HISTAMINE LEVELS IN FROZEN AND SMOKED FISH IN NUNGUA MARKET BY ARABA INCOOM B.Sc. FOOD SCIENCE AND TECHNOLOGY (HONS) THIS THESIS IS PRESENTED TO THE DEPARTMENT OF FOOD SCIENCE AND TECHNOLOGY, KWAME NKRUMAH UNIVERSITY OF SCIENCE AND TECHNOLOGY, KUMASI IN PARTIAL FULFILLMENT OF THE REQUIREMENT OF MASTER OF SCIENCE IN FOOD QUALITY MANAGEMENT JULY 2016 DECLARATION I HEREBY DECLARE THAT I HAVE WHOLLY UNDERTAKEN THE STUDY REPORTED HEREIN UNDER THE SUPERVISION OF DR. FRANCIS ALEMAWOR AND THAT EXCEPT WHERE REFERENCES HAVE BEEN DULY CITED, THIS DISSERTATION IS THE OUTCOME OF MY RESEARCH. NEITHER ALL NOR PARTS OF THIS THESIS HAVE BEEN PRESENTED FOR ANOTHER DEGREE ELSEWHERE. ARABA INCOOM _________________ _______________ (M. SC. STUDENT) SIGNATURE DATE FRANCIS ALEMAWOR, PhD __________________ _______________ (SUPERVISOR) SIGNATURE DATE PROF MRS I. ODURO __________________ _______________ (HEAD OF DEPARTMENT) SIGNATURE DATE II ACKNOWLEDGEMENT I am extremely grateful to God Almighty for His strength and wisdom during my study. My profound gratitude also goes to my supervisor Dr. Francis Alemawor for his patience and wiliness to direct me to get the best out of my research. May God richly bless you and increase you in every good thing. I also owe thanks to Dr. Emmanuel DeGraft Johnson for sharing his expertise in statistics which aided me in data analysis. A special thanks to Mr. Isaac Ofosu who gave invaluable suggestions at the beginning of my study. I also thank Mr. Clifford Frimpong, the Head of Department of the Food and Agric laboratories of Ghana Standards Authority for the concern shown in the progress of the studies. I thank the staff of the Mycotoxins and Histamine Laboratory for their unwavering support they gave me during my analysis. Special thanks go to my Mother, Ms. Esther Danby Cobbina and my Siblings; Mrs. Hilda Amoah and Mr. Ekow Incoom for their fervent prayers and support. They always expressed interest in my progress and gave me hope in challenging times. My utmost gratitude also goes to my wonderful Husband, Dr. Bright Adu, who assisted me in every step and encouraged me to learn. He has been a strong support and I am so much of a better person now than ever, God richly bless you. III TABLE OF CONTENT DECLARATION.......................................................................................................................... II ACKNOWLEDGEMENT .......................................................................................................... III TABLE OF CONTENT .............................................................................................................. IV LIST OF FIGURES ................................................................................................................. VIII LIST OF TABLES ...................................................................................................................... IX LIST OF ABBREVIATION........................................................................................................ X ABSTRACT ................................................................................................................................. XI CHAPTER ONE ........................................................................................................................... 1 1 INTRODUCTION ............................................................................................................... 1 1.1 Background ................................................................................................................... 1 1.2 Problem Statement ........................................................................................................ 2 1.3 Justification ................................................................................................................... 3 1.4 Overall Objective .......................................................................................................... 3 1.4.1 Specific Objective ................................................................................................. 3 CHAPTER TWO .......................................................................................................................... 4 2 LITERATURE REVIEW .................................................................................................... 4 2.1 Spoilage of fish ............................................................................................................. 4 2.2 Microbial flora in fresh fish .......................................................................................... 5 2.3 Bacterial deterioration of fish ....................................................................................... 6 IV 2.4 Histamine production in fish ........................................................................................ 7 2.5 Bacteria responsible for histamine production in fish .................................................. 8 2.6 Regulations on histamine levels in fish ...................................................................... 10 2.7 Scombroid poisoning .................................................................................................. 10 2.7.1 Treatment ............................................................................................................ 11 2.8 Toxicological aspects of histamine............................................................................. 11 2.8.1 Absorption, distribution, metabolism and excretion ........................................... 11 2.8.2 Toxicological responses in humans..................................................................... 13 2.9 Effects of storage time and temperature on histamine levels in fish .......................... 14 2.10 Effect of histamine on sensory characteristics of fish ................................................ 15 2.11 Control of histamine formation .................................................................................. 15 2.11.1 Refrigeration........................................................................................................ 15 2.11.2 Heading, gutting and skinning ............................................................................ 16 2.11.3 Heat processing ................................................................................................... 16 2.11.4 Modified atmosphere packaging ......................................................................... 17 2.11.5 Freezing and chilled storage ................................................................................ 17 2.11.6 Gamma irradiation............................................................................................... 18 2.11.7 Protective cultures of bacteria ............................................................................. 18 2.12 Fish consumption patterns .......................................................................................... 19 2.13 Selected species of scombroid fish ............................................................................. 20 V 2.13.1 The Atlantic Mackerel (Scomber scombrus) ....................................................... 20 2.13.2 Atlantic Horse Mackerel (Trachurus trachurus) ................................................ 20 CHAPTER THREE .................................................................................................................... 22 3 MATERIALS AND METHODS ....................................................................................... 22 3.1 Study area ................................................................................................................... 22 3.2 Materials ..................................................................................................................... 23 3.3 Methods ...................................................................................................................... 24 3.3.1 Experimental design, sampling and data collection ............................................ 24 3.4 Procedure for histamine Analysis ............................................................................... 26 3.4.1 Overview ............................................................................................................. 26 3.4.2 Extraction ............................................................................................................ 26 3.4.3 Preparation of standard solutions ........................................................................ 26 3.4.3.1 Preparation of stock solution of histamine (histamine 1000 mg/kg) ............... 26 3.4.3.2 Preparation of working standard ...................................................................... 27 3.4.3.3 Preparation of calibration standard solution .................................................... 27 3.4.4 HPLC analysis ..................................................................................................... 27 3.5 Statistical analysis....................................................................................................... 28 CHAPTER FOUR ....................................................................................................................... 29 4 RESULTS AND DISCUSSION ........................................................................................ 29 4.1 Study samples ............................................................................................................. 29 VI 4.2 Histamine concentrations in frozen Atlantic Mackerel