PHYTOCHEMICAL SCREENING AND PROXIMATE COMPOSITION

OF METHANOL LEAF EXTRACT OF TELFAIRIA OCCIDENTALIS

By

Umar Nashena Christopher

BCH/18U/2770

DEPARTMENT OF BIOCHEMISTRY, FACULTY OF LIFE SCIENCE, MODIBBO

ADAMA UNIVERSITY YOLA, ADAMAWA STATE.

Supervised by

Prof. M. I Bello

October, 2024.

i
 DECLARATION
I, Umar Nashena Christopher hereby declare that this project report was written by me and it is a
record of my own research work. It has not been presented before in any previous application for
a degree. All references cited have been duly acknowledged. This is followed by the name and
signature of the student and date

_________________________ _____________________

Umar Nashena Christopher Date

ii
 DEDICATION

This Project work is dedicated to my dear parents Mr and Mrs Umar Christopher my siblings,
uncles Mr samaila manga, Mr Edward Umar and aunty Mrs Queen Manga for their support and
prayers through my time of study.

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 APPROVAL PAGE
This project report entitled “Phytochemicals Screening and proximate composition of methanol
leaf extract of telfaria occudentalis meets the regulations governing the award of Bachelors of
Technology of the Modibbo Adama University, Yola and is approved for its contribution to
knowledge and literary presentation.

__________________________ ___________________

Prof. M. I Bello Date

Supervisor

____________________________ _____________________

Date

External Examiner

_________________________ _____________________

Prof. Hauwa A. Umaru Date

Head of the Department

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 ACKNOWLEDGEMENTS
All thanks be to God almighty for giving me the grace and ability to overcome all the challenges
encountered in this program with good health. I also appreciate the effort of my project
supervisor Prof. M.I Bello who devoted his time and energy in checking, reading and making
necessary corrections to this project without number to ensure that this work is error free the
research owes him much gratitude

I extend my gratitude to the head of department Prof. Hauwa A. Umaru for proving all necessary
resource for the success of my study. I also appreciate all my lectures for their support, guidance
and encouragement toward the journey of my project.

I wish to express my sincere gratitude and appreciation to my family for their sacrifice and
endless support in making sure I got all what I needed for the success of my research.

Finally my gratitude to my able friends and classmates, thank you all for being there for me I
pray God bless and grants you your heart desires.

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 Abstract
These study sought to undertake the phytochemical analysis (using standard methods of analysis)
and proximate composition nd mineral composition of methanol leaf extract of Telfairia
occidentalis consumed in jimeta bekaji, Adamawa state Nigeria. The preliminary phytochemical
analysis showed that the leaf of T. occidentalis contains flavonoids, tannins, saponins, alkaloids,
phenolic and glycosides is absent. Quantitative analysis showed that; Saponins ( 7.18± 0.02)
Tannins (2.77 ± 0.10) Flavonoids (16.9 ± 0.1) Steroids (0.22 ± 0.01) Terpenoids (1.06 ± 0.01)
Phenols (7.95 ± 0.013) Alkaloids ( 5.51 ± 0.11) were percentage respectively. Result on
proximate analysis showed; Moisture (10.99 ± 1.34%) Fiber (11.56 ± 0.68%) lipid ( 6.46 ±
0.07%) Ash ( 8.31 ± 0.21%) Protein (21.14 ± 0.32%) Carbohydrates (53.10 ± 0.68%). This
findings provide evidence that methanol extract of Telfairia occidentalis is a potential source of
natural bioactive compounds and are good source of carbohydrates and energy.

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 Table of Contents
DECLARATION ............................................................................................................................ ii

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

APPROVAL PAGE ....................................................................................................................... iv

ACKNOWLEDGEMENTS ............................................................................................................ v

Abstract .......................................................................................................................................... vi

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

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

1.1 Background of the Study ..................................................................................................... 1

1.2 statement of the problem. .......................................................................................................... 3

1.6 significance of the study ........................................................................................................... 4

CHAPTER TWO ............................................................................................................................ 5

2.0 Literature review ............................................................... 5

CHAPTER THREE ........................................................................................................................ 9

3.0. MATERIALS AND METHODS .............................................. 9

3.1.3 method.................................................................................................................................. 10

3.1.3 collection of plant material and extraction........................................................................... 10

3.2.2 proximate analysis .............................................................................................................. 10

3.2.3 phytochemicals screening. ................................................................................................... 13

3.3.1 Qualitative phytochemical ................................................................................................... 13

