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ANALYSIS OF CARBONYL COMPOUND FORMATION ON MAILLARD

REACTION USING RARE SUGAR D-SORBOSE AND THREONINE

BACHELOR THESIS

Composed by

FAHMI ILMAN FAHRUDIN

FOOD TECHNOLOGY STUDY PROGRAM

DEPARTMENT OF AGRICULTURAL SCIENCES

FACULTY OF ANIMAL AND AGRICULTURAL SCIENCES

DIPONEGORO UNIVERSITY

SEMARANG, INDONESIA

2017

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ANALYSIS OF CARBONYL COMPOUND FORMATION ON MAILLARD

REACTION USING RARE SUGAR D-SORBOSE AND THREONINE

Composed by

FAHMI ILMAN FAHRUDIN

Student Reg. Number 23020113140048

As One of Requirements for 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

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ENDORSEMENT LETTER

Title of Bachelor Thesis : ANALYSIS OF CARBONYL

COMPOUND FORMATION ON

MAILLARD REACTION USING RARE

SUGAR D-SORBOSE AND THREONINE

Name of Student : FAHMI ILMAN FAHRUDIN

Student Reg. Number : 23020113140048

Study Program/Department : FOOD

TECHNOLOGY/AGRICULTURAL

Faculty : ANIMAL AND AGRICULTURAL

SCIENCES

Has been examined by the examiner team

and declared to pass on……………………

Main Supervisor

Ahmad N. Al-Baarri S.Pt., M.P., Ph.D.

Secondary Supervisor

Prof. Dr.Ir. Anang M. Legowo, M.Sc

Head of Final Exam Program

Dr. Ir. Antonius Hintono, MP

Head of Study Program

Dr. Yoyok Budi Pramono

Dean

Prof. Dr. Ir. Mukh Arifin, M.Sc

Head of Department

Dr. Ir. Didik Wisnu Widjajanto, M.S

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ORIGINALITY STATEMENT OF BACHELOR THESIS

I, the undersigned below:

Name : Fahmi Ilman Fahrudin

Student Reg. Number : 23020113140048

Study Program : Food Technology

Hereby declare as follows:

1. Scientific work entitled:

Analysis of Carbonyl Compound Formation on Maillard Reaction

Using Rare Sugar D-Sorbose and Threonine, 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, July 2017

Author

Fahmi Ilman Fahrudin

Knowledged by

Main Supervisor

Ahmad N. Al-Baarri, S.Pt., M.P., Ph.D.

Secondary Supervisor

Prof. Dr. Ir. Anang M. Legowo, M.Sc.

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RINGKASAN

FAHMI ILMAN FAHRUDIN. 23020113140048. Analisis Senyawa Karbonil

pada Reaksi Maillard antara Gula D-sorbose dan Asam Amino Treonin.

(Pembimbing : Ahmad N. Al-Baarri and Anang M. Legowo)

Polisakarida merupakan senyawa yang umumnya terdapat pada bahan

pangan dan juga termasuk sumber nutrisi tubuh. Bahan utama pembentuk gula D-

sorbose adalah polisakarida yang melalui reaksi enzimatik menggunakan D-

tagatose 3-epimerase. D-sorbose yang terbentuk memiliki tingkat reaktifitas yang

tinggi dibandingkan dengan gula alami lainnya. Reaksi Maillard merupakan reaksi

yang terjadi antara gula dan asam amino dengan perlakuan panas, reaksi ini biasa

terjadi pada proses pengolahan pangan seperti pemanggangan dan pengovenan.

Reaksi Maillard memiliki dampak positif dan negative yang timbul dari perlakuan

suhu tinggi dan dapat mempengaruhi kesehatan.

Penelitian ini menggunakan sorbose (Sor) dan Threonine (Thr) sebagai

model reaksi Maillard untuk menghasilkan MRPs (Maillard Reaction Products)

dan mendapatkan dampak positif tanpa menimbulkan dampak negatif dari reaksi

Maillard. Penelitian ini bertujuan untuk menunjukan intensitas kecoklatan, indeks

kecoklatan, pengukuran spectrum, aktivitas antioksidan dan korelasi antara

intesitas kecoklatan dan aktivitas antiosidant MRPs yang dihasilkan dari Sor-Thr

pada suhu 50℃.

