Study on Optimization, Bioactive Compounds, and Stability in Calabura Fruit Extract

By Grace 1-4209-006

A Thesis submitted to the Faculty of LIFE SCIENCES

in partial fulfillment of the requirements for BACHELOR’S DEGREE

in

FOOD TECHNOLOGY

Swiss German University EduTown BSD City

Tangerang 15339 Indonesia

July 2013

STATEMENT BY THE AUTHOR

I hereby declare that this submission is my own work and to the best of my knowledge, contains no material previously published or written by another person, nor material which to a substantial extent has been accepted for the award of any other degree or diploma at any educational institution, except where due acknowledgement is made in the thesis

Grace

Student Date

Approved by:

Ir. Abdullah Muzi Marpaung, M.P.

Thesis Advisor Date

Hery Sutanto, S.Si., M.Si.

Thesis Co-Advisor Date

Irvan Kartawiria, M.Sc.

Dean of Life Sciences Faculty Date

ABSTRACT

THE STUDY ON OPTIMIZATION, BIOACTIVE COMPOUND, AND ITS STABILITY IN CALABURA FRUIT EXTRACT

By Grace

Ir, Abdullah Muzi Marpaung, M.P., Advisor Hery Susanto, S.Si., M.Si., Co-advisor

SWISS GERMAN UNIVERSITY

Nowadays natural antioxidants found in fruits and vegetables have gained increasing interest among consumers and scientists. Calabura fruit (Muntingia calabura L.) as underused fruit in Indonesia was also studied for its potent as a high source of antioxidant. This research was aimed to find optimum conditions to extract calabura fruit, to identify bioactive compounds in calabura fruit, and to study the stability of calabura fruit extract. There were three factors involved during optimization of the extraction process. They are blanching time (0-60 minutes), extraction time (30-90 minutes), and extraction temperature (60-80^oC). The research showed only blanching time and extraction time that had significant effect to the antioxidant activity and total phenol content by following linear model. Based on the models, the optimum conditions for extraction process was achieved when blanching time was 60 minutes, extraction time was 90 minutes, and extraction temperature was 80^oC. For bioactive compounds identification, from 10 of the most significant compounds, the base peak showed that there were 8 compounds belong to phenolic group, and 2 compounds belong to fatty acid. Antioxidant activity and total phenol content of calabura fruit extract were also found to be quite stable when stored without light and oxygen exposure at 5^oC for four weeks storage time.

Keywords: Calabura fruit, extraction, antioxidant activity, total phenol content,

© Copyright 2013 by Grace All rights reserved

DEDICATION

This thesis is dedicated to my beloved family and friends

ACKNOWLEDGEMENTS

First of all, the writer would like to express gratitude to God for His love and blessings especially during the research, preparation, and completion of this thesis entitled “THE STUDY ON OPTIMIZATION, BIOACTIVE COMPOUND, AND ITS STABILITY IN CALABURA FRUIT EXTRACT”.

The writer could not be able to finish this thesis without the help, prayer, and guidance from several parties, including:

1. Mr. Muzi Marpaung, as thesis advisor who has given the writer a lot of support and knowledge regarding the statistical analysis.

2. Mr. Hery Sutanto, as thesis coadvisor who has guided the writer during the research and has helped in the identification process of calabura fruit extract.

3. Mr. Irvan Kartawiria, as dean of Life Sciences Faculty who has approved and given a great opportunity for the writer to complete this research.

4. Mr. Tabligh Permana and Ms. Silvya Yusri, as laboratory assistant who have given the writer a lot of guidance, advises, and ideas regarding the thesis.

5. Mr. Hardyanto Lim and Mrs. Juliana Ngoei, as writer’s parents who have given the writer endless love and support throughout the thesis.

6. Ms. Ratna Sari Wijaya and Mr. Irianto Wijaya, as writer’s sister and brother who have accompanied the writer through all hardships.

7. Frebriansyah Hermawan, Debby Rosalina Chandra, Aida Febriani, Cindy Chandry, and Felix Chandra for the fun time that encourage the writer to complete the thesis.

8. Calvin Aledhean for the support, love, and patience towards one another.

9. Patrisia, Melissa Florence Mustopo, Hebert Hensel, Rinanti Pritanova, and Felix Sebastian as the fellow friends under supervision of Ir. Muzi Marpaung.

10. All Food Technology 2009 students and all people who have given a big contribution for this thesis completion.

hoped that this thesis may give useful information and knowledge for further development in science. Any suggestion and critics are gratefully accepted to improve the quality of this thesis.

BSD, June 2013

Grace

TABLE OF CONTENTS

Page

STATEMENT BY THE AUTHOR……… 2

ABSTRACT……… 3

DEDICATION……… 5

ACKNOWLEDGEMENTS……… 6

TABLE OF CONTENTS……… 8

LIST OF TABLES……….………….. 11

LIST OF FIGURES……….……….... 12

LIST OF APPENDICES……….…… 14

CHAPTER I – INTRODUCTION……….………. 15

1.1 Background………. 15

1.2 Research Objectives………... 16

1.3 Research Problems………. 16

1.4 Significance of Study………. 17

1.5 Hypothesis……….. 17

1.6 Scope of Research……….. 17

CHAPTER II – LITERATURE REVIEW………. 18

2.1 Calabura (Muntingia calabura)……….………. 18

2.2 Pharmacological Properties of Calabura……… 19

2.2.1 Antioxidant Activity………... 19

2.2.2 Anticancer Activity………. 20

2.2.3 Antinoiceptive Activity……….. 20

2.2.4 Hypotensive Activity……….. 20

2.2.5 Cardioprotective Activity………... 21

2.2.6 Antibacterial Activity………. 21

2.3 Calabura Phenolic Compounds.……….……… 21

2.4 Phenolic Compounds as Antioxidant………. 23

2.5 Stability of Phenolic Compounds………... 25

2.5.2 Effect of Light……… 26

2.5.3 Effect of Temperature………. 26

2.5.4 Effect of pH……… 26

2.6 Extraction……… 27

2.7 Response Surface Methodology – Box Behnken Design………... 27

CHAPTER III – RESEARCH METHODOLOGY……… 30

3.1 Time and Venue……….. 30

3.2 Materials and Equipment……… 30

3.2.1 Materials……….……… 30

3.2.2 Equipment………..………. 30

3.3 Research Procedure……….………... 3.3.1 Calabura Fruit Extraction……….……….. 31 33 3.3.2 Identification of Bioactive Compounds………. 35

3.3.3 Stability Test of Calabura Fruit Extract.………. 36

3.4 Analysis Procedure……….……… 36

3.4.1 Determination of Total Phenol Content……….. 36

3.4.2 Determination of Antioxidant Activity……….. 37

3.5 Design Experiment………. 38

3.6 Statistical Analysis……….…… 39

3.7 Verification of Equation Model………. 39

CHAPTER IV – RESULTS AND DISCUSSION……… 40

4.1 Extraction of Calabura Fruit………... 40

4.2 Effect of Extraction Conditions on Total Phenol Content………. 41

4.3 Effect of Extraction Conditions on Antioxidant Activity………. 45

4.4 Selection and Verification of the Optimum Process………...……... 50

4.5 Correlation between Total Phenol Content and Antioxidant Activity…... 51

4.6 Identification of Bioactive Compounds in Calabura Fruit Extract………. 53

4.7 Stability Test of Calabura Fruit Extract……….. 60

5.2 Recommendation……… 65

REFERENCES……… 66

APPENDICES……….… 78

CURRICULUM VITAE………. 98