CHAPTER ONE

1.0 INTRODUCTION

1.1 PHYTOCHEMICALS

The chemicals that are produced by plants are called as phytochemicals. These are produced

by the plant’s primary and secondary metabolism. These phytochemicals are important for

the plants to thrive or thwart other plants, animals, insects and microbial pests and pathogens.

They also help plants and protect them from disease and damage caused by environmental

hazards like pollution, UV, stress and draught. They are used as traditional medicine and as

poisons from ancient days (Vishnu Balamurugan et al., 2019).

Phytochemicals are not the essential nutrients they are rather than the essential nutrients

because there is no proof for them to cause any possible health effects in humans are not still

established. it is known that they have roles in the protection of human health. More than

4,000 phytochemicals have been catalogued and are classified by protective function,

physical characteristics and chemical characteristics. The phytochemicals are generally

classified into the following types; they include carotenoids and polyphenols which include

phenolic acids, stilbenes/ lignans. Which further have classifications like flavonoids are

further classified into flavones, anthocyanins, iso-flavones and flavanols (Markham, 1982).

Medicinal plants are a gift to us from the nature as they provide a number of health benefits

to us. In India these medicinal plants are used for about centuries for their properties and are

still used to this date. India has a variety of traditional medical systems like Ayurveda,

siddha, unani and a huge class of ethno-medicine (Ashi, 2003).

This knowledge of medicine was disappeared due to the western culture that has been on us

on the past and is reappearing again as their importance have been realized and lack of side

effects are also an important aspect in these types of traditional medicine. In wide-ranging

dietary phytochemicals are found in fruits, vegetables, legumes, whole grains, nuts, seeds,

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fungi, herbs and spices (Mathai, 2000). Broccoli, cabbage, carrots, onions, garlic, whole

wheat bread, tomatoes, grapes, cherries, strawberries, raspberries, beans, legumes, and soy

foods are common sources. Phytochemicals accumulate in different parts of the plants, such

as in the roots, stems, leaves, flowers, fruits or seeds. Many phytochemicals, particularly the

pigment molecules, are often concentrated in the outer layers of the various plant tissues.

Medicinal plants are very important in health care of individuals and communities in many

developing countries. Medicinal plants are believed to be much safer and are used in

treatment of various ailments. The plants provide the basic nutrients needed for the growth of

animals and humans like proteins, carbohydrates, fats, vitamins and oils minerals. The

phytochemicals are majorly classified as primary and secondary metabolites. The primary

metabolites are responsible for the basic development of the plant which includes the sugars,

amino acids, proteins, nucleic acids, chlorophyll, etc.

These compounds are known as secondary plant metabolites and have biological properties

such as antioxidant activity, antimicrobial effect, modulation of detoxification enzymes,

stimulation of the immune system, decrease of platelet aggregation and modulation of

hormone metabolism and anticancer property. There are more than thousand known and

many unknown phytochemicals. It is well-known that plants produce these chemicals to

protect themselves, but recent researches demonstrate that many phytochemicals can also

protect human against diseases (Narasinga, 2003).

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1.2 ANTIOXIDANTS

Antioxidants are naturally occurring plant substances that protect the body from damage

caused by harmful molecules called free radicals. Antioxidants help prevent oxidation, which

can cause damage to cells and may contribute to aging. They may improve immune function

and perhaps lower the risk for infection, cardiovascular disease, and cancer. Antioxidants

exist as vitamins, minerals and other compounds in foods. A diet containing plenty of fruits

and vegetables, whole grains and nuts can supply all the antioxidants your body needs. Diets

rich in antioxidants can be very beneficial (Kumar et al., 2008).

Antioxidants are compounds that inhibit oxidation. Oxidation is a chemical reaction that can

produce free radicals, thereby leading to chain reactions that can damage the cells of

organisms. Antioxidants, such as thiols or ascorbic acid (vitamin C) end these chain

reactions. To balance the oxidative state, plants and animals maintain complex systems of

overlapping antioxidants, such as glutathione and enzymes (e.g., catalase and superoxide

dismutase), produced internally, or the dietary antioxidants vitamins C and E (AOAC., 1984).

Antioxidant defences induction or endogenous ROS/RNS levels reduction is a rapid and clear

oxidative stress indicator. Indeed, ROS/RNS production and accumulation is a common

denominator in many disorders and environmental insults, at the same time that can cause

serious cell damage leading to physiological dysfunction and cell death in almost all aerobes.

Antioxidant therapy has long been investigated as a means of reducing the extent of injury

resulting from an ischemic stroke with varying degrees of success. (Svenja et al., 2021).

A few of the better known antioxidants include carotenoids (a form of vitamin A) — the

substance that gives fruits and vegetables their deep rich colours. Apricots, broccoli,

pumpkin, cantaloupes, spinach and sweet potatoes are good choices. Foods containing

vitamins C and E are also good sources of antioxidants, as well as selenium and zinc.

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Most Commonly Known Antioxidants & their food sources are:

 Carotenoids (a form of vitamin A) the substance that gives fruits and vegetables their

deep rich colours. May be effective allies against prostate cancer Apricots, peaches,

broccoli, pumpkin, cantaloupes carrots, spinach and sweet potatoes.

 Vitamin C enhances the immune response and protects against infection.

Citrus fruits like oranges and lime etc, green peppers, broccoli, green leafy vegetables,

strawberries and tomatoes

 Vitamin E May help prevent the oxidation of LDL or “bad” cholesterol which

contributes to plaque build-up in the arteries. Nuts & seeds, whole grains, green leafy

vegetables, vegetable oil and liver oil

 Selenium Fish & shellfish, red meat, grains, eggs, chicken and garlic

The above information was extracted from (AOAC., 1984).

Antioxidant enzymes made by the body:

superoxide dismutase (SOD), catalase, glutathione peroxidase

Benefits of Antioxidants

Protect Against Heart Disease, Protect Against Cancer, Protect Against Cancer, Boost

Immunity, Fight Aging.

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Fig 1: Image of some major sources of antioxidants (Mary, 2017).

5
1.3 Zingiber officinale (Ginger)

Zingiber officinale commonly known as ginger is one of the most commonly used spice

worldwide with medicinal value. It has been used extensively in various traditional and folk

medicinal systems around the world. Marinating or usage of ginger in food preparation is

useful in maintenance of the health as well prevention of food spoilage and reports indicate

that the antimicrobial effects contribute towards the observed effects. The ginger oil as a very

good antibacterial, antifungal property and prevents food borne diseases when used in food

preparation (Warrier, 1989).

Ginger is also reported to prevent rancidity, thereby increasing the shelf life of lipid

containing foods. The phytochemicals in ginger oil also possess free radical scavenging,

antioxidant and anti peroxidative effects. These properties are attributed to the plethora of

biologically active compounds present in the fresh as well dried ginger oils. The antioxidant

and lipid peroxidation inhibition properties of ginger prevent peroxidative damage, indicating

the benefits of ginger in prevention of microbial food spoilage, free radical-induced damage

and rancidity (Govindarajan, 1982a, b; Warrier, 1989).

