterilized cellophane bag.

Procedure

First, the researchers collected the leaves of Lemongrass

(Cymbopogon citratus). Second, the leaves were washed and cleaned

through tap water. Third, air dried for 72 hours. Next, the leaves were cut into

small pieces. Then, leaves were put inside the sterilized cellophane bag.

Finally, the researchers submitted the plant sample to the Department of

Chemistry, College of Science and Mathematics, MSU-Iligan Institute of

Technology for phytochemical screening.

8
 First, the researchers collected the leaves of Lemongrass
(Cymbopogon citratus).

Second, the leaves were washed and cleaned

through tap water.

Third, air dried for 72 hours.

Next, the leaves were cut into small pieces.

Then, leaves were put inside the sterilized cellophane bag.

Finally, submit the plant sample to the Department of

Chemistry, College of Science and Mathematics, MSU-Iligan
Institute of Technology for phytochemical screening.
Figure 1. Flowchart of the Study

Figure 1. Flowchart of the Study

9
 Chapter 4

Results and Discussions

This chapter presents the analysis and results of the data gathered by

the researchers. The data is presented in a table form with an appropriate

interpretation follows to clarify numerical data.

Table 1. Phytochemical Screening of Lemongrass

(Cymbopogon Citratus) Leaves Crude Extract

Secondary Metabolites Occurrence

Alkaloids ++

Anthraquinones -

Cyanogenic-glycoside -

Flavonoids +++

Saponins -

Steroids +++

Tannins +++

Table 1 shows the bioactive secondary metabolites present in

Lemongrass (Cymbopogon citratus) leaves, in which the amount level of

alkaloids, flavonoids, and steroids, tannins are higher, while the occurrence of

anthraquinones, cyanogenic-glycoside and saponins were not observed.

10
 Based on recent studies, Alkaloids have diverse structures and show a

range of pharmaceutical activities including antihypertensive effects,

antiarrhythmic effect, antimalarial activity, anticancer actions, antibacterial and

antifungal activities, cancer chemoprevention and anti-oxidants (Saxena et al.

2013).

Recent research has focused on flavonoids because of their extensive

biological and pharmacological functions. Flavonoids have been reported to

exert multiple biological properties including antimicrobial, cytotoxicity, anti-

inflammatory as well as antitumor activities but the best-described property of

almost every group of flavonoids is their ability to function as potent

antioxidants capable of shielding the organism from reactive oxygen species

and free radicals. The molecular makeup of flavonoids determines how

effective they are as antioxidants. The location of hydroxyl groups and other

characteristics of flavonoids' chemical structure play a crucial role in the

antioxidant and free radical scavenging abilities of these compounds. (Singh

et al., 2013).

Flavonoids as naturally occurring secondary metabolites in plants are

thought to have positive effects on human health. Plants with flavonoids have

been reported to have antiviral, antitumor, and antioxidant properties. These

compounds are considered as powerful antioxidants because they exhibited

inhibitory properties against carcinogenesis is a number of vivo studies.

Flavonoid has been confirmed to effectively scavenge most oxidizing

molecule such as singlet oxygen and various free radicals linked in chronic

diseases (Baang et al., 2017).

11
 Steroids possess many interesting medicinal, pharmaceutical and

agrochemical activities like anti-tumor, immunosuppressive, hepatoprotective,

antibacterial, plant growth hormone regulator, sex hormone, anti-helminthic,

cytotoxic and cardiotonic activity (Patel et al., 2015).

Tannins in medicine especially in Asian (Japanese and Chinese)

natural healing, the tannin-containing plant extracts are used as astringents,

against diarrhea, as diuretics, against stomach and duodenal tumors, and as

inflammatory, antiseptic, antioxidant and haemostatic pharmaceuticals.

Tannins are employed as caustics for cationic dyes (tannin dyes) in the

dyestuff business as well as in the creation of inks (iron gallate ink). Wine,

beer, and fruit juices are clarified by tannins in the food sector. Additional

industrial applications for tannins include the dyeing of textiles, antioxidants in

the brewing and wine industries, and coagulants in the manufacture of rubber.

Recently the tannins have attracted scientific interest, especially due to the

increased incidence of deadly illnesses such as AIDS and various cancer

(Saxena et al., 2013).

