ETHNO MEDICINAL, NUTRITION AND THERAPEUTIC VALUES OF “MARUGBO”

(BLACK SOUP) IN THE ILAJE AND IKALE COMMUNITIES 0F ONDO STATE,

NIGERIA

MBAEGBU, AUGUSTA CHINWE

MATRIC NUMBER 150402020

AUGUST, 2021

1
ETHNO MEDICINAL, NUTRITION AND THERAPEUTIC VALUES OF “MARUGBO”

(BLACK SOUP) IN THE ILAJE AND IKALE COMMUNITIES 0F ONDO STATE,

NIGERIA

BY

MBAEGBU, AUGUSTA CHINWE

MATRIC NUMBER 150402020

A PROJECT REPORT SUBMITTED TO THE DEPARTMENT OF BIOLOGICAL

SCIENCES, SCHOOL OF SCIENCE, OLUSEGUN AGAGU UNIVERSITY OF

SCIENCE AND TECHNOLOGY, OKITIPUPA, IN PARTIAL FUFILMENT OF THE

REQUIREMENTS FOR THE AWARD OF BACHELOR OF TECHNOLOGY (B.TECH)

DEGREE IN BOTANY.

SEPTEMBER, 2021

2
 DECLARATION

I, MBAEGBU, Augusta Chinwe with matriculation number: 150402020 hereby declare that

this research work was carried out by me. All sources of information are clearly acknowledged

by means of references.

----------------------------------------------------------

Signature/Date

i
 CERTIFICATION

This is to certify that this research work carried out by MBAEGBU, Augusta Chinwe

(Matriculation number: 150402020) and submitted to the Department of Biological Sciences,

School of Science, Olusegun Agagu University of Science and Technology, Okitipupa having

met the standard required by the institution.

DR. D. O. AWORINDE ----------------------------------------------

Project Supervisor Signature/Date

DR. E. E. NMEMA ---------------------------------------------

Ag, Head of Department, Signature/Date

Biological Sciences.

ii
 DEDICATION

This project is dedicated to God Almighty my creator, my strong pillar, my source of inspiration,

wisdom, knowledge and understanding and to my caring parents, family members and friends.

iii
 ACKNOWLEDGEMENTS

Special thanks to God Almighty, the creator of heaven and earth for granting me the grace to

complete this project report successfully. My sincere gratitude goes to my Project Supervisor Dr.

D. O. Aworinde for his understanding, patience, idea and technical competence that led to the

successful completion of this research. I want to appreciate the Ag, Head of Department Dr. E.

E. Nmema for her motherly care and advice.

I express sincere gratitude to my parents (Mr. and Mrs. Mbaegbu) and my siblings for their

prayers and support.

Finally, my appreciation goes to all the academic staff of Biological Sciences Department

Lecturers for their academic impact in my life. Thank you all and God bless you.

iv
 Table of Contents
DECLARATION..............................................................................................................................i

CERTIFICATION...........................................................................................................................ii

DEDICATION...............................................................................................................................iii

ACKNOWLEDGEMENTS............................................................................................................iv

LIST OF TABLES..........................................................................................................................vi

LIST OF FIGURES.......................................................................................................................vii

ABSTRACT.................................................................................................................................viii

CHAPTER ONE..............................................................................................................................1

1.0 INTRODUCTION.....................................................................................................................1

CHAPTER TWO.............................................................................................................................7

2.0 LITRATURE REVIEW.............................................................................................................7

CHAPTER THREE.......................................................................................................................10

3.0 MATERIALS AND METHOD...............................................................................................10

3.1 MATERIALS..........................................................................................................................10

3.1.1 Solvents.................................................................................................................................10

3.1.2 Plant Preparation...................................................................................................................10

CHAPTER FOUR.........................................................................................................................12

4.1 RESULTS................................................................................................................................12

4.2.1 PROXIMATE ANALYSIS..................................................................................................18

4.2.1.1 Moisture content................................................................................................................18

4.2.1.2 Ash value...........................................................................................................................18