1.3.1 Taxonomical Classification Zingiber officinale Roscoe

Kingdom: Plantae-Plants

Division: Magnoliophyta-Flowering plants

Class: Liliopsida-Monocotyledons

Subclass: Zingiberidae

Order: Zingiberales

Family: Zingiberaceae - Ginger family

Genus: Zingiber P. Mill. - Ginger

Species: Zingiber officinale Roscoe - Garden ginger.

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1.3.2 History of Zingiber officinale

Ginger (Figure 1), the root of the plant Zingiber officinale roscoe that belongs to the family

Zingiberaceae, is globally one of the most commonly used spice and medicinal agent. The

plant is known as Sringavera in Sanskrit and it is speculated that this term may have given

way to Zingiberi in Greek and then to the Latin term Zingiber (Vasala, 2004).

Historical evidence indicates that the plant was originally indigenous to the South-East Asia

(today’s Northeast India) but is today also found growing in other parts of the world

(Govindarajan, 1982a, b; Warrier, 1989). During the medieval times, ginger was exported

from India to other parts of the world. Today, ginger is cultivated in the other tropical

countries like Nigeria, Sierra Leone, Indonesia, Bangladesh, Australia, Fiji, Jamaica, Nepal,

Haiti, Mexico and Hawaii and today, India and China are the leading providers to the world

market (Govindarajan, 1982a, b; Warrier, 1989).

Ginger as commonly known, consists of the fresh or dried roots of Zingiber officinale.

(Ghosh et al., 2011). The English botanist William Roscoe (1753-1831) gave the plant the

name Zingiber officinale in an 1807 publication. The ginger family is a tropical group

especially abundant in Indo-Malaysia, consisting of more 1200 plant species in 53 genera.

The genus Zingiber includes about 85 species of aromatic herbs from East Asia and tropical

Australia. The name of the genus, Zingiber, was derives from a Sanskrit word denoting

"horn-shaped," in reference to the protrusions on the rhizome 2, 3. (Ghosh et al., 2011). The

first written record of ginger comes from the Analects of Confucius, written in China during

the Warring states period (475-221 BC). In it, Confucius was said to eat ginger with every

meal. in 406 AD, the monk Faxian wrote the ginger was grown in pots and carried on

Chinese ships to prevent scurvy. During the Song Dynasty (960-1279), ginger was being

imported into china from southern countries (Suarno et al., 1991).

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Ginger was introduced to the Mediterranean by the Arabs, and described by writers like

Diocorides (40-90) and Pliny the elder (24-70 AD) IN 150 AD, Ptoleny noted that ginger was

produced in Ceylon (Sri Lanka). Raw and preserved ginger was imported into Europe during

the Middlw Ages. In 14th century England, a pound of ginger cost as much as a sheep (Suarno

et al., 1991).

Figure 2: Image of Zingiber officinale (Suarno et al., 1991)

Ginger has been cultivated for thousands of years as a spice. It is an important cash crop in

India and is grown primarily in the states of Kerala, Karnataka and Northeast India

(Govindarajan, 1982a, b; Vasala, 2004). Of the Indian varieties, the Cochin and Calicut

ginger, have a lemon-like bye note and are popular (Govindarajan, 1982a, b; Vasala, 2004).

When compared to the Indian varieties, the Chinese ginger is low in pungency and is

principally exported as preserves in sugar syrup or as sugar candy (Govindarajan, 1982a, b;

Vasala, 2004).

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The yield and oil characteristic and content vary with cultivar and environmental factors.

There are many local varieties grown over the world. More than 400 accessions of ginger are

maintained at the Indian Institute for Spice Research in Calicut, Kerala, India (Vasala, 2004).

The following Indian cultivars are results of selection by the Indian Institute for Spice

Research with high yield and high oil content (Vasala, 2004). With respect to the African

varieties, the Jamaican ginger is highly popular basically due to its delicate aroma and fine-

textured powder, while the Nigerian and Sierra Leone dried ginger possess camphoraceous

and coarser odour and are rich in both aroma and pungency factors (Govindarajan, 1982a, b;

Vasala, 2004).

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1.4 Tetrapleura tetraptera

Tetrapleura tetraptera (Mimosaceae) is a perennial, single-stemmed plant with dark green

leaves. It is found in the rain forest belt of West Africa. The plant has many ethno-medicinal

and nonmedicinal uses such as anti-ulcer, anti-microbial, anti-convulsant, emulsifying,

contraceptive, and as a nutritive agent. Due to the foaming ability of the fruit, it is used in the

production of black soap. The dry fruit has a pleasant aroma that is insect-repellant in nature.

The use of the fruit as spices in foods is as a result of its medicinal value. There is wide

acceptance of medicinal plants in preference to synthetic drugs nowadays, because they are

cheaper and of little or no adverse effect.

Thus, there is great demand for natural antioxidants, because of lack of any undesirable

effect. The ethnomedicinal activity of the fruit extract of T. tetraptera have been investigated

by many authors with little or no work on the stem bark of the plant. The objective of the

present investigation therefore, was to evaluate, comparatively, the antioxidant potential of

different concentrations of the ethanolic extracts of the stem bark and fruit of T. tetraptera.

(Famobuwa et al., 2016).

Figure 3: Image of the pods of T. tetraptera (Margaret S., 1988).

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1.4.1 Botanical Classification of Tetrapleura tetraptera

 Kingdom - plantae

 Division - angiosperm

 Class - Eudicots

 Order - Fabales

 Family - Mimosaceae

 Genus - Tetrapleura

 Species - Tetrapleura tetraptera

1.4.2 Botanic Description

Tetrapleura tetraptera is deciduous; it reaches 20-25 m in height, with a girth of 1.5-3 m. The

bole is slender and older trees have very small, low, sharp buttresses. In the forest, the crown

is fairly small, thin and rounded, becoming flat when old, but it tends to spread when in the

open. Bark fairly smooth, grey-brown, very thin; slash reddish, strong smelling, fairly thick.

Twigs and young foliage virtually glabrous or minutely hairy.

Leaves are sessile, glabrous or minutely hairy with a common stalk 15-30 cm long, slightly

channelled on the upper surface. The pinnae are in 5-9 pairs, 5-10 cm long, mostly opposite

but sometimes alternate; 6-12 leaflets on each side of the pinna stalk, always alternate, 12-25

mm long, 6-12 mm broad, slightly elongated, elliptic or slightly obovate, rounded at both

ends, the apex sometimes very slightly notched, the base usually unequal, practically

glabrous, with slender stalks about 2 mm long; lateral nerves indistinct, running at a wide

angle to the prominent midrib.

