CHAPTER ONE

INTRODUCTION
1.1 Background
Nigeria is blessed with rich repertoire of plant foods. In this family are fruits of different types and value,

often consumed for their nutrients or for their medicinal purposes. Fruits are capable of contributing

appreciable quantities of nutrients, including protein, needed by both children and adults if properly

processed (Franke et al., 2005)

Over the years, it has been shown that fruits possess vitamins and antioxidants of great nutritional and

therapeutic values (Oyeniran, 2015). These antioxidants help in lowering incidence of degenerative

diseases such as cancer, arthritis, arteriosclerosis, heart disease, inflammation, brain dysfunction and

acceleration of the ageing process (Kaur and Kapoor, 2001; Wilcox et al., 2004; Laguerre et al.,2007).

Antioxidants are able to quench reactive free radicals, such as hydrogen peroxide, superoxide radical,and

hydroxyl radical. Thus, they can prevent or slow down body cell damage caused by free radicals (Liu et

al., 2018). In fact, date fruits for example, have high antioxidant properties due to their high content of

vitamins such as ascorbic acid and tocopherols, and phytochemicals such as polyphenols and carotenoids

(Al-Farsi et al., 2018; Al-Shwyeh, 2019; Hussain et al., 2020). Phenolic compounds exhibit an extremely

high antioxidant activity (Maqsood et al., 2020).

It is known that natural sources of antioxidants such as fruits are more advantageous to health than the

synthetic counterparts/supplements (Leong and Shui, 2002; Liu, 2003; Lim et al., 2007). Information

abound in literature on the vitamins antioxidant properties of many fruits and this promote their use

internationally as components of functional foods to promote health. Such information is dearth on most

indigenous Nigerian fruits and this limits their use for medicinal purposes. It is therefore necessary to

evaluate the vitamins and antioxidant properties of some selected fruits in Lafia, Nasarawa state, Nigeria,

1
namely: Orange (Citrus sinensis), Banana (Musa paradisiaca), and Apple (Malus pumila) to diversify

their use as nutritional and medicinal purpose that contain active ingredients which promote health and

reduce the risk of disease.

1.2 Problem Statement

Currently, evidence on characteristics of scientific research conducted to determine antioxidant content in

fruits and their research design consistency is still not widely known, especially in the area of study.

Therefore, the current research seek to add to few existing literatures that were carried out to determine

antioxidant content in fruits and to make future recommendations to promote higher research design

consistency and, thus, improve research quality in this research area.

1.3 Aim of the Research

The aim of this research was to determine vitamins and antioxidant properties from selected fruits in lafia

metropolis, Nasarawa State, Nigeria.

1.4 Objectives of study

The objective of this study was to ;

i. determine vitamins and antioxidant properties from Orange (Citrus sinensis), Banana (Musa

paradisiaca), Apple (Malus pumila)

1.5 Significance of study

There is no enough study carriedout on the vitamins and antioxidant properties of fruits in Lafia Local

Government Area of Nasarawa State, Nigeria. This study will therefore be used as a starting point for

further studies about the content of vitamins in fruits and the health benefit of fruits as the study is

intended to determine vitamins and antioxidant properties in selected fruits in Lafia market. The

2
generated data and information can be used as a baseline information for future researches on the

investigation of the content of vitamins and the antioxidant behavior of common fruits in Lafia market.

1.6 Scope of the study

The study is limited to the township market (modern market) in Lafia metropolis Nasarawa state. The

research is centered on determining vitamins and antioxidant properties from three types of fruits sold in

the market. These fruits include; Orange (Citrus sinensis), Banana (Musa paradisiaca), and Apple (Malus

pumila).

3
 CHAPTER TWO
LITERATURE REVIEW
2.1 Fruits phytochemicals and Human health

There is an array of evidence that consumption of fruits is important for human health as they are richly

endowed with health-improving nutrients (Radovich, 2011). Burton-Freema (2010) postulated that

antioxidant-rich fruits increases the antioxidant capacity of the blood, thus decreasing the risk of

developing diseases such as cancer, diarrhea, coronary atherosclerosis, and gastrointestinal tract diseases.

Results from epidemiological and laboratory studies conducted by Eckert (2008), Harding et al., (2008)

and Rouanet (2010) supported the hypothesis that consumption of fruits will prevent and significantly

reduce cancer, diabetes, and heart diseases.

