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ABSTRACT

The effect of Ethylenedianinetetraacetic acid (EDTA) on lead
content of blood has not been worked on in vitro. This led to
carrying out this research. The aims of this research is to ascertain
if EDTA has any effect on blood lead levels if the sample is collected
using an EDTA tube and also to find out if there is any significant
difference in the blood lead levels of the same sample when
collected with an EDTA tube and a non-EDTA tube. Hundred (100)
males who are commercial painters and who have painted for
more than 5 years were recruited for the research. Fifty (50) males
who had nothing to do with lead and lead products were also
recruited to serve as the control group. The blood lead levels of the
test group were significantly higher (p < 0.05) than that of the
control group. The haematological indices were significantly higher
(p < 0.05) in the blood in plainon-EDTA tubes than that in the tubes
containing EDTA. In the haemoglobin concentration, there was a
significant difference (p < 0.05) between that of the fresh blood
and the sequestered blood. The lead concentration was
significantly higher (p < 0.05) in the native blood than the
sequestered blood. From the investigations the EDTA in the tubes
had an effect on the lead already in the blood and the effect was
noticed on the haematological parameters.

TABLE OF CONTENTS

Title page …………………………………………………………………………. i
Approval ………………………………………………………………………….. ii
Certification …………………………………………………………………….. iii
Dedication ………………………………………………………………………. iv
Acknowledgements …………………………………………………………. v
Table of content ……………………………………………………………… vi
List of table’s ………………………………………………………………….. ix
List of figures …………………………………………………………………. x
Abstract ………………………………………………………………………… xi
Chapter One: Introduction ……………………………………………… 1
1.2 Justification ……………………………………………………………………. 3
1.3 Aims and objectives……………………………………………………… 4
Chapter Two: Literature review ……………………………………….. 5
2.1 Occupational lead exposure …………………………………………… 5
2.2 Mechanisms of toxicity ……………………………………………….. 8
2.3 Effects of lead on the heam biosynthesis and erythropoiesis 10
2.4 Effects of lead on the nervous system ……………………………. 15
2.5 Toxicity of other body organs ……………………………………….. 20
2.6 Ethylenediamine tetraacetic acid (EDTA) ………………………… 23
2.7 The EDTA molecule …………………………………………………………. 23
2.71 Uses of EDTA …………………………………………………………… 25
2.72 EDTA as a chelating agent ……………………………………….. 26
2.721 History of EDTA as a chelating agent ……………………….. 27
2.722 Uses ………………………………………………………………………. 29
2.723 Dosage …………………………………………………………………… 29
Chapter Three: Materials and methods …………………………… 31
3.1 Sample collection ………………………………………………….. 31
3.2 Method …………………………………………………………………….. 32
3.21 Packed Cell Volume ……………………………………………….. 32
3.22 Erythrocyte Sedimentation Rate …………………………….. 33
3.23 Total White Cell Count ………………………………………….. 34
3.24 Differential White Cell Count …………………………………. 35
3.25 Haemoglobin Concentration …………………………………. 36
3.26 Lead Assay ……………………………………………………………… 36
3.5 Analysis ……………………………………………………………………. 37
3.6 Statistical analysis …………………………………………………….. 37
Chapter Four: Results …………………………………………………. 38
Chapter Five: Discussion and Conclusion …………………………….. 39
5.1 Discussion ……………………………………………………………………….. 41
5.2 Conclusion ……………………………………………………………………….. 44
5.3 Recommendation ……………………………………………………………. 45
References ………………………………………………………………………….. 46
Appendix 1 ………………………………………………………………………… 57
Appendix 2 …………………………………………………………………………. 61
Appendix 3 ………………………………………………………………………… 67