4.2.1.3 Protein................................................................................................................................18

4.2.1.4 Crude Fibre........................................................................................................................18

4.2.1.5 Crude Fat...........................................................................................................................19

4.2.1.6 CHO...................................................................................................................................19

v
CHAPTER FIVE...........................................................................................................................20

5.0 DISCUSSION..........................................................................................................................20

5.1 CONCLUSION........................................................................................................................21

REFERENCES..............................................................................................................................22

LIST OF TABLES

Table 4.1 Enumeration of Recipes of Marugbo (black soup) in Ikale and Ilaje Community.. 12

Table 4.2 Phytochemical Constituents in present in marugbo ..…………………………….. 13

Table 4.3 Proximate Analysis of the Medicinal Plant Components ……………………….. 14

Table 4.4 Micronutrient Components of Powdered Plant Samples……………...………….. 15

vi
Table 4.5 …………………………………………………………………………………….. 16

LIST OF FIGURES

Figure 4.1 Proximate Analysis of the Medicinal Plant Component………………………….. 14

Figure 4.2 Micronutrient Components of Powdered Plant Samples………………………….. 15

vii
 ABSTRACT

The inclusion of leafy vegetables in human diets has been shown to be protective against

incidence of chronic, degenerative and age-related diseases, due to the presence of antioxidants,

in Nigeria, green leafy vegetables are traditionally cooked and eaten as a relish with starch

staple. Clerodendrium volubile (marugbo) is an under-utilized, tropical, non-convectional leafy

vegetable grown in Nigeria. This study is to know the Ethno medicinal, nutrition, and therapeutic

value of Marugbo (black soup) and some other leafy components and condiments were

determined using standard analytical procedures, proximate analysis unravelled a high

percentage of carbohydrate (52.25%), crude protein (1.75%), crude fibre (14%), ash (15.5%),

and fat (4.5%). Micronutrients components of powdered plants show high level iron

((38.9mg/100g), calcium (34.88mg/100g), magnesium (21.5mg/100), copper (0.74mg/100),

manganese (0.74mg/100g). The phytochemicals analysis shows presence of tannins, saponin,

cardiac glycosides, phenols, alkaloids, steroids ad absent of anthraquinone. Presence of tannins

and other phytochemical components in this soup can serve as antioxidant, to fight inflammation,

protect against heart disease and saponin to reduce cholesterol level, and strengthen the immune

system.

viii
 CHAPTER ONE

1.0 INTRODUCTION

Clerodendrum volubile, an understudied indigenous plant, belongs to the family

Lamiaceae and it is one of the widely distributed vegetables in the warm temperate and

tropical regions of the World (Burkill, et al., 1985). The plant is popularly known as

“Marugbo” or “Eweta” among the Ikale, Ilaje and Apoi people found in Southern-senatorial

district of Ondo State, South West Nigeria. “Obnettete”, as the plant is known among the

Itsekiri and Urhobo tribes in Niger-Delta, is a green climbing shrub reported to have height

of 3m and possesses numerous flowers. These are averagely about 1.5cm in length (Burkill,

et al., 1985), (Erukainure et al., 2011). The leaf of Clerodendrum volubile is commonly

consumed as vegetables mostly blended with other vegetables as spice with sweet aroma and

taste. Locally, the leaves can be blended either fresh or dried and applied as spices in

cooking (Adefegha et al., 2011). Commonly referred to as “Eweta” by the Ikales’, the leaves

of Clerodendrum volubile have great nutritional value as well as herbal and medicinal value.

The plant has been reported to contains very huge quantity of iron and zinc; elements which

are important in many enzymes for their functions and for maintenance of fresh skin. The

presence of phenolic compounds and other phytochemicals has also been observed by

(Adefegha et al., 2013). When consumed, the leaves are often noted for stimulating lost

appetite as well as replenishing vitality for mothers of new born babies (Adefegha et al.,

2011).