Further epidemiological studies have proposed an inverse correlation between high intake of fruits and

many degenerative and aging diseases (Ares et al., 2009). Improvement in diabetes mellitus, digestive

problems, immune disorder, cataract, bronchitis, asthma, and other respiratory syndromes have all been

reported upon regular intake of fruits. Butt and Sultan (Butt and Sultan, 2011) also suggested that humans

relying on fruits have a 10–15% lower risk of developing cataracts than those who consumed lower

proportions of fruits (Jaganath and Crosier, 2008) indicated that a proper balance between oxidants and

antioxidants is necessary to maintain health as alterations to this balance often leads to pathological

responses that result in functional disorders and diseases. The health functions of fruits are therefore

based on their phytochemical content, which in their various chemical forms, possess antioxidative,

antiviral, anti- inflammatory, antimicrobial, anti-carcinogenic, antiangiogenic, antibiotic, and

antithrombotic properties (Hounsome and Hounsome, 2011)

Baskar et al., (2011) emonstrated that regularly, free radicals are synthesized in the body either naturally

or through external factors such as environmental stress, thus resulting in various degenerative diseases

4
that harm the body. Though the body has an inbuilt defense mechanism to scavenge these radicals, there

is a need for the utilization of additional antioxidants available in fruits.

Figure 1. Chemical structure of carotene.

Dietary antioxidants are defined as food compounds that significantly reduce the deleterious effects of

ROS, RNS, or both, on the normal physiological function in humans (Wootton-Beard and Ryan 2011).

ROS (oxygen ions, free radicals, and peroxides) and RNS (nitrous anhydride, peroxynitrite, and nitrogen

dioxide radicals) cause oxidation, nitration, halogenation, and deamination of biomolecules of all types,

including lipids, proteins, carbohydrates, and nucleic acids, with the formation of toxic and mutagenic

products (Castrol and Freeman, 2001) Pathogenesis of non-communicable and non-nutritionally related

human diseases such as brain stroke, diabetes mellitus, rheumatoid arthritis, Parkinson’s disease,

Alzheimer’s disease and cancer have been associated with the oxidative damage of these oxidants to cells

of which fruits phytochemicals are counter-intuitive. Examples include lycopene: the principal carotenoid

in tomatoes, a very efficient quencher of singlet oxygen in the biological system. Another example is

5
vitamin E, a major lipid-soluble chain-breaking antioxidant in humans, that protects the DNA, low-

density lipoproteins, and polyunsaturated fatty acids from free radical- induced oxidation (Jaganath et al.,

2006).

2.1.1 Vitamin C

Bruno et al., (2006) stated that the roles of vitamin C include the regulation of cell growth, cell signaling,

apoptosis, antioxidants, and as cofactors for enzymes. Vitamin C occurs mainly in fruits and it is reduced

by heat during processing; hence, its nutrient density in raw fruits is higher than in processed forms. [114]

Vitamin C scavenges reactive oxygen species (ROS) and reactive nitrogen species (RNS) and also

regenerates α-tocopherol and coenzyme Q from α-tocopherol and coenzyme Q radical. This resultant

action helps in maintaining the antioxidant activities of α-tocopherol and coenzyme Q (Li, 2011). The

studies by Lee et al., (2001) and Chen et al., (2008) postulated that ascorbate induces lipid hydroperoxide

decomposition to genotoxins in the absence of redox-active metal ions and also leads to a reduction in the

growth of aggressive tumor xenografts. As documented in studies conducted on animal species,

consumption of FAV rich in vitamin C will greatly help protect the body against cardiovascular disorders,

gastrointestinal disorders, cancer, skin infections, and diabetes through the reduction of insulin glycation

and an increase in glucose homeostasis (Abdel-Wahab et al., 2002)

2.1.2 Vitamin E

Consisting of eight different types: α-, β-, γ- and δ-tocopherols and the α-, β-, γ- and δ-tocotrienols,

vitamin E can be obtained from vegetable oils, nuts, and seeds of different fruits (Lee et al., 2001).

Experimental model studies in vitro and in vivo have shown the antioxidative, pro-antioxidative, anti-

inflammatory, modulation of cell signaling, antiproliferation, antiangiogenesis, and apoptosis induction

effects of vitamin E. Other works have also shown that α-tocopherol is the major form in human tissues.

