b.sc Project

i

DETERMINATION OF LYCOPENE, VITAMIN A, VITAMIN C, PHENOL, FLAVONOID AND ANTIOXIDANT IN MUSA ACUMINATA (Colla), CAPSICUM ANNUMM (Var. grossum L.), PERSEA AMERICANA (var.

guatemalensis L.) AND PRUNUS DULCIS (var. sativa. L)

BY

ERHIAWARIE, BRIGHT AKPOJOTOR

DELTA STATE UNIVERSITY, ABRAKA, NIGERIA

MAY, 2015

ii

DETERMINATION OF LYCOPENE, VITAMIN A, VITAMIN C, PHENOL, FLAVONOID AND ANTIOXIDANT IN MUSA ACUMINATA (Colla), CAPSICUM ANNUMM (Var. grossum L.), PERSEA AMERICANA (var.

guatemalensis L.) AND PRUNUS DULCIS (var. sativa. L)

BY

ERHIAWARIE, BRIGHT AKPOJOTOR FOS/SLT/09/10/172792

A RESEARCH PROJECT REPORT SUBMITTED TO THE DEPARTMENT OF BOTANY, FACULTY OF SCIENCE, DELTA STATE UNIVERSITY,

ABRAKA

IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF BACHELOR OF SCIENCE (B.Sc.) DEGREE IN BOTANY, DELTA STATE

UNIVERSITY, ABRAKA, NIGERIA.

MAY, 2015

iii

CERTIFICATION

This is to certify that this project work was carried out by Erhiawarie, Bright

Akpojotor and supervised by Dr (Mrs) N.E. Edema in the Department of Botany,

Faculty of Science, Delta State University, Abraka, Nigeria.

____________________ ___________________

DR (MRS) N.E. EDEMA DATE (PROJECT SUPERVISOR)

____________________ ____________________

PROF (MRS) O.M. AGBOGIDI DATE (HEAD OF DEPARTMENT)

iv

DEDICATION

This project work is dedicated to God Almighty for His love, protection and

guidance all through my academic years.

v

ACKNOWLEDGEMENTS

I sincerely wish to express my profound gratitude to God Almighty my provider, my deliverer, my all in all and my everything for keeping me alive till now and providing all that was needed throughout the period of my studies.

I wish to acknowledge Dr (Mrs) N.E. Edema my supervisor for her patience, understanding, motherly advice and constructive criticism in this project work.

I also wish to thank my Head of Department, Prof (Mrs) O.M. Agbogidi and my course adviser, Dr E. Harrison and other lecturers in the Department of Botany, Delta State University, Abraka for the role they played during my academic programme.

Also, I give gratitude to my beloved parents, Chief and Mrs Lagos Erhiawarie and my wonderful siblings like Felicia, Lawrence, Alexandra and Kevwe for their prayers, financial and moral support throughout the period of my studies

I also wish to give gratitude to the family of Mr and Mrs Onoriode Odemakpore, Mr and Mrs Isokariari Damiete and the family of Mr and Mrs Cynathra Atigogo for their financial and moral support throughout the period of my studies.

I also acknowledge the work of the Director, Teezed Business Enterprise, Mr M.O. Jessa who painstakingly typed and made corrections and guidance during this project work.

Finally, I give gratitude to my friends and course mates who in one way or the other contributed to the success of my academic programme. Especially the likes of Ebiserikumo, Bola Enaifoghe, Gregory Okaludo, Josiah Ogedegbe and others too numerous to mention.

vi

TABLE OF CONTENTS Title Page - - - - - - - - - i Certification - - - - - - - - - - ii Dedication - - - - - - - - - - iii Acknowledgement - - - - - - - - - v Table of Contents - - - - - - - - - v Abstract - - - - - - - - - - ix CHAPTER ONE 1.1 Background of the Study - - - - - - - 1 1.2 Classification of Fruits - - - - - - - 3 1.3 Aims and Objectives of Study - - - - - - 4 1.4 Statement of Problem - - - - - - - - 4 1.5 Significance of the Study - - - - - - - 4 CHAPTER TWO: LITERATURE REVIEW 2.1 Lycopene - - - - - - - - - 7 2.2 Vitamin A - - - - - - - - - 8 2.3 Vitamin C - - - - - - - - - 9 2.4 Flavonoids - - - - - - - - - 10 2.5 Phenol - - - - - - - - - - 11 2.6 Anti-oxidants - - - - - - - - - 15 2.7 Musa acuminata - - - - - - - - 13 2.8 Prunus dulcis - - - - - - - - - 15 2.9 Persea americana - - - - - - - - 20 2.10 Capsicum annumm - - - - - - - 24 CHAPTER THREE: MATERIALS AND METHODS 3.1 Determination of Total Antioxidant Capacity - - - - 27 3.2 Determination of Vitamin C - - - - - - - 27 3.3 Determination of Flavonoids - - - - - - - 28 3.4 Determination of Phenols - - - - - - - 28 3.5 Determination of Lycopene - - - - - - - 29 3.6 Determination of Vitamin A - - - - - - - 29 CHAPTER FOUR Results - - - - - - - - - - - 31 CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATION 5.1 Discussion - - - - - - - - - 38 5.2 Conclusion - - - - - - - - - 39 5.3 Recommendation - - - - - - - - 40 References - - - - - - - - 41 Appendix - - - - - - - - - 46

vii

ABSTRACT Fruits are important sources of vitamins, which are essential components of human diet. This project is designed to investigate the identification and separation of lycopene, vitamin A, vitamin C, phenol, flavonoid and antioxidant in Musa acuminata, Capsicum annumm, Persea americana and Prunus dulcis. The results of this study showed that Persea americana (pear) is high in vitamins C when compared to the popular Citrus aurantifolia (Orange). Persea americana had the highest value of 350mg/100g for Ascorbic acid; 0.13mg/100g for flavonoid and 322.88mg/100g for antioxidant. Capsicum annumm recorded 300mg/100g as the second highest in vitamin C. Musa acuminata has the lowest values in ascorbic acid (252mg/5g), phenol (32.3mg/1g) and total antioxidant capacity (321.55mg/5g). From this study, it is recommended that fruits high in antioxidant properties should be consumed to reduce senescence and improve human health status. Further investigation is also required to isolate and characterize the individual components from these plants which are actually responsible for their antioxidant activities and develop their application for food and pharmaceutical industries.