Flowers are pinkish-cream turning to orange and are densely crowded in spike like racemes

5-20 cm long, usually in pairs in the upper leaf axils; individual flowers with slender stalks

and 10 short stamens, the anthers carrying a gland at the apex. Fruit is very persistent,

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hanging at the ends of branches on stout stalks 25 cm long. It is shiny, glabrous, dark purple-

brown, usually slightly curved, 15-25 cm long by about 5 cm across, with 4 longitudinal,

wing like ridges nearly 3 cm broad. Two of the wings are woody, the other 2 filled with soft,

sugary pulp, oily and aromatic.

The seeds, which rattle in the pods, are small, black, hard, flat, about 8 mm long, embedded

in the body of the pod, which does not split open. In Nigeria, the tree is deciduous in

December. Flowering begins towards the end of February and is over in early April. The

indehiscent pods are ripe from September to December. When the pods fall, their smell

attracts small animals, who probably disperse the seeds. Agroforestry Database (Orwa et al.,

2009).

1.5 AIMS AND OBJECTIVES

Aim

The aim of this study is to determine the phytochemical content and antioxidant activities of

methanol fruit extract of Zingiber officinale and Tetrapleura tetraptera.

Objectives

 To collect samples of Zingiber officinale and Tetrapleura tetraptera from the garden

 To transport the collected samples to the laboratory for evaluation.

 To determine the phytochemical components of Zingiber officinale and Tetrapleura

tetraptera.

 To determine the antioxidant of Zingiber officinale and Tetrapleura tetraptera.

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 CHAPTER TWO

2.0 LITRATURE REVIEW

2.1 BACKGROUND STUDY

Man’s acquaintance with the medicinal properties of plants is of great antiquity. Even the

higher mammals are said to be aware of the curative aspects of plant kingdom. Plants have

been used in a number of systems of medicines in our country as well as in other countries.

India is well known as the ‘Emporium of Medicinal Plants’.

The use of plants to treat various diseases in India dates back to the times of Rig Veda (3500

to 1800 B.C.). Later, the monumental Ayurvedic works like Charaksamhita and

Sushrutasamhita followed by other Ayurveda and Siddha treatises have incorporated nearly

700 plant drugs entering into several medicinal preparations used in the management of

health care. In fact, these systems have been in practice even in remote areas of our country

for centuries

2.2 Zingiber officinale

Abuse of antibiotic feed additives could poison animals, deposit undesirable residues in

animal products, cause microbial resistance to drugs and pollute the environment (Butaye et

al., 2003; Demir et al., 2005; Amlund et al., 2012). Due to these side-effects, in 2006, the

European Union banned the use of antibiotic growth promoters in animal feed (Mayer, 2020).

Since then, researchers and farmers began examining alternatives to antibiotics to provide

benefits of antibiotics without side-effects (Verstegen and Williams, 2002). These non-

antibiotic feed additives (natural growth promoters) include plant parts or extracts and live

beneficial microbes (Wenk, 2000; Verstegen and Williams, 2002).

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Examples of non-antibiotic growth promoters include acidifiers, microbials, feed enzymes,

immunity modulators, prebiotics, probiotics, feed cleaners, vitamins, micro-nutrients,

anticoccidials, anthelmintic, antioxidants, minerals and phytobiotics (Verstegen and

Williams, 2002; Menegat et al., 2019). Phytobiotics or phytogenics are various plant-derived

products, in powder or liquid oil forms, with pungent or sweet-smelling aroma, obtained from

leaves, barks, fruits, flowers, seeds, nuts, roots and woody parts of plants, added to feed to

improve livestock performance through amelioration of feed properties, improving health and

quality of food from the animals. They also demonstrate antimicrobial, antifungal,

coccidiostatic, anthelmintic, antiinflammatory, antiviral, antioxidant or sedative activity.

Some have flavouring and appetizing effects by increasing palatability of feed and enhance

animal immune system (Windisch et al., 2008).

They also enable beneficial gastrointestinal microbes to flourish. Content and concentration

of active substances in phytobiotics differ extensively dependent on plant, plant part, place of

origin, season of harvest, storage conditions and processing techniques. Active secondary

metabolites in phytogenics include isoprine derivatives, flavonoids and glucosinolates

(Windisch et al., 2008). These additives must be used in specified amount and form and

properly screened to assure expected results. Suitable candidates must be proven, cost-

effective, fit for the circumstances, available at farm level and at quantity needed for long-

term use (Verstegen and Williams, 2002; Karaskova, et al., 2015). Also, long-term

experimental use is needed to prove their efficacy and safety (Karaskova et al., 2015).

Zingiber officinale pod extract and powder singly or in combination with other non-antibiotic

growth promoters have been tested on performance, blood chemistry, and anti-microbial

activity in albino rats and broiler chickens (Nweze et al., 2011; Adeyemo, 2014; Olorunleke

et al., 2016; Kana et al., 201). Similar studies on rabbits are rare. Assessing hepatotoxic

effect of 10 days oral administration of ethanolic extract of Aidan on male rabbits, Odesanmi

14
et al., (2009) reported that Aidan treatment increased serum AST, total protein, direct

bilirubin and alkaline phosphate but decreased ALT as dose increased without obvious

pathological lesions in the liver. This study therefore evaluates the response of rabbit bucks to

diets containing Aidan pod powder as feed additive.

Nigerian ginger is mainly produced utilizing traditional methods, using manual labor in order

to plant, harvest, and process the ginger. Recently, there have been efforts made in order to

mechanize the process and remove human elements from production. More than 90% of the

ginger is sun-dried, but with the incorporation of dryer machines, the processing time is

reduced from 4-10 days to 4-5 hours. These drying machines can significantly increase the

quality of the products, but with the burden of extra costs. With Nigeria’s expansion into the

international market, quality control has been a consistent issue, and measures are being

taken in order to raise the quality of the products.

Emmanuel, (2008) opined that Nigeria has the potential to expand production in a medium to

long-term investment strategy that can develop into self-sufficient industry (FAO, 2010).

Ginger oil has been reported to possess antimicrobial effects and studies by Natta and co-

workers (2008) have shown that the essential oil of ginger extracted by hydro-distillation

possess high antibacterial effects on food pathogens (S. aureus, B. cereus and L.

monocytogenes), with a minimum concentration to inhibit B. cereus and L. monocytogenes of

6.25 μg/ml (Natta et al., 2008). Subsequent studies have shown that the oil extracted from the

leaf and rhizome were moderately active against the Gram-positive Bacteria Bacillus

licheniformis, Bacillus spizizenii and Staphylococcus aureus, and the Gram-negative bacteria

Escherichia coli, Klebsiella pneumoniae and Pseudomonas stutzeri (Sivosathy et al., 2011).

