ASSESSMENT of the LEVELS of THIOCYANATE in PROCESSED and UNPROCESSED RED and BROWN FINGER MILLET ( Eleusine Coracana) GROWN in MOGOTIO AREA, BARINGO COUNTY, KENYA
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i ASSESSMENT OF THE LEVELS OF THIOCYANATE IN PROCESSED AND UNPROCESSED RED AND BROWN FINGER MILLET ( Eleusine coracana) GROWN IN MOGOTIO AREA, BARINGO COUNTY, KENYA By Silva Jerono Chebet (B.ED SC) I56/CE/22219/2010 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF THE DEGREE OF MASTER OF SCIENCE IN CHEMISTRY IN THE SCHOOL OF PURE AND APPLIED SCIENCES OF KENYATTA UNIVERSITY NOVEMBER 2014 ii DECLARATION I hereby declare that this is my original work and has not been submitted for the award of a degree in this or any other university. Signed…………………………… date…………………………. Chebet Jerono Silva I56/CE/22219/2010 Kenyatta University This thesis has been submitted with our approval as the university's supervisors. Signed………………………………date…………………………. Dr. Mildred Nawiri Department of Chemistry Kenyatta University Signed………………………………date…………………………. Prof. Jane Murungi Department of Chemistry Kenyatta University. iii DEDICATION I dedicate this work to my beloved husband Daniel Cherono and our children Ian Kipchumba and Dennis Kipkemboi and my parents David Chebon and Eunice Talaa. iv ACKNOWLEDGEMENTS I would like to thank the almighty God for enabling me to come this far. I wish to express my sincere gratitude to my supervisors Prof. Jane Murungi and Dr. Mildred Nawiri for their constant encouragement, support, guidance and advice throughout the research process. I thank Kenyatta University for offering me admission to the course. I am also grateful to the lecturers and technical staff of Chemistry Department, Kenyatta University for their support during this study. Thanks to the University's Chemistry Laboratory, for the equipment and chemicals offered. I am grateful to the National Commission of Science and Technology (NACOSTI) for funding the research project. Many thanks go to my course mates and peers for their supportive ideas and advice throughout the course. I would also like to express my gratitude to my aunt Mrs. Margaret Sergon who kept reminding me to enroll for a masters Degree. Finally I would like to thank my husband for the financial support and our children for their patience and understanding during my absence. v TABLE OF CONTENTS TITLE PAGE…………………………………………………………………………….i DECLARATION………………………………………………………………………..ii DEDICATION…………………………………………………………………………..iii ACKNOWLEDGEMENTS ............................................................................................... iv TABLE OF CONTENTS…………………………………………………………………v LIST OF FIGURES ........................................................................................................... ix LIST OF TABLES .............................................................................................................. x ABBREVIATIONS AND ACRONYMS .......................................................................... xi ABSTRACT……………………………………………………………………………..xii CHAPTER ONE…………………………………………………………………………..1 INTRODUCTION………………………………………………………………………...1 1.1 Background information……………………………………………………………1 1.2 Problem statement and justification .......................................................................... 8 1.3 Hypotheses ................................................................................................................ 8 1.4 Objectives ................................................................................................................. 9 1.4.1 General objective ............................................................................................... 9 1.4.2 Specific objectives……………………………………………………………..9 1.5 Significance of the study and expected output……………………………………...9 1.6 Scope and Limitations of the study ......................................................................... 10 CHAPTER TWO………………………………………………………………………...11 LITERATURE REVIEW ................................................................................................. 11 vi 2.1 Goiter prevalence .................................................................................................... 11 2.2 Millet ....................................................................................................................... 13 2.2.1 Finger millet ......................................................................................................... 14 2.2.2 Uses of Finger millet ............................................................................................ 15 2.2.3 Antinutrients in finger millet ............................................................................... 16 2.3 Goitrogens and cyanogenic glycosides ................................................................... 17 2.3.1 Goitrogens ............................................................................................................ 17 2.3.2 Biosynthesis, catabolism and detoxification of cyanogenic glycosides .......... 18 2.3.3 Glucosinolate hydrolysis and products ............................................................ 20 2.4 Thiocyanate ............................................................................................................. 22 2.4.1 Thiocyanate content in food plants ...................................................................... 23 2.4.2 Role of thiocyanate in goiter development .......................................................... 25 2.5 Food processing and thiocyanate levels .................................................................. 28 2.5.1 Heat .................................................................................................................. 28 2.5.2 Fermentation .................................................................................................... 30 2.5.3 Sprouting .......................................................................................................... 31 2.5.4 Soaking ............................................................................................................ 33 2.6 Method of analysis of thiocyanate .......................................................................... 34 2.6.1 Principle of acid hydrolysis ............................................................................. 34 2.6.2 UV-VIS spectroscopy ...................................................................................... 35 vii CHAPTER THREE .......................................................................................................... 39 METHODOLOGY ........................................................................................................... 39 3.1 Research design ...................................................................................................... 39 3.2 Sample collection .................................................................................................... 39 3.3 Chemicals and Reagents ......................................................................................... 39 3.4 Cleaning of apparatus ............................................................................................. 40 3.5 Preparation of the standard solution ....................................................................... 40 3.6 Sample preparation ................................................................................................. 40 3.6.1 Preparation of the dried grains ......................................................................... 40 3.6.2 Sprouting of grains ........................................................................................... 41 3.6.3 Soaking of grains ............................................................................................. 41 3.6.4 Preparation of fresh flour ................................................................................. 41 3.6.5 Fermentation of flour ....................................................................................... 41 3.6.6 Cooked flour (porridge) preparation ................................................................ 42 3.6.7 Preparation of the green and dry leaves ........................................................... 42 3.7 Extraction and determination of thiocyanate ...................................................... 42 3.8 Method validation ................................................................................................... 43 3.9 Data analysis ........................................................................................................... 43 CHAPTER FOUR ............................................................................................................. 44 RESULTS AND DISCUSSION ....................................................................................... 44 viii 4.1 Introduction ............................................................................................................. 44 4.2 Method validation ................................................................................................... 44 4.3 Levels of thiocyanate in finger millet grains .......................................................... 46 4.4 Levels of thiocyanate in finger millet flour ............................................................ 49 4.5 Levels of thiocyanate in the green and dry leaves of finger millet ......................... 53 CHAPTER FIVE .............................................................................................................. 55 CONCLUSIONS AND RECOMMENDATIONS ........................................................... 55 5.1 Conclusions ............................................................................................................