1

CHAPTER ONE

INTRODUCTION

1.1 Background of the Study

Fruits are of great nutritional values. They are important sources of vitamins,

which carry essential components of human diet.

Fruits are important in human diet since they contain carbohydras, proteins,

as well as vitamins, minerals and trace elements (Akaneme, 2008). Regular intake

of fruits is indispensable for good health, fitness and feeling of well-being. In

addition, millions of people throughout the developing countries of the world were

reported to have inadequate food supply or nutrient deficiencies in their diets, which

led to problems due to starvation and malnutrition of various types (Tlili and

Lenucci, 2011). There has been an increase in the awareness on the food value of

fruits as a result of exposure to other cultures and acquiring proper food education

(Amusa, et al., 2003).

Pepper (Capsicum annumm. var. bola), is one of the most varied and widely

used food in the world. There are a vast number of varieties of pepper and every

variety is indicated with the original language of the local cultures, which differs

2

from town to town and from region to region. For these reasons the classification of

pepper is not simple (Basher and Abu-Goukh, 2003). Nigeria is known to be one of

the major producers of pepper in the world accounting for about 50% of the African

production (Ureigho, 2010). The pepper grown in Nigeria is in high demand because

of its pungency and good flavour (Akbugwo and Ugbogu, 2007).

Pepper is a fruit is now recognized as rich sources of antioxidants, in which

vitamin A, C and Lycopene are some of the most abundant antioxidants identified

in them (ArvouetGrand et al., 2000; Badr-Sherif et al., 2011). Since the protective

roles of dietary, antioxidants cant be over-emphasized against multiple diseases

such as cancer, anaemia, diabetes, carbohydrates diseases, etc. these antioxidants

perform their functions by contracting the oxidizing activities of highly reactive

oxygen species thereby, preventing the oxidative modification of low density

protein, nucleic acids, proteins, etc. (Briviba and Sies, 2000). Therefore, there have

been recent increases recorded in the demand in the consumption, particularly

among the urban community. This is due to the increased awareness on the food

value of fruits (e.g apple, pepper, banana, guava, pear and almond), especially as

antioxidants (Amusa, et al., 2003).

3

Epidemiological studies indicate a strong inverse correlation between the

consumption of fruits and the incidence of degenerative diseases. There is

considerable evidence for the role of antioxidant constituents of fruits in the

maintenance of health and prevention of diseases.

Phenolic compounds have the ability to prevent the oxidation of low-density

lipoprotein (LDL) owing to their antioxidant properties, attributable to the free

radical scavenging properties of their constituents hydroxyl groups. The inhibition

of low-density lipoprotein oxidation has been associated with a lower incidence of

coronary diseases. Among the several classes of plant phenolics, some have been

reported in pear fruits: phenolic acids, flavonoids and anthocyanin (Burkill, 2003).

1.2 Classification of Fruits

Fruits are classified into three main groups:

Simple Fruits: These are fruits containing one or more carpels they take roots from

a single ovary with or without accessory parts. E.g. Drupes, Nuts and Legumes.

Aggregate Fruits: These are fruits which one flower contains more than a few,

divided ovaries, which fuses together as it develops. E.g. Pineapple.

4

Multiple Fruits: These are fruits that consist of matured ovaries of several to many

flowers more or less united into a mass. Multiple fruits are almost invariably

accessory fruits. E.g. Blackberry, Raspberry and Strawberry.

1.3 Aims and Objectives of Study

The aim and objective of this study was to investigate the content of vitamin

C, vitamin A, phenol, flavonoids, lycopene and antioxidant properties of some fruits

collected from small market, Abraka.

The sampled fruits were Banana (Musa acuminata), Pepper (Capsicum

annumm), Pear (Persea americana) and Almond (Prunus dulcis).

1.4 Statement of Problem

1. The demand on lycopene, vitamin A, vitamin C, Phenol, Flavonoid and

Antioxidant have increased significantly, with increased of consumer

awareness about cancer.

1.5 Significance of the Study

This research will be useful in creating awareness of vitamin A, Vitamin C,

phenol, flavonoids, lycopene and total antioxidant properties in fruits used as food

5

e.g. Banana, Pepper, Pear and Almond that can inhibit the chain reaction caused by

free radicals which is responsible for many diseases in human body.

6

CHAPTER TWO

LITERATURE REVIEW

Nowadays the major interest of carotenoids, which are found in plants, is not

only due to their pro-vitamin A activity but also their antioxidant action by

scavenging oxygen radicals and reducing oxidative stress in the organism. There are

many studies showing strong correlation between carotenoids intake and a reduced

risk of some diseases, as cancer, aterogenesis, bone calcification, eye degeneration

and neural damages.

Lycopene is the compound responsible for red colour in pepper, watermelon

and other fruits, and it is also used as a colour ingredient in many food formulations.