CHAPTER ONE

1.1 INTRODUCTION
The heavy metal lead is ubiquitous, and its deposits are scattered
throughout the world. “The most common ore is galena” (lead
sulphide-87% lead). “It has been mined and disseminated throughout
the environment from where it has gradually become incorporated into
the structural tissue of plants, animals and humans” (Pracheta et al.,
2009). Lead is a soft, silvery gray metal, which rapidly becomes dull in
air, and is soft, malleable and ductile, melting at 327.5 °C. It is highly
resistant to Corrosion, but is soluble in nitric and hot sulphuric acids.
Lead (Pb), with atomic number of 82 and atomic weight of 207.19 has a
specific gravity of 11.34. It has four naturally occurring isotopes with
atomic weights 208, 206, 207, and 204, in decreasing order of
abundance. “The usual valence state in inorganic lead compounds is +
2” (Lead, 2001).
The common occurrences of lead include naturally occurring
ores. Lead ores comprise 0.002% (15g/t) of the earth’s crust. They
include galena (lead sulfide), anglesite (lead sulfate), cerussite (lead
carbonate), mimetite (lead chloroarsenate) and pyromorphite (lead
chlorophosphate).
1.11 INORGANIC LEAD
This is the form of lead found in old paint, soil, dust and various
consumer products. The color varies, depending on the chemical form,
and the most common forms are white lead (a lead carbonate
compound), yellow lead (lead chromate, lead monoxide) or red lead
(lead tetra oxide). Lead acetate has a sweetish taste.
1.12 ORGANIC LEAD
Tetra-ethyl lead is the form of lead used in leaded gasoline.
Organic forms of lead are extremely dangerous, as they are absorbed
through the skin and are highly toxic to the brain and central nervous
system, much more so than inorganic lead. The combustion of organic
lead – when it is added to petrol as a fuel additive – results in the
release of lead into the atmosphere.
Lead is a metal of antiquity and has been used for many
purposes for thousands of years. A variety of forms of lead are used
industrially, the most common being lead oxide (PbO) and red lead
oxide (lead tetra oxide; Pb3O4). “Lead oxides are used in the plates of
electric batteries and accumulators and in other substances” (Stellman
and Osinsky, 1997). About 40 to 50% of all lead production is used to
make lead-acid batteries (both lead metal and lead oxides are used). A
further 20% of production is used as the metal, for example, cable
sheathing, solder, ammunition, alloys, weights, ballast, and low-melting
alloys. “Lead is a major component of many alloys such as bronze and
solders” (Massaro, 1997). It is used for tank linings, piping, and building
construction. Lead-based pigments, such as white lead, have a long
tradition of being used in paints, although they have virtually all been
substituted in many countries by other pigments in the last 20 years for
obvious toxicological reasons. Lead compounds are used in glassware
and ceramics and as stabilizer in plastics. Red lead is used to make
television tubes. “About 10% is converted into alkyl-lead compounds
and used as antiknock additives in gasoline, although this use is in
decline as more and more countries move to limit the concentration of
such additives in gasoline” (Massaro, 1997). Some of the uses for lead
in ancient times (such as lead sheet for lining roofs) remain today.
“Even though worldwide lead exposure has decreased
tremendously in the past decades” (Pirkle, 1994; Hwang, 2004; Gulson,
2006), “lead is still ubiquitous and often found in flaking lead-based
paints” (Mathee, 2009), candies (CDC 2002; Medlin 2004), cosmetics
(Al-Ashban, 2004), traditional lead-glazed ceramics used in the
preparation and serving of food (Sabouraud, 2009), herbal remedies
(Obi, 2006), canned foods and beverages (lead solder) (Maduabuchi,
Nzegwu et al. 2006), toys (Greenway and Gerstenberger 2010), and
“can affect communities residing near smelters” (Hegde, 2010) and
“battery recycling plants” (Fuller 2009).
1.2 JUSTIFICATION
This research was designed to investigate the effect of
anticoagulant on lead activity in the blood. Bearing in mind that
one of the treatment for lead toxicity is by administering a drug
edentate disodium calcium intramuscularly, of which part of which
is an anticoagulant. It becomes necessary to carry out this
research in order to determine if collecting blood samples in
anticoagulant for lead investigation inhibits the activity of the
lead already in the blood.
1.3 AIMS AND OBJECTIVES.
The aims of the research are to investigate
1. Blood levels of lead and its effect on hematological
parameters among workers that work with lead in Enugu.
2. To determine by comparison if the anticoagulant has any
effect on the lead activity of the lead already in the blood.
3. To determine whether any effect is significant or not.

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