EXTRACT
By
SUDONO MAHADIKA 11305022
BACHELOR’S DEGREE in
FOOD TECHNOLOGY
FACULTY OF LIFE SCIENCES & TECHNOLOGY
SWISS GERMAN UNIVERSITY The Prominence Tower
Jalan Jalur Sutera Barat No. 15, Alam Sutera Tangerang, Banten 15143 – Indonesia
August 2017
Revision after Thesis Defense on 25th July 2017
Sudono Mahadika 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.
Sudono Mahadika
____________________________________________
Student Date
Approved by:
Della Rahmawati, S.Si, M.Si
____________________________________________
Thesis Advisor Date
Nina Artanti, Ph.D.
____________________________________________
Thesis Co-Advisor Date
Dr. Dipl. -Ing. Samuel P. Kusumocahyo
____________________________________________
Dean Date
Sudono Mahadika ABSTRACT
PROFILE OF α-GLUCOSIDASE INHIBITOR FROM Kalakai (Stenochlaena Palustris) EXTRACT
By
Sudono Mahadika
Della Rahmawati, S.Si M.Si, Advisor Nina Artanti, Ph.D., Co-Advisor
SWISS GERMAN UNIVERSITY
AGI are components that prevent the occurrence of sugar breakdowns into simple sugar (monosaccharides). AGI commonly consumed as oral medicine to treat diabetes. From the previous studies, several tropical plants in Indonesia have the ability to inhibit α-glucosidase.
This study focuses on the potential of Kalakai (Stenochlaena palustris) to be developed into food-safe product. The aim of this study is to select the best extraction method that produce highest yield of AGI activity and later being characterized based on polarities and phenolic content properties. Aqueous (cooked) extract with 12.0 mg/ml concentration from Stenochlaena palustris known to give significant inhibition (approximately 68%) towards α- glucosidase enzyme. Analyzed using Folin-Ciocalteau assay, methanol fraction (19.7 mg/g) and ethyl acetate fraction (12 mg/g) of aqueous extract gave a moderate amount of phenolic contents. However, the AG-inhibition % of both fractions are inferior to AG-inhibition level of the extract itself. While having only a small amount of polyphenols (6.8 mg/g), aqueous extract give a potent AG-inhibition activity. Thus, showing that inorganic materials such as minerals possibly play roles in determining the AG-inhibition level rather than polyphenols inside Stenochlaena palustris, because of the property of water being an inorganic solvent.
Keywords: Stenochlaena palustris, tropical ferns, α-glucosidase inhibitor, Diabetes Mellitus, extraction, fractionation in organic solvent, polarity determination, Total Phenolic Content
Sudono Mahadika
© Copyright 2017 by Sudono Mahadika
All rights reserved
Sudono Mahadika DEDICATION
I dedicate this thesis work for my family, friends, colleagues, my nation the Republic of Indonesia, for the future of antidiabetics developments and for all the sufferers of diabetes,
praying for their recoveries.
Sudono Mahadika ACKNOWLEDGEMENTS
I take this opportunity to express my gratitude to all those who provide possibilities for me towards the completion of this thesis work.
First and foremost, I would like to thank God, the creator, the Alpha and the Omega, the powerful sentient being above us, who had given me blessings and guidance to pass through any obstacles.
I would like to give the biggest gratitude to my lovely family, especially to both of my parents that never stop to support me mentally and financially through all the hardships that I encounter while doing this thesis work.
I would also express many thanks of Life Sciences family, from the dean, staffs, lecturers until the students from each department who has always been supportive to me and helping me to grow even better each and every day.
A special gratitude given to my kind and understanding advisor Mrs. Della Rahmawati, M.Si., S.Si., to my co-advisor Mrs. Nina Artanti, Ph.D. and also to Mrs. Maria DPT Gunawan-Puteri STP., M.Sc., Ph.D. who have always been very helpful towards the completion of this thesis research.
Thankyou also to Swiss German University and LIPI Puspitek which has already provided accommodation, supervision and necessary information that helps in completing the thesis research.
Lastly, I would to thank all of my friends and colleagues, especially LS faculty intake batch 2013 family, who together being part of big LS and SGU community shares the same vision, the same goal to graduate as bachelors with double degree. Every hardship that we have been through, every smile, every cry all of it shared together has made us very strong.
I believe that these words are not even enough to reply the love and supports I have felt from all of my surroundings. However, thank you for everything!
Sudono Mahadika
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 Problems……….. 12
1.3 Research Objectives……… 12
1.4 Significance of Studies……… 13
1.5 Research Questions………. 13
1.6 Hypothesis………... 13
CHAPTER 2 – LITERATURE REVIEW………... 14
2.1 Stenochlaena palustris……… 14
2.2 Maceration in organic solvent……… 15
2.3 AGI ( α-Glucosidase Inhibitor)……….. 15
2.4 Analysis of Total Phenolic Content using Folin-Ciocalteau assay………… 16
2.5 Fractionation of extract based on polarity……….. 17
2.6 Liquid-Liquid Extraction………... 17
2.7 Planar Chromatography – TLC……….. 18
2.8 Characterization of extract’s component……… 19
2.9 Functional Food……….. 19
CHAPTER 3 – RESEARCH METHODS……….. 20
3.1 Venue and Time……….. 20
3.2 Materials and Equipment……… 20
3.2.1 Raw Material………... 20
Sudono Mahadika
3.2.3 Equipment………... 20
3.3 Design of Experiment ……… 21
3.4 Experimental Procedure……….. 27
3.4.1 Extraction Process of Stenochlaena palustris………. 27
3.4.2 Reagent preparation for AG-inhibition assay……….. 29
3.4.3 AG-inhibition enzyme solution production………. 30
3.4.3 Fractionation……… 30
3.5 Analytical Procedure………... 31
3.5.1 AG-inhibition Assay………... 31
3.5.2 Preliminary bioactive component isolation (TLC)……….. 32
3.5.3 Total phenolic content analysis………... 33
3.5.4 Liquid Chromatography and defining the active compound ……….. 34
CHAPTER 4– RESULTS AND DISCUSSIONS..……….. 35
4.1 Screening of the best extraction method………. 35
4.2 Characteristic of aqueous (cooked) extract fractions ………. 38
4.3 TLC result of bioactive component separation………... 42
4.4 LC-MS reading of aqueous extract………. 44
CHAPTER 5 – CONCLUSION AND RECCOMENDATIONS 46 5.1 Conclusions……… 46
5.2 Recommendations……… 47
GLOSSARY……… 48
REFERENCES……… 49
APPENDICES………. 52
CURRICULUM VITAE……….. 74