A great interest has recently been focused on lycopene due to its preventive activity

against several pathologies such as cardiovascular disease, hepatic fibrogenesis,

solar light induced erythema, human papillomavirus persistence and some cancer

types such as prostate, gastrointestinal and epithetial. Lycopene has also been

reported to play a role in lung function as well as in foetal growth. Finally, it is also

important to consider the synergies action of carotenoids with other bioactive

compounds present in fruits and vegetables.

7

The parameters that are being worked on in this review are lycopene, vitamin

A, vitamin C, flavonoid, phenol and antioxidants.

2.1 Lycopene

Structure of Lycopene

Lycopene is a member of the carotenoid family of chemical substance.

Lycopene, similar to other carotenoids, is a natural fat. Soluble pigment (red, in the

case of lycopene) found in certain plants and microorganisms, where it serves as an

accessory light gathering pigment and to protect these organisms against the toxic

effects of oxygen and light. Lycopene may also protect humans against certain

disorders, such as prostate cancer and perhaps some other concern and coronary

heart disease.

Carotenoids are with principal pigments responsible for the colours of fruits.

Lycopene is responsible for the red colour of pepper. In addition to pepper and

tomato based products, such as ketchup, pizza sauce, pepper paste.

8

Lycopene is an acrylic isomer of beta-carotene. Beta-carotene, which contains

beta-ionone rings at each end of the molecules is formed in plants, including pepper,

via the action of enzyme lycopene beta-cyclase. Lycopene is a 40 carbon atom, open

chain polysioprenoid with conjugated double bonds.

2.2 Vitamin A

Chemical structure of retinol, one of the major forms of vitamin A

Vitamin A is a group of unsaturated nutritional organic compounds that

includes retinol, retinal, retinoic acid and several provitamin. A carotenoids among

which beta carotene is the most important. Vitamin A has a multiple functions: it is

important for growth and development, for the maintenance of the immune system

and good vision. Vitamin A is needed by the retina of the eye in the form of retinal

which combine with protein opsin to form rhodopsin, the light-absorbing molecule

necessary for both low-light (scotopic vision) and colour vision.

9

Vitamin A is found naturally in many foods like Pepper, Pear, Almond and

Banana.

2.3 Vitamin C

Structure of Vitamin C

Vitamin C or L-ascorbic acid or simply ascorbic (the anion of ascorbic acid),

is an essential nutrient for human and certain other animal species. Vitamin C refers

to a number of vitamins that have vitamin C activity in animals, including ascorbic

acid and its salts and some oxidized forms of the molecule like dehydroascorbic acid.

Ascorbate and ascorbic acid are both naturally present in the body where either of

these is introduced into the cells, since the forms interconvert according to pH.

In this research, we found out that a cup of chopped red bell pepper contains

nearly three times more vitamin C than an orange-190mg.

10

2.4 Flavonoids

Molecular structure of the flavone backbone (2-phenyl-1,4-benzopyrone)

Flavonoids are a class of plant secondary metabolites thought to provide

health benefit through cell signalling pathways and antioxidants effects. These

molecules are found in a variety of fruits.

Flavonoids are important antioxidants and promote several health effects.

Aside from antioxidant activity, these molecules provides the following beneficial

effects to humans:

Anti-viral

Anti-cancer

Anti-inflammatory

Anti-allergic

11

Almost all fruits, vegetables and herbs contain a certain amount of flavonoids

e.g. Musa acuminata, pear, Prunus dulcis and red bell pepper.

The best way to ensure a good intake of flavonoids is to consume plenty of

fresh fruit and vegetables on a daily basis. From my research, fruits like pear, carrots,

paw-paw, red pepper, etc. are rich in flavonoids.

2.5 Phenol

Structure of phenol

Phenols is a type of phytochemical called polyphenol. Other types of

polyphenols include flavonoids and stilbenes. Phenolic acids are found in a variety

of plant-based food, the seeds and stems of fruits and vegetables contain the highest

concentrations.

Phenolic acids are easily absorbed through the walls of your intestinal tract,

and they may be beneficial to health because they work as antioxidants that prevents

12

cellular damage due to free radical oxidation reactions. They also promotes anti-

inflammatory conditions in your body when you eat them regularly.

2.6 Anti-oxidants

The structure of the antioxidant

A substance that reduces damage due to oxygen, such as that caused by free

radicals. Well-known antioxidant include enzymes and other substances, such as

vitamin C, vitamin E and beta-carotene, which are capable of counteracting the

damaging effects of oxidation. Antioxidants are also commonly added to food

products such as fruits and vegetables. Antioxidants may possibly reduce the risk of

13

cancer. Antioxidants clearly slow the degeneration of age-related mocular

degeneration.

All these parameters were tested in Musa acuminata, Capsicum annumm,

Persea americana and Prunus dulcis.

2.7 Musa acuminata (Banana)

Plate 1: Musa acuminata

Musa acuminata is an edible fruit botanically a berry, produced by several

kinds of large herbaccous flowering plants in the genus Musa. The fruit is variable

in size, colour and firmness, but is usually elongated and carved, with soft flash rich

in starch covered with a rind which may be green or yellow when ripe.

14

The fruit grown clusters is hanging from the top of the plant. Almost all

modern edible parthenocarpic (seedless) Musa acuminatas come from two wild

species- Musa acuminata and Musa balbisana.

The Musa acuminata plant is the largest herbaceous flowering plant. All the

above-ground parts of a Musa acuminata plant grow from a structure usually called

a corm. Plants are normally tall and flairy study, and are often mistaken for trees

but what appears to be a trunk is actually a false stem or pseudosten. Musa

acuminata grown in a wild variety of soils, as long as the soil is at least 60cm deep,

has good drainage and is not compacted. The leaves of Musa acuminata plants are

composed of a stalk (petiole) and a blade (lamina).