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3.3.2 Quantitative Analysis of Phytochemicals ............................................................................ 13

3.3.3 Method of Data analysis. .................................................................................................... 14

CHAPTER FOUR ......................................................................................................................... 15

4.1. Results. ................................................................. 15

Table. 2 qualitative phytoconstituent of telfaria occudentalis methanol leaf extract ................... 16

Table. 3 quantitative phytochemical of T. Occidentalis. .............................................................. 17

CHAPTER FIVE .......................................................................................................................... 18

5.1.2. Conclusion .......................................................................................................................... 19

5.1.3. Recommendations ............................................................................................................... 19

viii
 CHAPTER ONE

1.0 INTRODUCTION

1.1 Background of the Study

Phytochemical screening and proximate composition analysis are essential tools for
understanding the bioactive compounds and nutritional value of plant materials.
Methanol leaf extract of T. occidentalis has been reported to possess antioxidant and
antimicrobial properties .This study aims to bridge this knowledge gap by investigating
the phytochemical constituents and nutritional content of methanol leaf extract of T.
occidentalis. Telfairia occidentalis, commonly known as fluted pumpkin, is a tropical
plant widely cultivated in West Africa for its nutritional and medicinal value. The leaves
of T. occidentalis have been used in traditional medicine for various ailments, including
fever, inflammation, and infectious diseases. Despite its potential, there is limited
scientific information on the phytochemical composition and nutritional content of T.
occidentalis leaves.

Fluted pumpkin (Telfapiria occidentalis); a member of Cucurbitaceous family is one of

the commonly consumed leafy and seed vegetables in Nigeria. Cucurbitaceae is a plant
family commonly known as gourds or cucurbits and includes crops like cucumbers,
squashes (including pumpkins), luffas melons and water melons. The family is
predominately distributed around the tropics, where those with edible fruits were amongst
the earliest cultivated plants in the world. The fruit is often a kind of berry called a pepo.
Renner, et al (2007). Phytogenetics of cucumis (curcurbitaceae). Journal of Food,
Agriculture and Environment. The cucurbita species are from primarily Africa and South
America. Jeffrey, C. (2005). Cucurbitaceae. In: Miline- Redhead, E. and Polhill, R. M.
(Editors). Flora of tropical East Africa Crown Agents for oversea Governments and
Administrators, London, United Kingdom. The fluted pumpkin is no longer known in the
wild, but most likely originated in West Africa’s high rainfall forest belt. The largest
diversity in plant populations can currently be found in Imo state and surroundings areas
in South-eastern Nigeria. Plants have been recognized for their several life saving and
therapeutic properties. Nearly 70% of the world’s population (mainly in the developing
countries) relied mostly on traditional medicine therapies as their basic way of health care

1
(Bewaji et al., 1983). Even presently rural population still considers herbal medicines as
the most preferred therapeutic source. Ambrose and Faith et al., (2015) reported that
consumption of crude extract of ginger (Zingiber officinale) contain an active component
which possess the ability to decrease serum lipid profile and lower the risk of
atherosclerosis and reduce the level of cardiovascular risk factors

In Nigeria, the consumption of the leaf of Telfairia occidentalis as a leafy vegetable in the
diet or as an infusion in medicinal preparation is being promoted in view of the various
medicinal properties such as antianeamic, antidiaebetic, antioxidant, antimicrobial
activities and as a purgative leafy vegetable (Oboh et al., 2006). According to Oboh,
(2005) Telfairia occidentalis prevents against garlic – induced oxidative stress. The plant
also prevents the occurrence of abdominal pain, small intestine obstruction, dermatitis,
asthma and increase of bleeding which are caused by garlic Several works reporting
compositional evaluation and functional properties of various types of edible wild plants
in use in developing countries abound in the scientific literature (Ekop, 2007). However,
much still needs to be done on the chemical composition of edible leafy vegetables grown
in Nigeria. This study therefore investigated the phytochemical compositions, mineral
and proximate compositions,

