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Food fortification is an important tool for improving the health of the population. Motivated by this, the
National Agency for Food and Drug Administration and Control (NAFDAC) has in the past few years
embarked on a campaign to create this awareness. There has been varying degree of compliance to
this, and so it is of utmost importance to know the food products that contribute significantly to the
dietary vitamin B intake. This study investigates the amount of four Vitamin B compounds, namely
nicotinamide, pyridoxine, riboflavin and thiamin, present in different packaged food items available in
Lagos metropolis of Nigeria. The food items analyzed include dairy products, fruit juices and cereal
products. These foods were chosen because of their widespread consumption in the area. The analysis
was done using a high performance liquid chromatographic technique with a UV detector. The
separation was carried out on a C18 column, using a mobile phase made up of 70% buffer (a solution of
sodium salt of hexane sulphonic acid) and 30% HPLC grade methanol. The identification was based on
retention time match against a standard while the quantitation was based on the peak area match
against those of a standard. The calibration curves for the standards were linear with a linear
regression coefficient close to unity.
INTRODUCTION
Vitamins in general, play a very important role in our Deficiency diseases (McCormick, 1988a, b, c; 1994;
health, even though they only make up a very small part Carpenter and Lewin, 1985; FAO/WHO, 1974) do not
of the food we eat each day (Olaniyi, 2000). Diets which only occur in poverty stricken communities. In affluent
do not contain adequate amounts of these vitamins often societies, deficiency diseases also occur often as the
result in deficiency diseases including blindness and result of poor choice of food or unhealthy eating habits,
mental retardation, depending on the particular vitamin. often coupled with different lifestyles. It is estimated that
For example, thiamine functions as a co-enzyme in the one in every three people in the world is at risk for one or
phosphogluconate pathway and a structural component more micronutrient deficiencies, thereby impacting on
of nervous system membranes (McCormick, 1996; 1997). their nutritional status. Our nutritional status has an
Nicotinamide is essential for the metabolism of carbo-hy- important impact on our health, productivity and quality of
drates and for non redox adenosine diphosphate-ribose life in general. It is also commonly thought that women of
transfer reactions involved in DNA repair (Berger, 1985; childbearing age, young children, the elderly, and those
McCormick, 1996; 1997). Pyridoxine plays an essential recovering from illness are most at risk of developing
role in amino acid transamination (McCormick, 1996; micronutrient deficiencies.
1997). Riboflavin functions as a co-enzyme for a wide va- In order to prevent and address the problem of micro-
riety of respiratory enzymes (McCormick, 1996) etc. nutrient deficiencies, several strategies are used, one of
which is food fortification in which specific and scienti-
fically identified micronutrients are added to certain food
vehicles. The type and amount of micronutrients added
*Corresponding author. E-mail: chimezie@email.com. are determined by the nutritional status, and therefore
030 Afr. J. Pharm. Pharmacol.
nutritional needs, of the population while the food to be deionization was carried out by means of a Millipore deionizer. A
fortified (food vehicle) depend on the eating habits of the buffer solution comprising of hexane sulphonic acid sodium salt
(BDH, England), deionized water, and glacial acetic acid (M and B,
population. England) was used in sample preparation and separation of the
The increasing interest in good eating habits has vitamins. The buffer solution was prepared by weighing 1.8822g of
provided a greater awareness of the vital role that micro- hexane sulphonic acid sodium salt into a 2l volumetric flask and
nutrients play in growth and health. The National Agency dissolved with a minimum amount of deionized water. This was
of Food and Drug Administration and Control (NAFDAC), followed by the addition of 1500 ml of deionized water to the
in conjunction with the micronurient Initiative (MI) has solution; 20 ml of glacial acetic acid was then added to keep the
medium acidic, and finally making up with deionized water, which
contributed immensely to the creation of this awareness helped to dissolve the water soluble vitamins. The mobile phase,
in Nigeria. However, since there is still varying degree of which comprised of methanol (BDH England) and buffer (made of
compliance to this, it is therefore of utmost importance to hexane sulphonic salt of sodium), was properly sonicated prior to
know the food products that contribute significantly to the analysis. The food samples were obtained from a local market in
dietary micronutrient intake. In the present work, the Lagos metropolis.