The Musa acuminata fruits develop from the Musa acuminata heart in a large

hanging cluster, made up of tiers (called hands), with up to 20 fruits to a tier. The

hanging cluster is known as a bunch, comprising 3-20 tiers and can weigh 30-50kg.

The fruit has been described as a teathery berry. There is a protective outer

layer (a peel or skin) with numerous long, thin strings (the phloem bundles), which

non length wise between the skin and the edible inner portion.

15

Musa acuminata are naturally slightly radioactive, more so than most other

fruits, because of their potassium content and the small amount of the isotope

potassium-40 found in naturally occurring potassium. A study has shown that ripe

Musa acuminatas florese when exposed to ultraviolet light. This property is

attributed to the degradation of chlorophyll leading to the accumulation breakdown

product is stabilized by a propionate ester group.

Musa acuminata are excellent sources of vitamins and contains moderate

amount of manganese and dietary fiber.

2.8 Prunus Dulcis (Almond)

Plate 2: Prunus dulcis

16

The Prunus dulcis is a species of tree native to the Middle East and South

Asia. "Prunus dulcis" is also the name of the edible and widely cultivated seed of

this tree. Within the genus Prunus, it is classified with the peach in the subgenus

Amygdalus, distinguished from the other subgenera by the corrugated shell

(endocarp) surrounding the seed.

The fruit of the Prunus dulcis is a drupe, consisting of an outer hull and a hard

shell with the seed (which is not a true nut) inside. Shelling Prunus dulcis refers to

removing the shell to reveal the seed. Prunus dulcis are sold shelled (i.e., after the

shells are removed), or unshelled (i.e., with the shells still attached). Blanched

Prunus dulcis are shelled Prunus dulcis that have been treated with hot water to

soften the seedcoat, which is then removed to reveal the white embryo.

The Prunus dulcis is a deciduous tree, growing 410 m (1333 ft) in height,

with a trunk of up to 30 cm (12 in) in diameter. The young twigs are green at first,

becoming purplish where exposed to sunlight, then grey in their second year. The

leaves are 35 inches long, with a serrated margin and a 2.5 cm (1 in) petiole. The

flowers are white to pale pink, 35 cm (12 in) diameter with five petals, produced

singly or in pairs and appearing before the leaves in early spring.(Lim, T. 2000).

17

Prunus dulcis grows best in Mediterranean climates with warm, dry summers and

mild, wet winters. The optimal temperature for their growth is between 15 and 30C

(6085F) and the tree buds have a chilling requirement of between 300 and 600

hours below 7.2C (45F) to break dormancy.

Prunus dulciss begin bearing an economic crop in the third year after planting.

Trees reach full bearing five to six years after planting. The fruit matures in the

autumn, 78 months after flowering.

The Prunus dulcis fruit measures 3.56 cm (12 in) long. In botanical terms,

it is not a nut, but a drupe. The outer covering or exocarp, fleshy in other members

of Prunus such as the plum and cherry, is instead a thick, leathery, grey-green coat

(with a downy exterior), called the hull. Inside the hull is a reticulated, hard, woody

shell (like the outside of a peach pit) called the endocarp. Inside the shell is the edible

seed, commonly called a nut. Generally, one seed is present, but occasionally two

occur.

The Prunus dulcis is native to the Mediterranean climate region of the Middle

East, eastward as far as the Indus River in Pakistan (Cushnie and Lamb 2005). In

India and Pakistan, it is known as Badam. It was spread by humans in ancient times

18

along the shores of the Mediterranean into northern Africa, Asia and southern

Europe and more recently transported to other parts of the world, notably California,

United States (Faleyimu and Oluwalana, 2008).

The wild form of domesticated Prunus dulcis grows in parts of the Levant;

Prunus dulcis must first have been taken into cultivation in this region. The fruit of

the wild forms contains the glycoside amygdalin, "which becomes transformed into

deadly prussic acid (hydrogen cyanide) after crushing, chewing, or any other injury

to the seed." (Halliwell, 2005).

Wild Prunus dulcis are bitter, the kernel produces deadly cyanide upon

mechanical handling, and eating even a few dozen in one sitting can be fatal.

Selection of the sweet type, from the many bitter types in wild, marked the beginning

of Prunus dulcis domestication. How humans selected the sweet type remains a

mystery (Idowu, et al., 2003). It is unclear as to which wild ancestor of the Prunus

dulcis created the domesticated species. Ladizinsky suggests the taxon Amygdalus

fenzliana (Fritsch) Lipsky is the most likely wild ancestor of the Prunus dulcis in

part because it is native of Armenia and western Azerbaijan where it was apparently

domesticated.

19

While wild Prunus dulcis species are toxic, domesticated Prunus dulcis are

not; Jared Diamond argues that a common genetic mutation causes an absence of

amygdalin, and this mutant was grown by early farmers, "at first unintentionally in

the garbage heaps, and later intentionally in their orchards" (Iqbal, et al., 2001).

Zohary and Hopf believe that Prunus dulcis were one of the earliest domesticated

fruit trees due to "the ability of the grower to raise attractive Prunus dulcis from

seed. Thus, in spite of the fact that this plant does not lend itself to propagation from

suckers or from cuttings, it could have been domesticated even before the

introduction of grafting" (Zohary, 2000). Domesticated Prunus dulciss appear in the

Early Bronze Age (30002000 BC) such as the archaeological sites of Numeria

(Jordan) (Iqbal, et al., 2001), or possibly a little earlier. Another well-known

archaeological example of the Prunus dulcis is the fruit found in Tutankhamun's

tomb in Egypt (c. 1325 BC), probably imported from the Levant (Jacob, 2003). Of

the European countries that the Royal Botanic Garden Edinburgh reported as

cultivating Prunus dulciss, Germany is the northernmost, though the domesticated

form can be found as far north as Iceland.