15
Studies with the gram-positive bacteria, Bacillus subtilis (NCIM 2162), Staphylococcus

aureus (NCIM 2602), Micrococcus luteus (NCIM 2704), and gram-negative bacteria,

Escherichia coli (NCIM 2576), Pseudomonas aeruginosa (NCIM 2200), Proteus vulgaris

(NCIM 2813), Klebsiella pneumoniae (NCIM 2957) have also shown ginger oil to be

effective (Sayyad and Chaudhari, 2010). The results indicate that the antibacterial effects

were as follows: Bacillus subtilis > Staphylococcus aureus > Escherichia coli = Proteus

vulgaris > Pseudomonas aeruginosa > Micrococcus luteus > Klebsiella pneumoniae

(Sayyad and Chaudhari, 2010). Ginger oil has also been shown to possess antibacterial

effects on the growth of psycrotrophic food-borne bacteria (Fabio et al., 2003).

On a comparative note, recent studies by Sasidharan and Menon, (2010) have shown that the

fresh ginger oil was more effective than the dry ginger in inducing the antimicrobial effects

on Aspergillus niger, Candida and Pseudomonas aeruginosa, weaker towards Saccharomyces

cerevisiae and inactive against Bacillus subtilis, Pencillium spp and Trichoderma spp; while

the dry ginger oil was more active towards Pseudomonas aeruginosa , on par with standard

towards Candida, weaker than standard against Bacillus subtilis, Aspergillus niger,

Pencillium spp, Saccharomyces cereviseae. Fresh ginger oil had an MIC value of <1 μg/mL

against Aspergillus niger and Candida albicans and. dry ginger oil had an MIC value of less

than 1 μg/mL against Pseudomonas aeruginosa, Pencillium spp and Candida albicans.

Fresh ginger is more abundant in oxygenated compounds and the observed variance in the

antimicrobial effects is possibly due to this (Sasidharan and Menon, 2010). Nanasombat and

Lohasupthawee, (2005) studied the antibacterial effects of the fresh ginger oil in standardized

twenty five bacterial strains (twenty serotypes of Salmonella and five species of other

enterobacteria) commonly involved in the spoilage of food and those to be associated with

food borne illness and observed that the antibacterial effects were as follows Serratia

16
marcescens> Klebsiella pneumoniae = S. Typhimurium DT104 (8748A-1) > Escherichia coli

(DMST 4212) > S. Anatum (DMST 7108) = S. Choleraesuis ssp. Choleraesuis (DMST 8014)

= S. Derby (DMST 8535) = S. Enteritidis (DMST 10633) = S. Hadar = S. Newport (DMST

7101) = S. Newport (DMST 7101) = S. Orion (SAP 08991/02) = S. Rissen (DMST 7097) = S.

Senftenberg (DMST 7113) = S. Typhimurium (DMST 0562, non-DT104 strain) =

Citrobacter freundii (DMST 1959) > S. Virchow (DMST 10635) = Salmonella Agona

(DMST 10338).

The essential oil extract of ginger showed significant reduction in total vial count,

Staphylococcus, E. coli and Salmonella counts at dilution of 1: 150 and 1: 250 than the

dilution of 1:500. The dilution at 1:150 is the best dilution for the effective reduction of

bacterial counts for both Gram positive cocci (Staphylococcus) and Enterobacteriaeceae (E.

coli and Salmonella) which are the main contaminants seen in the poultry meat. But, the

aqueous extract of ginger was not effective in reduction of microbial counts (Sudharshan et

al., 2010).

2.3 Tetrapleura tetraptera

It is used extensively in soups of nursing mothers to prevent post-partum contractions and

gasintestinal disorders especially stomach ulceration (Atawodi et al., 2014; Nwawu and Alah,

1986; Noamesi et al., 1992). In cold weather, it is used to prepare pepper soup and the aroma

is believed to drive away snakes (Abii and Amarachi, 2007). The fruit has wide application in

Nigeria folk medicine. It is used extensively in the management of an array of human

ailments including diabetes mellitus, arthritis, hypertension, epilepsy, asthma, etc. (Abii and

Amarachi, 2007; Akin-Idowu et al., 2011).

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The fruit is said to contain caffeic acid which serves as HIV replication inhibitor and also

inhibit antitumor and inflammatory characteristic (Adesina, 1982). Although much

researches have been done on this plant Tetrapleura tetraptera, particularly on its dry fruits to

assess the phytochemicals, minerals and nutrients contents (Abii and Amarachi, 2007;

AkinIdowu et al., 2011; Adewunmi, 2001; Adebayo et al., 2000; Adesina, 1982; Antwi-

Boasiko and Animapauh, 2012; Essien et al., 1994; Noamesi et al., 1992), reports on the

essential oils and fatty acids composition of the fruit has been scarce.

This work examines the essential phytochemistry and antioxidants composition of the dry

fruits, to ensure full exploitation of its therapeutic properties and health benefits.

Molluscicidal activity of Tetrapleura tetraptera was also conducted on methanolic extractive

of the fruit at a concentration of 7.5 – 60 mg/litre, which caused a slowing of the heart of

intact Biomphelaria glabrata which was found to be dose and time dependent. Also the water

extract of the fruit was applied at a concentration of 15, 20 and 25mg/litre in selected water

contact sites in three villages near Ile-Ife, Nigeria over a period of 24 months. Snail surveys

were carried out in control and treated locations to assess the effectiveness of the extract.

The difference in snail numbers between pre-treatment snail population and post treatment snail

population was statistically significant (P < 0.004), an indication that the extract was able to

control the snails at the treated sites only. It was also found out that Lymnaea natalensis was not

detected in any of the treated sites (Adewunmi, 1984). It is important that the effect of this plant

molluscicide be shown on non-target organisms before it could be recommended for further

development and the chicken happened to be one of these non-target animals. It was equally

shown that Tetrapleura tetraptera had little or no toxic effect on the weight gain and/or blood

values of the domestic chicken (p>0.05).

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Blood values analysis for RBC and WBC when compared with exotic breed values recorded in

temperate countries raises the need to carry out research into normal blood values of Nigerian

domestic chicken and animals. The results indicated that Tetrapleura tetraptera fruit aqueous

extract possesses anti-inflammatory and hypo-glycaemic properties. These findings lend

pharmacological credence to the suggested folkloric uses of the fruit of the plant in the

management and/or control of arthritis and other inflammatory conditions and adult-onset type 2

Diabetes mellitus in Yoruba speaking communities of South-West Nigeria (Ojewole et al., 2004).

Adewunmi, (2004/2005) in his recent scientific appraisal of Tetrapleura tetraptera itemised

the potential uses of this plant with some of the corresponding phytochemicals; as

molluscicide, anti-ulcer, antimicrobial, anticonvulsant, emulsifying property, birth control,

intuitive activities. Amoako-Atta et al., (2004/2005) upgraded the value of the indigenous

biological food resources that abound in Ghana.

In 1999, the Centre for Biodiversity Utilization and Development (CBUD) was established.

