THE CREATION OF BROMELAIN LOZENGES FROM PINEAPPLE (ANANAS COMOSUS)

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THE CREATION OF BROMELAIN LOZENGES FROM PINEAPPLE (ANANAS COMOSUS)

By

Stefanus Pangestu

A Bachelor’s Thesis submitted to the Faculty of LIFE SCIENCES

In Partial Fulfillment of the Requirements for BACHELOR’S DEGREE

WITH A MAJOR IN PHARMACEUTICAL ENGINEERING

Swiss German University EduTown BSDCity

Tangerang 15339 INDONESIA Telp. +62 21 3045 0045

Fax. +62 21 3045 0001 E-mail: info@sgu.ac.id

www.sgu.ac.id

August 2012

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THE CREATION OF BROMELAIN LOZENGES FROM

PINEAPPLE (ANANAS COMOSUS) Page 2 of 153

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.

_______________________________________ ________________

Stefanus Pangestu Date

Approved by:

________________________________________ __________________

Ir. Paulus Gunawan, M. M Date

________________________________________ __________________

Dr. rer nat. Maruli Pandjaitan Date

______________________________________ _________________

Chairman of the Examination Steering Committee Date

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ABSTRACT

THE CREATION OF BROMELAIN LOZENGES FROM PINEAPPLE (ANANAS COMOSUS)

By

Stefanus Pangestu

SWISS GERMAN UNIVERISTY Bumi Serpong Damai

Ir. Paulus Gunawan M.M, Advisor

Pineapple has a proteolytic enzyme called bromelain. Recent discovery, bromelain has the potential to treat HIV/AIDS patients. The problem is that pineapple has very short shelf life and very easy to spoil. Bromelain in table form has relatively longer shelf life and can be consume anytime and anywhere. In this research, the bromelain tablet was produced with wet granulation, drying process and tablet compression.

Another problem with drying is bromelain is heat sensitive and can lose activity so excipients such as binder, filler, disintegrant, lubricant and preservative was added at the granulation process before the drying process. The addition of excipients can protect the bromelain but at the same time, it reduce the amount of active pharmaceutical ingredient by diluting the bromelain with excepients by several times.

The best possible formulation needed to be discovered in order to make a good tablet and at the same time it protect the bromelain enzyme activity in the tabler form.

Keywords: Bromelain Extraction, Specific Activity Preservation, Tabletting Process, Hardness Testing.

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DEDICATION

I dedicate this thesis to my family,

Whose supports helps me in many aspects of my life

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ACKNOWLEDGMENTS

The author wishes to first and foremost thanks to God Almighty for it is only by His grace, love and guidance that I can finish this thesis on time.

I would also like to thank Mr. Paulus Gunawan as my advisor for his teaching, guidance and wisdom that give a great contribution in this research and allow me to finish this thesis on time. I am also grateful for his topic that inspired me to create new product rather than following other’s footstep.

I am also grateful to Mr. Maruli Pandjaitan, as my co-advisor, for his recent discovery about HIV/AIDS and Hepatitis C that inspired me to take his research to the new level, the tableting process.

I also give my gratitude to Mr. Tabligh Permana, that give me a lot of helpful information, help me a lot in the laboratory analysis and guide me when I lost. Mr.

Irvan S. Kartawiria, I thank you for your assistance in tablet processing and granulation process.

I would like to give thanks to my family especially my parents, whose hard works make me capable to study at SGU, help to raise me until now and make me what I am right now. I also want to give my gratitude to my aunt that help me to support my thesis by financially and encouragement that I can finish my thesis on time.

I also want to thank to my friend, Anthony, Arti, Benedictus, Denny, Diana, Fanny, Jessica that help me to taste my formulation. I would like to thank you to Stefanie for lending me her laptop (since I have problem with my old laptop) that contribute greatly for statistic process and thesis writing. I would like also to thank to all my friend that cheer me up during my laboratory time.

All my friend, Adiel, Andre, Rio, Aradea, Hendra, Jonathan, Luna and Otto that help me to support and stop me when I am playing too much video games. I would also

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like to thank you to all the player of Eclipse Ragnarok which help me to explain about enzyme and the activity.