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Accounting for more than 90% of the literature on vitamin E studies, α-tocopherol is the most studied

vitamin E isoform (Sen et al., 2007) α-tocopherols, and other forms of vitamin E play crucial roles in

protection against lipid peroxidation, scavenging of peroxyl radicals, reaction with ROS and RNS and

reduction in the synthesis of ROS from NADPH oxidase (Traber and Atkinson, 2007). The pro-oxidant

effect of α- tocopherol also includes its ability to reduce redox-active metals such as copper and iron.

When present in the human skin, vitamin E serves as a vital line of dermal antioxidant protection. Its

isoforms such as tocopherols and tocotrienols can protect the skin against disease conditions such as

dermatitis, UV-irradiation induced skin injury, and chemically induced oxidative stress in animal models

(McVean and Liebler, 1997). Thus, vitamin E obtained from fruits has found application as cosmetics due

to its protective dermal effects (Sen et al., 2007)

2.1.3 Carotenoids

Carotenoids consist of a group of lipophilic pigmented compounds that is made up of over six hundred

fat-soluble plant pigments. They are chiefly responsible for colors such as yellow, red, and orange present

in FAV and from which the compounds are derived. Major carotenoids present in human diets are α-, β-

carotene, lutein, lycopene, zeaxanthin, astaxanthin, and β-cryptoxanthin, with these compounds playing

an active role in the protection of plants from the damaging and scourging effects of exposure to sunlight

(Li, 2011). Carotenoids function in the body as a precursor of vitamin A, thus preventing vitamin A

deficiency in the body. Carotenoids further undertake antioxidant activities by scavenging reactive

species, oxides, and radicals, suppressing inflammatory responses in both in vivo and in vitro systems, as

well as assisting in the modulation of cell signaling and induction of apoptosis (Stahl and Sies, 2005).

This is apart from their roles in cardiovascular protective activities, obesity, cancer, and gastrointestinal

disorders (Nishino et al., 2009).

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2.2 Antioxidant-rich fruits

Orange, plum, pineapple, lemon, date, banana, apple, and grapefruit have been identified with high

antioxidant properties (Berrino, 2008). Other fruits associated with a high amount of antioxidants include

dog rose, sour cherry, blackberry, strawberry, raspberry, cloudberry, and rowanberry. Among these fruits,

berries account for the highest antioxidant content and dog rose has the highest compared to others such

as crowberry, wild berry, black currant, sour cherry, wild blackberry, wild strawberry, cultivated

blackberry, and cowberry/cranberry. Berries have a high content of phytochemicals such as flavonoids,

tannins, stilbenoids, phenolic acids, and lignans (Amakura et al., 2000)

Table 1. Antioxidant contents of some fruits in mg/100 g fresh food.

Fruits Flavanols Β- Lycopene Anthocyanins Flavan-3-ols Proanthocyanidins

carotene
Apple 2.0–26.0 - - 1.8–17.0 2.3–7.3 7.5–141.0
Avocado - - - - 0.1 0.02–7.4
Banana 25.2–58.8 - - - 0.1 4.0
Mango - - - - - 12.8
a
Orange - 173211 - - - -
Papaya - 471a - - - -
Pear 0.3–4.5 - - - 0.9 1.1–42.3
Pineapple - - - 11.6 c - -
a
Watermelon - 616– 11,378 - - -
1040a
a
1 g/serving of edible portion; b µg/100 g dry basis; c mg/100 g dry basis.
Source: Da Silva et al., (2014)
Table 1 shows the antioxidant contents of fruits in mg/100 g of fresh food. The citrus family consists of

fruits that are made up of high nutritional and antioxidant properties. Rich in ascorbic acid, citrus fruits

8
also contain flavanone glycosides (hesperidin, narirutin, and naringenin), limonoids, flavones (sinensetin

and nobiletin), and phenylpropanoids such as hydrocinnamates (Da Silva et al., 2014). Lako et al., (2007)

who investigated the phytochemical flavonols, carotenoids, and the antioxidant properties of a wide

selection of fruit, vegetables, and other readily available foods, showed that green leafy vegetables had

the highest antioxidant capacity followed by fruits and root crops. Anyasi et al., (2015) also reported that

banana (‘Muomva-red cultivar’) contains a high content of total polyphenols which is an indication that

the banana could be a source of bio-nutrients with great medicinal and health functions.