This Dutch-funded programme has emerged as a Centre of Excellence with the mission to co-

ordinate the process of identification of potential products of Ghana’s biodiversity and to

subsequently support and facilitate their production, processing and marketing. It has pursued

this process of domestication and product chain development with five lesser-known food

resources – snails, indigenous leafy vegetables, Tetrapleura tetraptera known for its

medicinal and food value called “Prekese”, a commercial product of Natu-Bi Preserve

Establishment, a fruit juice canning company has made soft drinks called “Prekese” fruit (T.

tetraptera) which is proclaimed/served as medicinal drink especially for the cure of

hypertension; grass-cutter (Thryonomis swenderianus, a popular bush meat) and Telfairia

occidentalis.

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It is worthy of note that Institutional arrangements already involved forty partner institutions

across Ghana, and over 1,500 agricultural producers have taken up the production of the

commodities promoted by the Centre throughout Southern Ghana. A review of plants used

for poison fishing in tropical Africa was done by Neuwinger, H.D., (2004) in which 325 fish-

poisoning plants, spread among 71 plant families with 183 genera were presented. The

closely related groups of Caesalpiniaceae, Mimosaceae and Papilionaceae clearly dominate.

It was also remarkable that a great proportion were Euphorbiaceae. The plants most used are

Tephrosia vogelii, Mundulea sericea, Euphorbia tirucalli, Gnidia kraussiana, Adenia lobat,

Balanites aegyptiaca, Swartzia madagascariensis, Neoratanenia mitis, Tetrapleura

tetraptera and Strychnos aculeate. It was shown that many fishing poisons play an important

part in the preparation of arrow poisons and in traditional medicine as fishing with the aid of

plant poisons has a long tradition all over the world and is still used in many places in the

world today.

A very recent work was done by Aderibigbe et al., (2006) in which Aridanin isolated from

Tetrapleura tetraptera fruit was investigated for anticonvulsant, analgesic and hypothermic

activities in mice. The results suggested that aridanin could be acting as a central nervous

system (CNS) depressant and that its anticonvulsant property may be mediated through the

membrane stabilizing property while the analgesic and hypothermic actions were mediated

through opioids, cholinergic and 5-HT receptors respectively. In a dosely related work of

Aderibigbe et al., (2006), aridanin was evaluated for neuropharmacological activity on

novelty-induced behaviours such as locomotory, exploratory, stereotyped and hexobarbitone-

induced sleeping time in mice. The results suggested that aridanin has a strong sedative and

central depressant action but lacks psychopharmacological activities.

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 CHAPTER THREE

3.0 FIELD WORK

3.1 PLANT COLLECTION
A 1.5 kilogram of Zingiber officinale rhizomes and a twenty-five ripe fruits of Tetrapleura

tetraptera, were bought from Oyingbo Local Government Area of Lagos, Nigeria. The fruits

and roots of this specimens, were botanically identified and authenticated at the herbarium of

the Department of Botany, University of Lagos, Nigeria. The herbarium number for Zingiber

officinale was 8696 and the one for Tetrapleura tetraptera, was 8703 both were identified on

the fourth of November, 2020, at exactly 11:25am.

3.2 LABORATORY WORK

The plants were transported to the laboratory where the evaluation of the contents present in
them were to be work on.

3.2.1 Qualitative Analysis Preparation of the extracts

The Zingiber officinale was piled and chopped into smaller units and the Tetrapleura

tetraptera was grounded, both were oven dried at a room temperature in the laboratory for

three to four weeks (1 month). The dried samples were individually milled into a powder and

the resulting powder was screened with a sieve having 0.5Mm mesh size. 258.2g of the

samples was weighed and was infused into 1litre of methanol solution the mixture was

vigorously stirred and allow to stand for 72hours. The solution was now filtered using a

watsman’s filter and the solvent was then removed and concentrated at 40degree Celsius

using rotary evaporator, the extracts were stored in a refrigerator at 4degree Celsius until

required for usage. The yield of the dried extract, 79.07g was calculated using the expression

as follow;

Yield (%) = weight of dried extract ×100

Weight of sample used

21
3.2.2 Qualitative analysis of phytochemical compounds

Qualitative analysis was carried out to ascertain the presence of the different phytochemical

compounds contained in the extract. Preliminary phytochemical analysis of the extracts was

carried out with reference to the standard methods of Sofowora (1998) and Evans (2009).

Test for Alkaloids

The extracts were dissolved individually in dilute hydrochloric acid and filtered. The filtrates

were used to test the presence of alkaloids using dragendroffs test.

Dragendroff’s Test: Filtrates were treated with Dragendroff’s reagent (solution of Potassium

Bismuth Iodide). Formation of red precipitate indicates the presence of alkaloids.

Test for Flavonoids

Lead acetate test: Extracts were treated with few drops of lead acetate solution. Formation

of yellow colour precipitate indicates the presence of flavonoids in the extracts.

Test for Steroids

2ml of acetic anhydride was added to 5mg of the extracts each with 2ml of H2SO4. A colour

was changed from violet to blue or green in some samples indicates the presence of steroids

in the extract.

Test for Terpenoids

Salkowski s test: 5mg of the extract of the leaves, flowers and seeds was mixed with 2ml of

chloroform and 3ml concentrated H2SO4 was carefully added to form a layer. An appearance

of reddish brown colour in the inner face indicates the presence of terpenoids.

22
Test for Phenols

Ferric chloride test: 5ml extracts were treated with few drops of ferric chloride solution.

Formation of bluish black colour indicates that the presence of phenol.

Test for Saponins

0.5mg of the extract was shaken with 5ml of distilled water. Formation of frothing

(appearance of creamy mass of small bubbles) shows that the presence of saponins.

Test for Tannins

A small quantity of extract was mixed with water and heated on a water bath. The mixture

was filtered and Iron (III) chloride was added to the filtrate. Formation of a dark green colour

indicates the presence of tannins.

Test for reducing sugar

Benedict’s test

Equal volume (2ml each) of Benedict’s solution and aqueous extract were mixed in a test

tube and heated in boiling water bath for 10min the changes in colour to yellow, green and

red indicates the presence of reducing sugar.

3.3 QUANTITATIVE PHYTOCHEMICAL SCREENING

Estimation of Alkaloids

Total alkaloid was measured using methods as described by Shamsa et al., (2007) with slight

modification. 1ml plant extract, 5 ml pH 4.7 phosphate Buffer was added and 5 ml BCG

solution and the mixture was shake with 4 ml of chloroform. The extracts were collected in a

10-ml volumetric flask and then diluted to adjust volume with chloroform. The absorbance of

the complex in chloroform was measured at 470 nm against blank prepared as above but

without extract. Atropine is used as a standard (using calibration curve) and compared the

assay with Atropine equivalents.

23
Preparation of Standard Curve Accurately measure aliquots (0.4, 0.6, 0.8, 1 and 1.2 mL) of

atropine standard solution and transfer each to different separatory funnels. Add 5 mL pH 4.7

phosphate buffer and 5 mL BCG solution. Shake mixture with 1, 2, 3 and 4 mL of

chloroform. The extracts were collected in a 10 mL volumetric flask and then diluted to

volume with chloroform. The absorbance of the complex in chloroform was measured at 470

nm against blank prepared as above but without atropine.