Proximate and nutrient analyses have been reported to play important role in

assessing nutritional significance of edible plant and vegetables (Pandey et al., 2006).

1
 Clerodendrum volubile has continued to be an important plant in South Western

Nigeria where it is widely consumed mainly as vegetables in soup. Evidence abounds that

the plant is majorly grown as food. The increase in cultivation and consumption of

Clerodendrum volubile may be an evidence of its rich nutritional properties. The basic

nutritional value of this plant can actually be assessed by its nutrients content as

determination of plants potentials, as a therapeutic agent or food, demands knowledge of its

overall nutritional worth and composition.

Although, the plant is essentially grown as food, it has continued to be an important

medicinal plant (Erukainure et al., 2010). Its medicinal value may be the likely reason for

much of the recent attention and increased consumption of the plant as well as its spread into

new areas. Over the time, a wide variety of claims have been reportedly made for its

efficacious medicinal properties as a treatment for many ailments ranging from its ability to

relieve pain and swelling (Fred et al., 2012) to general healing properties for clinical

conditions such as oedema, rheumatism, dropsy, gout and arthritis (Neeta et al., 2007); thus

its consumption will improve the health of the consumers.

Clerodendrum volubile is also cultivated as ornamental crop. “Marugbo soup” often

is served as a delicacy to important dignitaries or visitors especially on unique occasions or

celebration.

African Indigenous Green Leafy Vegetables (AIGLV) are inexpensive sources of

proteins, carbohydrate, minerals, vitamins and dietary fibres (Ogunwa TH, et al. 2015). Some of

the AIGLV indigenous to the Some of the AIGLV indigenous to the Yoruba ethnic group of

South West Nigeria include: Amunututu {(Basella alba (green) and rubra (red))}, Odu

(Solanum nigrum), African Glossy Night Shade (Solanum scabrum), Ewuro or Bitter leaf

2
(Vernonia amygdalina), Ebolo (Crassocephalum rubens), Isapa or Roselle (Hibiscus sabdariffa),

Wild leafy Marugbo (Clerodendrum volubile), Eggplant leaf (Solanum macrocarpon), Efinrin or

African Basil (Ocimum Gratissimum L.), Igbo (Detarium microcarpum), Gbure or Water leaf

(Talinum triangulare), Snake tomato (Trichosanthes cucumerina), Sokoyokoto or Spinach

(Celosia argentea L.), Tete or Amaranth (Amaranthus hybridus), Fluted pumpkin (Telfaria

occidentalis) and Ila or Okra (Abelmoschus esculentus L.).

Every food substance consumed by humans has either a therapeutic, nutritional or toxic

effect on the body. Plants and their products have been in used for as old as the history of man

for their therapeutic purposes. In the past decades, pharmacologists and organic chemists have

synthesized a large number of interesting chemical substances from plants, which have been of

great help in the practice of pharmacology and traditional medicine.

In eastern Nigeria, most of the foods consumed as soup are prepared with different plant

parts (such as the leaves, seeds, and fruits) which serve for different purposes as spices,

condiments, thickeners etc. According to Ikechuku and Emmanuel, (2010) Thickening agents, or

thickeners, are substances which, when added to an aqueous mixture, increase its viscosity. They

provide body, increase stability and improve suspension of added ingredients (the strength of the

food materials).

The food use of these thickeners calls for more research to provide data on the nature of

their chemical compositions and properties of their constituents so as to ascertain their actual

nutritional values, health and other medicinal importance. Examples of these soup thickeners

include Defarium microcarpum (Ofor), Brachystegia nigerca (Achi), and Irvingia gabonensis

(Bush mango). These plant seeds have been reported for their nutritional and medicinal

importance. The existence of ethnomedicinal claims for Clerodendrum species in folk medicines

3
and diverse traditional systems of medicines, across Asian and African continents, for the

management of several life-threatening ailments like jaundice, syphilis, typhoid, asthma,

cataract, malaria, pyreticosis, hypertension as well as diseases of skin, blood and lung cancer has

drawn attention of researchers to these plants (Neeta and Tejas, 2007).