2.3 Determination of the antioxidant capacity of fruits

The concentration of antioxidants in fruits varies in their degree of activity and mode of action, hence the

difficulty in their analysis. Various methods exist for the determination of the antioxidant capacity of

fruits (Lako et al., 2007). However, there are known factors that affect the determination of the

antioxidant capacity in fruits and such factors include geographical location, agricultural and farming

practices, the season of cultivation, particle size, amount of extraction steps, sample to solvent ratio, and

temperature of the process (Floegel, 2011). Sample preparation is therefore a critical step in the total

determination of antioxidants in fruits. Established protocols for the analysis and determination of

antioxidant capacity in fruits include oxygen radical absorbance capacity (ORAC), Trolox equivalent

antioxidant capacity (TEAC), 2,2-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS), 1,1-diphenyl-

2-picrylhydrazyl (DPPH), ferric reducing ability of plasma (FRAP), and vitamin C equivalent antioxidant

capacity (VCEAC). The ORAC method is one of the most widely used methods for evaluating

antioxidant capacity (AOC) due to known and unknown antioxidants present in tested foods (Haytowitz,

2010). Peroxyl radical-scavenging activities of water-soluble and lipid-soluble antioxidants in samples

are evaluated by the hydrophilic ORAC (H-ORAC) and lipophilic ORAC (L-ORAC) methods,

respectively.

9
2.4 Review of empirical Studies

Lim et al., (2006) in their study titled “Antioxidant Properties of Guava Fruit” analysed two varieties of

guava fruits for total phenol contents, ascorbic acid contents and antioxidant activities. The antioxidant

activities were assessed based on the ability of the fruit extracts in 50% ethanol to scavenge DPPH,

reduce Fe(III) to Fe(II) and to bind to Fe(II) ion. The results were compared to several other local fruits as

well as orange. It was found that the guava fruit contains relatively high amounts of antioxidant. It also

has high primary, but low secondary antioxidant potential. Storage at 4 oC has the effect of increasing

ascorbic acid content, and the non-peeled fruit has higher total phenol and ascorbic acid contents

compared to the peeled fruit. The length and width of the seedy guava were also monitored over a period

of 17 weeks to define specific stages of fruit ripening.

Similarly, Andargie et al., (2023) determined the contents of ascorbic acid and the antioxidant activity of

eight types of common fruits commercially available in Addis Ababa, Ethiopia using iodometric titration

and UV-Vis spectrophotometric methods, respectively. The contents of ascorbic acid were found in the

range 15.41 mg/100 g in grapes to 78.01 mg/100 g in papaya. The antioxidant activity was found highest

in papaya (89.41) and lowest in water melon (31.28) in mg of ascorbic acid equivalent per 100 g of the

edible part of fruit juice. The calibration curve of ascorbic acid was in the range 50 - 3.125 mg/L with R2

= 0.9994. The results also revealed the strong correlation (r = 0.863) between ascorbic acid contents in

fruits and their antioxidant activity. Using the optimized conditions, UV-Vis spectrophotometric method

with the 2,2-diphenyl-1-picrylhydrazil assay allows determination of antioxidant activity of fruits with

limit of detection 0.32 mg/L and limit of quantification 0.96 mg/L. The precision was in the acceptable

10
range (relative standard deviation, RSD < 20%) for determination of ascorbic acid and < 10% for

antioxidant activity determination. The recovery of ascorbic acid was between 85% and 113% for

selected fruits showing good accuracy of the results.

In another study “ Determination of Variation of Vitamin ‘C’ Content of Some Fruits and Vegetables

Consumed in Ugbokolo After Prolonged Storage, ” Anebi et al., (2016) investigated the variation of

vitamin C in some fruits: guava, (Psidium guajava), orange (Citrus sinensis) pineapple, (Ananas

comosus) and apple (Malus pumila), and vegetables: tomato (Lycopersicum esculentum), okra

(Abelmoschus esculentus), red pepper (Capsicum annuum) and green pepper (Capsicum annuum) at

prolonged storage by titrimetric method using dichlorophenolindophenol (DCPIP) titrant. The vitamin C

content of orange was found to be (74.67 and 51.79mg/100g sample), guava (69.6 and 52.8mg),

pineapple (53.42 and 27.93mg), apple (27.3 and 7.29mg), tomato (27.93 and 9.93mg), okra (11.41 and

5.77mg), red pepper (81.53 and 28.26mg) and green pepper (27.62 and 10.65mg). The amount of vitamin

C in fresh red pepper, orange and guava was found to be the highest and that of fresh okra was found to

be lowest. It was observed that content of vitamin C decreased at prolonged storage. In conclusion, red

pepper, orange and guava prove useful for vitamin C deficiency.

