ternatea): A Review
By
Clarence Sukmana 11605018
BACHELOR’S DEGREE In
FOOD TECHNOLOGY
FACULTY OF LIFE SCIENCES AND TECHNOLOGY
SWISS GERMAN UNIVERSITY The Prominence Tower
Jalan Jalur Sutera Barat No.15 Alam Sutera Tangerang, Banten 15143 - Indonesia
July 2020
Revision after Thesis Defense on 14th July 2020
Clarence Sukmana 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 acknowledgment is made in the thesis.
Clarence Sukmana
_____________________________________________
Student
July 24, 2020 Date
Approved by:
Dr. Ir. Abdullah Muzi Marpaung, M.P
_____________________________________________
Thesis Advisor
July 25, 2020 Date
Dr. Hery Sutanto, S.Si., M.Si
_____________________________________________
Thesis Co-Advisor
July 25, 2020 Date
Dr. Dipl.-Ing. Samuel P. Kusumocahyo
_____________________________________________
Dean
July 25, 2020 Date
Clarence Sukmana ABSTRACT
Isolation of Anthocyanins From Butterfly Pea (Clitoria ternatea): A Review
By
Clarence Sukmana
Dr. Ir. Abdullah Muzi Marpaung, M.P, Advisor Dr. Hery Sutanto, S.Si., M.Si, Co-Advisor
SWISS GERMAN UNIVERSITY
Because of the potential in Clitoria ternatea petals anthocyanin, its popularity among researchers increases. Without the availability of complete isolated ternatin in the market, researchers may find difficulties in studying single compound ternatin for various functionality. Besides, ternatin is a complex anthocyanin, which requires modification in isolation. Thus, appropriate anthocyanin isolation information, especially ternatin isolation, is crucial. The anthocyanin isolation steps include extraction, purification, and separation using the chromatographic method. The separation using thin-layer chromatography is low-cost and enables good separation but yields small quantity and require a longer time. Column chromatography able to gives higher yield than others, but the separation quality and quantity vary to the stationary phase used while the high- performance liquid chromatography (HPLC) can provide fast and excellent separation, but costly and small yields. Additionally, ternatin isolation requires additional fragmentation step using thin-layer chromatography or column chromatography for optimum result. By the time, multiple-step preparative HPLC used for the fragmentation of the ternatin mixture. The isolation from various sources shows a variation of the anthocyanins content in Clitoria ternatea petal extract. It predicted due to the different acylation and glycosylation pace of different plants, which yields in a variety of ternatin compositions. This pace may caused by different plant environment conditions.
Keywords: anthocyanin, chromatography, Clitoria ternatea, isolation, ternatin
Clarence Sukmana
© Copyright 2020 by Clarence Sukmana
All rights reserved
Clarence Sukmana DEDICATION
I dedicate this works for the future of the country I loved: Indonesia
Clarence Sukmana ACKNOWLEDGMENTS
Foremost, thank God Almighty for His blessing and help unto me. Without His love and guidance, I would not be able to achieve anything, including this thesis.
In my journey towards this degree, I have found a teacher, a friend, an inspiration, a role model, and a pillar of support in my Guide, Dr. Ir. Abdullah Muzi Marpaung, M.P and Dr. Hery Sutanto, S.Si., M.Si. They were there, giving their support and guidance and gave me invaluable guidance, inspiration, and advice in my search for knowledge. They have given me all the freedom to pursue my research while silently ensuring that I stay on track and not deviate from the core of my research. Without their guidance, this thesis would not have been possible, and I would always thank them for their help. In addition to my advisor, I would like to thank the Department of Food Technology lecturers and laboratory assistants for their time in providing valuable suggestions and comments.
My sincere thanks to my family and my dear friends: Frida, Johanssen, Astrid, Nia, Michaella, and others who gave me the most excellent source of strength during difficult times. We spend much time discussing and emphasizing deadlines and joking to lighten up the burden.
Last but not least, to my classmates who share the same burden through this thesis, their motivation makes me go further.