12
 Chapter 5

Summary, Findings, Conclusions, and Recommendations

The findings, conclusions, and recommendations of the study are

summarized in this chapter.

Summary

The study focused on the determination of phytoconstituents found in

Lemongrass (Cymbopogon citratus) leaves crude extract. The objectives of

the study presented in two (2) questions. First, focus of the study was on the

presence of bioactive secondary metabolites in lemongrass (Cymbopogon

citratus). Second, was on its potential activity as an antioxidant.

Findings

Based on the analysis and interpretation of the following are the

findings:

1. The occurrence of bioactive secondary metabolites of Lemongrass

(Cymbopogon citratus) leaves crude extract obtained higher amount were

alkaloids, flavonoids, steroids, and tannins, while the occurrence of

anthraquinones, cyanogenic-glycoside and saponins were not observed.

2. The potential antioxidant activity of Lemongrass (Cymbopogon

citratus) was then observed through its occurrence of bioactive secondary

metabolites.

13
Conclusions

Based on the findings, the results show through phytochemical

screening that the existence of the higher amount of bioactive secondary

metabolites in Lemongrass (Cymbopogon citratus) leaves were the

components potent as available sources of natural antioxidant and its

positivity in fatty acid which are involved in the production of soap, detergents,

and cosmetics can be used as an antifungal, antibacterial as well as

antimicrobial activities.

Recommendations

On the basis of the study's findings, the following advice was given.

For the communities and students, they should be well educated in

investigating medicinal plants as a possible source of active antioxidant. To

the future researchers, this study may be needed to determine its

phytochemicals and its proper application to the human body.

14
 References

Asaolu, M.F., Oyeyemi, O.A., Olanlokun, J.O. 2009. Chemical Compositions,

Phytochemicals Constituents and In Vitro Biological Activity of Various

Extracts of Cymbopogon Citratus. Available at

http://docsdrive.com/pdfs/ansinet/pjn/2009/1920-1922.pdf. Retrieved

on July 23, 2018.

Danlami, U., Rebecca, A., Machan, D.B., Asuquo, T.S. 2011. Comparative

Study on the Antimicrobial Activities of the Ethanolic Extracts of Lemon

Grass and Polyalthia Longifolia. Available at

http://imsear.li.mahidol.ac.th/bitstream/123456789/151013/1/japs2011v

1n9p174.pdf. Retrieved on July 26, 2018.

Ekpenyong, C., Akpan, E., Daniel, N. 2014. Phytochemical Constituents,

Therapeutic Applications and Toxicological Profile of Cymbopogon

Citratus Stapf (DC) Leaf Extract. Available at

file:///C/Users/User/Downloads/LEMON%20GRASS/LEMON%20GRA

SS%2012.pdf. Retrieved on July 23, 2018.

Nambiar, V., Matela, H., 2012. Potential Functions of Lemon Grass

(Cymbopogon citratus) in Health and Disease. Available at

http://ijpba.info/ijpba/index.php/ijpba/article/viewFile/816/548. Retrieved

on July 23, 2018.

15
Salome, C., Emeka, U.C., Ikechukwu, O., Sinye, B., Calister, U., Godswill, O.

2012. Formulation and Evaluation of Cymbopogon citratus Dried Leaf-

Powder Tablets. Available at http://www.academicjournals.org/AJPP.

Retrieved on July 25, 2018.

Saxena, M., Saxena, J., Nema R., Singh, D., Gupta, A. 2013. Phytochemistry

of Medicinal Plants. Available at

http://www.phytojournal.com/vol1Issue6/Issue_march_2013/26.pdf.

Retrieved on July 24, 2018.

Soares, M., Alves, R., Pires, P., Oliveira, M.B., Vinha, A. 2013. Angolan

Cymbopogon citratus Used for Therapeutic Benefits: Nutritional

Composition and Influence of Solvents in Phytochemicals Content and

Antioxidant Activity of Leaf Extracts. Available at

https://www.researchgate.net/profile/Ana_Vinha2/publication/25517648

8_Angolan_Cymbopogon_citratus_used_for_therapeutic_benefits_Nutr

itional_composition_and_influence_of_solvents_in_phytochemicals_co

ntent_and_antioxidant_activity_of_leaf_extracts/links/004635273c7c55

2b54000000/Angolan-Cymbopogon-citratus-used-for-therapeutic-

benefits-Nutritional-composition-and-influence-of-solvents-in-

phytochemicals-content-and-antioxidant-activity-of-leaf-extracts.pdf.