In another research by Fabusola et al., (2020) methanol extract of pawpaw and pineapple were analysed

to determine its vitamin content, total phenol content, mineral compositions, reducing power (FRAP) and

antioxidant activities of crude extracts (FTC) with the ability to scavenge 2, 2‐ diphenyl ‐ 1‐

picrylhydrazyl (DPPH.) radical. The result showed that the total phenolic concentrations were (29.27±

0.55mg/100 GAE) for pawpaw (58.16± 27.05 mg/100g GAE) for pineapple. The extract showed a potent

DPPH radical scavenging activity with different concentrations of 20 µg/mL – 100 µg/mL. Pawpaw had

maximum inhibition at 60 µg/mL (29.4%) compared to pineapple at 40 µg/mL (33.6%). The extracts

contained appreciable amounts of vitamin C (174.900±147.510 mg/100g) for pawpaw and vitamin E

11
(1.940±1.940 mg/100g) for pineapple . Minerals experiment was carried out for major elements for which

pawpaw had the highest amount of calcium (295.22±0.16 mg/100g) and magnesium (9.49±0.01

mg/100g). Trace minerals were carried out for which pineapple had the lowest amount of selenium

(2.07±0.01 mg/100g) and copper (0.24±0.02 mg/100g). This therefore suggests that pawpaw and

pineapple could be a good source of antioxidants to ameliorate conditions in diseases whose pathogenesis

implicates oxidative stress.

12
 CHAPTER THREE
MATERIALS AND METHODS
3.1 Description of Sample Area

The study was conducted in Lafia Local Government Area (LGA), the capital city of Nasarawa State,

Nigeria. The LGA is located between longitude 8 0.001 - 90.001 East of the greenwich meridian and

latitudes 80.001 - 90.001N of the equator. In terms of vegetation, the LGA lies largely within the Guinea

Savanah Ecological Zone. It’s wet season starts from May to October while the dry season starts from

October to March. The LGA has annual rainfall of about 1233 mm and average temperature of about

320C (NADP, 2014). The LGA shares boundary with Nassarawa Eggon to the North, Doma to the West,

Obi to the South and finally Quanpan in Plateau State to the East (Nasarawa State Ministry of

Information, 2008).

3.2 Sample Collection

The fruits samples (Orange, Banana, Apple) used in this study were purchased from Lafia Modern

market, Nasarawa State, Nigeria. Upon arrival, the fruit were rinsed with distilled water and labelled for

vitamins and antioxidant screenings.

3.4 Estimation of Vitamins (AOAC, 1990)

3.4.1 Estimation of Vitamin A (Beta-Carotene)

To 10g aliquot of orange, banana or apple sample was added 50ml of acetone: petroleum ether (1:1v/v).
After two hours, the mixture was filtered and the volume of the filtrate measured. An equal volume of

13
50% NaCl was added to wash the filtrate. It was shaken and transferred into a separating funnel. The
lower layer was removed and the supernatant collected and washed with equal volume of 10% K 2CO3 and
separated. The upper layer was washed with 20ml distilled water, separated carefully and its absorbance
was read at 390 nm using 1:1 v/v acetone/ petroleum ether as blank. To get the concentration of vitamin
A in mg/g , the following relationship was employed as shown in equation 3.7.

Conc. of vitamin A (mg/g) =

Where X is concentration extrapolated from the standard curve.

3.4.2 Determination of Vitamin C

To a 10g quantity of each sample was added 80 ml ethanol and 20ml of distilled water; it was covered
and shaken in an orbital shaker for 2 hours. Then, it was filtered and the volume of the filtrate measured.
The filtrate (5 ml) was measured into a conical flask, 50ml of distilled water and 2.5ml of 1M H 2SO4
were added. 1ml of 10% starch indicator was added and titrated with 0.05M iodine solution till a blue-
black colour appeared. To get concentration of vitamin C in mg/g, the following relationship was
employed as presented in equation 3.7

Conc. of vitamin C (mg/g) =

Where,
T.V is Titre value.
One ml of 0.05M iodine solution liberates 0.00886g of Vitamin C

Determination of Vitamin E
Sample (1g) was weighed into a conical flask and 5ml acetone was added and allowed to stand for
10minutes. 2ml of distilled water and 5ml of petroleum ether were added to the filtrate (oily layer). 5ml
of the oily layer was collected and its absorbance was read at 450nm. A standard curve was prepared
using vitamin E standard treated same as sample. To get the concentration in mg/g of vitamin E, the
following relationship was employed as presented in equation 3.7

Where X is concentration extrapolated from the standard curve.