Clerodendrum volubile has often been termed as magic leaf due to its high efficacy when

used for management and treatment of numerous ailments such as diabetes, ulcer and other

common diseases (Burkill, 1985). Its medicinal value may be the most probable reason for much

of the recent attention and increased utilization of the plant as well as it’s spread into new areas.

We have reported the nutritional value of this plant in our previous article (Ogunwa et al., 2015).

There have been reports on the contributions of free radicals to several human diseases such as

cancer, development of AIDS as well as heart diseases (Kumpulainen and Salonen, 1999;

Elekofehinti et al., 2012; Ejelonu et al., 2013). In fact, oxidative stress plays a major role in the

growth of chronic and degenerative disorders (Willcox et al., 2004; Pham-Huy et al., 2008).

Fortunately, antioxidants can combat such disorders either by inhibiting the formation of free

radicals (scavenging them) or increasing the rate of decomposition. The discovery that synthetic

antioxidants are carcinogenic with harmful effect on liver and lungs has given birth to a rising

interest toward natural antioxidants of herbal sources (Gokhan et al., 2011). The fact that plants

constituents with antioxidant capacity are capable of providing protective actions against stress

induced oxidative damage in biological systems, has been strongly supported by in vitro and

epidemiological studies on medicinal vegetables and plants (Ness and Powles, 1997; Umesh and

Veeru, 2012).

Despite the fact that many plants have been investigated for phytochemical components

and anti-oxidant effects, research is encouraged in this area because discoveries are always

4
acceptable for new plants with these potentials especially those with high therapeutic potential

and low toxicity risk. The search for natural antioxidant phytochemicals has been intensified

because of their capacity to reduce the impact of free radical reactions, thereby giving protection

from diseases (Terao and Piskula, 1997). Hence, the act of discovering new anti-oxidant

bioactive compounds from natural sources is still attractive since they are used in folk medicine

because they are relatively cheap, easily available with high efficacy and have reduced

occurrence of side effects.

The choice to investigate the anti-oxidant properties of aqueous extract of Clerodendrum

volubile leaves was made based on the revelation that local traditional healers in Ikale land,

located in Southern-Senatorial district of Ondo State, Nigeria, employed this part of the plant in

the treatment of many oxidative stress mediated diseases and their complications. Therefore, this

research work was done to evaluate the phytochemical components and antioxidant potential of

Clerodendrum volubile leaves to corroborate the traditional claims for its pharmacological

properties.

There have been reports on the contributions of free radicals to several human diseases

such as cancer, development of AIDS as well as heart diseases (Kumpulainen and Salonen,

1999; Elekofehinti et al., 2012; Ejelonu et al., 2013). In fact, oxidative stress plays a major role

in the growth of chronic and degenerative disorders (Willcox et al., 2004; Pham-Huy et al.,

2008). Fortunately, antioxidants can combat such disorders either by inhibiting the formation of

free radicals (scavenging them) or increasing the rate of decomposition. The discovery that

synthetic antioxidants are carcinogenic with harmful effect on liver and lungs has given birth to a

rising interest toward natural antioxidants of herbal sources (Gokhan et al., 2011).

5
 The choice to investigate the anti-oxidant properties of aqueous extract of Clerodendrum

volubile leaves was made based on the revelation that local traditional healers in Ikale land,

located in Southern-Senatorial district of Ondo State, Nigeria, employed this part of the plant in

the treatment of many oxidative stress mediated diseases and their complications. Therefore, this

research work was done to evaluate the phytochemical components and antioxidant potential of

Clerodendrum volubile leaves to corroborate the traditional claims for its pharmacological

properties.

Despite the vast nutritional and medicinal significance of marugbo leaves, there is

dearth of information on its amino acid and, only little detail is available on the nutritional

composition of this plant. Therefore, the objective of this research work was to:

To determine the chemical components of the plant species with a view to further

establishing the nutritive values.