Retrieved on July 31, 2018.

16
 LIST OF APPENDICES

APPENDIX TITLE

A Research Plan (1A) 18

B Approval Form (1B) 19

C RRI / Industrial Settings Form (1C) 20

D Application Form – Individual Project 21

E Certification as Individual Project 22

F Certification as Official Project Adviser 23

G Certificate of Parental Consent 24

H Laboratory Test Results Form 25

I Documentation 26

17
 APPENDIX A

2019 NATIONAL CHILDREN’S SCIENCE CONGRESS Research Plan (1A)

1. Name ARIANNA TRISHA C. ABIVA

2. Category Individual Category, Life Science

3. School Buenavista SPED Elementary School

4. School Address Buenavista, Agusan del Norte

5. Fair Division Agusan del Norte

6. Mobile No. 09096287908

7. Title of the Project Phytoconstituents of Lemongrass

(Cymbopogon citratus) Leaves Crude
Extract Potent as an Antioxidant

8. Project Adviser/Author Mrs. FLOR E. CARIÑOSA

9. This year’s experiment began at August 01, 2018

& ended September 12, 2018

10. Where did you conduct your lab?

[ / ] Research Institution [ / ] School

[ ] Field [ ] Home

11. Name, address & phone no. of lab work site (s)

Department of Chemistry, College of Science and Mathematics,

MSU-Iligan Institute of Technology, 9200 Iligan City.

Telephone No. (63) 221-4050 to 56 loc. 123

Website: http://www.msuiit.edu.ph

18
 APPENDIX B

APPROVAL FORM (1B)

1) a. Pupil’s Acknowledgment: I understand the risks and possible dangers

to me of the proposed Research Plan (1A). I will adhere to all
International Rules when conducting this research.

ARIANNA TRISHA C. ABIVA ________________ September 12, 2018

Research Presenter’s Printed Name Signature Date

b. Project Adviser’s Approval: I have read the Research Plan (1A) and
attachment prior to experimentation and reviewed the checklist for sponsor
with the pupils. I agree to sponsor the pupils above and assume
reasonable responsibility for compliance with all International ISEF Rules
as they pertain to the Research Plan (1A).

FLOR E. CARIŇOSA ________________ September 12, 2018

Project Adviser/Author’s Printed Name Signature Date

2) SRC/IRB Approval:
2a. Required for projects that need. 2b. Required for research conducted at all
prior SRC/IRB approval BEFORE Regulated Research Institutions with no
experimentation (i.e., see Item #8 prior fair SRC/IRB Approval.
on Form 1A)
This project was conducted at a regulated
The SRC/IRB has carefully studied
research institutions (not home or high school,
this project’s Research Plan (1A) and
etc.), was reviewed and approved by the proper
Attachment, and all the required forms are
institutional board before experimentation and
included. My signature indicates approval of
complies with the ISEF Rules. Attach (1C) and
the Research Plan (1A) before the pupils
required institutional approvals (e.g.,
begins experimentation.
IACUC, IRB).

____________________________ ____________________________
SRC/IRB Chair’s Printed Name SRC/IRB Chair’s Printed Name

3) Final ISEF Affiliated Fair SRC Approval.

________________ ________________
SRC Approval after Experimentation and Shortly before Competition
_______________ _______________
at the 2019 National Children’s Science Congress
Signature Date of Signature Date of
ApprovalI certify that this project adheres to the approved Research Plan (1A) and Attachment and
Approval
complies with all ISEF Rules.
(Must be (Must be
____________________________ __________________________ _________________
prior to prior to
Division SRC Chair’s Printed Name Signature Date of Approval

experimentation.) experimentation.) 19
 APPENDIX C

Regulated Research Institution / Industrial Settings Form (1C)

This form must be completed by the scientist supervising the student

research conducted. Research institution (e.g., university lab, medical center, etc.) or
industrial setting

Name of the Researchers: ARIANNA TRISHA C. ABIVA

Title Project: Phytoconstituents of Lemongrass (Cymbopogon

citratus) Leaves Crude Extract Potent as Antioxidant

a. Only to use the Equipment b. / to perform experiments

If the following question must be answered.