Estimation of Steroids

Total steroid was measured using methods as described by Salomi et al., (2019) with slight

modification. 1ml of test extract of steroid solution was transferred into 10ml volumetric

flasks, Sulphuric acid (4N, 2ml) and iron (III) chloride (0.5% w/v, 2 ml), were added,

followed by potassium hexacyanoferrate (III) solution (0.5% w/v, 0.5 ml). The mixture was

heated in a water-bath maintained at 70±2 0C for 30 minutes with occasional shaking and

diluted to the mark with distilled water. The absorbance was measured at 780 nm against the

reagent blank. Total steroids in extracts was expressed in terms of cholesterol equivalents

(mg of CHO/g of extract).

Estimation of Flavonoids

Aluminium chloride colorimetric method (Chang et al.,2002) with some modifications was

used to determine flavonoid contents. Plant extract (1mL) in methanol was mixed with 1ml of

methanol, 0.5 mL aluminium chloride (1.2%) and 0.5 mL potassium acetate (120 mM). The

mixture was allowed to stand for 30 min at room temperature; then the absorbance was

measured at 415 nm. Quercetin was used as standard. Flavonoid content is expressed in terms

of quercetin equivalent (mg g-1 of extracted compound).

24
Estimation of Total Phenols

The Follins method described by pearson (1979) was used to determine the phenol contents.

1 ml of the extract was placed in a test tube, 2.5ml of 10% Folin-Ciocalteu’s reagent

dissolved in water and 2.5 ml of 7.5% of NaHCO3 aqueous solution were added. The

samples were thereafter incubated in a thermostat at 45oC for 45 min. The absorbance was

determined using spectrophotometer at wave length = 765 nm.

The samples were prepared in triplicate for each analysis and the mean value of absorbance

was obtained. The same procedure was repeated for the standard solution of Zingiber

officinale and garlic acid and the calibration line was construed. Based on the measured

absorbance, the concentration garlic acid equivalent expressed in terms of (mg of GA/g of

extract).

Estimation of Tannins

Tannin contents was determined by the Folin-Denis colorimetric method described by Kirk

and Sawyer (1998). 5g sample was dispersed in 50mls of distilled water and shaken. The

mixture was allowed to stand for 30min at 28oC before it was filtered through whatman No.

42 grade of filter paper. 2mls of the extract was dispersed into a 50ml volumetric flask.

Similarly, 2ml standard tannin solution (tannic acid) and 2ml of distilled water were put in

separate volumetric flasks to serve as standard and reagent was added to each of the flask

and the 2.5ml of saturated Na2C03 solution added. The content of each flask was made up to

50ml with distilled water and allowed to incubate at 28oC for 90 min. Their respective

absorbance was measured in a spectrophotometer at 765nm using the reagent blank to

calibrate the instrument at zero.

25
DNS reducing sugar assay

This assay is based on the protocol described by Hussain et al., (2018) and miller (1978) with

slight modification. 1ml of the plant samples was taken and 3ml of DNSA was added and

boiled for 10min and absorbance was taken at 540nm. Glucose was used as standard

(100mg/ml). DNS reagent was prepared by mixing 1.6 g NaOH and 1.0 g dinitrosalicylic acid

(Sigma) in 70 mL dH2O and the mixture was heated in boiling water to dissolve. Once

dissolved, 3.0 g Na2K tartrate (Sigma) was added to the solution and swirled until dissolved

followed by further addition of dH2O to make up to 100 mL. The reagent was stored dark at

room temperature. 1ml of the plant samples is taken and 3ml of DNSA was added and boiled

for 10min and absorbance was taken at 540nm. Glucose was used as standard (100mg/ml).

3.4 IN VITRO ANTIOXIDANT ASSAYS

Reducing power assay (Oyaizu, 1986)

The reducing powers was determined according to the method with slight modifications.

Reaction was carried out in a mixture containing 1 ml of sample (25-100 µg/ml), 2.5 ml of

0.1 M sodium phosphate buffer (pH 6.6) and 2.5 ml of 1%, w/v potassium ferro cyanate

[K3Fe(CN)6] by incubating at 50°C for 20 min. After addition of 2.5 ml trichloroacetic acid

(10%, w/v), the mixture was centrifuged at 5000rpm for 10 min. The upper layer (5 ml) was

mixed with 0.5 ml of fresh FeCl3 (0.1%, w/v), and the absorbance at 700 nm was measured

against a blank. Gallic acid was used as the control.

DPPH radical scavenging assay (Shimada et al., 1992)

Briefly 0.1 Mm solution of DPPH in ethanol was prepared; 1ml of the solution was added to

1 ml of extract in water at different concentrations (25-100 μg/ml). The mixture was shaken

vigorously and allowed to stand at room temperature for 30 min. Then the absorbance was

measured at 517 nm by using a UV-Visible Spectrophotometer. Lower absorbance of the

26
reaction mixture indicated higher free radical scavenging activity. The percent DPPH

scavenging effect was calculated using the following equation:

DPPH Scavenging effect (%) = [(A0-A1)/A0] x 100

Where A0 was the absorbance of the control and A1 was the absorbance in the presence of the

standard sample or extract. The IC50 value represented the concentration of the compounds

that caused 50% inhibition of DPPH radical formation.

NITRIC OXIDE (GRIESS REAGENT)

Sodium nitroprusside (10mM) in phosphate buffer saline was mixed with different

concentrations (25-100 µg/ml) of each extract, and incubated for 30min, after incubation

period, 0.5ml of Griess reagent (1% sulfanilamide, 2% H2PO4 and 0.1% N-(1-naphthyl)

ethylenediamine dihyrochloride was added. Absorbance was taken at 550nm and ascorbic

acid was used as standard. IC50 which is an inhibitory concentration of each extract required

to reduce 50% of nitric oxide formation was determined. The same reaction mixture without

the extract but equivalent amount of methylated spirit was used as control.

Nitric oxide (inhibition %) = A (control) – A (sample) x 100

A (control)

27
 CHAPTER FOUR

4.0 RESULTS

4.1 QUALITATIVE AND QUANTITATIVE ANALYSIS

Table 1: Result of phytochemical screening (Qualitative Analysis) for the methanol

extracts of Zingiber officinale and Tetrapleura tetraptera rhizomes (Qualitative Analysis).

Specimens Reducing Saponin Phenol Flavonoid Tannin Alkaloid Steroid Terpernoid

sugar
Tetrapleura + - + + + + + +
tetraptera
Zingiber + - + + + + + +
officinale

Key: + present

- Absent

Table 2: Percentage (%) results of phytochemical parameters of the methanol extract of

Tetrapleura tetraptera pods and Zingiber officinale rhizomes (Quantitative Analysis).

