application of spray dried butterfly pea root extract to

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APPLICATION OF SPRAY DRIED BUTTERFLY PEA ROOT EXTRACT TO LOWERING ACETHYLCHOLINESTERASE LEVELS ON ANIMAL

MODELS

By

Cosmas Oktavianus Santosa 14212059

BACHELOR’S DEGREE in

FOOD TECHNOLOGY

LIFE SCIENCES AND TECHNOLOGY

SWISS GERMAN UNIVERSITY EduTown BSD City

Tangerang 15339 Indonesia

August 2016

Revision After Thesis Defense On July 26

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Cosmas Oktavianus Santosa STATEMENT BY THE AUTHOR

I hereby declare that this submission is my own work and to the best of my knowledge, it 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.

Cosmas Oktavianus Santosa

____________________________________________

Student Date

Approved by:

Dr. rer nat Maruli Pandjaitan

____________________________________________

Thesis Advisor Date

Ir. Abdullah Muzi Marpaung, MP.

____________________________________________

Thesis Co-Advisor

Date

Dr. Dipl.-Ing Samuel P. Kusumocahyo

____________________________________________

Dean Date

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APPLICATION OF SPRAY DRIED BUTTERFLY PEA ROOT EXTRACT Page 3 of 47

ON ACETYLCHOLINESTERASE LEVELS ON ANIMAL MODELS

Cosmas Oktavianus Santosa ABSTRACT

APPLICATION OF SPRAY DRIED BUTTERFLY PEA ROOT EXTRACT ON ACETHYLCHOLINESTERASE LEVELS ON ANIMAL MODELS

By

Cosmas Oktavianus Santosa Dr. rer nat Maruli Pandjaitan, Advisor Ir. Abdullah Muzi Marpaung, MP., Co-Advisor

SWISS GERMAN UNIVERSITY

Butterfly Pea or Clitoria ternatea is a decorative plant found throughout south east asia and india. Though mostly used as a decoration it has been used for a variety of medical symptomps. One of these symptom is Alzheimer’s which is marked with the increase of Acetylcholinesterase enzyme level in the brain and blood. While it is proven that the extract does inhibit the Acetylcholinesterase production only small amounts of the extract can be made at a time. A new method of extraction using a spray dryer is used to create larger amounts of the Extract but with lower concentrations compared to the traditional technique of freeze drying extraction. To learn if the Lower concentration can still effectively treat Alzheimer’s this experiment was done. The Alzheimer’s disease in this experiment are induced using high amounts of scopolamine. While there were significant difference before and after the treatment the decrease of the acetylcholinesterase levels are insignificant between the groups.

The results prompted a scientific guess whereas the decrease of acetylcholinesterase levels in the rats are naturally occurring phenomenon. At the same time the data shows potential that the treatment and the dosage maybe the key to getting better results

Keywords: Butterfly Pea Root, Clitoria Ternatea, Alzheimer’s Disease, Acetylcholinesterase enzyme, Root Extraction

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Cosmas Oktavianus Santosa DEDICATION

I dedicate this work to those who’ve suffered through Alzheimer’s disease and did not give up.

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Cosmas Oktavianus Santosa ACKNOWLEDGEMENTS

First and foremost I would like to thank god and his grace for if it was not in his will I will not be able to finish this thesis.

I would also like to thank all that have helped me in the writing of this thesis report:

1. Dr.rer.nat Maruli Pandjaitan as my thesis advisor, for giving me his time, knowledge, and patience along the writing of this thesis.

2. Ir. Abdullah Muzi Marpaung, MP. as my thesis co-advisor, for polishing my thesis work and his statistical knowledge.

3. My friends in the faculty of life sciences and technology that gave me their advice and views on the problem at hand.

4. Mr. Endang Haerudin for taking care of the mice at IRatco dramaga bogor.

5. My family that gave me moral support along the way.

6. The SGU staff that cannot be mentioned one by one.

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Cosmas Oktavianus Santosa TABLE OF CONTENTS

Page

STATEMENT BY THE AUTHOR ... 2

ABSTRACT ... 3

DEDICATION ... 5

ACKNOWLEDGEMENTS ... 6

TABLE OF CONTENTS ... 7

LIST OF FIGURES ... 9

LIST OF TABLES ... 10

CHAPTER 1 - INTRODUCTION ... 11

1.1 Background ... 11

1.2 Research Objectives ... 12

1.3 Research Problems ... 12

1.4 Significance of Study ... 13

1.5 Research Questions ... 13

1.6 Hypothesis... 13

CHAPTER 2 – Literature Review ... 14

2.1 Alzheimer’s Disease and The Cholinergic System ... 14

2.2 Acetylcholine and Acetylcholinesterase... 15

2.3 Butterfly Pea ... 16

2.4 Butterfly Pea Root... 16

2.5 Ellman’s Reagent ... 17

2.6 Prostigmin... 17

2.7 Scopolamine ... 18

2.8 Animal models ... 18

2.9 Spray drying ... 18

CHAPTER 3 – RESEARCH METHODS ... 19

3.1 Time and Venue ... 19

3.2 Materials and equipment ... 19

3.3 Research Method ... 20

3.3.1 Animal Models and Adaptation ... 20

3.3.2 Extraction of Butterfly Pea Root Extract ... 21

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3.4 Primary Research ... 22

3.4.1 Adaptation ... 22

3.4 Dosage Preparation ... 23

3.5 Analytical Procedure ... 23

3.5.1 Phenol analysis ... 23

3.5.2 Acetylcholinesterase Enzyme Testing ... 24

3.6 Statistical Analysis ... 27

CHAPTER 4 – RESULTS AND DISCUSSIONS ... 28

4.1 Butterfly Pea Root Extraction and Spray Drying ... 28

4.2 Adaptation Results ... 29

4.3 Treatment Results ... 30

CHAPTER 5 – CONCLUSION AND RECCOMENDATIONS ... 32

5.1 Conclusion ... 32

5.2 Recommendations ... 32

REFERENCES ... 33

CURRICULUM VITAE ... 47