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Bangladesh J. Sci. Ind. Res. 52(2), 153-158, 2017

Determination of vitamin A content from selected Nigerian fruits using spectropho-

tometric method

S. O. Aremu1* and C. C. Nweze2

1
Department of Biological Sciences, University of Agriculture, PMB 2373 Makurdi, Nigeria
2
Department of Biochemistry, Nasarawa State University, PMB 1022 Keffi, Nigeria

Abstract
The vitamin A content of the fresh fruits was determined by the extraction and quantitative determination of the pro-vitamin A carotenoid-
β-carotene. The Retinol Equivalent (RE) was then obtained using standard conversion formula. The result indicated that the fruit with the
minimum vitamin A content and percentage among the six samples was found in mango 301.61 ± 1.03 (7%), which was followed closely by
watermelon 350.12 ± 19.01 (8%), guava 504.1 ± 0.75 (11%), tomato 542.86 ± 20.20 (12%), pawpaw 683.93 ± 5.15 (16%) and the highest
content was found in carrot 2054.10 ± 1.28 (46%) in (RE/100 g). Statistical calculations and analysis showed that there was significant
difference in vitamin A content among the fruits (p<0.05). Thus, besides major nutrients fruits and vegetables contribute very good amount of
micro nutrient-vitamin A from β-carotene.
Keywords: Fruits; Vegetables; β-carotene; Vitamin A; Spectrophotometer

Introduction

Vitamin A is not a single compound but exists in several lycopene, lutein, beta cryptoxanthin,zeaxanthin and
forms: retinol (an alcohol), retinal (an aldehyde), retinoic astaxanthin. Carotenoids have extensive applications as
acid and other related compounds, all known as retinoids. It anti-oxidants in dietary supplements and as colours in foods
helps prevent diseases such as lung and breast cancer and beverages as well as pigments in poultry and fish. The
(Lacroix and Lippman, 1980; Prakash et al., 2000) and can carotenoids used as food ingredients include astaxanthin,
be used therapeutically in the treatment of retinitis beta-apocarotenal, canthaxanthin, beta-carotene, lutein,
pigmentosa, leukemia (Rego et al., 2000) and skin disorders zeaxanthin and lycopene. Carotenoids are important for
(Heller and Shiffman, 1985) such as psoriasis and acne. human health, but its structure ultimately determines its
β-carotene which is a pigment-carotenoid is precursor of potential biological function(s). The essential role of
vitamin A. Vitamin A is needed in the retina of the eye in the beta-carotene and others as the main dietary source of
form of a specific metabolite, the light absorbing retinal, vitamin A has been known for many years (Carlier et al.,
that is absolutely necessary for both low light (scotopic 1993). More recently, protective effects of carotenoids
vision) and colour vision (Tang et al., 2005). against serious disorders such as cancer (Donaldson, 2004;
Kantoff, 2006), heart disease (Lonn and Yusuf, 1999; Sesso
Vitamin A functions in different roles, as an irreversible et al., 2003) and degenerative eye disease(Mozaffarieh et
oxidized form of retinoic acid, which is an important al., 2003) have been recognized and have stimulated
hormone-like growth factor for epithelial and other cells intensive research into the role of carotenoids as
(Duester, 2008). antioxidants and as regulators of the immune response
Among the various classes of pigments in nature, the system. Carotenoids which are alpha and beta carotene are
carotenoids are among the most widespread and most the most common and important pro vitamin A carotenoid
important ones, especially due to their varied functions. because it can be represented as two connected retinal
These are fat-soluble pigments found mostly in plants, groups (Carolyn, 1997), gamma carotene and beta
fruits, flowers, algae and photosynthetic bacteria, but they cryptoxanthin.
also occur in some non-photosynthetic bacteria, yeasts and Despite the availability of fruits and other sources of
molds. The most abundant carotenoids in natural consumed vitamin A, World Health Organization (WHO, 2005)
foods are beta-carotene, alphacarotene, gamma-carotene,

