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PLASMA
ASCORBIC ACID LEVELS OF PREGNANT WOMEN IN PORT HARCOURT
ABSTRACT
The values of
plasma ascorbic acid were determined in fifty blood samples collected from
pregnant women who were attending the antenatal clinic of the University of
Teaching Hospital, Port Harcourt, Nigeria. The mean plasma ascorbic acid level
was 0.767 ± 0.253mg/100mls, with a range of 0.420 – 1.585mg/100ml of plasma.
This is similar to the result got by Ibeziaka in 1981 working on forty pregnant
women at University College Hospital, Ibadan, Nigeria. The results showed no
correlation between the plasma ascorbic acid of these women and their body
weight.
TABLE OF CONTENTS
Title
Page - - - - - - - - - - i
Declaration - - - - - - - - - ii
Dedication - - - - - - - - - iii
Acknowledgement - - - - - - - - iv
Abstract - - - - - - - - - - v
Table
of Contents - - - - - - - - vi
CHAPTER
ONE: INTRODUCTION AND LITERATURE REVIEW
1.1
Introduction
1.2
Ascorbic Acid
1.3
Methods of Assessing Ascorbic Acid
Status in man
1.4
Ascorbic Acid Status of Human
CHAPTER
TWO: METHODOLOGY
2.1 Subjects
2.2 Collection of Blood
2.3 Analysis
CHAPTER THREE
RESULTS
CHAPTER FOUR: DISCUSSION AND
CONCLUSION
4.1 Discussion
4.1 Discussion
4.2 Conclusion
Bibliography
Appendix
Questionnaire
Formulae
for Mean and Standard Deviation Reading
A
typical Protocol
TABLES
AND FIGURES
Figure 1: Structure of Ascorbic Acid and Dehydroascorbic Acid
Tables:
I.
Plasma Ascorbic Acid of 50 Pregnant
Women in Port Harcourt
II.
Plasma Ascorbic Acid in the Second and
Third Trimesters of Pregnancy
III.
Comparism of Plasma Ascorbic with
Different Age Groups
IV.
Comparism of Plasma Ascorbic Acid with
Total Body Weight
V.
Plasma Ascorbic Acid Level of Pregnant
Women in Port Harcourt Compared with the Results of other workers
VI.
Different Levels of Plasma Ascorbic
Acid of the 50 Pregnant Women
CHAPTER ONE
INTRODUCTION AND LITERATURE
REVIEW
1.1 INTRODUCTION
Plasma
ascorbic acid is the amount of free ascorbic acid in the plasma. In other words
it is the non-cell bound or non-protein bound ascorbic acid in the blood it is
expressed in mg per 100ml of plasma. Plasma ascorbic acid level is a correct
index of body reserve and intake of the vitamin. This plasma varies in
different individuals depending on intake, metabolism, rate of excretion and
body needs of the vitamin. In the light of recent experiments, most workers
accept the normal Plasma ascorbic acid level to be within 0.40 – 1.6mg/100mls
(Dodds, 1980).
Javert
and Stander (1943), Martin (1957), Rivers (1971) and Baker (1975) reported a
low level of Plasma ascorbic acid in pregnant women and that this level further
decreases as pregnancy progresses. However, some investigators have reported no
significant change (Hoch and Marrack, 1948). Women with an intake above
80mg/day maintain essentially the same average plasma level throughout
pregnancy, while those with lower intake show progressively lower levels in each
trimester (Martin, 1957). This is probably due to transfer of ascorbic acid
from the maternal blood to the fetus.
At
plasma levels less than 0.20mg/100ml, clinical signs of scurvy occur (Hodges,
1978), although dietary induced scurvy is rarely reported in tropical
countries. However, low intakes contribute to morbidity especially when the
stress of pregnancy is combined with that of infections.
Therefore,
since it has been known that ascorbic acid is one of the nutrients most
commonly inadequate in the edicts of pregnant women (Rivers and Mason, 1971)
and the fact that there is tendency for plasma level to decrease in pregnancy
probably due to need by the developing fetus, the primary aim of this study was
to evaluate the Plasma ascorbic acid levels of pregnant women in Port Harcourt
and to compare these with the reports of other workers.
1.2 LITERATURE REVIEW
Albert
Szent – Gyorgi isolated vitamin c in 1932 and was awarded Nobel Prize for it. Haworth
(1933) discovered the molecular structure of L – ascorbic acid while Reichstein
(1933) synthesized it. Chemically, L – ascorbic acid is a white crystalline
compound with a molecular weight of 176 and melting point of 1920c. It
is a powerful reducing agent and this is the main role in the tissues. 
0
= c
O = C
HoC
O = C
HoC
O = C
HC HC
HOCH HOCH
CH2OH CH2OH
Ascorbic
Acid Dehyroascorbic
Acid
Fig
1: Structure of Ascorbic Acid and Dehydroascorbic Acid.
The physiological function of ascorbic
acid is still unclear. Probably the most clearly established functional role of
the vitamin is in maintaining the normal intercellular material play a part in
growth, wound healing, synthesis of polysacharrides, collagen and in the
maintenance of capillaries.
