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PLASMA ASCORBIC ACID LEVELS OF PREGNANT WOMEN IN PORT HARCOURT




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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
4.2   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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