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Bioavailability of Iron from Basella Alba and Amaranthus Hybridus Leaves Supplemented Diet in Iron Deficient Anaemic Albino Rats

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Bioavailability of Iron from Basella Alba and Amaranthus Hybridus Leaves Supplemented Diet in Iron Deficient Anaemic Albino Rats BIOAVAILABILITY OF IRON FROM BASELLA ALBA AND AMARANTHUS HYBRIDUS LEAVES SUPPLEMENTED DIET IN IRON DEFICIENT ANAEMIC ALBINO RATS. BY Ceaser Antiya, MOSES DEPARTMENT OF BIOCHEMISTRY FACULTY OF SCIENCE AHMADU BELLO UNIVERSITY ZARIA JANUARY, 2016 i BIOAVAILABILITY OF IRON FROM BASELLA ALBA AND AMARANTHUS HYBRIDUS LEAVES SUPPLEMENTED DIET IN IRON DEFICIENT ANAEMIC ALBINO RATS. BY Ceaser Antiya MOSES, B.SC BIOCHEMISTRY (A.B.U) 2011 MSc/SCIE/44812/2012-2013 A DISSERTATION SUBMITTED TO THE SCHOOL OF POSTGRADUATE STUDIES, AHMADU BELLO UNIVERSITY, ZARIA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE AWARD OF MASTERS DEGREE IN BIOCHEMISTRY DEPARTMENT OF BIOCHEMISTRY FACULTY OF SCIENCE AHMADU BELLO UNIVERSITY, ZARIA NIGERIA JANUARY, 2016 ii DECLARATION I hereby declare that the work in this dissertation entitled ―BIOAVAILABILITY OF IRON FROM BASELLA ALBA AND AMARANTHUS HYBRIDUS LEAVES SUPPLEMENTED DIET IN IRON DEFICIENT ANAEMIC ALBINO RATS” has been performed by me in the Department of Biochemistry under the supervision of Mr. O. A. Owolabi and Prof. E. Oyinke. The information herein derived from literature has been duly acknowledged in the text and a list of references provided. No part of this dissertationwas previously presented for another degree or diploma at any university to the best of my knowledge. …………………………………. ……………………… …………………… Moses Ceaser Antiya Signature Date iii CERTIFICATION This dissertation entitled ―BIOAVAILABILITY OF IRON FROM BASELLA ALBA AND AMARANTHUS HYBRIDUS LEAVES SUPPLEMENTED DIET IN IRON DEFICIENT ANAEMIC ALBINO RATS” by Ceaser, Antiya MOSES meets the regulations governing the award of the degree of MASTER of Science Ahmadu Bello University, and is approved for its contribution to knowledge and literary presentation. Mr. O. A. Owolabi ………………………. ……………………………… Chairman, Supervisory Committee Signature Date Prof. E. Onyike ………………………. ……………………………… Member, Supervisory Committee Signature Date Prof. M. N. Shuaibu ……………………….. ..…………………………....Head of Department Signature Date Prof. K. Bala ……………………….. ……………………………… Dean, School of Postgraduate Studies Signature Date iv DEDICATION This work is dedicated to my late Father, Mr. Moses Antiya v ACKNOWLEDGEMENT I must first of all express my gratitude to God for his unfailing love, grace and mercy to me; I give him Glory for his faithfulness, provision, and divine health he has granted me throughout my period of study in Zaria. My profound gratitude to my project supervisors, Mr. O. AOwolabi and Prof. E. Onyike for their limitless effort and suggestions, constructive criticism, corrections, close supervision and inspiration during the course of this work. It was your assistance, in all ramifications that made this work a success. My deep appreciation goes to my Head of Department, Prof. I.A Umar and the entire academic and nonacademic staff of the Department of Biochemistry for their individual and collective support. I will like to specially appreciate Dr. D. B James for her guidance and encouragement during the course of my studies. I will also like to acknowledge the staff of animal house Department of Pharmacology, Faculty of Pharmaceutical Science for allowing me to use the animal house for the course of this study. I sincerely want to appreciate my dear friends OkoloIjeoma, BindaAndongma, Ugoji Cynthia, Daniel Abba, AyubaDanburam, Ann Chinedu, Femi Omogoye, Mike Drenkat, KambaBayo, Samson Melah, Sale Gelte, Gideon Rimamfate, Josiah Angut, IfeomaUche, Ebuka Samuel,Chidi Anthony, Atobiloye Ahmed andAbdulmuttalab Baba-Ali. You people made the project experience a pleasant one. Finally, I am grateful to the entire Moses Antiya family for your prayers, emotional and financial supports that made my studies a huge success. vi ABSTRACT Iron deficiency anaemia is the most common form of micronutrient deficiency especially in the developing countries.The bioavailability of iron from blanched and unblanched Basella alba and Amaranthus hybridus leaves supplemented diet in iron deficient anaemic albino rats was investigated by determining vitamin C, antinutrients, speciation pattern of iron and effect of the leaves supplemented diet on serum ferritin and some haematological parameters using standard methods. Forty eight young albino rats were divided into eight groups. After induction of anaemia by feeding with iron deficient diet and deionized water, diet supplementation was carried out for 21 days. Amaranthus hybridus has a significantly (p<0.05) higher content of vitamin C (35 mg/100g), phytate (2.06 g/100g) and oxalate (89 mg/100g) than Basella alba. Basella alba recorded a significantly (p<0.05) higher levels of Fe, Zn, Cu, Ca and Mg than Amaranthus hybridus. Blanching significantly (p<0.05) decreased the vitamin C, phytate, oxalate and mineral content of both Basella alba and Amaranthus hybridus. Iron speciation pattern in the leaves showed that exchangeable and the residual forms of iron predominates over reducible and exchangeable in both leaves, which increased in B.alba but decreased in A.hybridus after blanching.After diet supplementation for 21 days, diet supplemented groups recorded significant (p<0.05) increase when compared with the anaemic rats. Groups supplemented with 10% unblanched A. hybridus recorded a significant increase in serum ferritin (138.40 ng/ml), red blood cell count (RBC) (11.97 x 106µl) and packed cell volume (PCV) (40.67%) when compared to the anaemic rats. Similarly, diet supplementation with 10% unblanched and blanched Basella alba and Amaranthus hybridus leaves respectively significantly (p<0.05) increased hemoglobin vii concentration, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration but recorded no significant (p>0.05) increase in the mean corpuscular volume when compared with the anaemic rats. The results suggest that Basella alba and Amaranthus hybridus supplemented diet increase bioavailability of iron in iron deficient anaemic albino rats with unblanched vegetables exhibiting greater potentials than the blanched. viii TABLE OF CONTENTS Page Cover Page ………………...…………..……………………………………………………….i Fly Page ……………………………………………………………………….………..…..ii Title Page ………………………………………………………….………….……………..iii Declaration Page ……………………………………………………………………………….....iv Certification Page ………..…………………………………………..…………………………….v Dedication ……….………….……………………………………...……………………………..vi Acknowledgment .……………………………………………...……………………….......vii Abstract ..………….……………………………………………..……………………….viii Table of Contents …….…………………..………………………………………………………..x List of Tables …....………………………...…………………………………………………..xvii List of Figures ……...……………………....…………………………………………………..xviii List of Plates …….…………………………..………………………………………………..…xix List of Appendix ………………………..……………………………………………………....xx List of Abbreviation ………………………….………………………………………………....xxii CHAPTER ONE 1.0 INTRODUCTION ………………….....…………………...………...…………………...1 1.1 Statement of Research Problem ……..….……………………………….........................3 1.2 Justification …..……….……………..………………………….….……….....................4 1.3 Aim and Objectives …………………..…………………..………..…………………..…5 ix 1.3.1 Aim .……………………………………………………………...……………………..5 1.3.2 Specific Objectives .…….………………………..……….…………….……………....5 CHAPTER TWO 2.0 LITERATURE REVIEW ….…………………………….……………………………….6 2.1 Basella alba and Amaranthus hybridus ...………..…………………………………….…6 2.1.1 Origin and distribution ……………………………………….………...……………….…6 2.1.2 General description ………………………………………………………………………..7 2.1.3 Botanical information ……………………………………………………………..9 2.1.4 Classification …..…………..……..………………….…………………………………...10 2.1.5 Chemical composition …………………...…………………………………………...….11 2.1.6 Uses …………………………………………………………..…………………………..12 2.1.6.1 Culinary uses ...…………………...……………………………………………..12 2.1.6.2 Medicinal uses ………..…………………………..…………...………………………...13 2.1.6.3 Industrial uses ………...………………………………………………………………....14 2.2 Iron ………………………………………………………...……...…………………….15 2.2.1 Occurrence ………………………………………………………………………………15 2.2.2 Iron requirement and distribution in the body …………...…..……………...................15 2.2.3 Dietary iron ………………………….………………………..……………………….16 2.2.4 Iron absorption …………………………………………..…………......................17 2.2.4.1 Non-haem iron absorption …...……………………………...…..………....................18 2.2.4.2 Haem iron absorption……………...……………..……..………………………...……..19 2.2.5 Iron Deficiency …………...………………….……………………………………20 x 2.2.6 Major causes of iron deficiency anaemia …………….…..…...……………………..21 2.2.6.1 Blood Loss ……………………………………………………………………………21 2.2.6.2 The Maternal– Fetal Bridge of Iron Deficiency ……………………………………21 2.2.6.3 Malaria ………………..………………………………………………..................23 2.2.6.4 Hookworm ……..…….……………………………………………………….………23 2.2.7 Some haematological parameters and their role in diagnosis of iron deficiency anaemia …...………………………………………………………………24 2.2.7.1 Haemoglobin Concentration …...……………………..…………..……………………..24 2.2.7.2 Serum ferritin concentration ……...…..……...……………………..……………………25 2.2.7.3 Total Iron Binding Capacity and Transferrin Saturation ………….……………………27 2.2.7.4 Serum Transferrin Receptor ……….…...………..………………........…………………..27 2.2.7.5 Serum Iron Concentration …..…….…………………………………………………….28 2.2.8 Prevention of Iron Deficiency ……….………………………………......………………28 2.2.8.1 Dietary Diversification ..……………..…...…………………………….……………….28 2.2.8.2 Food Fortification …….……………………..……………………………………….28 2.2.8.3 Supplementation ………………………………………….......…………………….29 2.3 Blanching …………..……..……………………………………………..……………....30 2.4Antinutrients ………………………………………………………………………..…30 2.4.1 Antinutrtional factors…………………………………………………………………….30 xi 2.4.2
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