* Corresponding author e-mail: [email protected]

154 Vitamin A content from selected Nigerian fruits 52(2) 2017

database on vitamin A deficiency (1995-2005) showed that course plenty of the fresh fruit, which can be eaten as it is or
approximately one third of children under the age of five in a form of juice (Sunita and Attar, 2008).
around the world is suffering from vitamin A deficiency and
it is estimated to claim the lives of 670,000 children under Guava (Psidium guajava) is a tropical fruit with pear shape
the age of five annually (Black et al., 2008). The highest and it is of two types the green and pinkish flesh with small
prevalence is in developing countries of which Southeast seeds inside. It belongs to the family of myrtaceae. And it
Asia and Africa continents are not spared. originates from America. They are usually eaten raw or
processed into juice (De boer and Cotingting, 2014).
Fruit is part of plant which is developed from a fertilized
ovary with one or two seeds (Mauseth, 2003). Most fruits Carrot (Daucus carota), they belong to the umbrelliferae
have been discovered to be functional foods because they family named after the umbrella like flower clusters plant.
are rich in micronutrients and other phytochemicals. Carrot roots have crunchy texture, sweet, minty aromatic
taste and are eaten raw in salads or processed into juice
Tomato (Solanum lycopersicum) is a savoury, red and edible (Ensminger and Esminger, 1986). The carrot can trace its
fruit. It originated from South America and spread around ancestry back to thousands of years originally having been
the world following the Spanish colonization of America cultivated in central Asia and Middle East. Recently there
and its many varieties are now widely grown, often in has been renewed interest in examining bio-availability
greenhouses in cooler climates. The tomato fruit is factors of vitamin A by using more quantitative stable
consumed in diverse ways, raw, as an ingredient in many isotope techniques for measuring whole-body stores in
dishes and sauces and in drinks (Burton-Freeman and Sesso, response to controlled intakes spread around the world
2014). Tomato is also used in diverse ways including raw in following some Spanish research work (Adams and Green,
salads and processed into purees, ketchups and tomato 1994; Furr, 1989; Haskell, 1998; Maron, 2014). Also by
soups. following post absorption carotenoids in the
triacylglycerol-rich lipoprotein fraction (Van and Van, 1998;
Water melon (Citrullis lanatus) is a member of the Castenmiller and West, 1998).
Cucurbitaceae family; its close relatives include pumpkin,
squash and cantaloupe that also grow on vines on the Thus, the present study aims at determining the pro vitamin
ground. Their shapes may be roundish, oblonged, or A carotenoid (beta carotene) content in selected Nigerian
spherical and it has feature of thick green rinds that are often fruits which include tomato, water melon, mango, pawpaw,
spotted or stripped with an internal deep red-pink colour pink guava and carrots using the spectrophotometric
when ripened. Their seeds are black, brown with a few method because it is the precursor of vitamin A (i.e. it can be
seedless variety. It has extremely high water content, converted into vitamin A) and it is eaten in both raw and
approximately 92%, giving its flesh a crumbly and subtly cooked form by humans in daily life (Bendich and Higdon,
crunchy texture and making it a favourite thirst-quenching 2004) and hence determine the vitamin A by getting the
fruit. retinol equivalent (RE/100g) of the beta carotene content of
these fruits by using the standard conversion formulae.
Mango (Magnifera indica) is a tropical tree cultivated in
many regions of India now distributed widely across the Materials and methods
Globe in many continents. Its Outer skin is smooth and is
green when unripe but turns into golden yellow or orange Sample collection and preparation
red when ripening depending on the cultivar. Internally, its The six different fresh ripe fruits were purchased from five
pulp is Juicy and yellowish in colour with numerous soft different zones called Groups in Keffi local government
fibrils radiating from the husk enveloping a single large area of Nasarawa state, Nigeria. Group I: Keffi market;
kidney - shaped seed. The flavour is pleasant and rich and Group II: Nasarawa State University, Keffi (NSUK) fruits
tastes sweet with mild tartness. shop; Group III: Dadinkowa market; Group IV: Kofa Hausa
Pawpaw (Carica papaya) has an oblong shape, normally market; Group V: Karu market. The ripe fruits included
greenish-yellow, yellow or orange in colour. It has sweet tomato (Lycopersum esculentum), pawpaw (Carica papaya),
taste when ripe, but tart or bland when unripe. It grows well watermelon (Citrullis lanastus), mango (Magnifera indica),
in tropical regions because of its higher humidity. It is native guava (Psidium guajava) and carrot (Daucus carota). Each
to Mexico, countries of Central America, Thailand, Africa paste of these six different samples was prepared from well
and Asia and grows well in Australia. The fruits, seeds, stem washed fruit with tap water in order to remove any dirt or
and leaves are all useful. One can buy pawpaw in all forms, adhering particles. Each fruit was peeled off to remove the
in a form of fermented papaya, herbal teas and tablets and of bark and the seeds removed. The flesh was then crushed
 Aremu and Nweze 155