1. How did the researcher get the idea for their project?
(e.g. was the Project assigned picked from a test, an original idea etc. )

The researcher obtained the idea from the elders and related literature.

2. Where you made aware of the ISEF rules before experimentation?

Yes.

3. Did the researher work on the project as well as a part of a research

team?

Yes, with the presence of the analyst and research adviser/author.

4. What are the specific procedures did the researcher actually perform and
how independently the student work?

Pls. List and Describe (do not list procedures student only observed)

Research projects dealing with human subject, vertebrate animals or DNA

required review and approval by an institutional regulatory board (IRE / IACUC)

_____________________________________ ________________ ________________

Scientist’s Printed Name Signature Position

_____________________________________________ _____________
Institution Date Signed

_____________________________________________ _____________
Address Email/Phone
20
 APPENDIX D

Republic of the Philippines

Department of Education
Caraga Administrative Region
Division of Agusan del Norte
Buenavista District – I
BUENAVISTA SPED ELEMENTARY SCHOOL

APPLICATION FORM – INDIVIDUAL PROJECT

Region: CARAGA

Level: [ ] Division [ ] Regional [ x ] National

Science Fair Regional: [ x ] Life Science [ ] Physical Science

Field of Study/Discipline: Elementary

Name of the researcher: Address:

ARIANNA TRISHA C. ABIVA Pob. 1, Buenavista, Agusan del Norte

School Address: Buenavista SPED Elementary School

Title of Investigatory Project: Phytoconstituents of Lemongrass (Cymbopogon

citratus) Leaves Crude Extract Potent as an
Antioxidant

Attach one copy of accomplished OFFICIAL ABSTRACT FORM: (Not

more than 250 words, one page only

ARIANNA TRISHA C. ABIVA

Printed Name & Signature
of the research presenter

FLOR E. CARIÑOSA
Printed Name & Signature
of Project Adviser/Author

Date signed: January 28, 2019

NOTED:
RIZALITA C. ESPERA
District Science Coordinator
21
 APPENDIX E

Republic of the Philippines

Department of Education
Caraga Administrative Region
Division of Agusan del Norte
Buenavista District - I
BUENAVISTA SPED ELEMENTARY SCHOOL

CERTIFICATION AS INDIVIDUAL PROJECT

THIS IS TO CERTIFY that the project submitted by ARIANNA TRISHA

C. ABIVA of Buenavista SPED Elementary School entitled

“Phytoconstituents of Lemongrass (Cymbopogon citratus) Leaves

Crude Extract Potent as an Antioxidant” is an official entry to the 2019

National Children’s Science Congress, Science Investigatory Project

Event, Life Science – Individual Category.

Given this on the 28th day of January, 2019 at the Buenavista SPED

Elementary School, Poblacion 5, Buenavista, Agusan del Norte.

FLOR E. CARIÑOSA
Project Adviser

NOTED:

RIZALITA C. ESPERA
District Science Coordinator

22
 APPENDIX F

Republic of the Philippines

Department of Education
Caraga Administrative Region
Division of Agusan del Norte
Buenavista District - I
BUENAVISTA SPED ELEMENTARY SCHOOL

CERTIFICATION AS OFFICIAL PROJECT ADVISER

This is to certify that FLOR E. CARIÑOSA is the official project

adviser/author of the project entitled “Phytoconstituents of Lemongrass

(Cymbopogon citratus) Leaves Crude Extract Potent as an Antioxidant”

as an official entry of Buenavista SPED Elementary School to the 2019

National Children’s Science Congress.

Given this on the 28th day of January, 2019 at the Buenavista SPED

Elementary School, Poblacion 5, Buenavista, Agusan del Norte.

NOTED:

RIZALITA C. ESPERA
District Science Coordinator
23
 APPENDIX G

IPSF-Form I-D

CERTIFICATE OF PARENTAL CONSENT

We, Mr. & Mrs. JOSE R. ABIVA, JR. who are legal parents/guardians
Name of Parents/Guardian

of ARIANNA TRISHA C. ABIVA with residence at Poblacion 1, Buenavista,

Name of Project Investigator

Agusan del Norte, do hereby certify that we give our full Support for our
son/daughter to participate in the 2019 National Children’s Science
Congress under the following conditions:

1. Our Child will be chaperoned/escorted by his/her project adviser;

a. The chaperone will see to the safety, behavior and physical

upkeep of our child as far as humanly possible;

b. Considering the above conditions and the benefits that our child
will derive from his/her participation in this activity, and realizing
that every precaution will be undertaken by the adviser/
chaperone and your office, we the undersigned parent/guardian
of the above name project investigatory do hereby give our
consent for him/her to participate in the 2019 National
Children’s Science Congress. We shall bring him/her to the
point official debarkation.