20

2.9 Persea americana (Pear)

Plate 3: Persea americana

Persea americana is a tree native to Mexico and Central America, classified

in the flowering plant family Lauraceae along with cinnamon, camphor and bay

laurel. Avocado or alligator pear also refers to the fruit, botanically a large berry that

contains a single seed.

Avocados are commercially valuable and are cultivated in tropical and

Mediterranean climates throughout the world. They have a green-skinned, fleshy

body that may be pear-shaped, egg-shaped, or spherical. Commercially, they ripen

21

after harvesting. Trees are partially self-pollinating and often are propagated through

grafting to maintain a predictable quality and quantity of the fruit.

The tree grows to 20 m (66 ft), with alternately arranged leaves 1225 cm

(4.79.8 in) long. The flowers are inconspicuous, greenish-yellow, 510 mm (0.2

0.4 in) wide. The pear-shaped fruit is 720 cm (2.87.9 in) long, weighs between

100 and 1,000 g (3.5 and 35.3 oz), and has a large central seed, 56.4 cm (2.02.5 in)

long.

The subtropical species needs a climate without frost and with little wind.

High winds reduce the humidity, dehydrate the flowers, and affect pollination. When

even a mild frost occurs, premature fruit drop may occur, although the 'Hass' cultivar

can tolerate temperatures down to 1 C. The trees also need well-aerated soils,

ideally more than 1 m deep. Yield is reduced when the irrigation water is highly

saline. These soil and climate conditions are available in southern and eastern Spain,

Portugal, Morocco, Crete, the Levant, South Africa, Colombia, Peru, parts of central

and northern Chile, Vietnam, Indonesia, parts of southern India, Sri Lanka,

Australia, New Zealand, the Philippines, Malaysia, Central America, the Caribbean,

22

Mexico, California, Arizona, Puerto Rico, Texas, Florida, Hawaii, Ecuador, and

Rwanda. Each region has different cultivars.

A typical serving of avocado (100 g) is moderate to rich in several B vitamins

and vitamin K, with good content of vitamin C, vitamin E and potassium (right table,

USDA nutrient data). Avocados also contain phytosterols and carotenoids, such as

lutein and zeaxanthin.

Avocados have diverse fats. For a typical avocado:

About 75% of an avocado's energy comes from fat, most of which (67%

of total fat) is monounsaturated fat as oleic acid.

Other predominant fats include palmitic acid and linoleic acid.

The saturated fat content amounts to 14% of the total fat.

Typical total fat composition is roughly (rounded to digits): 1% -3,

14% -6, 71% -9 (65% oleic and 6% palmitoleic), and 14% saturated

fat (palmitic acid).

A 2013 epidemiological NHANES study funded by the Hass Avocado

Board showed that American avocado consumers had better overall diet

quality, nutrient levels, and reduced risk of metabolic syndrome; why

23

they also had better diet quality and how the confluence of these factors

contributed to health benefits was not determined.

High avocado intake was shown in one preliminary study to lower

blood cholesterol levels. Specifically, after a seven-day diet rich in

avocados, mild hypercholesterolemia patients showed a 17% decrease

in total serum cholesterol levels. These subjects also showed a 22%

decrease in both LDL (harmful cholesterol) and triglyceride levels and

11% increase in HDL (helpful cholesterol) levels. In a study of obese

patients on a moderate fat diet (34% of calories), additional

consumption of one avocado (136 g) per day over 5 weeks produced a

significant reduction of circulating LDL, an effect the authors attributed

to the combination of avocado monounsaturated fats, dietary fiber and

the phytosterol, beta-sitosterol.

24

2.10 Capsicum Annumm (Pepper)

Plate 4: Capsicum annumm

Capsicum annumm is a species of the plant genus Capsicum native to southern

North America and northern South America. This species is the most common and

extensively cultivated of the five domesticated Capsicums. The species encompasses

a wide variety of shapes and sizes of peppers, both mild and hot, ranging from bell

peppers to chili peppers. Cultivars are descended from the wild American bird

pepper still found in warmer regions of the Americas. In the past some woody forms

of this species have been called C. frutescens, but the features that were used to

25

distinguish those forms appear in many populations of C. annumm and there is no

consistently recognizable C. frutescens species.

Although the species name annumm means annual (from the Latin annus

year), the plant is not an annual and in the absence of winter frosts can survive

several seasons and grow into a large perennial shrub. The single flowers are an off-

white (sometimes purplish) color while the stem is densely branched and up to 60

centimetres (24 in) tall. The fruit is a berry and may be green, yellow or red when

ripe. While the species can tolerate most climates, C. annumm is especially

productive in warm and dry climates.

Hot peppers are used in medicine as well as food in Africa and other places

around the world.

English botanist John Lindley described C. annumm on page 509 of his 1838

'Flora Medica' thus:

It is employed in medicine, in combination with Cinchona in intermittent and

lethargic affections, and also in atonic gout, dyspepsia accompanied by flatulence,

tympanitis, paralysis etc. Its most valuable application appears however to be in

26

cynanche maligna (acute diphtheria) and scarlatina maligna (malignent Scarlet

fever, used either as a gargle or administered internally.)

