[Bioscience]

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ACKNOWLEDGEMENT

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DECLARATION

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TABLE OF CONTENT

ACKNOWLEDGEMENT2

DECLARATION3

ABSTRACT5

CHAPTER 017

1 Introduction7

2 Mineral Deficiencies8

3 Anaemia9

4 Solanum torvum11

5 Minerals19

6 Iron22

7 Ferritin35

Structure35

Metabolism35

In Human36

In Plants (Phyto-ferritin)37

8 Phenolic acids42

9 Techniques44

9.1 Digestion Methods46

9.2 ICP-MS and ICP-OES47

9.3 Electrophoresis51

9.4 High performance liquid chromatography (HPLC)51

9.5 Transmission Electron Microscopy (TEM)54

10 Aims and Objectives55

CHAPTER 257

Methods and Material57

Abstract

Iron-deficiency, anaemia, is a common nutritional disorder affecting people all over the world. Ferritin is an iron storage compound with a mass of 450 ka; it is found in plants (in plastids) & in animals (largely in the cytoplasm), which have been studied extensively in recent years in order to characterise the large protein of different species. The ferritin in plants has been utilised as a natural iron supplement in various countries for the treatment of iron-deficiency anaemia. In Asia, & Africa Solanum Torvum (swatz) has been utilised as an effective, herbal for treating iron-deficiency; though its ferritin has not been characterised, & so there is no evidence backing this up. The objective of this research was to conclude if Solanum Torvum (swatz) ferritin has the potential to aid mineral iron absorption in iron-deficient humans, as an alternative natural supplementation strategy. The study consists of a comprehensive examination of those Fe-containing compounds found in Solanum torvum. The goal of the study are to determine Fe content by Inductively Coupled Plasma-Atomic Emission Spectrometry (ICP-AES) & to separate iron containing compounds utilising high performance liquid chromatography (HPLC) techniques. The characterisation, separation, & identification of ferritin were addressed utilising mass spectroscopy (MS), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), & Electron microscopy (EM), & tryptic digest were also utilised, & the well-characterized horse spleen ferritin was analysed to ensure that all methods had worked. SDS-PAGE results showed 3 bands; 25, 26.5 & 28 kDa, of which the latter two were characteristic of soybean seed ferritin & pea seed ferritin. Matrix-assisted laser desorption ionisation-Time of flight spectrometry Mass (MALDI-TOF MS) of the extracted Solanum torvum ferritin after tryptic digest demonstrated all the three peptide masses. These results furnish a good start to the characterisation of Solanum torvum ferritin, & somewhat to back up the use of Solanum Torvum for iron-deficiency anaemia treatment, though further analysis is required to determine the full amino acid sequence.

Chapter 01

1 Introduction

Normal physiological functions are maintained when an adequate simply of minerals is available, which in most instances, all the necessary essential minerals, & other dietary requirements could be obtained in a balanced diet (Agunod 2009 400). Though, research has revealed that it is often the case that food group's for instance fresh vegetables & fruits, while being rich ...