All the staff and lecturers of the Swiss German University, especially from the faculty of Life Science for your contribution to teach me the basic information that help me to finish the thesis. Thank you to all of Phamaceutical Engineering student that accompanying me through this 4 years of campus life that made a precious memory

Jakarta, June 2009

Stefanus Pangestu

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TABLE OF CONTENTS

STATEMENT BY THE AUTHOR ... 2

ABSTRACT ... 3

DEDICATION ... 4

ACKNOWLEDGMENTS ... 5

CHAPTER 1 – INTRODUCTION ... 16

1.1 Background Information ... 16

1.2 Research Problem ... 19

1.3 Significance of Study ... 19

1.4 Research Purpose ... 20

1.5 Thesis Scope ... 21

CHAPTER 2 – LITERATURE REVIEW ... 22

2.1 Pineapple ... 2222

2.2 Enzyme ... 27

2.3 Bromelain ... 34

2.4 Fruit Blending (Using Juice) ... 37

2.5 Wet Granulation ... 38

2.6 Drying of Solid ... 39

2.7 Tableting Process ... 40

2.7.1 Factor that affect tablet compression ... 41

2.7.1.1 Product Variation ... 41

2.7.1.2 Powder Flow and Feed Rate ... 41

2.7.1.3 Particle Size Problem ... 42

2.7.1.4 Variation in Tablet Hardness ... 42

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2.8 Excipient ... 43

2.8.1 Direct Interaction between Active Ingredient and Excipient ... 43

2.8.1.1 Effect of pH... 44

2.9 Functional Assay ... 44

2.9.1 Protein Content ... 44

2.9.2 Enzyme Activity Determination ... 45

2.9.3 Moisture Content ... 45

2.9.4 Sugar Testing (Brix) ... 46

2.9.5 Hardness Testing ... 46

CHAPTER 3 – METHODOLOGY ... 47

3.1 Time and Venue ... 47

3.2 Research Methodology ... 47

3.2.1 Laboratory Research ... 47

3.2.2 Literature Study ... 48

3.3 Material and Equipment ... 48

3.3.1 Raw Material ... 48

3.3.2 Equipment ... 48

3.3.3 Chemicals ... 50

3.4 Research Design... 51

3.4.1 Filtration Process ... 52

3.4.2 Granulation Process ... 56

3.4.3 Protein Content Analysis ... 58

3.4.4 Enzyme Activity Analysis ... 61

3.4.5 Brix Test... 63

3.4.6 Drying Process ... 64

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3.4.7 Angle of Repose ... 65

3.4.8 Moisture Content Analysis ... 66

3.4.9 Hardness Testing ... 67

3.4.10 Statitical Analysis ... 68

CHAPTER 4 – RESULT & DISCUSSION... 699

4.1 Pineapple Juice Filtration ... 69

4.1.2 Enzyme Specific Activity ... 70

4.2 Wet Granulation ... 70

4.2.1 Optimum Mixing Time ... 72

4.2.2 Moisture Content Analysis ... 77

4.2.4 Specific Enzyme Activity Analysis ... 79

4.3 Drying Process ... 80

4.3.1 Moisture Content ... 80

4.3.3 Specific Enzyme Activity Analysis ... 83

4.4 Hardness Analysis ... 85

4.4.1 Hardness Analysis before Drying Process ... 85

4.4.2 Hardness Analysis after Drying Process ... 86

CHAPTER 5 – CONCLUSION AND RECOMMENDATION ... 929

5.1 Conclusion ... 89

5.2 Recommendation ... 89

GLOSSARY ... 9290

REFERENCES ... 91

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APPENDICES ... 96

Appendix 1: Standard Curve Enzyme Activity ... 96

Appendix 2: Protein Content Standard Curve ... 97

Appendix 3: Volume Activity, Protein Content and Specific Activity of Concentrated Pineapple Juice... 98

Appendix 4: Moisture Content after Granulation Process ... 99

Appendix 5: Volume Activity, Protein Content and Specific Activity of Bromelain Preliminary ... 101

Appendix 6: Brix Test Result 5% Binder ... 103

Appendix 7: Volume Activity, Protein Content and Specific Activity of Bromelain after Heating... 106

Appendix 8: Moisture Content after Heating... 111

Appendix 9: Hardness Testing before Heating Process ... 112

Appendix 10: Hardness Testing after Heating Process... 113

Appendix 11: Statistical Analysis ... 114

CURRICULUM VITAE ... 149