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UNIVERSITI PUTRA MALAYSIA
BABAK HATAMI BAROOGH
FSTM 2012 16
CORRELATION OF ACRYLAMIDE WITH PRECURSORS AND COLOUR IN FROZEN PAR-FRIED FRENCH FRIES
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CORRELATION OF ACRYLAMIDE WITH PRECURSORS AND COLOUR IN FROZEN PAR-FRIED FRENCH FRIES
By
BABAK HATAMI BAROOGH
Thesis Submitted to the School of Graduate Studies, Universiti Putra Malaysia, in Fulfillment of the Requirements for the Master of Science
April 2012
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I DEDICATE THIS THESIS TO MY PARENTS AND ALL MY TEACHERS AND LECTURERS
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Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfillment of the requirements for the degree of Master of Science
CORRELATION OF ACRYLAMIDE WITH PRECURSORS AND COLOUR IN
FROZEN PAR-FRIED FRENCH FRIES
By
BABAK HATAMI BAROOGH
April 2012
Chairman : Prof Jinap Selamat, PhD
Faculty : Food Science and Technology
The aim of this study was to verify the occurrence of acrylamide formation after final
frying and to study the correlation of acrylamide content with amino acids, reducing
sugars and moisture content in frozen French fries samples, also to investigate
acrylamide formation and reducing sugar content as main precursor in different
production date and production plant of frozen fried French fries samples. Acrylamide
analysis was based on gas chromatographic mass spectrometric technique (GC-MS) that
included extraction and derivatization before analysis. Amino acids and reducing sugars
determination has been done using high performance liquid chromatography (HPLC). It
was discovered that final frying generates 76.04-348.55 µg/kg of acrylamide in French
fries samples by wide range of reducing sugar content in the form of fructose and
glucose (0.11-1.30 and 0.096-1.294 mg/g, respectively) and fifteen free amino acids
(0.01-9.17 mg/g) that had been detected in frozen French fries sample before final
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frying. Although, asparagine content (2.76-9.17 mg/g) was higher among other free
amino acids, however; threonine and glutamine showed the higher correlation with
acrylamide content. In addition, among reducing sugars, there was a significant
correlation between glucose and acrylamide content and acrylamide and color
development (�a). It can be concluded correlation of acrylamide and precursors in
frozen par fried French fries samples is not same as fresh fried potatoes. In second part,
it has been investigated that there was not a significant difference between means of
acrylamide content and reducing sugars of different brand (P > 0.05) and was no
significant difference (P > 0.05) between reducing sugars and acrylamide content of two
harvest years (2008-2009), although; acrylamide content in 2009 was higher than 2008.
This part of study indicated that climate change from 2008 to 2009 has no significant
single effect on acrylamide content in this product. Acrylamide content formed after
frying frozen French fries sample of twelve month production within these two years
are significantly different (P < 0.05). The highest content of acrylamide (348.55µg/kg)
was found in samples produced in January that were significantly different among other
months. The lowest amount of acrylamide (76.04µg/kg) was found in samples produced
in September.
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Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagi memenuhi keperluan untuk ijazah Master Sains
KORELASI AKRILAMIDA DENGAN PREKURSOR DAN WARNA DALAM
KENTANG BEKU SEPARUH MASAK YANG DIGORENG
Oleh
BABAK HATAMI BAROOGH
April 2012
Pengerusi: Professor Jinap Selamat, PhD
Fakulti: Sains Makanan dan Teknologi
Tujuan kajian ini adalah untuk mengesahkan berlakunya pembentukan akrilamida
selepas menggoreng dan mengkaji perkaitan antara kandungan akrilamida dengan asid
amino, gula penurun dan kandungan lembapan di dalam sampel kentang goreng beku,
juga untuk menyiasat pembentukan akrilamida dan kandungan gula penurun sebagai
prekursor utama pada tarikh pengeluaran dan kilang pengeluaran kentang goreng beku
yang berbeza. Analisis akrilamida berdasarkan teknik kromatografi gas spektrometrik
(GC-MS) yang melibatkan pengekstrakan dan derivatization sebelum analisis. Asid
amino dan gula penurun ditentukurkan mengikut kaedah kromatografi cecair
berprestasi tinggi (HPLC). Ia telah ditemui bahawa menggoreng menjana 76.04-348.55
�g / kg akrilamida dalam sampel kentang goreng oleh pelbagai kandungan gula penurun