Deficiency of ascorbic acid in
susceptible species leads to scurvy which is characterized by systemic
lassitude and fatigue, loss of certain secretary functions, various hemorrhagic
manifestations, articular and skeletal abnormalities, neurologic impairment,
cardiovascular malfunctioning and in some instance antiiuresis causing edema.
Faulty protein synthesis explains poor wound healing (Wiltshire, 1919; Crandon,
1940).
Ascorbic acid in the diet is primarily
of plant origin. Although some animals can and do synthesize enough ascorbic
acid for their own needs. Man do not synthesize ascorbic acid although some
workers speculated synthesis of ascorbic acid in the breast and placenta
(Rajalakshihmi, 1965).
Dictary sources in African countries
such as Nigeria include edible vegetables as onions, water leaf, tomato,
pepper, fruits such as mangoes, oranges, canned fruit juices and other
miscellaneous sources as cassava, cocoyam (B. Reiff, 1959).
Daily allowances of ascorbic acid in
various nations vary from 30 – 120mg/day as official recommendations and some
private individuals advocate 2500mg/day or more Food and Agricultural
Organization (FAC) and World Health Organization (WHO) established lower
allowances for ascorbic acid because of the findings that clinical symptoms of
scurvy can be relieved by giving 10mg of ascorbic acid daily.
1.3 METHODS OF ASSESSING ASCORBIC
ACID: STATUS IN MAN
The
different methods for assaying ascorbic acid have been complicated by the need
to eliminate interference with the assay procedures by other substances. The
experimental techniques that have been developed can correctly be divided into
chemical methods. Radioactive tracer determinations, Enzymatic techniques,
Column, Paper and thin Layer separation techniques (Chromatography) Gas
chromatography. Another method which is non – scientific and rather crude is
assessment of incidence of scurvy.
CHEMICAL METHODS: Majority
of the numerous chemical methods are based on two procedures, the formation of
an osazone between dehydro ascorbic acid and 2, 4 – dinitrophanylhydrazine (Roe
and Kuether, 1943), or the reducing
potential of ascorbic acid which
can be measured with a number of indicators, most commonly 2, 6 - dichlorophenol indophenol (Bassey, 1938). The
hydrazine technique is relatively specific (Roe and Kuether, 1943), but
estimates not only the biologically active components of ascorbic but L –
dehydroascorbic acid, diketogulonic acid (Schaffert and Kingsley, 1955). On the
other hand indophend is only reduced by ascorbic acid and not by the more
oxidized metabolites of the vitamin, but is susceptible to other reducing
agents.
RADIOACTIVE TRACER DETERMINATION
This method involves the use of labeled radioactive
substance. This method can be used experimentally to measure metabolic body
pool but it is a complicated process and requires sophisticated equipment’s,
also it is not practical in evaluating individual patients.
ENZYMATIC TECHNIQUES: The
ideal measurement for any substance which is capable of influencing enzymatic
activity is a measure of its biological activity. Thus the functional activity of ascorbic acid
in the hydroxylation of Lysine and praline can be used to assess the vitamin
status; but this has been considered an insensitive method (Bates, 1979).
CHROMATOGRAPHY: This
could be paper or gas chromatography. In any case thin layer separation is
applied. This method is also not accurate. In this study, the2, 4 –
dinitrophenylhydrazine method method by Roe and Kuether (1943) was used plasma
has been chosen as the biological fluid because of the following reasoys:
Plasma method of assey is simple plasma ascorbic
acid status. In other words plasma ascorbic acid reflects tissue reserve well
for it to have been used to assess ascorbic acid status. In other plasma ascorbic
acid levels show a strong positive correlation with lencocytoe levels and
therefore are a good index of ascorbic acid status (Evans, Crrieand Compell,
1982).
Plasma assay requires less blood and
technically it is less difficult than lencocyte ascorbic acid measurement.
However, lencocytes ascorbic acid level is a better measure of tissue ascorbic
acid, although the lencocytes are contaminated with platelets. It is also known
that plasma concentration may fall too low to be measured while lencocyte level
may still be within normal range. Thus, lencocytes provide a measure of
availability of the vitmin for storage, while plasma level is a good index of
tissue content.
1.4 ASCORBIC ACID STATUS OF HUMANS
The
ascorbic acid reserves have been found to differ in different population. It
differs from countries to countries probably depending on intake. Even in the
same country there are differences within groups such as elderly, females,
pregnant women, children and lactating mothers. These different concentrations
are put up by different workers, in the light of recent experiments; most
workers accept normal range to be 0.50 – 1,6mg/100ml plasma.
In pregnancy, it is generally agreed
that plasma ascorbic acid decreases (Javert and Stander, 1943; Martin 1957;
Mason and Rivers, 1971; Baker, 1975), although some investigators have reported
no significant change (Hock and Marrack, 1948; Dawson, 1969). Women with an
intake greater than 80mg/day maintain essentially the same average plasma
ascorbic acid level throughout pregnancy, while those with lower intakes show
progressively lower levels in each trimester (Martin, 1975).
Urinary
excretion of a test of ascorbic acid is reduced during pregnancy and lactation
(Toverud, 1939). The studies indicating a drop in blood levels and decreased
urinary excretion of ascorbic acid during pregnancy help to establish the fact
that the developing fetus is parasitic on the mother in respect to ascorbic
acid
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