6
 CHAPTER TWO

2.0 LITRATURE REVIEW

Little research has been done on the nutritional potentials of these Nigerian indigenous

soups despite the high level of literature on the leaves, hence, this study was carried out to

increase the knowledge and know the Ethno medicinal value, nutritional value and therapeutic

value of marugbo in Ikale and Ilaje community of Ondo state, Nigeria.

This is to increase the knowledge base as well as promote consumption of these soups

which are rich naturally in micronutrient and can help reduce occurrences of some micronutrient

deficiencies.

Countries like India and China have long been recognized to have fairly well-established

practices and policies for traditional medicines that have gained worldwide acceptance and are

being practiced alone side modern medicines. In America, herbal remedies and dietary

supplements with established scientific validation are sold as over-the-counter medications

(OTC) and are popular among medical professionals. The European Agency for the evaluation of

medicinal products has recently approved guidelines that specify the test procedures and

acceptance criteria for herbal drugs, herbal drug preparations and herbal medicinal products

(EMEA, 1999).

Medicinal plants constitute a source of valuable foreign exchange for many developing

countries and the global market for herbs and medicinal plants runs into several billion dollars

per annum. Countries like Bulgaria, Germany, Poland, and others in Africa and Asia are

recognized as major exporters of plant-based medicinal products and raw materials for overseas

medicinal plant industries (Hoareau et al., 1999). Plethora of literature has indicated that the

7
global market for herbal medicines currently stands at over US$ 43-60 billion annually and is

growing steadily (Enwonwu, 2003). Over US$ 2.4 billion Traditional Chinese Medicines were

sold and 400 million US$ worth of Traditional Chinese Medicines were exported out of China in

1993; about 60 million US$ was realized from herbs in 1996 in Malaysia (Elujoba, 2005).

Medicinal plants and other natural substances are an integral component of ethno-veterinary

medicine and human health notably in the areas of cardiac, cancer (Cragg & Newman, 2005) and

microbial (Ríos and Recio, 2005) chemotherapies.

The recent development and therapeutic success of plant derived antimalarial, artemisinin,

amidst increasing microbial resistance to conventional drugs is worthy of note. Unfortunately,

research, development and commercial production of medicinal plant-based products particularly

in the developing world are hampered greatly by the absence of essential infrastructure in both

the public (such as universities and other governmental institutions) and private sectors,

compounded by lack of governmental interest and financial support. Malaysia is one of such

countries whose commendable efforts in natural medicinal products R & D are encouraging and

worthy of emulation by other developing nations. The Malaysian Natural Products Society

formed in 1994 oversees and coordinates activities relating to medicinal plant research and

development and aim to eventually release the Malaysian Pharmacopoeia. The Malaysian

government through relevant establishments and agencies is playing a leading and commendable

role of financing and encouraging research and development of herbal and complementary

alternative medicines. In conjunction with drug industries and other stakeholders, the people and

government of Malaysia have laid a very good foundation for the development of this sector and

have recorded great successes evidenced by the increasing number of good quality herbal

products in the market. The designation of Malaysia as WHO’s centre for regulatory control in

8
1996 is an eloquent testimony of their effort and determination to achieve an internationally

acceptable standard in the pharmaceutical industry.

(Ajao, et al., 2018). Found that despite the fact that C. volubile is an underutilized indigenous

vegetable, it has been studied ethnopharmacologically to some extent. Its folkloric usage such as

diabetics, analgesic and hepatoprotective agent has been proven scientifically, although more

work needs to be done on analgesic and hepatoprotective activities of the plant such as isolation

of the metabolite responsible for these actions. However, detailed study needs to be done on the

mechanism of action of all the isolated compound through in vivo studies. The antimicrobial

activity of the plant has only been done on just one Fungi ‘Aspergillus flavus’, there is need to

explore other virulent fungi and bacterial to unravel the potential of this plant in the treatment of

pathogenic diseases. Due to the effectiveness of the extract of C. volubile on breast cancer cells

and ROS scavenging there is possibility it could also be effective on other cancer cells since the

Etiology of all cancers are the same. Therefore, research in this direction should be probed

further. Furthermore, the toxicological reports in this review did not credit any therapeutic side

effect to the plant if its consumption is not abused. Overall, proper awareness needs to be created

on the ethnopharmacological importance of this underutilized vegetable so that it can be properly

harnessed in combating malnutrition and diseases.