Bahan yang digunakan pada penelitian ini adalah D-sorbose, Threonine,

ABTS atau 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid). Sorbose dan

Threonine digunakan untuk memproduksi MRPs menggunakan perlakuan

pemanasan pada suhu 50°C selama 48 jam. MRPs dengan intensitas coklat

diperiksa dengan pengukuran spektral yang menggunakan spektrofotometer pada

420 nm dan aktifitas antioksidan dianalisis dengan menggunakan panjang

gelombang 734nm dengan metode ABTS, sedangkan indeks browning

menggunakan colorimeter untuk menentukan nilainya. Korelasi antara intensitas

kecoklatan dan aktivitas pemulungan dianalisis dengan menggunakan GraphPad

Prism 6.0 untuk mendapatkan nilai signifikansi 60% sehingga mendapatkan

korelasi antar data yang dihasilkan.

Selama proses reaksi Maillard berlangsung untuk menghasilkan MRPs,

Sor-Thr menunjukkan aktivitas yang lebih baik daripada Sor. Intensitas

kecoklatan, perubahan warna, dan aktivitas antioksidan meningkat seiring dengan

meningkatnya durasi perlakuan pemanasan. Berdasarkan pengukuran spektral,

MRPs dari Sor-Thr awalnya terdeteksi pada 48h pada saat perlakuan pemanasan

berlangsung. Korelasi antara intensitas kecoklatan dan aktivitas oksidan dilakukan

sebagai korelasi positif non linier dan korelasi signifikan. Karena proses reaksi

glikasi yang berlangsung secara terus menerus disarankan harus adanya kontrol

dengan memberi perlakuan khusus lama pemanasan dan suhu yang digunakan

pada gula dan asam amino.

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SUMMARY

FAHMI ILMAN FAHRUDIN. 23020113140048. Analysis of Carbonyl

Compound Formation on Maillard Reaction using Rare Sugar D-Sorbose And

Threonine. (Supervisor : Ahmad N. Al-Baarri and Anang M. Legowo)

Polysaccharides are commonly used in various foods processing, including

the sources of food nutrients, food additives, and source of biologically active

molecules. Polysaccharides are also found as a main resource to produce rare

sugar D-sorbose through the enzymatic reaction using D-tagatose 3-epimerase.

Through the various foods processing including heat treatment, fructose may be

glycated to amino groups result the Maillard Reaction Product or MRPs that may

contains negative impact for health. This research was used sorbose (Sor) and

Threonine (Thr) as a model of glycation reaction to generate the MRPs. This study is

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 Sor – Thr at 50°C.

The materials on this research are D-sorbose, Threonine, ABTS or 2,2’-

azino-bis (3-ethylbenzthiazoline-6-sulfonic acid). Sorbose and Threonine were

used to producing MRPs using heating treatment at 50°C for 48 hour. The

browning intensity MRPs was investigate with spectral measurement used

spectrophotometer at 420 nm and scavenging was analyzed at 734nm with ABTS

method, 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 about 83%.

During Maillard reaction process to generate MRPs, Sor-Thr were

showing better performance than Sor. The browning intensity, color development,

and scavenging activity were improved along with the increasing of duration

heating treatment. Based on spectral measurement, MRPs from Sor-Thr was

initialy detected at 48h of heating treatment. The correlation between browning

intensity and scavenging activity were assigned as a positive non-linear

correlation and significant correlation. Since the further process of glycation

should be controlled using duration of heating treatment of sugar-amino it be

advised.

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PREFACE

Praise and great gratitude to Almighty God, Allah s.w.t., who always gives

His gracious mercy and tremendous blessing that has helped the author to

successfully finish this bachelor thesis with the title, “Analysis of Carbonyl

Compound Formation on Maillard Reaction Using Rare Sugar D-sorbose And

Threonine”. 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. Dr. Yoyok Budi Pramono and Dr. Ir. Bambang Dwiloka, M.S., as examiner

commitee 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,

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Diponegoro University. Thank you very much for the time, knowledge, 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-Sorbose 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 Adi Fahrudin and Husmiati

Yusuf, 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 Fikran Aranda Fahrudin,

the author’s brother,

7. Special thanks and appreciation dedicated to Fina Fitriana Budiman who

always been supported in many things during the up and down of the author’s

study.