14
2.2.6.5 Determination of Vitamin K
A volume of 20 ml of petroleum ether was added to 1 g of the sample and was allowed to stand for 1 hr
with intermittent shaking every 10 min. It was centrifuged for 5 min then 3 ml of supernatant was put in a
flask and evaporated to dryness. Water, 2 ml and 1 ml of 0.04% 2, 4-dinitrophenylhydrazine was added,
the mixture was then boiled in a water bath for 15 min, cooled and made up to 10 ml with ammonium
hydroxide. It was mixed properly and absorbance measured at wavelength of 635nm. Vitamin K
concentration was extrapolated from the standard curve.

3.5 Quantitative Phytochemical Analysis

3.5.1 Determination of alkaloids
The powdered root sample (30.0 g) was weighed into a 250 beaker and 150.0 mL of 20% absolute

ethanol and 10% acetic acid were added and allowed to stand for 4 minutes. The mixture was agitate on a

shaker for 1 hour and allowed to stand overnight, filtered and the extract concentrated on a water bath.

Acetone 20 ml was further added and agitated for another 30 min and then filtered. The filtrate 10 ml was

collected in a cuvette and the absorbance was measured by UV-visible spectrophotometer at 540 nm. The

actual concentration of the alkaloid in the sample was calculated by equation 3.1

A−B
Conc. of alkaloid (mg/100) = X 100
W

Where, A = Absorbance B = Blank W = Weight of sample

Determination of flavonoid
Approximately 0.25 mg of powdered root sample was dissolved in 1 ml distilled water; 1 ml of 5%
NaNO2 solution, 0.150 ml of freshly prepared aluminum chloride (AlCl 3) and 1 M NaOH solutions were
added. The mixture was allowed to stand for 5 min and absorbance measured at 510 nm. The result was
expressed as quercetin equivalents (QE).

Determination of saponin
Powdered root sample 2 g was weighed into a beaker and 2 ml of isobutyl alcohol was added. The
mixture was filtered through Whatman No. 1 filter paper into a beaker containing 40% magnesium

15
carbonate (MgCO3) solution. One ml of the solution was transferred into volumetric flask, 2 ml of iron
(III) chloride (FeCl3) solution was added and made up to mark with distilled water. This was allowed to
stand for 30 min for the colour development and absorbance was read at 380 nm.

3.5.2 Determination of tannins

A quantity of 30.0 g of the sample was weighed into a plastic bottle and 50ml of distilled water was

added and shaken for 3 hours in a vibrator. The sample was filtered into a 250ml volumetric flask and

made up to mark. A volume, 5ml of the filtrate was dispensed into a test tube and mixed with 2 ml of

0.1M FeCI2 in 0.1NH4CI and 0.008 M potassium ferrocyanide, the absorbance were measured by Uv-

visible spectrophotometer at 325 nm. The concentration of tannins in the sample was calculated

according to equation 3.2.

A−B
Concntration of Tannin in (mg/100) = X 100
W

Where, A = Absorbance B = Blank W = Weight of sample

3.4.5 Determination of Anthraquinone.

A quantity 30.0g of the powdered root sample was weighed into a 250ml beaker and 150.0 mL of 20%
absolute ethanol and 10% acetic acid was placed in a 250 ml beaker. The mixture was allowed to agitate
on a shaker for 1 hour and allowed to stand overnight. The samples was filtered and the extract was
concentrated with 20ml of acetone, the extract was placed on a shaker for another 30 min. The filtrate
was filtered, and 10 ml was placed in a cuvette and the absorbance was measured by UV-visible
spectrophotometer at 350 nm. The concentration of anthraquinone was calculated using equation 3.4
A−B
Conc. of tannin (mg/100) = X 100
W
Where, A = Absorbance B = Blank W = Weight of sample