9
 CHAPTER THREE

3.0 MATERIALS AND METHOD

3.1 MATERIALS

Water bath, Crucible, Burette, Desiccator, Petri dishes, conical flask, Test tube, Bunsen burner,

Oven, Weighing balance, Filter paper, Glass funnel

3.1.1 Solvents

100ml of petroleum ether, 2% sodium hydroxide, 2% sulphuric acid , 45% NAOH ,

Hydrochloric acid , 0.1% Ferric chloride, Alcohol, Lead acetate, Chloroform, Methylene

chloride, Hydrogen tetra oxosulphate (v) acid, 2% 3,5, dinitrobenzoic acid, Dichloromethane,

Ammonia, Distilled water, Litmus paper, Meyer’s reagent, Wagners reagent , Drangendurffs

reagent.

3.1.2 Plant Preparation

The plant leaves were purchased locally from the people of Ikale land located in Okitipupa Local

Government, of Ondo State, Nigeria. The samples were identified at the Biological Department,

Olusegun Agagu University of Science and Technology in Nigeria with herbarium specimens.

Which was deposited at the Herbarium of the University. The leaves were detached from their

stalks carefully washed with tap water then. They were subsequently rinsed with distilled water

so as to remove sand and other impurities. All the recipes were air-dried in order retain the active

components in the plant and it was oven dried at 40c so that the plants will not be denatured and

it was grounded into powdered. Phytochemical screening and other analysis of the sample

followed standard procedure after was carried out on the powdered samples.

10
 PROCEDURE (Cardenolide and Flavonoid)

1g of sample was measured using a weighing scale and 20ml of alcohol was added and

was placed on the Bunsen burner for 5minutes. It was filtered and allowed to cool down

before carrying out the analysis.

 PROCEDURE (Alkaloid and Anthraquinone)

1g of sample was measured using a weighing scale and 20ml of Hcl was added and

placed on the Bunsen burner for 5 minutes in other to extract the bioactive component, it

was filtered and allowed to cool down.

 PROCEDURE (Tanin, Phenols and Saponin)

1g of samples was measured using a weighing scale and 20ml of water was added and

placed on Bunsen burner in other to extract the bioactive component. It was filtered and

allowed to cool down before the analysis is carried out.

 PROCEDURE (Steroids)

1g of sample was measured using weighing balance and 20ml of (Dichloromethane) was

added and placed on Bunsen burner for 5 minutes to extract the bioactive component. It

was filtered and allowed to cool down.

11
 CHAPTER FOUR

4.1 RESULTS

Table 4.1: Enumeration of Recipes of Marugbo (black soup) in Ikale and Ilaje Community.

INDEGINOUS NAME BOTANICAL NAMES OTHER INGREDIENTS

Marugbo Clerodendron volubile Lapalapa funfun (Jatropha
curcas)
Efirin wewe Ocimum gratissimum Posa (Monodora myristica)
Ailu Secamone afzelli Atale pupa (Zingiber officinale)
Paki/ ege Manihot esculentum Iru (Pakia biglobosa)
Ata wewe Capsicum annum Efunle (Aerva lanata)
Iyere Piper guineense Iteji (Gongronema latifolium)
Egusi Cucumeropsis mannii Ekikan (Spondia mombin)
Akintola Chromolaena odorata Eyin aripo (Diodia sarmentosa)
Ugwu Telferia occidentalis Aidan (Tetrapleura tetraptera
Owu Gossypium bardadens Lefi funfun (Ricinus communis)
Ewe tea Cymbopogon ciratus Ita ode (Indigofera pulchra)
Ayuu Allium sativum Eragiri (Lagenaria breviflorus)
Alubosa Allium cepa
Ewuro Vernonia amygdalina
Osan wewe Citrus aurantifolia
Ogbo Parquetina nigrescense
Adenden odo Harungana madagascariensis
Eyin yarinbo Diodia sarmentosa