In ayurvedic medicine, C. annumm is classified as follows:

Gunna (properties) ruksh (dry), laghu (light) and tikshan (sharp)

Rasa dhatu (taste) katu (pungent)

Virya (potency) ushan (hot)

27

CHAPTER THREE

MATERIALS AND METHODS

3.1 Determination of Total Antioxidant Capacity

Procedure

A 5g of fruit sample was crushed, and 10ml of distilled water was added to

sample and homogenised. A clear supernatant of sample was taken. A measurement

of 1ml supernatant (liquid from sample reagent solution) and 1ml of reagent solution

was added and boiled for 90minutes at about 800c. Samples were allowed to cool

and readings taken at 695nm on a uv-spectrometer. Reading gotten for sample and

standard ascorbic acid were put into formula for antioxidant capacity and the final

reading were calculated.

3.2 Determination of Vitamin C

Procedure

A 5g of fruit sample was pulverised (grinded) and mixed with 20ml of distilled

water. A ml of clear sample supernatant (liquid) was taken. 4ml of oxalic E.D.T.A.

and 1ml of orthosphroic acid was added to sample. 1ml of 5% H2S04 was added to

sample mixture and allowed to stand for 15 minutes for colour development and

28

readings were taken at about 760nm on a UV Spectrophotometer. Readings were

compared against standard ascorbic acid on a graph to get actual readings in mg/kg.

3.3 Determination of Flavonoid

Procedure

A 10g of fruit sample was pulverized, and 20ml of 0% methanol was added

to pulverized (grinded) sample, which was homogenized and mixed, then a clear

supernatant (pure liquid) was weighed out into a pre-weighed crucible and hearted

to dryness at about 1050C and the crucible was weighed.

The difference between empty crucible and dry crucible and was taken as final

reading per 10grams

3.4 Determination of Phenol

Procedures

A 1g of fruit sample was boiled with 50ml of ether for 15 minutes, then 5ml

of sample was weighed out and added to 10ml of distilled water. Again, 2ml of

ammonium hydroxide (ammonia) was added and 5ml of ethanol was added and the

mixture was allowed to stand for 30 minutes.

29

The UV-spectrophotometer readings were taken at 505nm. The readings were

plotted against reading of standard solution on a graph to get the final reading in

mg/kg.

3.5 Determination of Lycopene

Procedure

A 100g of fruit sample is ground to paste and 30ml of 95% ethanol was added

and stirred for 6 minutes. The solid is isolated via a vacuum pump filtration and

dichloromethane was added to separate the lycopene via a separating funnel. The

liquid is heated with 30ml of dichloromethane for 5minutes and filtered. Extracts

was mixed with 5ml of 5% sodium chloride. The mixture was heated to dryness and

the weight was subtracted from an already pre-weighed. The result was taken as the

weight in mg/100g.

3.6 Determination of Vitamin A

Procedure

A 5g of fruit sample was pulverized (grinded) and 10ml of chloroform was

added. Later, 5ml of supernatant (clear liquid from sample mixture) was added to

5ml of 30% antimony tri-chloride was added and readings were taken at 415nm.

30

Readings were plotted against a graph of vitamin A table and reading was taken in

1.(mg).

31

CHAPTER FOUR RESULTS AND DISCUSSION

The results of this study are presented in Tables 1-6 Table 1 shows Ascorbic acid content of some fruits. Persea americana (Pear)

had the highest value of ascorbic acid and the least was found in Musa acuminata.

Table 2 shows the vitamin A contents of the four fruits. The highest value of

vitamin A was in Capsicum annumm.

Table 3 shows the phenolic content of four different fruits. The highest value

for phenolic acid content was recorded in Persea americana, while the least value

was Musa acuminata.

Table 4 shows the flavonoid content of six different fruits. The highest value

was recorded for Musa acuminata. While the least value was recorded for Capsicum

annumm.

Table 5 shows the lycopene content of four fruits. The least value of lycopene

content was recorded for Persea americana, while the highest value was for

Capsicum annumm.

Table 6 shows the antioxidant content of some fruits. The highest value of

antioxidants was found in Prunus dulcis (Almond).

32

Table 1. Mean Values of ascorbic acid in four fruits (mg/5g)

S/N Fruit Mean Value Botanical Name Common Name

1 Musa acuminata Banana

252 1.4

2 Capsicum annumm Pepper

300 5.5

3 Persea Americana Pear

350 0.7

4 Prunus dulcis Almond 262 1.4

33

Table 2. Values of Vitamin A in four fruits (mg/5g)



6.61 0.20


21.08 0.02

3 Persea americana Pear

4.95 0.03

4 Prunus dulcis Almond 5.40 0.02

34

Table 3. Values of phenol in four fruits (mg/1g)



32.2 0.14


54.0 0.07


64.0 0.14


35

Table 4. Values of flavonoids in four fruits (mg/10g)



0.46 0.07


0.12 0.07


0.13 0.07


36

Table 5. Values of lycopene in four fruits (mg/100g)

S/N Fruit Mean Value Botanical Names Common Names


0.07 0.021


0.31 0.007


0.01 0.007


37

Table 6. Values of antioxidant in four fruits (mg/5g)


1 Musa acuminate Banana

321.55 0.1060


382.63 0.2616


322.86 0.0141


Discussion

The determination of vitamin C, vitamin A, phenol, flavonoid, lycopene and

antioxidant properties of some fruits were carried out. Persea americana (Pear) had

the highest values of 350mg/5g for Ascorbic acid; 0.13mg/10g for flavonoid and

322.88mg/5g for antioxidant.

This shows that Persea americana is rich in vitamin C when compared to

other three (3) fruits evaluated. Vitamin C is one of the most important dietary

antioxidant. It is also considered the most important water soluble antioxidant in

38

extracellular fluid. It is capable of neutralizing reactive oxygen species in aqueous

phase before peroxidation is initiated (Padayatty et al., 2003).

