i BROWNING INTENSITY, COLOR DEVELOPMENT, SCAVENGING ACTIVITY OF MAILLARD REACTION PRODUCT FROM FRUCTOSE LYSINE AND GLUCOSE LYSINE MODEL SYSTEM BACHELOR THESIS Composed by FINA FITRIANA BUDIMAN FOOD TECHNOLOGY STUDY PROGRAM DEPARTMENT OF AGRICULTURAL SCIENCES FACULTY OF ANIMAL AND AGRICULTURAL SCIENCES DIPONEGORO UNIVERSITY SEMARANG, INDONESIA 2017
ii BROWNING INTENSITY, COLOR DEVELOPMENT, SCAVENGING ACTIVITY OF MAILLARD REACTION PRODUCT FROM FRUCTOSE LYSINE AND GLUCOSE LYSINE MODEL SYSTEM Composed by FINA FITRIANA BUDIMAN Student Reg. Number 23020113140038 As in partial fulfillment of the requirements for the Bachelor Degree in Food Technology Study Program Faculty of Animal and Agricultural Sciences Diponegoro University FOOD TECHNOLOGY STUDY PROGRAM DEPARTMENT OF AGRICULTURAL SCIENCES FACULTY OF ANIMAL AND AGRICULTURAL SCIENCES DIPONEGORO UNIVERSITY SEMARANG, INDONESIA 2017
iii ENDORSEMENT LETTER Title of Bachelor Thesis :BROWNING INTENSITY, COLOR DEVELOPMENT, AND SCAVENGING ACTIVITY OF MAILLARD REACTION FROM FRUCTOSE LYSINE AND GLUCOSE LYSINE MODEL SYSTEM. Name of Student :FINA FITRIANA BUDIMAN Student Reg. Number :23020113140038 Study Program/Department :FOOD TECHNOLOGY/AGRICULTURAL SCIENCES Faculty :ANIMAL AND AGRICULTURAL SCIENCES Has been examined by the examiner team and declared to pass on Main Supervisor Secondary Supervisor Ahmad N. Al-Baarri S.Pt., M.P., Ph.D. Head of Final Exam Program Prof. Dr.Ir. Anang M. Legowo, M.Sc Head of Study Program Dr. Ir. Antonius Hintono, MP Dean Dr. Yoyok Budi Pramono Head of Department Prof. Dr. Ir. Mukh Arifin, M.Sc Dr. Ir. Didik Wisnu Widjajanto, M.S
iv ORIGINALITY STATEMENT OF BACHELOR THESIS I, the undersigned below: Name : Fina Fitriana Budiman Student reg. number : 2302011314038 Study program : Food Technology Hereby declare as follows: 1. Scientific work entitled: Browning intensity, color development, scavenging activity of Maillard reaction product from Fructose Lysine and Glucose Lysine model system, and the research related to this scientific work is the result of my own work. 2. Each core or citation from other people in the form of publications in this scientific work has been recognized in accordance with standard procedures. 3. I also recognize that this scientific work has been done under supervision of: Ahmad N. Al-Baarri, S.Pt., M.P., Ph.D and Prof. Dr. Ir. Anang M. Legowo, M.Sc. Semarang, May 2017 Author Fina Fitriana Budiman Knowledged by Main Supervisor Secondary Supervisor Ahmad N. Al-Baarri, S.Pt., M.P., Ph.D NIP. 19740601 200112 1 002. Prof. Dr. Ir. Anang M. Legowo, M.Sc. NIP. 19600417 198609 1 001
v SUMMARY FINA FITRIANA BUDIMAN. 23020113140038. 2017. Browning intensity, color development, scavenging activity of Maillard reaction product from Fructose Lysine and Glucose Lysine model system. (Supervisor : Ahmad N. Al-Baarri, and Anang M. Legowo, M.Sc.) Polysaccharides occurs abundantly in the environment as food resources and may be produced commercially to generate fructose. This research was used fructose (Fruc) and lysine (Lys) as a model of glycation reaction to generate the MRPs. This study was aimed to demonstrate the browning intensity, browning index, spectra measurement, scavenging activity, and the correlation between browning intensity and scavenging activity of the MRPs generated from Fruc Lys using low temperature at 50 C. Materials used in this present study were D-fructose, D-Glucose, Lysine, ABTS or 2,2 -azino-bis (3-ethylbenzthiazoline-6-sulfonic acid). Fruc and Lys were used to produce MRPs using heating process at 50 C for 48 hour. Browning intensity, scavenging activity, and spectral measurement used spectrophotometer, while browning index used colorimeter for determining the value. The correlation between browning intensity and scavenging activity was analyzed using GraphPad Prism to obtain the significance value of correlation. Result from this research demonstrated that the browning intensity, color development, and scavenging activity were improved along with the increase in duration of heating process. Based on spectra analysis, MRPs from Fruc-Lys was initially detected at 48 h of heating treatment. The correlation between browning intensity and scavenging activity were assigned as a positive and significant correlation. In conclusion, Maillard reaction were able to be detected at 48 h of heating process. Since the further process of glycation should be controlled, the use of limited duration on the heating process of sugar-protein compound is advised.