12
Table 4.2: Phytochemical Constituents in present in marugbo
PHYTOCHEMICALS

Anthraquinone Tannins Saponin Cardiac Phenols Alkaloids Steroids

glycosides
- + + + + + +

13
Table 4.3: Proximate Analysis of The Medicinal Plant Components

PARAMETERS (%)

Crude Crude fibre Ash Moisture Fat CHO

protein content
1.75% 14% 15.5% 12% 4.5% 52.25%
Table 3: Medicinal Plant Components Parameters (%)

Proximate Analysis of the Medicinal Plant Com-

ponent
0.6
52%
0.5

0.4

0.3
parameter

0.2 16%
%

14%
12%
0.1
5%
2%
0
Crude protein Crude Fibre Ash Moisture content Fat CHO
Medicinal Plant Components

Figure 4.1

14
Table 4.4: Micronutrient Components of Powdered Plant Samples.
PARAMETERES (mg/100g)

Mn Mg Cu Fe Ca
0.74mg/100g 21.5mg/100g 0.74mg/100g 38.9mg/100g 34.88mg/100g

MICRONUTRIENT COMPONENTS OF POWDERED

PLANT SAMPLES
45
40 38.9
34.88
35
30
Parammter

25
mg/100g

21.5
20
15
10
5
0
0.74
Mn Mg 0.74
Cu Fe Ca
Micronutrient Component

Figure 4.2

15
SN PHYTOCHEMICALS TEST RESULT INFERENCE
1. Alkaloid 5ml of the HCL There was formation +
extract was measured of cream coloured Alkaloid is
NH3 was added and precipitate when present
tested to litmus paper. Meyer’s reagent was
Added there was
formation of reddish-
brown precipitation
when Drangedurffs
reagent was added.
There was formation
of yellow colour
when Wagner
reagent was added.

2. Tannin 1ml of sample Blue black +

extracted with water precipitate was There was
extract was measured formed. presence of
into a test tube and tannin.
was diluted with 5ml
of water and a few
drops of ferric chloride
(0.1%) was added.
3. Saponin 1ml of water extract There was formation +
was measured in a test of foam. There was
tube and diluted with presence of
5ml of water and was saponin.
shaken vigorously.
4. Anthraquinone 5ml of the HCL There was no _
extract was measured formation of rose- Anthraquinone
in a test tube and few pink colour. is absent.
drops of chloroform
was added, the
supernatant (upper
layer) was taken away
and 10% ammonia
was added.

16
 5. Steroids 2ml of the DCM The upper layer +
extract was measured turns red and the There was
in a test tube and 2ml lower layer turns presence of
of concentration yellow. steroids.
H2SO4 was added.
6. Phenols 1ml of the water Blue black +
extract was measured precipitate was Phenols are
and was diluted with formed. present.
5ml of water then
0.1% of Ferric
chloride was added.
7. Cardiac glycosides 5ml of alcohol extract There was formation +
was measured into a of dirty brown Cardenolide is
test tube and lead precipitate (Keddes present.
acetate was added test). There was
chloroform was added formation of brown
and the supernatant ring (Killanis test)
taken away.
Methylene chloride
was added and it was
divided into two test
tube and two test was
carried out.
Keller killanis test
(3ml Ferric chloride
reagent and
concentration H2SO4)
Keddes test (1ml of
2% 3,5 dinitro benzoic
acid) was carried out.

Table 4.5

17
4.2.1 PROXIMATE ANALYSIS

4.2.1.1 Moisture content

The weight of the crucible was measured using a weighing balance and zeroed and 2g of plant

sample was measured using weighing balance and was kept in oven (100c) for 4hours and then

one hour and another one hour until the result is constant. The percentage of the moisture content

was determined by subtracting the new value from the initial value multiply by hundred to give

the result which is the moisture content.