Specimens Reducing Saponin Phenol Flavonoid Tannin Alkaloid Steroid Terpernoid

sugar (mg/100g) (mg/100g) (mg/100g) (mg/100g) (mg/100g) (mg/100g) (mg/100g)
(mg/100g)
Tetrapleura 42.71 - 87.67 36.69 63.77 33.63 31.30 17.49
tetraptera
37.10 - 94.93 36.24 69.05 33.19 30.89 17.73
Zingiber 18.49 - 85.87 43.60 62.46 102.83 18.12 17.40
officinale
17.99 - 85.17 45.69 61.95 103.10 18.53 17.08

28
 4.2 ANTIOXIDANT ESSAY RESULTS

Table 3: Results of Anti-oxidant activity of Tetrapleura tetraptera fruits and Zingiber

officinale rhizomes (REDUCING POWER).

Specimens 25µg/ml 50µg/ml 75µg/ml 100µg/ml

Tetrapleura tetraptera 0.139 0.228 0.288 0.356

0.141 0.230 0.290 0.359

Zingiber officinale 0.127 0.144 0.151 0.226

0.129 0.141 0.153 0.227

ASCORBIC ACID 0.242 0.382 0.481 0.624

0.233 0.379 0.485 0.626

Table 4: Results of Anti-oxidant activity of Tetrapleura tetraptera fruits and Zingiber

officinale rhizomes (DPPH) (Antioxidant Essay Results).

Specimens 25µg/ml 50µg/ml 75µg/ml 100µg/ml

Tetrapleura tetraptera 36.67 56.17 67.62 83.54
36.14 54.92 66.37 83.18
Zingiber officinale 20.04 43.47 56.89 74.42
20.39 42.40 57.60 73.52
ASCORBIC ACID 45.71 62.29 78.64 86.39
46.58 61.47 79.57 87.61

29
 Table 5: Results of Nitric oxide Tetrapleura tetraptera fruits and Zingiber officinale
rhizomes

Specimens 25µg/ml 50µg/ml 75µg/ml 100µg/ml

Zingiber officinale 23.31 45.73 63.70 78.47

22.06 45.20 63.17 78.83

Tetrapleura tetraptera 31.67 49.29 64.23 82.74

32.56 48.22 63.70 83.81

ASCORBIC ACID 47.65 63.51 76.89 85.44

48.13 62.67 75.25 84.38

Table 6: Results of Total Anti-Oxidant Capacity Tetrapleura tetraptera pods and

Zingiber officinale rhizomes

TOTAL ANTIOXIDANT CAPACITY

Specimens Total Antioxidant Capacity
mg/100g
Tetrapleura tetraptera 51.23

50.57

Zingiber officinale 65.20

65.77

30
4.3 STATISTICAL RESULTS
Table 7: Statistical data analysis of results of Tetrapleura tetraptera fruits and Zingiber
officinale rhizomes

Row stats of Grp

Tetrapleura tetraptera Zingiber officinale
Mean SD Mean SD
Reducing Sugar 39.91 3.97 18.24 0.35
Phenol 91.30 5.13 85.52 0.49
Flavonoid 36.47 0.32 44.65 1.48
Tannin 66.41 3.73 62.21 0.36
Alkaloid 33.41 0.31 102.97 0.19
Steroid 31.10 0.29 18.33 0.29
Terpenoid 17.61 0.17 17.24 0.23
TAC 50.90 0.47 65.49 0.40

150
Z in g ib e r o ffic in a le
T e tr a p le u ra te tra p te ra

100
m g /1 0 0 g

50

0
r

C
id
id

in
o

id
a

id

A
g

lo
o

ro

T
u

n
h
S

te
o

lk

e
P

T
v
g

rp
S
A
la
in

e
F
c

T
u
d
e
R

P h y t o c h e m ic a ls

Fig 4: Bar charts results of the phytochemical screening.

31
Table 8: The Mean and Standard Deviation of Reducing Power of Zingiber officinale

rhizomes and Tetrapleura tetraptera pods (Antioxidant)

Row stats of Reducing Power

Zingiber officinale Tetrapleura tetraptera ASCORBIC ACID

Mg/100g Mean SD Mean SD Mean SD

25 0.128 0.001 0.140 0.001 0.238 0.006

50 0.143 0.002 0.229 0.001 0.381 0.002

75 0.152 0.001 0.289 0.001 0.483 0.003

100 0.227 0.001 0.358 0.002 0.625 0.001

Table 9: The Mean and Standard Deviation of DPPH data analysis of Zingiber
officinale rhizomes and Tetrapleura tetraptera pods (Antioxidant)

Row stats of DPPH

Zingiber officinale Tetrapleura tetraptera ASCORBIC ACID

Mg/100g Mean SD Mean SD Mean SD

25 20.22 0.25 36.41 0.37 46.15 0.62

50 42.94 0.76 55.55 0.88 61.88 0.58

75 57.25 0.50 67.00 0.88 79.11 0.66

100 73.97 0.64 83.36 0.25 87.00 0.86

32
Table 10: The Mean and Standard Deviation of Nitric Oxide data analysis of Zingiber

officinale rhizomes and Tetrapleura tetraptera pods (Antioxidant)

Row stats of NITRIC

Zingiber officinale Tetrapleura tetraptera ASCORBIC ACID

Mean SD Mean SD Mean SD

25 22.69 0.88 32.12 0.63 47.89 0.34

50 45.47 0.37 48.76 0.76 63.09 0.59

75 63.44 0.37 63.97 0.37 76.07 1.16

100 78.65 0.25 83.27 0.76 84.91 0.75

0 .8
R e d u c in g P o w e r a c t iv ity

A S C O R B IC A C ID
T e tr a p le u ra te tra p te ra
0 .6
Z in g ib e r o ffic in a le

0 .4

0 .2

0 .0
5 0 5 0
2 5 7 0
1

C o n c e n t r a t io n ( µ g /m l)

Figure 5: Simple graph chart showing the antioxidant (REDUCING POWER) values of

the Zingiber officinale roots and Tetrapleura tetraptera pods.

Value expressed as mean + standard error of mean (SEM)

33
 D P P H S c a v e n g in g a c t iv ity 100
A S C O R B IC A C ID

80 T e tr a p le u ra te tra p te ra
( % In h ib it io n )

Z in g ib e r o ffic in a le
60

40

20

0
5 0 5 0
2 5 7 0
1

C o n c e n t r a t io n ( µ g /m l)

Figure 6: Simple graph chart showing antioxidant (DPPH) values of the constituent in

Zingiber officinale roots and Tetrapleura tetraptera pods

Value as expressed as mean + standard error of mean (SEM).

N itr ic o x id e s c a v e n g in g a c t iv ity

100
A S C O R B IC A C ID

80 T e tr a p le u ra te tra p te ra
( % In h ib it io n )

Z in g ib e r o ffic in a le
60

40

20

0
0
5

0
2

C o n c e n t r a t io n ( µ g /m l)

Figure 7: Simple graph chart showing antioxidant (NITRIC OXIDE) values of the

constituent in Zingiber officinale rhizomes and Tetrapleura tetraptera pods.