Determination of total phenols

16
The total phenol content was analysed using Folin-Ciocalteu method as reported by Singleton et al., 1999.
For each standard gallic acid solutions and samples as well as the blank (distilled water), 0.1 mL was
pipetted into a 10 mL volumetric flask and 5.0 ml of distilled water added. This was followed by the
addition of 0.5 ml of 2M Folin-Ciocalteu reagent. Each sample was thoroughly mixed and then left to
stand for 5 minutes after which 1.5 mL of 20% sodium carbonate solution was added. The solution was
made up to the 10 mL mark with distilled water and mixed thoroughly. The resulting solution was then
incubated at room temperature for 2 hours in the dark cupboard after which absorbance readings were
taken at 760 nm UV-VIS spectrophotometer. Triplicate absorbance readings were taken for each of the
duplicate determinations for each sample. The results for the samples were expressed as concentration of
gallic acid equivalent (mg GAE/100 g).

17
 CHAPTER FOUR

RESULT AND DISCUSSION

Table 4.1. Vitamins contents of orange, apple and banana

Fruits Orange (mg/100g) Apple (mg/100g) Banana (mg/100g)

Vitamin A 2.81±0.01 2.08±0.02 1.30±0.01

Vitamin C 1.68±0.01 1.47±0.01 1.06±0.00
Vitamin E 1.04±0.03 0.82±0.02 0.54±0.02
Vitamin K 0.27±0.00 0.24±0.02 0.18±0.00

Data were expressed as mean ± SD of three replicates

Table 4.2. Phytochemical constituents of orange, apple and banana

Parameters Orange (mg/100g) Apple (mg/100g) Banana (mg/100g)
Alkaloids 0.00 0.00 0.00
Flavonoids 0.15±0.01 5.10±0.03 3.05±0.02
Tannins 0.00 0.00 0.00
Saponnins 0.00 0.00 0.00
Anthraquinones 0.00 0.00 0.00
Polyphenol 2.59±0.02 7.02±0.05 4.07±0.05
Data were expressed as mean ± SD of three replicates.

18
4.2 Calibration Curves of Vitamins

19
20
21
4.3 Discussion

The result of vitamins and antioxidants are presented in Table 1. The result of analysis of samples of

fruits shows Orange has the maximum vitamin C content, followed by Apple with Banana having the

least content of the vitamin. This finding is in line with the report of Anebi et al., (2016) who reported in

their study “Determination of Variation of Vitamin ‘C’ Content of Some Fruits and Vegetables

Consumed in Ugbokolo.” that Orange has maximum vitamin C content and apple has low vitamin C

content. Vitamin C is well known for its antioxidant properties and it helps inhibiting infection, and

toxicity.It is also required for the prevention of scurvy and maintenance of healthy skin. It was also

observed that Orange still has the maximum content for all other vitamins, that is, Vitamin A , E, and K,

followed by Apple and then Banana. Vitamin A supports vision, immune function and skin health while

vitamin K on the other hand is vital for blood clotting and bone health. Apart from their (vitamin A and

E) vitamin roles, thay also act as antioxidants, neutralizing free radicals that can damage cells. Further

analysis as regard antioxidant revealed that same fruit (Orange) has higher content of lycopene while

Banana has low lycopene content.

22
 CHAPTER FIVE

CONCLUSION AND RECOMMEDNDATION

5.1 Conclusion

Conclusively, Orange has a high quantity of vitamins and antioxidant such as ascorbic acid and lycopene.

Apple also has high antioxidant potential when compared to Banana. It can be ascertained that the fruits

in question can be exploited as a source of natural antioxidant and vitamins. This knowledge of

antioxidant and nutraceutical potential of these fruits will be useful in selecting plants as nutritional

supplements as well in developing antioxidant based drugs.

5.2 Recommendation

Based on the findings of this study, the populace are encouraged to consume a sufficient quantity of fruit

such as Orange, Apple, Banana and other fruits varieties for they offer vitamins and antioxidants

contributing to overall health.

23
Reference

Anebi O. P, Ugbe A. F, Igwe C. P, Odumu O. F., (2016). Determination of Variation of Vitamin ‘C’
Content of Some Fruits and Vegetables Consumed in Ugbokolo After Prolonged Storage. IOSR
Journal of Environmental Science, Toxicology and Food Technology (IOSR-JESTFT). Volume
10, Issue 7 Ver. III (July 2016), PP 17-19

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