JOSE R. ABIVA, JR. JESSICA C. ABIVA

Name & Signature Name & Signature
of father/guardian of mother/guardian

Date signed: January 28, 2019

Noted by:

RIZALITA C. ESPERA
District Science Coordinator
24
 APPENDIX H

(LABORATORY TEST RESULTS FORM)

25
 APPENDIX I

(DOCUMENTATION)

First, the researchers collected the leaves

of Lemongrass (Cymbopogon citratus).

Second, the leaves were washed and cleaned through tap water.
26
 Third, air dried for 72 hours.

Next, the leaves were cut into small pieces.

27
 Then, leaves were put inside the sterilized cellophane bag.

Finally, the researchers submitted the plant sample to the Department

of Chemistry, College of Science and Mathematics, MSU- Iligan institute
of Technology, Iligan City for phytochemical screening.

28
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Abstract Plants have been a source of traditional medicines which exist for
thousands of years. Human has been depended to it and lives long because
of natural medicines. Over the years, the number of synthetic drugs has been
developed all over the world. And the number of diseases rises due to
commercialized synthetic drugs. This heightened the researcher’s ability to
investigate Lemongrass as a potent source of antioxidant. This study focused
on the determination of phytoconstituents found in Lemongrass
(Cymbopogon citratus) leaves crude extract. The result showed through
phytochemical screening that the occurrence of bioactive secondary
metabolites of Lemongrass (Cymbopogon citratus) leaves crude extract
obtained higher amount were alkaloids, flavonoids, steroids, and tannins,
while the occurrence of anthraquinones, cyanogenic-glycoside, and saponins
was not observed. The potential antioxidant activity of Lemongrass
(Cymbopogon citratus) was then observed through its occurrence of
bioactive secondary metabolites. Based on the findings, the researchers
found out that the existence of the higher amount of bioactive secondary
metabolites in Lemongrass (Cymbopogon citratus) leaves were the
components potent as available sources of natural antioxidant and its
positivity in fatty acid which are involved in the production of soap,
detergents, and cosmetics can be used as an antifungal, antibacterial as well
as antimicrobial activities.

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Chapter 1 Introduction Background of the Study Considering their usage in conventional

medicine, natural resources have long been a source of medicinal substances. An

astounding number of modern pharmaceuticals have been obtained from these

sources. This is due to control major diseases. The fundamental knowledge of modern

medicine comes from plants. Rich natural sources supply the fundamental chemical and

active structures needed to create synthetic fields (Preeti et al., 2010). Lemongrass

(Cymbopogon citratus) belongs to the family Poaceae, is a tall perennial grass with

slender sharp edge and green leaves that have a pointed apex. Warm temperate and

tropical climates are where it is native (Bleasel et al., 2002). Used as a pesticide and

preservative is lemon grass oil. Used as a pesticide and preservative is lemon grass oil.

Moreover, it functions as an antifungal (Shadab et al., 2001). In soap, creams, candles,

and detergents, it serves as a fragrance. Fever, a cold, a cough, and upset stomach can

all be treated with lemon grass tea. It has diuretic properties and water retention,

making it helpful in an individual with high blood pressure. It can also help to prevent

typhoid fever, cancer, blurring of vision, lower cholesterol levels, relieve menstrual

problems and nausea. It can be used as herbal medicine to treat nervous condition and

inflammation, chest infections, sores, muscle cramps and headache (Leite et al., 2000).

Alkaloids, saponins, tannins, anthraquinones, steroids, phenols, and flavonoids are all

present in lemongrass. These phytochemicals each have a variety of medicinal and

protective properties. (Nabiar and Matela, 2012). Objectives of the Study The main

purpose of this study was to determine the phytoconstituents found in Lemongrass

(Cymbopogon citratus) leaves crude extract. Specifically, this study aimed to: 1.