Flavonoid appear to function as biological response modifier. Flavonoid have

been demonstrated to have anti-inflammatory, anti-allegenic, anti-viral, anti-aging

and anti-carcinogenic activity (Havsteen, 2000). They are also known to exert

protection against heart diseases through the inhibition of cyclooxygenase and

lypolygnase activities in platelets and microphages (Amusa et al., 2003).

Phenolic acids are plant secondary metabolic widely spread throughout the

plant kingdom (Brumeton, 2000). Recent interest in phenolic acid sterms from their

potential as protective factors against cancer and heart diseases in plant because of

their potent phenolic properties (Breinholt, 2005).

The results of this study show that Musa acuminata recorded 252 mg/5g

(Table 1). It is the second highest in vitamin C among the other three fruits.

Fruits with total phenolic content and antioxidant capacity have been reported

to have the anti-diabetic, anti-hypertension and anti-cancer (Bardr-Sharit, 2011).

39

CHAPTER FIVE

SUMMERY, CONCLUSION AND RECOMMENDATION

5.1 Summary

This study investigated the determination of lycopene, vitamin a, vitamin c,

phenol, flavonoid and antioxidant in Musa acuminata (colla), Capsicum annumm

(var. grossum l.), Persea americana (var. guatemalensis l.) and Prunus dulcis (var.

sativa. L). The literature review discussed lycopene, vitamin C, A, flavonoid, phenol

and antioxidant. The fruits used were also reviewed, they are: banana, pear, pepper

and almond. The results of this study showed that Persea americana (pear) is high

in vitamins C when compared to the popular Citrus aurantifolia (Orange). Persea

americana had the highest value of 350mg/100g for Ascorbic acid; 0.13mg/100g for

flavonoid and 322.88mg/100g for antioxidant. Capsicum annumm recorded

300mg/100g as the second highest in vitamin C. Musa acuminata has the lowest

values in ascorbic acid (252mg/5g), phenol (32.3mg/1g) and total antioxidant

capacity (321.55mg/5g). From this study, it is recommended that fruits high in

antioxidant properties should be consumed to reduce senescence and improve human

health status. Further investigation is also required to isolate and characterize the

40

individual components from these plants which are actually responsible for their

antioxidant activities and develop their application for food and pharmaceutical

industries.

5.2 Conclusion

The results of the present study showed that Persea americana has the highest

Vitamin C properties. Average amount of antioxidant properties of fruits extract was

observed from Musa acuminata. Low antioxidant activity was observed from

Capsicum annumm. Different antioxidants are necessary because the vital role they

play in controlling oxidation and preventing chronic diseases in man. A combination

of different fruit is necessary, since a single fruit is not capable of providing

comprehensive effect of the antioxidants.

5.3 Recommendation

It is hereby recommended that it is necessary to consume fruits high in

antioxidant properties that may be efficient as preventive agent in aging and in some

diseases. As the plant extracts are quite safe and the use of synthetic antioxidant has

been limited because of their toxicity, therefore, these local fruits could be exploited

as antioxidant additive or as nutritional supplement.

41

However, further investigation is required to isolate and characterize the

individual components from these plants which are actually responsible for their

antioxidant activities and develop their application for food and pharmaceutical

industries.

42

REFERENCES

Akaneme, F.I. (2008). Identification and preliminary phytochemical analysis of herbs that can arrest threatened miscarriage in Orba and Nsukka town of Enugu State. African Journal of Biotechnology. 7: 6-11

Akbugwo, I.E. and Ugbogu, A.E. (2007). Physiochemical studies on oil from five selected Nigerian plant seed. Parkinson Journal of Nutrition. 6: 75-78

AMusa, N.A., Ashaye, O.A., and Oladapo, M.O. (2003). BiOdeterioration of the Africa Star Apple (Chrysophyllum albidum) in storage and the effect on its food value. African Journal of Biotechnology. 2: 56-57.

Arvouet Grand, A. Vennat, B. Pourrat, A. and Legret, P. (2000). Standardization of unextrait de propolis et identification des principaux constituant. Journal of Pharmacology Belgium. 49: 462-268

Badr-Sherif, E.A., Shaaban, M., Elkholy, V.M., Hela, M.H., and Safty, M.M.E (2011). Chemical composition and biological activity of ripe pumpkin fruits (Cuccurbita pepo L.) cultivated in Egyptian habitats. National Production Resources. 25(165): 1524-1534

Basher, H.A. and Abu-Goukh, A.A (2003). Compositional Changes during Guava ripening. Food chemistry. 80: 557-563.

Briviba, K., and Sies, H., (2000). Non enzymatic Antioxidant defense systems. In Frei, B.A. (ed). National Antioxidant in human health and disease Academic Press, San Diego, pp 10 1-128.

Burkill, H.M. (2003). The useful plants of West Tropical Africa. Royal Botanical Garden Kew. 2nd edition. Families M-R: 647

Cheeseman, K.H. and Slater, T.F. (2003). An introduction to free radical chemistry. British Medical Bulletin. 49: 48 1-93

43

Cook, N.K. and Samman, S. (2001). Flavonoids- Chemistry, metabolism, cardioprotective effect and dietary sources. Journal of Nutrition Biochemistry. 7: 66-76.

Cushnie, T.P.T and Lamb, A.J (2005). Antimicrobial activity of flavonoids. International Journal of Antimicrobial Agent. 26 (5): 343-356.

Duthie, G.G., and Brown, K.M. (2001). Reducing the risk of cardiovascular disease. In Goldberg, I. (ed). Functional food. New York, Chapman and Hall, pp 19-3 8.

Faleyimu, O.L. and Oluwalana, S.A. (2008). Tropical value of forest plant seeds in Ogun state, Nigeria. Medicinal value of Forest plant seeds.