T. occidentalis is known by different names such as "iroko" or "apiroko" in Yoruba,

"ubong" in Efik, "ugu" in Igbo, "umeke" in Edo, and "umee" in Urhobo. The leaves are
rich in iron and are used to cure anaemia. The oily seeds also have lactation promoting
properties and are widely consumed by nursing mothers. The roots are known locally as
potent human, fish and rodents poison. The plant also contains considerable amount of
anti-nutrients such as phytic acid, tannin and saponin which could also have some health
benefits. Alogun et al (2006). Ethnobotany of Telfairia occidentalis among Igbos of
Nigeria. Journal of Food, Agriculture and Environment. Previous studies usually focused
on how to improve its cultivation. Ehiagbonare, J. E. (2008). Conservation studies on
Telfairia occidentalis Hook. F. A. indigenous plant used in enthnomedicinal treatment of
anemia in Nigeria. African Journal of Agricultural Research. Increase its yield and use it
in controlling pests or nutritional purposes. Odiaka, et al (2004). Telfairia occidentalis
Hook. f, in G. J. H. Grubben and O. A. Denton (Editors), Plant Res. Trop. Afr. 2:

2
 Vegetables, (PROTA Foundation, Netherlands/Backhuys Publishers: Leiden,
Netherlands/CTA Wageningen, Netherlands.

1.2 statement of the problem.

• What are the major phytochemical constituents present in the methanol leaf extract of
Telfairia occidentalis?
• What is the proximate composition of the methanol leaf extract of Telfairia occidentalis
in terms of moisture, ash, crude fibre, fat, protein, and carbohydrate content?
• How do the phytochemical constituents and proximate composition of Telfairia
occidentalis methanol leaf extract compare to those of other solvents, such as water or
ethanol?
• What are the potential medicinal or therapeutic applications of the phytochemical
constituents identified in the methanol leaf extract of Telfairia occidentalis?
• How does the methanol leaf extract of Telfairia occidentalis compare to other plant-based
extracts in terms of its phytochemical profile and proximate composition?
• What are the optimal extraction conditions (e.g. solvent, temperature, time) for
recovering bioactive compounds from Telfairia occidentalis leaves?
• How does the phytochemical composition and proximate analysis of Telfairia
occidentalis methanol leaf extract vary across different regions, climates, or cultivation
practices?
1.3 Aim
The aim of this study is to investigate the phytochemical constituents and nutritional
content of Telfairia occidentalis leaf extract, which can provide insights into its potential
medicinal and nutritional applications.
1.4 objectives
1. To identify and quantify the major phytochemical classes present in the methanol leaf
extract of Telfairia occidentalis, such as alkaloids, flavonoids, and terpenes.
2. To determine the proximate composition of the methanol leaf extract, including moisture,
ash, crude fiber, fat, protein, and carbohydrate content.
3. To evaluate the nutritional potential of Telfairia occidentalis leaf extract as a source of
essential macronutrients and micronutrients.

3
 4. To investigate the potential medicinal applications of the phytochemical constituents
identified in the methanol leaf extract.
5. To compare the phytochemical profile and proximate composition of Telfairia
occidentalis methanol leaf extract with those of other solvents or plant-based extracts.
6. To provide a scientific basis for the traditional use of Telfairia occidentalis in folk
medicine and nutrition.
7. To contribute to the development of new pharmaceutical or nutraceutical products from
Telfairia occidentalis leaf extract.

1.6 significance of the study

This study will provide valuable information on the phytochemical composition and nutritional
content of T. occidentalis leaves, which will contribute to the development of novel
pharmaceutical and nutraceutical products. The findings will also provide a scientific basis for
the traditional use of T. occidentalis leaves in medicine and highlight its potential as a functional
food ingredient.