Determine the bioactive secondary metabolites found in Lemongrass (Cymbopogon

citratus). 2. Determine if Lemongrass (Cymbopogon citratus) leaves crude extract

potent as an antioxidant. Significance of the Study The result of the study is significant

to the communities to support them in investigating medicinal plants as a possible

source of active antioxidant. To the future researchers, this study might be needed to

determine its phytoconstituents and its proper application to the human body. Scope

and Limitations The study will be conducted at Buenavista SPED Elementary School,

located at Poblacion 5, Buenavista, Agusan del Norte on August 01, 2018 up to

September 12, 2018, with the supervision of the project adviser. The analysis of

phytoconstituents found in Lemongrass (Cymbopogon citratus) leaves crude extract

was referred from the Department of Chemistry, College of Science and Mathematics,

MSU-Iligan Institute of Technology, Iligan City. This science investigatory project was

limited only in using Lemongrass (Cymbopogon citratus) leaves crude extract potent as

an antioxidant. Definition of Terms To promote understanding and clarity on the

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concepts presented in this study, the following terms were defined operationally.

Antioxidant. It is a substance that inhibits oxidation, especially one used to counteract

the deterioration of stored food products. Bioactive compounds. These are

distinguished from essential nutrients. Crude extract. It is a chemical, typically a

physiologically active ingredient, that is created by separating it from the original

material using solvents or evaporation. Fatty acid. Particularly any of those appearing

as esters in fats and oils, it is a carboxylic acid made composed of a hydrocarbon chain

and a terminal carboxyl group. Free radical. It describes a non-charged molecule with

an unpaired valence electron that is often extremely reactive and short-lived.

Metabolite. It is a substance formed in or necessary for metabolism. Micronutrient. It is

a chemical substance or element that is necessary for the regular growth and

development of living things in very little amounts. Oxidative. It is relating to the process

or result of oxidizing or being oxidized.

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Chapter 2 Review of Related Literature This section provides the literature that has the

relevance of the study. Most of the biological effects ascribed top Cymbopogon citratus

extracts have been attributed to its primary bioactive constituents, derived from its

leaves, stem, and roots, and their secondary metabolites. Recently, a large number of

empirical studies have been carried out aiming to expand our understanding of

Cymbopogon citratus phytoconstituents including a number of recently conducted

studies. These investigations have shown that the chemical composition of

Cymbopogon citratus extracts varies according to the geographical origin, genetic

differences, part of the plant used, a method of extraction, age/stage of maturity, and

season of harvest. Despite these differences, a number of classes of compounds are

reproducibly found, including tannins, saponins, flavonoids, alkaloids phenols, and

anthraquinones (Ekpenyong et al., 2014). It shows that Lemongrass contains alkaloids,

saponins, tannins, anthraquinones, steroids, phenols, and flavonoids. These

phytochemicals each have a variety of medicinal and protective properties. For

instance, it was known that phenol modified erythrocyte membranes (Adesanya and

Sofowora, 1983). The cited literature focused on the phytoconstituents of Cymbopogon

citratus. Hot Water Extract had significantly higher DPPH radical scavenging ability,

Fe2+ chelating ability and OH* scavenging ability than Cold Water Extract.

Conclusively, heat treatment may affect antioxidant properties due to the release of

phenolic phytochemicals, hence contributing to the health promoting and disease

preventing abilities of Cymbopogon citratus (Oboh et al., 2010). Patel and Mehta 2006,

evaluated the compounds having antioxidant property in lemon Grass and found that

dry lemon grass contains more phenol and flavonoids than a fresh one. Hence we can

infer that the dried lemon grass has greater antioxidant potential than the fresh lemon

grass. The cited literature focused on the antioxidant activities of Cymbopogon citratus.

Cymbopogon citratus is used as traditional medicine in the treatment of the nervous

condition, gastrointestinal disturbances, fever, and hypertension. Coughs, elephantiasis,

the flu, gingivitis, headaches, leprosy, malaria, ophthalmia, pneumonia, and vascular

problems can all be treated with lemon grass. It is principally taken in the form of tea as

a remedy for digestive problems, diarrhea and stomach ache. Lemon grass has

historically been used as a carminative and an insect repellent (Dama et al., 2011).