dalam bentuk fruktosa dan glukosa (0.11-1.30 dan 0.096-1.294 mg / g, masing-masing)
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dan 15 asid amino bebas (0.01-9.17 mg / g) yang telah dikesan dalam sampel kentang
goreng beku sebelum goreng. Walaupun, kandungan asparagine (2.76-9.17 mg / g)
adalah lebih tinggi di kalangan asid amino bebas yang lain, bagaimanapun, threonin dan
glutamin menunjukkan korelasi yang lebih tinggi dengan kandungan akrilamida. Di
samping itu, antara gula penurun, terdapat hubungan yang signifikan antara glukosa dan
kandungan akrilamida dan akrilamida dengan pembangunan warna (�a). Dapat
disimpulkan korelasi akrilamida dan prekursor dalam kentang goreng beku tidak sama
seperti kentang goreng segar. Dalam bahagian kedua, tiada perbezaan yang signifikan
antara purata kandungan akrilamida dan gula penurun bagi jenama yang berbeza (P>
0.05) dan tiada perbezaan yang signifikan (P> 0.05) antara gula penurun dan kandungan
akrilamida bagi dua tuaian tahun (2008-2009), walaupun; akrilamida kandungan pada
tahun 2009 adalah lebih tinggi daripada 2008. Kajian juga menunjukkan bahawa
perubahan iklim dari 2008 hingga 2009 tidak mempunyai kesan yang ketara ke atas
kandungan akrilamida dalam produk ini. Kandungan akrilamida dibentuk selepas
menggoreng kentang goreng beku daripada dua belas bulan pengeluaran dalam tempoh
dua tahun ini berbeza secara signifikan (P <0.05). Kandungan tertinggi akrilamida
(348.55�g/kg) telah dijumpai dalam sampel pada bulan Januari yang menunjukkan
perbezaan yang ketara di kalangan bulan-bulan yang lain. Jumlah terendah akrilamida
(76.04�g/kg) telah dijumpai dalam sampel yang dihasilkan pada September.
.
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ACKNOWLEDGEMENTS
I want to thanks God for enabling me to complete my research.
I would like to express my deepest gratitude to my chair person of the supervisory
committee, Professor Dr. Jinap Selamat who was a great source of motivation, support
and scientific supervision throughout our research. You have worked in such a good
harmony that made my research life enjoyable, the way you helped me to shape my
ideas has eased up all the hard work. All your efforts are really appreciated. I am also
deeply appreciated her for arranging of the necessary funding and financial supporting.
Your endless support and encouragements made me crave for more and more research. I
am full of thanks to my supervisory committee members, Associate Professor Dr. Md.
Zaidul Islam Sarker and Dr. Alfi Khatib for their valuable contribution and suggestions.
Your enthusiastic discussions and ideas have opened up a whole new horizon in my
scientific thinking. My dear friends in Food Safety Lab, Dr. Parvaneh, Gissia,
Maimunah, Wendy and Diana, Thanks for your all cooperation and friendly chats we
shared. I always enjoyed your critics and cultural discussions. I am appreciated to Mrs
linawatti, Mrs. Liza, Mr. Halim, Mr. Azman, for their helps during laboratory
experiments. My deepest appreciation and thanks to my family members who had been
constantly supporting me. I love you all and thank you for loving me and being great
familly.
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I certify that an Examination Committee met on 26/04/2012 to conduct the final examination of Gisia Daniali on his Mst degree of Food Science thesis entitled “Determination of acrylamide in banana-based snack and the effect of precursors on the acrylamide formation” in accordance with Universiti Pertanian Malaysia (Higher Degree) Act 1980 and Universiti Pertanian Malaysia (Higher Degree) Regulations 1981. The committee recommends that the student be awarded the Master of Science degree. Members of the Examination Committee are as follows:
Lasekan Olusegun Associate Professor Dr. Faculty of Food Science and Technology Universiti Putra Malaysia (Chairman) Faridah Abas Associate Professor Dr. Faculty of Food Science and Technology Universiti Putra Malaysia (Internal Examiner) Seyed Hamed Mirhosseini Dr. Faculty of Food Science and Technology Universiti Putra Malaysia (Internal Examiner) Bahruddin Saad Professor Dr. School of Chemistry Universiti Sains Malaysia (External Examiner)
SEOW HENG FONG, PhD Professor and Deputy Dean School of Graduate Studies
Date:
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This thesis was submitted to the Senate of Universiti Putra Malaysia and has been accepted as fulfilment of the requirement for the degree of Master of Science. Members of the Supervisory Committee were as follows.