4
 CHAPTER TWO
2.0 Literature review
T.occidentalis is a dioecious, perennial, tropical vine grown for its leaves and edible seeds.
Phillip IB, et al (2009). Economic potentials of plantain and fluted pumpkin intercropping as a
poverty reduction strategy in South‑Western Nigeria. World J Agric Sci. It is a creeping
herbaceous vegetable with lobed leaves and twisted tendrils that extends over the
soil.Horsefall M, et al (2005). Equilibrium sorption study Al3+, Co2+, Ag+ in aqueous solutions
by fluted pumpkin (Telfairia occidentalis hook f) waste biomass. Fluted pumpkin can be grown
on mounds or on flatlands. In domestic gardens, it is often cultivated beside fences or adjacent to
a tree, thus enabling the suspension of the fruit from a branch. Nwauwa LO, et al (2010).
Efficiency of vegetable production under irrigation system in Ilorin metropolis: A case study of
fluted pumpkin (Telfairia occidentalis). It may also be cultivated along trellis of different kinds
including bamboo. T. occidentalis leaves possess high curative, industrial, and nutritional values.
According to Akanbi et al. Akanbi WB, et al (2007). Growth, herbage and seed yield and quality
of Telfairia occidentalis as influenced by cassava peel compost and mineral fertilizer. the leaves
are abundant in fat (18%), protein (29%), and minerals and vitamins (20%). The leaves are also a
rich source of phosphorus, calcium, zinc, iron, and copper. Phytochemical screening of fluted
pumpkin leaf extracts by Oyewale and Abalaka confirmed the presence of saponins, alkaloids,
tannins, and phenolics. Oyewole OA, et al (2012). Antimicrobial activities of Telfairia
occidentalis (fluted pumpkins) leaf extract against selected intestinal pathogens. Seeds of T.
occidentalis can be ground and added to soups or roasted, cooked, and eaten. The seeds contain
45% fat, 23% carbohydrates, 20.5% protein, 2.2% fibers, and 4.8% total ash. In addition, the
seeds contain phospholipid, glycolipid, and neutral lipid contents of 58%, 26%, and 15%,
respectively. Fluted pumpkin seed oil contains 61% unsaturated fatty acids. The healing
properties of T. occidentalis are popular in Nigerian folklore medicine, and these curative
properties have been evaluated by a number investigators. The present study was aimed at
reviewing systematically, existing publicly available literature on the pharmacological properties
of T.occidentalis.

Globally, there has been a growing interest in medicinal plants by researchers. Telfairia
occidentalis Hook F. (Fluted pumpkin) is cucurbitaceous vegetable grown in West Africa,
particularly in Nigeria for its leaves and seeds. The curative properties of fluted pumpkin are

5
popular in Nigerian folklore medicine, and several investigators have validated these therapeutic
effects using animal models. The aim of this work therefore, was to review publicly available
literature on the pharmacotherapeutic activities of T. occidentalis. Searches were performed on
PubMed, Scopus, ScienceDirect, and Google Scholar for related studies, with search dates set
between 1990 and 2018. A total of 499 articles were retrieved and analyzed, with 38 studies
ultimately retained. Studies contained in this review were carried out in Nigeria, across different
locations. 13 categories of pharmacological activities for fluted pumpkin were documented after
analyzing full texts of the articles retained. T. occidentalis offers myriad of healing properties
that can be explored by pharmaceutical industries. As the search for potent drugs from botanical
sources continues, there is need for future investigations to isolate and characterize
pharmacologically active agents that confer medicinal properties on fluted pumpkin, as well as
elucidate the structures of these agents and pathways by which they exert their healing properties.

The present study sought to undertake the phytochemical analysis (using standard methods of
analysis), proximate and mineral composition as well as in vitro antioxidant properties
(involving inhibition of DPPH and reducing power ability) of aqueous leaf extract of Telfairia
occidentalis consumed in Ekpoma, Edo state, Nigeria according to U Usunobun et al (2014)
journal of basic and applied science. The preliminary phytochemical analysis showed that the
leaf of T. occidentalis contains flavonoids, tannins, saponins, alkaloids and phenolics.
Quantitative analysis showed that phenol, flavonoids, tannins, alkaloids and saponins in
percentage were 0.19±0.12, 7.12±0.50, 0.12±0.08, 1.03±0.24 and 7.01±0.30 respectively. Result
on proximate analysis showed 89.01% dry matter, 10.99% moisture content, 21.14% crude
protein, 6.46% lipid, 11.56% crude fiber, 8.31% Ash content, and 53.10% carbohydrate. Result
on Mineral composition in mg/100g dry matter showed Calcium (61.03±0.04), Sodium
(51.49±1.32), Iron (25.75±1.65), Zinc (13.15±0.31), Potassium (801.21±0.45), Magnesium
85.11±1.22), Phosphorus (18.09±0.09), Manganese (21.27±0.32) and Copper (0.93±0.03). In
vitro antioxidant scavenging activity using 2, 2-diphenyl 1-picrylhydrazyl (DPPH) and Ascorbic
as standard and reducing power ability of the plant extract was found to be concentration
dependent with maximum inhibition and reducing power ability at 0.4 mg/ml which were
69%(lower than 89% for ascorbic acid) and 0.199 respectively. Our findings provide evidence
that aqueous extract of Telfairia occidentalis is a potential source of natural antioxidants. The

6
result of these findings also revealed that Telfairia occidentalis leaves are good source of
carbohydrates and energy. The leaves are also a good source of minerals