4.2.1.2 Ash value

The percentage ash value was determined when the moisture content was determined. The

residue was placed on the Bunsen burner for hours until it turns into ashes and it was weighed on

the weighing balance.

4.2.1.3 Protein

Protein was determined using the digestion process. 2g of the sample was measured and the ash

value determined. 10ml of HCL was poured into the ash powdered samples and one selinum salt

was added and was rinsed with distilled water and add 45% of NaoH was boiled filtered and

boric acide was added into an empty conical flask and was titrated until the colour turns red

when phenolphthalein was added.

4.2.1.4 Crude Fibre

2g of samples was weighed 10ml of 2% of sulphuric acid was added and placed on water bath to

boil and 10ml of distilled was used to rinse the residue and the residue was scraped into another

conical flask and 10ml of 2% sodium hydroxide was added placed on water bath and filtered and

was also rinsed with distilled water. The residue was poured into boroxillicate glass and placed

18
in oven at 100c for hours until it dries off, it was weighed and scraped into crucible and burn for

almost 6hours till it turns to ash and it was calculated.

4.2.1.5 Crude Fat

2g of samples was put in a test tube and 100ml of petroleum ether was added and was boiled on

water bath and it was filtered. The filtrate was dried in the oven at 70c using petri dish and was

put in the desiccator before re-weighed

4.2.1.6 CHO

The following test are carried out on the samples (crude fat, crude fibre, protein, ash value,

moisture content) and was subtracted from hundred to get the percentage of the carbohydrate.

19
 CHAPTER FIVE

5.0 DISCUSSION

The phytochemical are physiologically active compounds exhibiting and possessing great

potential for therapeutic uses. It is important to note that medicinal plants contain mixtures of

different chemical compounds that act individually, or synergistically to improve health (Gurib-

Fakim, 2006). Phytochemicals such as tannins and phytates are regarded as botanical chelators

and possess potential malaria suppressive effects through sequestration of iron (Etkins, 1996).

Moreover, saponins, polyphenols and phytosterols exhibit hypocholestrolemic effects on animals

and humans thereby contributing to the phenomenon of the low rate (incidence) of

atherosclerosis (Johns and Chapman, 1995; Johns, 1996. Phenols are very important

phytochemicals because of their ability to scavenge free radicals owing to their redox properties

which permit them to act as reducing agents, hydrogen donors and singlet oxygen quenchers

(Rice – Evans et al., 1997).

In this study, phytochemical screening of ethanolic leaf extract of C. volubile revealed the

presence of flavonoids, alkaloids, saponins, tannins, triterpenes, cardiac glycosides and steroid.

The quantitative estimation of the extract showed that it contained high concentration of

phenolics, A low concentration of saponins and tannins was observed. The high polyphenol

content of the C. volubile leaf is likely to be responsible for its various therapeutic properties

such as antioxidant activities (Kris-Etherton et al., 2002; Vaya et al., 2003), hypolipidemic

potential (Narender et al., 2006; Harnafi and Amrani, 2007) and vaso-relaxant activities

(Bernatova et al., 2002).

20
Saponin is known to possess serum cholesterol lowering activity (Topping et al., 1980). The

cholesterol lowering ability of saponin is based on the fact that it binds either bile acids or

cholesterol in the intestinal lumen. Binding with cholesterol will make it more easily re-

absorbed causing a reduction in enterohepatic circulation of bile acid in the liver (Potter et al.,

1979; Kritchevoky, 1997).

5.1 CONCLUSION

This study indicated that the soup marugbo (black soup) is of high nutritional value. It is good

source of carbohydrate, fibre, fat, protein and more. The soup is a great antioxidant due to the

presence of tannin and the good value of this soup is an indicator that the cultivation and the

utilization should be promoted. It is recommended that clerodendrum volubile cultivation should

be encouraged and further work should be done on the bio availability of these nutrient in human

body.

21
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