determination of dietary water-soluble vitamins namely
nicotinamide, pyridoxine, riboflavin, and thiamin (Figure Chromatographic conditions
1) in selected food products in Lagos metropolis of Nige-
Chromatographic separation and analysis of the vitamins were
ria is carried out using high performance liquid chromato- done using a reversed-phase Nova-Pack C18 (250 x 4.6 mm, 5 µm)
graphic method (HPLC). The food items analyzed include column (Waters,) at room temperature. The mobile phase composi-
dairy products, fruit juices, and cereal product, which tion used was 70% buffer (that is, sodium salt of hexane sulphonic
were chosen because of their widespread consumption in acid) and 30% methanol (HPLC grade). The analysis was carried
the area. The results show a relatively high level of these out in isocratic mode at a flow rate of 1ml/min, with column effluent
being monitored at 254 nm wavelength.
compounds in the analyzed samples.
Table 1. Analytical characteristics of standards. phonic acid, glacial acetic acid and deionized water). For
the solid samples, the samples were dissolved in buffer
2
Compound Mol. Wt. tr r solution and by vigorous shaking. Hexane sulphonic acid
Nicotinamide 122.1 3.86 0.995 in the buffer served as a deproteinating agent, removing
Pyridoxine 205.6 6.34 0.996 the bulky proteins in the food, hence fostering the proper
Riboflavin 376.4 11.56 0.995 dissolution of the vitamins. The solution was filtered and
Thiamin 337.3 17.98 0.982 the filtrate diluted to mark. The target com-pounds are all
water soluble hence also soluble in the buffer.
900000
2
800000 R = 0.9947
700000
600000
500000
area count
400000
300000
200000
100000
0
0 0.05 0.1 0.15 0.2 0.25 0.3
conc. [in mg/ml]
2500000
2
R = 0.9962
2000000
1500000
area count
1000000
500000
0
0 0.05 0.1 0.15 0.2 0.25 0.3
conc. [in mg/ml]
1000000
900000 2
R = 0.9948
800000
700000
600000
area count
500000
400000
300000
200000
100000
0
0 0.05 0.1 0.15 0.2 0.25
conc. [in mg/ml]
1600000
2
R = 0.9819
1400000
1200000
1000000
area count
800000
600000
400000
200000
0
0 0.05 0.1 0.15 0.2 0.25
conc. [in mg/ml]
NH2
N HO CH3
O
H3C N
NH H3C N S OH
1 H3C N
+
H3C N O N N -
Cl
(CHOH)
4
HOH
2C NH2 CH3
2
3
Figure 6. Standard vitamins. Nicotinamide (3.857 min), pyridoxine (6.345 min), riboflavin (11.563 min) and thiamin (17.9781min)
respectively analyzed at different concentrations employing the chromatographic conditions explained above.
compounds (FAO/WHO, 2002). Figure 8 compares the quate for complementing other sources of these com-
results from this study with the World Health Organi- pounds in the body.
zation’s recommended daily intake. Even though only the We can conclude that even though the food fortification
estimated average daily intake for Riboflavin andpyri- campaign is working, a more precise recommendation
doxine fell above the minimum requirement, some of the should be pursued to stop having such a huge variation
samples individually considered fell far above the recom- as discovered in this study. For instance nicotinamide in
mendation but quite a number fell below this recom- 12 samples taken at random ranged from 0 ug/ml to 854.05
mended daily intake. Nevertheless we can still say that ug/ml with a mean value of 182.22 µg/ml. Pyridoxine has a
the effort of the agency is working since these food sam- mean value of 51.17 µg/ml but ranged from 0 to 189.659
ples taken are not the only sources of these compounds’ µg/ml. Riboflavin with a mean value of 217.56 µg/ml ranged
intake by this population. They just serve as a supple- from 0 to 1433.055. Thiamine which ap-peared generally in
ment to traditional sources of vitamins B’s. Therefore the lowest concentration ranged from 0 to 121.055 µg/ml with a
concentrations in the analyzed samples are very ade- mean value of 13.33 µg/ml.
Anyakora et al. 035
1600
1400
1200
1000
Conc(ug/ml)
Nicotinamide
Pyridoxine
800
Riboflavin
Thiamin
600
400
200
0
1 2 3 4 5 6 7 8 9 10 11 12
Samples
25
Sample WHO recommended
20
15
mg
10
0
Nicotinamide Pyridoxine Riboflavin Thiamine
Figure 8. Comparison of the target compounds detected in the samples analyzed with the
WHO recommendation [assuming 50 ml (g) daily consumption
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