Finally, I admit that there are still many shortcomings in this thesis. Criticism and suggestions will be beneficial for the writer. Hopefully, this work will be useful material for further research if desired.
Tangerang, June 10, 2020
Clarence Sukmana
Clarence Sukmana TABLE OF CONTENT
Page
STATEMENT BY THE AUTHOR ... 2
ABSTRACT ... 3
DEDICATION ... 5
ACKNOWLEDGMENTS ... 6
TABLE OF CONTENT... 7
LIST OF FIGURES ... 9
LIST OF TABLES ... 10
CHAPTER 1 - INTRODUCTION ... 11
1.1 Background ... 11
1.2 Research Problem ... 12
1.3 Research Objectives ... 12
1.4 Significance of the Study ... 12
CHAPTER 2 - RESEARCH METHODOLOGY ... 14
2.1 Journal Reviews ... 14
2.2 Research Studies ... 14
2.2.1 Material and Equipment ... 14
2.2.2 Design of Experiment ... 15
2.2.3 Butterfly Pea Flower Pretreatment ... 15
2.2.4 Mechanical Press... 16
2.2.5 Extract Conditioning ... 16
CHAPTER 3 – CHEMISTRY OF ANTHOCYANIN ... 17
3.1 Chemical Structure of Anthocyanin ... 17
3.2 Anthocyanin Solubility ... 19
3.3 Anthocyanin Species in Aqueous Solution ... 19
CHAPTER 4 – ISOLATION OF ANTHOCYANIN ... 22
4.1 Anthocyanin Extraction Technique ... 22
4.1.1 Solid-Liquid Extraction ... 23
4.1.2 Modern Extraction ... 24
4.2 Anthocyanin Purification ... 25
4.3 Anthocyanin Isolation Technique ... 26
Clarence Sukmana
4.3.2 Isolation using Column Chromatography ... 34
4.3.3 Isolation using HPLC ... 35
CHAPTER 5 – ISOLATION OF ANTHOCYANIN IN Clitoria ternatea ... 37
5.1 Anthocyanin in Clitoria ternatea ... 37
5.2 Chemistry of Ternatin ... 38
5.3 History of Ternatin Isolation... 39
5.4 Isolation of Ternatin from Clitoria ternatea ... 45
5.5 Clitoria ternatea Source Variety to Isolation Result ... 46
CHAPTER 6 - CONCLUSION AND RECOMMENDATIONS ... 53
REFERENCES... 54
APPENDICES ... 61
CURRICULUM VITAE... 62
Clarence Sukmana LIST OF FIGURES
Figures Page
Figure 1. Design of experiment flowchart ... 15 Figure 2. The general structure and 6 most common anthocyanidins (Marpaung, 2017) ... 18 Figure 3. Structure of different anthocyanins species in aqueous solution (Trouillas et al., 2016) ... 20 Figure 4. Butterfly pea (Clitoria ternatea) flowers (Marpaung, 2017) ... 37 Figure 5. Delphinidin 3 – malonyl glucoside (G = D-glucose ; C = p-coumaric acid) (Terahara et al., 1998) ... 38 Figure 6. Ternatin biosynthesis pathway. (T = ternatin ; G = D-glucose ; C = p-coumaric acid ; p-GT = p-coumaric acid acylation ; GT = glucosylation) ... 50 Figure 7. TLC of Clitoria ternatea from various sources. ... 51
Clarence Sukmana LIST OF TABLES
Table Page
Table 1. Anthocyanins isolated from various source ... 28 Table 2. hRF values of anthocyanins and anthocyanidins from berry fruit (Filip et al., 2012) ... 33 Table 3. Chromatographic properties of Clitoria ternatea extract with different solvent (Norihiko Terahara et al., 1990a) ... 40 Table 4. Ternatin in Hydrophilic Butterfly Pea Extract (Shen et al., 2016) ... 44 Table 5. Type and relative amount of ternatins in Clitoria ternatea extract (Marpaung, 2017) ... 46 Table 6. TLC of Clitoria ternatea extract with Various mobile phase ... 48