Studies on extract leaves have demonstrated anti-inflammatory, vasorelaxing, a diuretic

and valuable remedy in treating ringworm as a local application (Gore et al., 2010).

Cymbopogon citratus is an Angolan medicinal plant use by local people for treating high

fever, stomach, gut problems and headache. Due to the unpleasant aroma of its

infusion, this plane is widely known and used in tropical and sub-tropical countries.

Page 1 of 2
Moreover, it has antidepressant and mood-enhancing properties. Additional studies

have revealed antioxidant, anxiolytic, sedative, hypolipidemic, and hypoglycemic effects

(Blanco et al., 2009). Numerous plant species have been discovered to have medicinal

uses (Ali-Shtayeh and Abu, 1999). Numerous plant species have been discovered to

have medicinal uses (Ali-Shtayeh and Abu, 1999). Several essential oils display

antibacterial (Ozcan et al., 2006), antifungal, and other therapeutic properties. Essential

oils are fragrant compounds that are produced from various plant parts by steam

distillation (Chee and Lee, 2007) and antiviral (Khan et al., 2005) activity. Lemon grass

(Cymbopogon citratus is widely used in traditional medicine in many countries around

the world. Among its attributable properties are those related to antibacterial and

antifungal activities (Inouye et al., 2001, Inoue et al., 2006; Bansod and Rai, 2008;

Revathi et al., 2012), along with having painkilling and anti-inflammatory qualities

(Negrelle and Gomes, 2007). The cited literature focused on the herbal and traditional

medicinal uses of Cymbopogon citratus. Cymbopogon citratus has been used over

many years to make caffeine-free tea and as an herbal drink, suggesting that it may be

a healthier alternative to caffeine-containing tea products. In comparison to other tea

brands consumed among Nigerians (Lipton tea, Nescafe, Green tea, and Top tea);

Cymbopogon citratus tea was a good source of antioxidants such as flavonoids, and

therefore a nutritionally acceptable and medicinally valuable beverage. While some

research has indicated that it contains tannins, coumarins, saponins, and

anthraquinones, which have been linked to modest harmful effects, the limited

bioavailability of these phytochemicals in people makes these effects less of a concern

may confer a measure of protection against toxicity. The average concentrations of anti-

nutrients such as phylate are not significantly higher in Cymbopogon citratus while

cardiac glycosides, cyanates, phlobatannins, and heavy metals like lead and mercury

are absent. Therefore, Cymbopogon citratus is considered to be safe for human

consumption on the basis of its phytochemical constitution, both nutritional and non-

nutritional (Akande et al., 2014). The cited literature focused on the use of Cymbopogon

citratus as a tea for protection against toxicity and diseases. In the United States of

America and some regions of Asia, where they are grown for commercial purposes as

industrial raw materials to cosmetic and insecticide manufacturers, Cymbopogon

citratus is a plant of enormous economic importance. It is used as a scent in many

product including soap, perfume candle, mosquito and other insect repellents

(Praditvarn and Samhandharaksa, 1990). The cited literature focused on the industrial

use of Cymbopogon citratus raw materials.

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Chapter 3 Methodology This chapter presents the materials used in the study,

procedure, and utilization of Lemongrass (Cymbopogon citratus) leaves for

phytochemical screening. Materials Used in the Study Lemongrass (Cymbopogon

citratus) leaves, basket, basin, knife, chopping board, strainer, and sterilized cellophane

bag. Procedure First, the researchers collected the leaves of Lemongrass

(Cymbopogon citratus). Second, the leaves were washed and cleaned through tap

water. Third, air dried for 72 hours. Next, the leaves were cut into small pieces. Then,

leaves were put inside the sterilized cellophane bag. Finally, the researchers submitted

the plant sample to the Department of Chemistry, College of Science and Mathematics,

MSU-Iligan Institute of Technology for phytochemical screening.

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Chapter 4 Results and Discussions This chapter presents the analysis and results of

the data gathered by the researchers. The data is presented in a table form with an

appropriate interpretation follows to clarify numerical data. Table 1. Phytochemical

Screening of Lemongrass (Cymbopogon Citratus) Leaves Crude Extract Secondary

Metabolites Occurrence Alkaloids ++ Anthraquinones - Cyanogenic-glycoside -

Flavonoids +++ Saponins - Steroids +++ Tannins +++ Table 1 shows the bioactive

secondary metabolites present in Lemongrass (Cymbopogon citratus) leaves, in which

the amount level of alkaloids, flavonoids, and steroids, tannins are higher, while the

occurrence of anthraquinones, cyanogenic-glycoside and saponins were not observed.