Feskanich, D., Ziegler, R.G., Michaud, D.S., Giovannucci, S.L., Speizer, F.E., Willett, W.C. and Colditz, G.A. (2000). Prospective study of fruit and vegetables consumption and risk of lung cancer among men and women. Journal of the National Cancer Institute. 92: 1812-1823.

Halliwell, B. (2005). How to catarize an antioxidant An update: Biochemistry Society Symposium. 61: 73-10 1

Havsteen, B. (2003). Flavonoids, a class of Natural products of high pharmacological potency. Biochemistry Pharmacology. 32(7): 1141-1 148.

Idowu, T.O, Onawunmi, G.O., Ogundaini, A.O and Adesanya, S.A. (2003). Antimicrobial constituents of clirysophyllum albidum seed cotyledeons. Journal of National Production and Medical. 7: 33-36

Iqbal, Z., Nadeem, Q.K., Khan, M.M., Akhtar, M.S and Waraich, F.N. (2001). In-vitro Anthredlinintic activity of Allium sativum, Zingiber officinale, cucurbita mexiana and fleas religiosa. International Journal of Agriculture Biology. 3(4): 454-445.

Jacob, R.A. (2003). The Integrated Antioxidant System. Nutrition Resource. 15(5): 755-766.

44

JimnezEscrig, A., Rincon, M., Pulido, R., and Saura-Calixto, F. (2001). Guava fruit (psidium guava L.) as a new source of antioxidant dietary fiber. Journal of Agriculture Food Chemistry. 49: 8489-5493.

Jyotsna M. R.K., Srivestava, S.V., Shukia C.S. (2007). Antioxidant in Aromatic and Medicine and plants, Science Technology Entrepreneur. 7:1-16.

Kessler, M. Ubeand, G. and Jung, L. (2003). Anti-and Pro-oxidant activity of rufin and guerretin derivatives. Journal of Pharmacy Pharmacology. 55: 131-142.

Leonard, S.S., Cutler, D., Ding, M., Vallayathan V. and Castranova, V. (2002). Antioxidant properties offruit and vegetable Juices: More to the strong than ascorbic acid. Annual Clinical Laboratory Science. 32: 193-200.

Lewis, O.B and Robert A. (2002). Dictionary of Agricultural Science: CRC Press. Lim, T.K and Khov, K.C (2000). Guava in Malaysia: Production, Pests and disease.

Kuala Lumpar: Tropical Press, Malaysia.

Olorunnisola, D.S., Amao, I.S., Ehigihe, D.O and Ajayi, Z.A.F. (2008). Anti-hypergylcerinie and Dalessandro (2011). Bioactive. Compounds and antioxidant activities during fruit ripening of watermelon cultivar. Journal of Science Food Agriculture. 81, 983

Padayatty, S.J., Katz, A. Wang, Y., Eck, P. Kwon, 0. Lee, J.H., Chen, S., Corpe, C., Dutta, A., Dutta, S.K. and Levine, M. (2003). Vitamin C as an antioxidant: Evaluation of its role in disease prevention, 22(1) 18-35.

Pamplona-Roger, G.D., (2005). Encyclopedia of food and their healing power. Editorial safeliz, Madrid

Paulovicsova, B., Turicinica, I., Turikova, T., Bologhova, M., and Matuskovic, J. (2009). Antioxidant properties of less common fruit species. Zootechnie si Biotechnologi 42: 21-27.

Pearson, D. (2001). Chemical analysis of foods. London, Churchill Livingstone 181.

45

Perkins,V.P. and Collins, J.K., (2006). Carotenoid changes of intake watermelons after storage. Journal of Agricultural food Chemistry. 54(16): 5868-5874.

Petel, V.R., Prakash, R. Patel and Sushil, S.K. (2010). Antioxidant activity of some selected medical plants in Western Region of Indian. Advances in Biological Research. 4(1) 23-26.

Pourmorad, F., Hosseinimehr, S.J. and Shahabimajd .0. (2006). Antioxidant activity, phenol and flavonoid content of some selected Iranian medicinal plants. African Journal of Biotechnology. 5(11): 1142-1145.

Rice-Evans, R. and Ames, B.N. (2008). Antioxidant defences and lipid peroxidation in human blood plasma. Proceedings Nation Academic of Science. 37: 57-71

Saha, P., Mazumder, U.K., Halder, P.K., Bala, A., Kar, B., and Ivaskar, S. (2011). Evaluation and hyperprotective activity of curbita maxima derialparts. Journal Herbal Medical Toxicology. 5(1): 17-22.

Savitree, M., Isana, P., Nittaga, S.R., and Worapon, S. (2004). Radical scavenging activity and total phenolic content of medicinal plant used in primary health care. Journal of Pharmaconosy Science. 9(1): 32-35

Souri, E., Amin, G., Farsam, H., Jalalzadeh, H. and Barezi, S. (2008). Screening of thirteen medicinal plant extract for antioxidant activity. Iranian Journal of Pharmacent Resource. 71: 149-154.

Taylor, J.E. (2000). Exotics. In: Seymour, G.B., Taylor, J.E., and Tucker, G.A (Eds); Biochemistry of fruit ripening pp 163-165.

Tlili, H. L., Lenucci, M.S. (2011). Bioactive compounds and antioxidant activities during fruit ripening of water melon cultivars. Journal of food composition and Analysis. 24:923-928.

Ureigho, U.N. (2010). Nutritive values of Chrysophyllum albidum linn Africa star apple as a domestic income plantation species. An International Multidisciplinary Journal. 4(2): 50-56.

46

Young, I. S and Woodside, J.V. (2001). Anti-oxidant in health and disease. Journal of Clinical Pathology. 54: 176-186

b.sc Project

Documents

Transcript of b.sc Project