Value as expressed as mean + standard error of mean (SEM).

34
 CHAPTER FIVE

5.0 DISCUSSION

Ginger is a rhizomatous plant grown throughout South-eastern Asia and China and in parts of

Japan, Austria, Latin America, Jamaica, and Africa. Ginger has been used as a spice and

medicine in the Indian subcontinent since ancient times. Its medicinal values have been

known for centuries. It is the most widely used condiment, flavoring, and garnishing agent.

The herb serves as a stimulant and carminative and is used in dyspepsia and colic. It is known

to have blood thinning and cholesterol lowering properties, due to which it is used in treating

heart diseases.

The major phenolic compounds and essential oils act as potent antioxidant and exhibit free

radical scavenging properties. The antimicrobial properties are due to the presence of

componentssuch as thymol, eugenol, 1, 8‐ cineole, α‐ and β‐pinenes, linalool, and α‐terpineol.

Ginger tea is considered a good home remedy for cold. The herb can also used to treat

arthritis, diarrhea, motion sickness, diabetes, bronchitis, and rheumatism. It is a remedy for

nausea due to seasickness, morning sickness, and chemotherapy. Overall, ginger is a versatile

herb with phenomenal phytotherapeutic and medicinal properties. It would be difficult to find

a place or nation on this globe that has not been benefited through this extraordinary aromatic

herb.

5.1 QUANTITATIVE DETERMINATION

The quantitative assay was performed for the phytochemicals (phenols, flavonoids, saponins,

alkaloids), antioxidant methanolic extracts using UV-Vis Spectrophotometer with standard

procedures. the highest phytochemical concentrations corroborate the findings by Nwoba

(2015) and Uyoh et al., (2013); who estimated the concentration of phenols, flavonoids,

saponins and alkaloids in the pulp and seeds of the same plant species.

35
Phytochemical screening of the Zingiber officinale revealed the presence of phenolic and

flavonoid. Flavonoids have been shown to have antioxidant, antibacterial anti-inflammatory,

antiallergic activity etc. The presence of phenolic and flavonoids in the drug extract is likely

to be responsible for the antioxidant activity. These compounds are reported to be antioxidant

or free radical scavengers. The gingerols, shogoals, which are a homologous series of

phenols. The essential oil, which is a mixture of monoterpenic and sesquiterpenic

compounds, might be contributed to its antioxidant potential of the drugs.

However, this present study has unveiled the concentrations of these major phytochemicals in

the Tetrapleura tetraptera pod, with less data in literature. Per gram of each extracts, alkaloid

seems to be the most abundant bioactive compound having the highest concentration in

methanolic extracts with flavonoids being the least. Alkaloids have been reported to function

as defensive elements against predators, especially mammals because of their general toxicity

and deterrence capability as well as analgesic, anti-inflammatory and adaptogenic activities

which help to alleviate pains, developed resistance against diseases and endurance against

stress (Kaur and Arora, 2015).

Flavonoids have also been referred to as nature's biological response modifiers because of the

strong experimental evidence of their inherent ability to modify the body's reaction to

allergies, virus and carcinogens (Vasantha et al., 2012). Also, Phenols are one of the major

groups of non-nutritive dietary components that have been associated with the inhibition of

cancer, atherosclerosis, and age-related degenerative brain disorder (Chang et al., 2006).

Saponins from plants sources are essential to man owing to their responsibility for some

pharmacological effects like anti-inflammatory, antimicrobial, antidiabetic and anticancer,

hypocholesterolemia and anticonvulsant in humans (Ali et al., 2011).

36
5.2 ANTIOXIDANT ACTIVITY

The antioxidant activity determination performed on the methanolic extracts of both Zingiber

officinal and the whole pod of Tetrapleura tetraptera are displayed in Tables 4.1-4.3

respectively. In both extracts, the samples exhibited much antioxidant property. The obtained

result is however lower than the 0.94 - 0.01 mg AAE/g, estimated by Silva et al., (2007), in

the methanolic extract of the same plants species. The difference in antioxidant activity

compared to the reported result may be due to the differences in varieties of Tetrapleura

tetraptera used (different varieties possess different chemical properties), different growing

medium (soil) or the differences in extraction techniques. However, this present study

compares the antioxidant property in the whole fruit methanolic plant extracts, with limited

facts in literature and is also relatively in accordance with the findings of Ojewole J.A.,

Adewunmi C.O., (2004) who worked on the same plant. Antioxidants function as reducing

agents, ultimately eliminating free radical intermediates and inhibiting further oxidation

(Silva et al., 2007).

Antioxidant activity is known to account for anti-inflammatory activity of plants.

Tetrapleura tetraptera has been reported to possess anti-inflammatory (Agarwal et al., 2009),

antioxidant activity (Badu et al., 2012) which may have contributed to the contraction and

increased rate of epithelialisation observed in this study. Tetrapleura tetraptera fruits have

been reported to contain many chemical compounds such as alkaloids, flavonoids and other

phenolic compounds (Adewunmi and Marquis, 1987).

37
5.3 CONCLUSION

The results obtained in the present study indicate that Zingiber officinale extract exhibits free

radical scavenging, reducing power. The overall antioxidant activity of Zingiber officinale

extract might be due to its flavonoid, polyphenolic and other phytochemicals constituents.

The findings of the present study suggested that Zingiber officinale could be a potential

source of natural antioxidant that could have great importance as therapeutic agents in

preventing or slowing the progress of aging and age associated oxidative stress related

degenerative diseases. Further research work is required to isolate phyto-constituents

responsible for antioxidant activity.

Also, this study has unveiled that the extract of Tetrapleura tetraptera fruit, which has been

reported to be commonly used in the traditional systems of medicine, contains an appreciable

amount of phytochemicals and also possesses antioxidant and metal chelating capacities. The

phytochemical screening tests revealed the presence of many phytochemicals including

phenols, flavonoids, alkaloids, tannins, terpenoids, steroids and saponins in the extract

(methanolic) of the whole pod of Tetrapleura tetraptera. It was also discovered from the

study that the pod had significant antioxidant activity and is therefore recommended for

antioxidant role. The presence of many active phytochemical compounds and antioxidants

capacity in the fruit of Tetrapleura tetraptera advocate its antioxidant role and ethno-

pharmacological uses in traditional medicine.

38
5.4 RECOMMEDATION

The effect of the isolation of each phytochemical and antioxidant present in the samples should

be further researched on so as to understand it importance to the human body.

Knowing the importance of how this plant extracts works in the human body when taken in.

Further study should be done on clinical chemistry on how this plants react with the physiology

of the body.

Consideration should be taken as regards to the caution of the use of this samples in foods and

drugs.

39
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