using food blender (Binatone product) to get a fine pulp or Results and discussion
paste.
Table I shows that the fruits are of the same species and was
β-carotene extraction and analysis grown in similar environmental conditions since the beta
carotene content obtained were very close in range. The
The β-carotene was determined by soaking 1 g of the result in Table II showed that vitamin A content varied from
sample (that is the paste or pulp of the fresh fruits) in 5 ml relatively lower amount found in mango and water melon,
of methanol for 2 h at room temperature under dark when compared to the relatively higher amounts found in
condition in order to get a complete extraction. The β guava, tomato, pawpaw and carrot which have the maximum
-carotene layer was separated using hexane through content of beta-carotene among the fruits assayed. From the
separating funnel. The volume was made up to 10 ml with data, it was observed that all the fruits assayed contained
hexane and then this layer was again passed through sodium good sources of Vitamin A. This agreed with the literature
sulphonate through a funnel in order to remove any and other work done which showed that all the fruits assayed
moisture from the layer. The absorbance of the layer was are rich in beta carotene and hence vitamin A (Williams and
measured at 436 nm using hexane as a blank (Ranganna, Caliendo, 1984; Bereau and Bushway, 1986; Ahamad et al.,
1999). The beta carotene was calculated using the formula: 2007).

Calculation To express the vitamin A deviation or the sample error mean

(SEM) of the six fruit ranged from 1.28 to 20.20 while its
Beta-carotene (μg/100g) = Absorbance(436 nm) x V x D x variance or standard deviation (%) ranged from 0.06 to 5.43.
100 x 100/W x Y There may be slight variation in the data when compared to
other work done which may be due to difference in sources
where: V = Total volume of extract; D = Dilution factor; W
of vitamin A: experimental condition, extraction procedures,
= Sample weight; Y = Percentage dry matter content of the
method used. Variation in ecological growth conditions like
sample.
variety and environmental aspects may also be contributing
Vitamin A determination factors. For example; Comparatively lower contents were
reported in carrots (11210 μg/100g) and tomato (1610 μ
To express the vitamin A activity of carotenoids in diets on g/100g) by Ahmad et al. (2007) in the determination of beta
a common basis, a joint FAO/WHO Expert Group carotene and those reported in which beta carotene content
(FAO/WHO) in 1967 introduced the concept of the Retinol in carrot was 10110 μg/100g and tomato 1930 μg/100g. The
Equivalent (RE) and established the following relationships variation observed in the tomato content of beta carotene
among food sources of vitamin A: may be due to the use of overripe samples in their own study,
because content of beta carotene drops by 77% during the
1 μg retinol = 1 RE method used. Variation in ecological
ripening process (Rigo et al., 1999) and may also be due to
growth conditions
varietal difference among other factors.
1 μg β-carotene = 0.167 μg RE like variety and
Like wise, the beta carotene content of the fruits assayed also
environmental aspects may also be
corresponds to those reported by Robinson et al. (1986). They