Jinap Selamat, PhD Professor Faculty of Food Science and Technology Universiti Putra Malaysia (Chairman) MD Zaidul Islam Sarker, PhD Associate Professor Faculty of Food Science and Technology Universiti Putra Malaysia (Member) Alfi Khatib, PhD Lecturer Faculty of Food Science and Technology Universiti Putra Malaysia (Member)
BUJANG BIN KIM HUAT, PhD Professor and Dean School of Graduate Studies Universiti Putra Malaysia
Date:
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I declare that the thesis is my original work except for quotations and citations, which have been duly acknowledged. I also declare that it has not been previously and is not concurrently submitted for any other degree at Universiti Putra Malaysia or at any other institutions.
BABAK HATAMI BAROOGH
Date: 26 April 2012
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TABLE OF CONTENTS
Page ABSTRACT iii ABSTRAK v ACKNOWLEDGEMENTS vii APPROVAL viii DECLARATION x LIST OF TABLES xiv LIST OF FIGURES xvi LIST OF ABBREVIATIONS xx
CHAPTER
1 INTRODUCTION 1
1.1 Background of study 1
1.2 Objectives 3
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2 LITERATURE REVIEW 4
2.1 Frozen par fried French fries 4
2.2 Climate change and potato crops 7
2.3 Chemistry of acrylamide 9
2.4 Occurrence of acrylamide in food 11
2.5 Metabolism and toxicology of acrylamide 13
2.6 Acrylamide dietary exposure 15
2.7 Acrylamide formation 15
2.7.1 Formation from acrolein 16
2.7.2 Formation from Maillard reaction 17
2.8 Factors affecting the acrylamide formation 19
2.8.1 Potato variety and storage condition 20
2.8.2 Potato tuber size and underwater weights 20
2.8.3 Climate changes in growing seasons 21
2.8.4 Amino acids and reducing sugars contents 21
2.8.5 Moisture content 22
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2.8.6 pH 22
2.8.7 Blanching 23
2.8.8 Frying oil 24
2.8.9 Frying temperature 24
2.8.10 Frying time 26
2.9 Analytical methods for acrylamide precursors detection 27
2.9.1 Amino acids analysis 27
2.9.2 Reducing sugar analysis 28
2.10 Analytical method for acrylamide detection 29
2.10.1 Determination by GC-MS 30
2.10.2 Determination by LC-MS/MS 31
3 MATERIALS AND METHODS 32
3.1 Chemicals and instruments 32
3.2 Collection and Preparation of samples 33
3.2.1 Frying of samples 33
3.3 Analysis of acrylamide 34
3.3.1 Acrylamide extraction 34
3.3.2 GC-MS analysis 35
3.3.3 Calibration, LOD and LOQ 36
3.3.4 Recovery test 39
3.4 Analysis of glucose, fructose and sucrose 39
3.5 Analysis of free amino acids 40
3.5.1 Protein hydrolysis 40
3.5.2 Sample preparation and derivatization 40
3.5.3 Amino acid determination 41
3.6 Moisture analysis 42
3.7 Color measurement 42
3.8 Statistical analysis 42
�4 RESULTS AND DISCUSSION 45
4.1 The content of acrylamide, amino acids, reducing sugars and moisture in frozen
French fries 45
4.2 Correlation of acrylamide content with amino acids, reducing sugars and moisture 47
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4.2.1 Correlation of acrylamide with amino acids 49
4.2.2 Correlation of acrylamide with sugars 56
4.2.3 Correlation of acrylamide with moisture content 57
4.3 Correlation of acrylamide with color development 57 4.4 acrylamide formation and reducing sugar content in different production date and plant in frozen French fries samples 65
4.4.1 Impact of production month on acrylamide formation in frozen French fries samples 68
4.4.2 Impact of production year on acrylamide formation in frozen French fries samples 71 4.4.3 Impact of production plant (brand) on acrylamide formation in frozen par French fries samples 76
5 SUMMARY, GENERAL CONCLUSION AND RECOMMENDATION FOR FUTURE RESEARCH 81
5.1 Conclusion 81
5.2 Recommendation 84
�REFERNCES 86 APPENDICES 96 BIODATA OF STUDENT 112 LIST OF PUBLICATIONS 113
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