8. Sincere thanks to my secondary family “Wacana Guys” during four year

college that always accompany; Affan Tian Dita, Taufiq Shaleh, Deshtyan

Erlangga and Beta Haedar.

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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, July 2017

Author,

Fahmi Ilman Fahrudin

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TABLE OF CONTENTS

RINGKASAN .......................................................................................................... v

SUMMARY ........................................................................................................... vi

PREFACE ............................................................................................................. vii

TABLE OF CONTENTS ......................................................................................... x

LIST OF FIGURE ................................................................................................. xii

LIST OF APPENDIX .......................................................................................... xiii

CHAPTER I INTRODUCTION .............................................................................. 2

CHAPTER II LITERATURE REVIEW .................................................................. 5

2.1. Maillard Reaction Products (MRPs) .......................................................... 5

2.2. Rare Sugar of D-sorbose ............................................................................ 6

2.3. Amino Acids Threonine ............................................................................. 7

2.4. Browning Intensity..................................................................................... 8

2.5. Color Development .................................................................................... 9

2.6. Spectrophotometric .................................................................................. 10

2.7. Radical Scavenging Activity.................................................................... 11

2.8. Correlation between Scavenging Activity and Browning Intensity ........ 12

CHAPTER III MATERIAL AND METHODS ..................................................... 13

3.1. Material .................................................................................................... 13

3.2. Methods.................................................................................................... 13

3.2.1. Preparation of MRPs Model .................................................................. 13

3.2.2.Browning Intensity ................................................................................. 14

3.2.3.Color Development ................................................................................ 14

3.2.4.Spectroscopic Measurements ................................................................. 15

Pages

xi

3.2.5.ABTS Radical Scavenging Activity ...................................................... 15

3.2.6.Correlation between Browning Intensity and Scavenging Activity....... 16

3.3. Data Analysis ........................................................................................... 16

CHAPTER IV RESULTS AND DISCUSSIONS ................................................. 17

4.1 Browning Intesity..................................................................................... 17

4.2. Color Development .................................................................................. 18

4.3. Spectroscopic Characteristics of Sor-Thr MRPs ..................................... 20

4.4 ABTS Radical Scavenging Activity ........................................................ 22

4.5. The Correlation between Browning Intensity and Scavenging Activity . 23

CHAPTER V .......................................................................................................... 26

CLOSING .............................................................................................................. 26

5.1. Conclusions .............................................................................................. 26

5.2 Suggestions .............................................................................................. 26

REFRENCE ........................................................................................................... 27

APPENDIX ............................................................................................................ 33

BIOGRAPHY ........................................................................................................ 39

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LIST OF FIGURE

Number Pages

1. Maillard Reaction Proccess …………………………………………………….6

2. Chemical chain of D-Sorbose .............................................................................. 7

3. The flow diagram of Maillard reaction products ............................................... 14

4. Browning intensity of rare sugar Sor (•) and Sor-Thr (∆). ................................. 17

5. Browning index of rare sugar Sor (•) and Sor-Threonine (∆) ............................ 19

6. Development of emission spectra 190-750 nm of Sor (a) and Sor-Thr (b) ....... 21

7. The scavenging activity of Sor (•) and Sor-Threonine (∆) ................................ 22

8. A significant correlation between browning intensity and scavenging activity of

Sor-Thr . ............................................................................................................ 24

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LIST OF APPENDIX

Number Pages

1. Output of Browning Intensity at Spectrophotometer ......................................... 33

2. Raw Data of Browning Index (Sor) ................................................................... 34

3. Raw Data of Browning Index (Sor-Thr) ............................................................ 35

4. Output of ABTS Radical Scavenging Activity Calculation (Sor) ..................... 36

5. Output of ABTS Radical Scavenging Activity Calculation (Sor-Thr) .............. 37

6. Output of GraphPad Prism Correlation between Browning Intensity and

Scavenging Activity .......................................................................................... 38