According to Temitope Onuminya, et al (2017). Faculty of Physical Sciences and Faculty of Life
Sciences, Univ. of Ilorin, Nigeria. A systematic survey of leafy vegetables in three different
markets in Lagos state, Nigeria was carried out to compare their proximate and phytochemical
compositions. The vegetable samples were oven dried for two weeks, pulverized into powdery
form and soaked in ethanol for extraction of bioactive constituents. The proximate and
phytochemical analyses were carried out following standard procedures. A total of sixteen
vegetable samples were collected, six are used as spices while others are taken either raw or
cooked as vegetables. The phytochemical analysis of the methanolic extracts revealed that the all
the leafy vegetables studied are rich in flavonoids, tannins (except in Laurus nobilis–bay leaf),
saponins (except in Amaranthus viridis-slender amaranth) and alkaloids (except Murraya
koenigii–curry leaf and Amaranthus hybridus–green amaranth); with considerable amount of
phenols, anthraquinones and phlobatannins while cardiac glycosides were recorded in 75% of the
samples studied. Analysis of the proximate composition of the various leafy vegetables
examined varied with each species however the ranges recorded across the samples are as
follows: moisture content (2.4–19.8%), crude fiber (9.8–22.4%), total ash (9.0-20.3%), crude
lipid (0.5–13.5%), carbohydrates (0.25–9.12%) and total nitrogen (32.5–71.1%). This result
indicates that the leafy vegetables are valuable reservoir of bioactive compounds of substantial
medicinal merit and could be a good source for nutrients supplement and as a source for drug
production. Therefore, if consumed in sufficient amount, these vegetables would contribute
greatly towards meeting human nutritional requirement for normal growth and adequate
protection against diseases arising from malnutrition.

According to LC Chuku et al (2021). Article no.JOCAMR.64445, Proximate and phytochemical

analyses were carried out on dried fruited pumpkin (Telfaira occidentalis) seeds . Standard
experimental procedure and statistical analysis using SPSS version 20.1 were carried out after
triplicate evaluation of the sample. Results of the proximate analysis indicated that the dried
seeds contains moderate percent of carbohydrate (11.43±0.92%) and fibre (15.71±0.74%), high
percentage protein (34.56±1.36%) and lipid (32.50±1.08%), ash (4.40±0.02%) and moisture
(1.40±0.01%) contents were low statistically. The qualitative and quantitative phytochemical

7
analysis showed that dried seeds of Telfairia occidentalis possessed very low (statistically
insignificant, p> 0.05) level of cyanogenic glycoside (0.001±0.01 µg/g) which makes it non toxic
and consumable. Tannin (0.488±0.012 µg/g) and oxalate (0.194±0.01 µg/g) where low in
percentage, while phylate (3.75±0.018 µg/g) and saponin (4.00±0.02 µg/g) levels were
statistically high (p< 0.05). The analysis revealed the sample as rich in protein and lipid, with
moderate amounts of carbohydrate and fibre. It also indicates absence of cynogenic glycoside
and phenolics.

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 CHAPTER THREE
3.0. MATERIALS AND METHODS
3.1 materials

3.1.1 Equipments

• Oven.
• Centrifuge.
• Chromatography.
• Spectrometer.
• Balance scale.
• PH meter.
• Water bath.
• Funnel
• Beaker
• Cotton wool
•

3.1.2 chemicals/Reagents

• Methanol.
• Hydrochloric acid
• sodium hydroxide.
• Potassium dichromate
• sulfuric acid.
• Acetic acid .
• Boric acid.
• %5 ferric acid.
• Chloroform

9
3.1.3 method.
3.1.3 collection of plant material and extraction.
In a black polythene bag, fresh leaves of Telfairia occidentalis was obtained from jimeta, bekaji
Gambia crescent no 14, Yola north l.g.a of adamawa state Nigeria.

3.2.1 Experimental design.

The leaf was properly rinsed and dried at room temperature (24°c). The leaf was pulverized
using mechanical grinder to obtain a smooth mixture. Methanol extracts of the plants were
prepared by soaking 1000g of the dry powdered leave of T. Occidentalis in 600ml of methanol
and then kept at room temperature for 24hours (for thorough extraction). At the end of the
24hours, the extracts were filtered first through a Whatmann filter paper No. 42 (125mm) and
then through cotton wool. The filtrate was kept in an oven at a very low temperature for 24hrs to
dry in other to get a smooth powder.