1 μg other pro-vitamin A carotenoids = 0.084 μg RE analyzed summer fruits and vegetables for their micronutrient
contributing factors. content, though there was still variation in tomato. Patricia et
al. (2004) worked on new data of carotenoid in raw salad
These equivalencies were derived from balance studies to vegetables and they reported much lower content of beta
account for the less-efficient absorption of carotenoids carotene in carrot (5340 μg/100 g). This may be due to their
(thought to be about one-third that of retinol) and their sample of carrot having higher moisture content as compared
bioconversion to vitamin A (one-half for β -carotene and to those analyzed in the present study (Ahamad et al., 2007).
one-fourth for other pro-vitamin carotenoids). It was However, the report about beta carotene content in tomato
recognized at that time that the recommended conversion (3500 μg/100g) was in fair agreement with the result of the
factors (i.e., 1:6 for vitamin A: β-carotene and 1:12 for present study. But one thing is similar in all the work carried
out and worthy of note in the present study which is that carrot
vitamin A: all other pro-vitamin carotenoids) were only
is the richest and well known source of beta-carotene and
average estimates for a mixed diet. β-carotene and 1:12 for
hence vitamin A among the selected fruits in Nigeria
vitamin A: all other pro-vitamin carotenoids) were only (Lieberman and Bruning, 1990; Rigo et al., 1999; Patricia et
average estimates for a mixed diet. al., 2004). This was clearly shown in Fig. 1.
156 Vitamin A content from selected Nigerian fruits 52(2) 2017

Table I. Result of Beta carotene (µg/100 g) in the six fresh tropical fruits

Group I Group II Group Group Group V

Fruits Scientific name (µg) (µg) III (µg) IV (µg) (µg)
Guava Psidium guajava 3015.27 3012.94 3025.15 3021.86 3017.97
Pawpaw Carica papaya 4043.45 4139.96 4072.13 4100.90 4097.01
Water melon Citrullis lanastus 2148.05 2090.78 2156.89 2021.84 2156.14
Tomato Lycopersum esculentum 3294.91 3317.22 3317.22 2995.03 3321.26
Mango Magnifera indica 1797.21 1803.48 1812.84 1807.19 1809.49
Carrot Daucusa carota 12288.02 12311.98 12300.00 12300.60 12299.40

Table II. Vitamin A content (RE/100g), IU and standard deviation in percentage) of the six tropical fresh fruits
*Vitamin A content Standard Vitamin A content
Fruits Scientific name
(µg or RE) Deviation (%) (IU)
Guava P . guajava 504.10 ± 00.75 0.15 151.23
Pawpaw C . papaya 683.93 ± 05.15 0.75 206.98
Water melon C . lanastus 350.12 ± 19.01 5.43 105.04
Tomato L . esculentum 542.86 ± 20.20 3.72 162.86
Mango M . indica 301.61 ± 01.03 0.34 90.48
Carrot D. carota 2054.10 ± 01.28 0.06 616.20
*Vitamin A content (μg or RE) is in Mean±SD of five determinations

Guava Pawpaw Water melon Tomato Mango Carrot

11%

16%
46%

8%

12%
7%

Fig. 1. Pie Chart representation of the Vitamin A content (IU) of six selected fruits obtained in
Karu Local Government Area of Nasarawa State

A test of significance (t-test) was carried out on the mean of difference (p<0.05 that is the p-value was less than 0.05) in
the vitamin A content of the fruits using the computer Vitamin A content among the fruits.
software - Statistical Package for Social Sciences version
21.0 (SPSS 21.0) and it showed that there was significant
 Aremu and Nweze 157

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