vi PREFACE Praise and great gratitude to Almighty God, Allah SWT, who always gives His gracious mercy and tremendous blessing that has helped the author to successfully finish this bachelor thesis with the title, Browning intensity, color development, scavenging activity of Maillard reaction product from Fructose Lysine model system. This bachelor thesis is as a requirement in accomplishing the Bachelor Degree of Agricultural Technology at Food Technology Study Program, Department of Agricultural Sciences, Faculty of Animal and Agricultural Sciences, Diponegoro University. The author would like to thank very much for those who have given the contribution, suggestion, and guidance, so that this bachelor thesis can be finished properly. The writer would like to deliver this thank to: 1. Ahmad N. Al-Baarri, S.Pt., M.P., Ph.D and Prof. Dr. Ir. Anang M. Legowo, M.Sc., as the main and secondary supervisor who had guided and assisted the author in writing and finishing the script. Thank you very much for your memorable advice, valuable experience and ideas, precious suggestion and guidance that you have shared to the author. 2. Examiner Committee, who have been gave a big chance to the author in delivering her bachelor thesis presentation. Thank you very much for all the valuable suggestion and great motivation for the author s next journey. 3. All the amazing lecturers in Food Technology Study Program, Department of Agricultural Sciences, Faculty of Animal and Agricultural Sciences, Diponegoro University. Thank you very much for the time, knowledge,
vii advice, great support and great motivation that have given to the author since study in this great campus. 4. Prof. Shigeru Hayakawa and Prof. Masahiro Ogawa in Kagawa University, Japan, for providing rare sugar D-Fructose and D-Glucose as one of the materials of this research. Thank you for your cooperation and support. 5. Food Technology second batch class, intake 2013, for the best support and great positive energy to the author during the study until finishing this research. 6. Greatest thanks and appreciation dedicated to Arif Budiman and Sri Lestari, the author s beloved parents, the most important people who never tired for always praying and supporting every steps of author to reach her dreams. The greatest thanks and appreciation also dedicated to Fani Setiawan Budiman, the author s brother. 7. Special thanks and appreciation dedicated to Fahmi Ilman Fahrudin, who always been supported in many things during the up and down of the author s study. 8. Sincere thanks to my secondary Family TELO during four year college that always accompany ; Nabila, Katlya, Asyifa, Saffira, Tito, Rifqi, Bayu, Yudhis, Brilly, Kukuh, dan Umar. 9. Deepest thanks to the author s previous school, especially for the great teachers of TK Kemala Bhayangkari, SDN IV Kutoharjo, SMPN 2 Rembang, SMA 1 Rembang and all the author s best friends in school, who
viii have shaped the author s character, attitude and big spirit of learning to develop all the potencies far beyond. Final words, the author would like to thank very much for those who cannot be mentioned the names individually. Hopefully this bachelor thesis can be useful for us and can be the references for the parties in need. Semarang, Indonesia, May, 2017 Author, Fina Fitriana Budiman
ix TABLE OF CONTENTS Pages ENDORSEMENT LETTER... PREFACE... LIST OF FIGURES... LIST OF APPENDIX... iii vi xi xii CHAPTER I INTRODUCTION... 1 CHAPTER II LITERATURE REVIEW... 4 2.1. Maillard Reaction Products (MRPs)... 4 2.2. Glycation Reaction... 5 2.3. Lysine - D-Fructose Maillard Reaction... 8 2.4. Browning Intensity... 9 2.5. Browning Index... 10 2.6. Spectroscopic Measurements... 11 2.7. ABTS Radical Scavenging Activity... 12 2.8. Correlation between Browning Intensity and Scavenging Activity. 13 CHAPTER III MATERIALS AND METHODS... 14 3.1. Material... 14 3.2. Method... 14 3.2.1.Preparation of MRPs Model... 14 3.2.2. Browning Intensity... 16 3.2.3.Color Development... 16 3.2.4.Spectroscopic Measurements... 16 3.2.5. ABTS Radical Scavenging Activity... 17 3.2.6. Correlation between Browning Intensity and Scavenging Activity... 17 3.3. Data Analysis... 18 CHAPTER IV RESULTS AND DISCUSSIONS... 19 4.1. Browning Intensity... 19 4.2. Color Development... 20 4.3. Spectroscopic Characteristics of Fruct Lys and Glc Lys MRPs 22 4.4. ABTS Radical Scavenging Activity... 24 4.5. Correlation between Browning Intensity and Scavenging Activity. 27
x CHAPTER V CLOSING... 29 5.1. Conclusions... 29 5.2. Suggestions... 29 REFERENCES... 30 APPENDIX... 35 BIOGRAPHY... 45
xi LIST OF FIGURES Figure Pages 1. Condensation of Amino Carbonyl... 5 2. Amadori Rearrangement Mechanism... 6 3. Degradation Strecker Aldehyde... 7 4. Aldehyde Compound Forming... 7 5. Chemical structure of D-Fructose and D-Glucose... 9 6. The flow chart of Maillard reaction..... 15 7. Browning intensity of Fruc, Fruc Lys, Glc, and Glc Lys... 19 8. Browning index of Fruc, Fruc Lys, Glc, and Glc Lys... 21 9. Development of emission spectra of Fruc, Fruc Lys, Glc, and Glc Lys... 23 10. The scavenging activity of Fruc, Fruc Lys, Glc, and Glc Lys... 25 11. Correlation between browning intensity and scavenging activity... 27
xii LIST OF APPENDIX Appendix Pages 1. Raw Data of Browning Intensity at Spectrophotometer... 35 2. Raw Data of Browning Index (Fruc)... 36 3. Raw Data of Browning Index (Fruc Lys)... 37 4. Raw Data of Browning Index (Glc)... 38 5. Raw Data of Browning Index (Glc Lys)... 39 6. Raw Data of ABTS Radical Scavenging Activity (Fruc)... 40 7. Raw Data of ABTS Radical Scavenging Activity (Fruc Lsy )... 41 8. Raw Data of ABTS Radical Scavenging Activity (Glc)... 42 9. Raw Data of ABTS Radical Scavenging Activity (Glc-Lys)... 43 10. Raw Data of GraphPad Prism Correlation between Browning Intensity and Scavenging Activity... 44