3.2.2 proximate analysis

• Determination of Moisture content

A dry clean petridish was placed in an oven at 80°C for about 30 minutes, cooled in a desiccator
and weighed as (w). A 1.4g of the samples was added to the petridish and weighed as (b). The
petridish and its content were placed in an oven adjusted to 70°C. After 1 hour, the petridish
containing the sample was removed and quickly transferred to a desiccator for cooling. The
petridish was put back into the oven and adjusted to 105°C for another 1 hour after which it was
removed, put in desiccators for cooling. This process was repeated and weighed until a constant
weight (c) was obtained.

The % moisture content was determined as follows;

% moisture content =

Where

W = weight of moisture can; b = weight of petridish + sample before drying; c = weight of

petridish + sample after drying.

• Determination of ash content

10
An empty crucible was first ignited in a muffle furnace for 1min and allowed to cool in
desiccators containing silica gel. A 1.4g of the sample was accurately weighed into the preheated
dish. The weight of the porcelain dish and the samples were noted. Afterwards, the dish was
heated with a Bunsen burner in a fume cupboard until smoking ceases and later transferred into a
muffle furnace at 550-570°C for about 18-24hours to burn off all organic matter. After ashing,
the crucible was removed from the furnace and placed in desiccator to cool at room temperature
and weighed. The percentage ash content of the sample was calculated thus;

% Ash =

Where;

W1 = weight of empty crucible; W2 = weight of crucible + sample before ashing

W3 = weight of crucible + sample after ashing.

• Determination of lipid.

1.4g of the sample was weighed into a thimble and was extracted with petroleum ether until it
siphons using the Soxhlet extraction method. The lipid was exhaustively extracted using
petroleum ether at 40 – 60°C for 6hrs. The sample in the thimble was removed and dried in air at
in air at 50°C for 5mins, cooled in a desiccator and weighed. The % lipid content was calculated
as follows;

% Lipid =
Where
W1 = weight of empty thimble; W2 = weight of thimble + sample
W = weight of sample used
• Determination of crude fiber

1.4g of the defatted sample was weighed into conical flask and 200mls of 1.25% of boiling
sulphuric acid was added within a minute. The content of the flask was filtered through a
buchner funnel prepared with wet 12.5cm filter paper. The sample was washed back into the
original flask with 200mls of 1.25% NaOH, and boiled for 30mins. All insoluble matter was
transferred to the crucible and treated till the sample was free from acid. The sample was again

11
washed in a muffle furnace at 550°C/hr. The crucible was then cooled in desiccator and
reweighed.

% Crude fiber =

Where

W = weight of sample; W1 = weight of crucible+ sample

W2 = weight of crucible+ filter paper after ashing.

• Determination of Crude protein

A 1.4g of the sample was weighed and transferred into Khedahl flask. Few chips of antibumping
granules, 2g of digestion catalyst and 10mls of conc. sulphuric acid were added at a 40°C angle
with a retort stand on an electro thermal heater. The flask was gently heated for frothing to occur
and subside, and then heat was increased to about 250°C. The digestion was carried out within 2-
6 hours by which time the entire sample was digested completely. The digest was cooled to room
temperature and diluted to 100mls with distilled water.

For distillation, 20mls aliquot of the digest was transferred into a round bottomed flask. This
flask was connected to a Liebig condenser through a monoarm steel head (Adaptor). The liebig
condenser was connected to a receiver flask through a receiver adapter. 10mls of 2% boric acid
and two drops of double indicator were pipetted into the distillation flask. 30mls of 40% sodium
hydroxide was injected into the distillation flask through a cork with the aid of a syringe. The
flask was heated for 10mins to digest the content. The distillate was collected in the boric acid
and then titrated with 0.1M HCL. The vol. of HCl added was recorded as the titre value. The %
Crude protein was calculated thus;

Where, 1.4 = N2 equivalent to 0.1NHCI used in titration

100 = Total volume of digest

• Determination of total carbohydrate.

The total carbohydrate content of the sample was estimated as the Nitrogen free extract (NFE).
The arithmetic different methods involve adding the total percentage value of crude volume.

12
3.2.3 phytochemicals screening.

3.3.1 Qualitative phytochemical

Phytochemical screening was performed using standard procedure to identify the constituents as
described by Harborne (1973), Nweze et al., (2004) and Senthilkumar and Reetha, (2009).