Based on recent studies, Alkaloids have diverse structures and show a range of

pharmaceutical activities including antihypertensive effects, antiarrhythmic effect,

antimalarial activity, anticancer actions, antibacterial and antifungal activities, cancer

chemoprevention and anti-oxidants (Saxena et al. 2013). Recent research has focused

on flavonoids because of their extensive biological and pharmacological functions.

Flavonoids have been reported to exert multiple biological properties including

antimicrobial, cytotoxicity, anti-inflammatory as well as antitumor activities but the best-

described property of almost every group of flavonoids is their ability to function as

potent antioxidants capable of shielding the organism from reactive oxygen species and

free radicals. The molecular makeup of flavonoids determines how effective they are as

antioxidants. The location of hydroxyl groups and other characteristics of flavonoids'

chemical structure play a crucial role in the antioxidant and free radical scavenging

abilities of these compounds. (Singh et al., 2013). Flavonoids as naturally occurring

secondary metabolites in plants are thought to have positive effects on human health.

Plants with flavonoids have been reported to have antiviral, antitumor, and antioxidant

properties. These compounds are considered as powerful antioxidants because they

exhibited inhibitory properties against carcinogenesis is a number of vivo studies.

Flavonoid has been confirmed to effectively scavenge most oxidizing molecule such as

singlet oxygen and various free radicals linked in chronic diseases (Baang et al., 2017).

Steroids possess many interesting medicinal, pharmaceutical and agrochemical

activities like anti-tumor, immunosuppressive, hepatoprotective, antibacterial, plant

growth hormone regulator, sex hormone, anti-helminthic, cytotoxic and cardiotonic

activity (Patel et al., 2015). Tannins in medicine especially in Asian (Japanese and

Chinese) natural healing, the tannin-containing plant extracts are used as astringents,

against diarrhea, as diuretics, against stomach and duodenal tumors, and as

inflammatory, antiseptic, antioxidant and haemostatic pharmaceuticals. Tannins are

employed as caustics for cationic dyes (tannin dyes) in the dyestuff business as well as

Page 1 of 2
in the creation of inks (iron gallate ink). Wine, beer, and fruit juices are clarified by

tannins in the food sector. Additional industrial applications for tannins include the

dyeing of textiles, antioxidants in the brewing and wine industries, and coagulants in the

manufacture of rubber. Recently the tannins have attracted scientific interest, especially

due to the increased incidence of deadly illnesses such as AIDS and various cancer

(Saxena et al., 2013).

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Chapter 5 Summary, Findings, Conclusions, and Recommendations The
findings, conclusions, and recommendations of the study are summarized in
this chapter. Summary The study focused on the determination of
phytoconstituents found in Lemongrass (Cymbopogon citratus) leaves crude
extract. The objectives of the study presented in two (2) questions. First, focus
of the study was on the presence of bioactive secondary metabolites in
lemongrass (Cymbopogon citratus). Second, was on its potential activity as an
antioxidant. Findings Based on the analysis and interpretation of the
following are the findings: 1. The occurrence of bioactive secondary
metabolites of Lemongrass (Cymbopogon citratus) leaves crude extract
obtained higher amount were alkaloids, flavonoids, steroids, and tannins,
while the occurrence of anthraquinones, cyanogenic-glycoside and saponins
were not observed. 2. The potential antioxidant activity of Lemongrass
(Cymbopogon citratus) was then observed through its occurrence of
bioactive secondary metabolites. Conclusions Based on the findings, the
results show through phytochemical screening that the existence of the
higher amount of bioactive secondary metabolites in Lemongrass
(Cymbopogon citratus) leaves were the components potent as available
sources of natural antioxidant and its positivity in fatty acid which are involved
in the production of soap, detergents, and cosmetics can be used as an
antifungal, antibacterial as well as antimicrobial activities. Recommendations
On the basis of the study's findings, the following advice was given. For the
communities and students, they should be well educated in investigating
medicinal plants as a possible source of active antioxidant. To the future
researchers, this study may be needed to determine its phytochemicals and
its proper application to the human body.

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