3.3.2 Quantitative Analysis of Phytochemicals

• Determination of Alkaloids (Harborne, 1973)

A 3ml of plant extract and 3ml of concentrated hydrochloric acid was added into a test tube. To
the mixture above, 3drops of Mayer’s reagent was added. Presence of green colour or white
precipitate indicates the presence of alkaloids

• Determination of Tannins

A 1.4g of the plant extract was weighed in a cornical flask. 10ml of distilled water was added
and shaken for 15minutes and then filtered into a volumentary flask. About 1ml of plant extrac
was pipette into a test tube and 2ml of 5% ferric chloride was added and observed for Brownish
green or a blue-black colouration.

• Determination of Saponins

To 1ml of the plant extract was pipette into a test tube, 5ml of distilled was added and shaken in
a graduated cylinder for 15minutes. Formation of 1cm layer of foam indicates the presence of
Saponins.

• Determination of Flavonoids (Sofowora, 1993; Harborne, 1973).

A 5ml of diluted ammonia solution were added to a portion of the filtrate of each plant
extract followed by addition of concentrated H2SO4. A yellow colouration observed in
each extract indicates the presence of flavonoids. The yellow colouration disappeared on
standing. Few drops of 1% aluminium solution were added to a portion of each filtrate. A
yellow colouration was observed indicating the presence of flavonoids. A portion of the
powdered plant sample was in each case heated with 10ml of ethyl acetate over a steam
bath for 3minutes. The mixture was filtered and 4ml of the filtrate was shaken with 1ml

13
 of dilute ammonia solution. A yellow colouration was observed indicating a positive test
for flavonoids.
• Determination of Total Phenols.

To 1ml of the extract, 2ml of distilled water followed by 5drops of 10% ferric chloride was
added. Formation of blue or green colour indicates the presence of phenols.

• Determination of cyanogenic glycosides.

To 2ml of plant extract, 1ml of glacial acetic acid and 5% ferric chloride was added
followed by 3drops of concentrated sulphuric acid. Presence of greenish blue colour
indicated the presence of glycosides.
• Determination of steroids
To 1ml of plant extract, equal volume of chloroform and 3 drops of concentrated
sulphuric acid was added. Formation of brown ring indicates the presence of
steroids.
• Determination of terpenoids.
5ml of aqueous extract of each plant sample was mixed with 2ml of Chloroform
in a test tube and then 3ml of concentrated H2SO4 was carefully added to the
mixture to form a layer. An interface with a reddish brown coloration indicates
that terpenoids constituent is present.

3.3.3 Method of Data analysis.

All data collected was subjected to descriptive and one way analysis of variance using Statistical
Package for Social Sciences (SPSS), Inc.20.0 software. All data will be represented in
mean±standard deviation (M±S.D). Confident level of determination (P=0.05).

14
 CHAPTER FOUR
4.1. Results.

Percentage proximate composition of the leaves as shown in Table 1 are as follows: moisture
content (10.99 ± 1.34 ), fiber content (11.56 ± 0.68 ), lipid content (46 ± 0.07), ash content (8.31
± 0.21 ), protein content (21.14 ± 0.32 )and carbohydrate (53.10 ± 0.68 ). The phytochemical
analysis conducted on T. occidentalis dried and powdered leaves (in Table 2 and 3) revealed the
presence of saponins , tannin, alkaloids, flavonoids, steroids, terpenoids and phenolics with
composition as follows: saponins (7.18± 0.02 ); tannin (2.77 ± 0.10); alkaloids (5.51 ± 0.11);
flavonoids (16.9 ± 0.01), steroids (0.22 ± 0.01) terpenoids (1.06 ± 0.01 ), phenols(7.95 ± 0.013).

Table. 1 proximate composition

Proximate Composition

%Moisture 10.99 ± 1.34

%Fiber 11.56 ± 0.68

% lipid 6.46 ± 0.07

%Ash 8.31 ± 0.21

%Protein 21.14 ± 0.32

%Carbohydrates 53.10 ± 0.68

Values are expressed Mean ± SEM (n = 3)

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Table. 2 qualitative phytoconstituent of telfaria occudentalis methanol leaf extract
Phytochemicals Present or negative

Saponins +

Tannins +

Alkaloids +

Flavonoids +

Steroids +

Terpenoids +

Glycosides _

Phenols +

Where; + = present and –= absent.

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Table. 3 quantitative phytochemical of T. Occidentalis.
T. Occidentalis Sample

Saponins 7.18± 0.02

Tannins 2.77 ± 0.10

Flavonoids 16.9 ± 0.1

Steroids 0.22 ± 0.01

Terpenoids 1.06 ± 0.01

Phenol 7.95 ± 0.013

Alkaloids 5.51 ± 0.11

Values are expressed Mean ± SEM (n = 3)

*Significantly higher compared to different constituent

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 CHAPTER FIVE
5.1 Discussion

The phytochemical analysis conducted on T. occidentalis dried and powdered leaves (Tables 2
and 3) revealed the presence of saponins, tannins, alkaloids, flavonoids, steroids, terpenoids and
phenolics. These phytochemical compounds are known to support bioactive activities in
medicinal plants. Tannins are known to be useful in the treatment of inflamed or ulcerated tissues
and have remarkable activity in cancer prevention (Ruch et al., 1989, Motar et al., 1985). Thus T.
occidentalis containing tannins may serve as a potential source of bioactive compound in cancer
prevention and treatment. Flavonoids are antiinflammatory, antitumor, anti-viral, antiplateletes
(Corkan et al, 1998, Pourmorad et al, 2006)). Flavonoids also are potent water soluble
antioxidants and free radical scavengers, which prevent oxidative cell damage and have strong
anticancer activity (Salah et al., 1995, Del-Rio et al., 1997, Okwu, 2004). It has been recognized
that flavonoids, which contain hydroxyl groups are responsible for the radical scavenging effects
of most plants. Alkaloids are beneficial chemicals to plants serving as repellant to predators and
parasites. Some alkaloids are useful against HIV infection as well as intestinal infection
associated with AIDS (McDevitt et al., 1998). Also, the plant extract was revealed to contain
saponins, known to produce inhibitory effect on inflammation (Just et al, 1998). Saponins are
known bioactive substances that can reduce the uptake of cholesterol and glucose in the gut
through intra-lumenal physiochemical interaction (Price et al., 1987). Saponins as a class of
natural products are also involved in complexation with cholesterol to form pores in cell
membrane bilayers (Francis et al., 2002) as such may be used as anticholesterol agents or
cholesterol lowering agent. The presence of these phenolic compounds in this plant contributes
to their antioxidative properties and thus the usefulness of these plants in herbal medicament.
Phenols also have a potential of combating oxidative stress syndrome, causative of some
neurodegenerative diseases and cardiovascular diseases. These properties bestow high medicinal
activities on T. occidentalis. Steroids which plays a role in regulating Human hormones and also
supports heart health. Terpenoids which have therapeutic benefits such as anti-inflamatory and
antimalaria.

Plant proteins are source of food nutrient especially for the less privileged population in
developing countries including Nigeria. Fibre cleanses the digestive tract by removing potential

18
carcinogens from the body and prevents the absorption of excess cholesterol. Fibre also adds
bulk to the diet and may therefore guard against metabolic conditions such as
hypercholesterolemia and diabetes mellitus. Thus, the benefit of consumption of T. occidentalis
leaves cannot be over-emphasized. The high ash content is a reflection of the mineral content of
this food material. The results therefore suggest a high deposit of mineral elements in the leaves
(Antia et al., 2006). Carbohydrates are essential for the maintenance of life in both plants and
animals and also provide raw materials for many industries (Ebun-Oluwa and Alade, 2007). The
plant is a good source of carbohydrate when consumed because it meets the recommended
dietary allowance (RDA) values (FND, 2002). The plant is considered a good source of protein
because it provides more than 12% of caloric value from protein (Pearson, 1976). Lipid increases
the palatability of food by absorbing and retaining flavours (Antia et al., 2006). A diet providing
1 - 2% of its caloric of energy as fat is said to be sufficient for humans as excess fat consumption
is implicated in certain cardiovascular disorders (Antia et al., 2006).The moisture content of the
dried T.occidentalis leaves was 10.99% which is relatively low, and would therefore hinder the
growth of spoilage microorganisms and thereby enhance the shelf life.

5.1.2. Conclusion

This study justifies the use of T. occidentalis leaves in treatment of certain diseases such as
diabetes, hypercholesterolemia, liver problems e.t.c, in which the participation of reactive
oxygen species have been implicated. This could be as a result of the phytochemicals, nutritional
ability of T. occidentalis leaf. Thus T. occidentalis leaf if consumed in sufficient amount could
contribute greatly towards meeting human nutritional requirement for normal growth and
adequate protection against diseases arising from malnutrition.

5.1.3. Recommendations

Further research on the effect and antinociceptive of methanol leaf extract of telfaria
occudentalis should be carried out for broad understanding.

19
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