CHAPTER ONE

1.0 INTRODUCTION

In the history of humanity, plants have always been present as a source of health. The knowledge

of the various healing properties of plants has been transmitted in an empirical way. However,

over time, man has been interested in knowing where the properties of plants come from. In the

process of knowledge generation, man has developed many methodologies to know the

structures of organic compounds responsible for the healing properties of plants. This is the birth

of phytochemistry that is defined as the science responsible for the study of the compounds

contained in plants. In this field, various techniques have been developed, ranging from the

preparation of the plant tissue sample to sophisticated techniques for the elucidation of organic

structures (Dreyfuss et al., 1994).

The genus Jatropha, which belongs to the tribe Joannesieae in the Euphorbiaceae family,

contains approximately 170 known species. The name Jatropha is derived from the Greek word

‘‘jatros’’ (doctor) and ‘‘trophe” (food), which implies its medicinal uses (Kumar and Sharma

2008). Jatropha species are widely used in traditional folklore medicine to cure various ailments

in Africa, Asia and Latin America and are also used as ornamental plants and energy crops

(Félix-Silva et el., 2014).

Jatropha species have been used as medicinal plants by native people in many tropical and

subtropical countries. For instance, Jatropha species are famous for the purgative effect of the

seed oil. This purgative effect has been directed to cure digestive system symptoms like diarrhea,

dysentery, vomiting, retching and stomachache. Additionally, some parts of Jatropha plants are

employed to heal skin-related ailments. The seed oil, leaf, latex, stem bark or root of Jatropha

plants are pounded and applied on infected skin such as eczema, itches, mouth blisters,

1
carbuncles, wounds and swellings. They are also believed to cure venereal diseases and urinary

discharge. Moreover, the roots of some Jatropha species have long been applied on people

suffering from leprosy and gonorrhea (Sabandar et al., 2012).

Several reviews have been conducted on the different species of the genus Jatropha covering

various aspects such as their ethnobotany, medicinal properties, phytochemistry, and toxicity

among others (Devappa et al., 2010). Phytochemical studies of the genus Jatropha have

increased in recent years due to the high potential of these species as natural sources of bioactive

compounds. Investigations of the chemical constituents of Jatropha plants resulted in the

isolation of a number of alkaloids, cyclic peptides, terpenes (monoterpene, sesquiterpenes,

diterpenes and triterpenes), flavonoids, lignans, coumarins, coumarino-lignoids, a non-

cyanogenic glucoside, phloroglucinols, ester ferulates, phenolics, deoxypreussomerins and fatty

acids (Sabandar et al., 2012). Moreover, extracts and isolated compounds from various species

of this genus have been found to possess properties of cytotoxicity, antimicrobial, anti-

inflammatory, antioxidant, insecticidal, larvicidal, cholinesterase inhibition, and toxicity

activities (Cavalcante et al., 2020).

In particular, among the various Jatropha species has been documented to exhibit promising

biological effects. For instance, its stem latex has been reported to possess coagulating features

by reducing clotting and bleeding times in experiments, thereby providing a scientific basis for

its use as a haemostatic agent (Oduola et al., 2020). Furthermore, Jatrophone, an active

compound isolated from Jatropha species has been reported to show a better anticancer effect

against hepatocellular carcinoma (Hep G2 1886) compared to standard anticancer drugs like

sorafenib and arsenic trioxide (Sukohar 2017).

2
Another important species of the genus J. curcas has also been appraised for its broad spectrum

of pharmacological activities. As example, extracts of this plant were found to display antiviral

activity on human immunodeficiency virus, (HIV) (Dahake 2013).While others reported

remarkable anti-inflammatory and antibacterial, cosmetic and wound healing properties (Warra

2012).

Psidium guajava (guava) is a well-known tropic tree which is abundantly grown for fruit. Many

countries have a long history of using guava for medicinal purposes. This plant finds applications

for the treatment of diarrhea, dysentery, gastroenteritis, hypertension, diabetes, caries and pain

relief and for improvement in locomotors coordination. Its leaf’s extract is being used as a

medicine in cough, diarrhea, and oral ulcers and in some swollen gums wound. Its fruit is rich in

vitamins A, C, iron, phosphorus and calcium and minerals. It contains high content of organic

and inorganic compounds like secondary metabolites e.g. antioxidants, polyphenols, antiviral

compounds, anti-inflammatory compounds. The phenolic compounds in guava help to cure

cancerous cells and prevent skin aging before time. The presence of terpenes, caryophyllene

oxide and p-selinene produces relaxation effects. Guava leaves contain many compounds which

act as fungistatic and bacteriostatic agents. Guava has a high content of important antioxidants

and has radio-protective ability. Quercetin is considered as most active antioxidant in the guava

leaves and is responsible for its spasmolytic activity. Its ethyl acetate extract can stop the germ

infection and thymus production. Guava possesses anti-viral, anti-inflammatory, anti-plaque and

anti-mutagenic activities. Guava extract shows antinociceptive activity and is also effective in

liver damage inflammation and serum production. Ethanolic extract of guava can increase the

sperm quality as well as quantity and can be used for the treatment of infertile males (Naseer et

al., 2017).

3
1.1 Statement of problem

Antimicrobial resistance happens when germs like bacteria and fungi develop the ability to

defeat the drugs designed to kill them. That means the germs are not killed and continue to grow.

Resistant infections can be difficult and sometimes impossible to treat. This Antimicrobial

resistance is an urgent global public health threat, killing at least 1.27 million people worldwide

and associated with nearly 5 million deaths in 2019. In Nigeria, more than 2.8 million

antimicrobial-resistance infections occur each year. More than 35,000 people die as a result,

according to CDC’s 2019 Antibiotic Resistance (AR) Threats Report. This serious global

problem has prompted the search for new organic molecules with antimicrobial properties

(Phytochemicals) and potentials to serve as sources of raw material for the synthesis of new

drugs.

1.2 Significance of study

Since inception it has been known that plant materials are used as food and that they contain

phytochemicals. These phytochemicals protect plants from harm; they are nonnutritive plant

chemicals that have protection or disease preventive properties. The need for a cheap, easily

available and less side effect of this source of material has therefore attracted me to investigate

its content and how it can be utilized as an alternative to solve some health issues.

4
1.3 Aim

To assess the phytochemical composition of the leaf extracts of Jatropha curcas and

Psidium guajava.

1.4 Objectives

I. To identify the phytochemicals present in the leaf extract of Jatropha curcas and Psidium

guajava

II. To determine the quantity of phytochemicals present in the leaf extract of Jatropha

curcas and Psidium guajava

III. To compare the quantity of phytochemicals present in the leaf extract of Jatropha curcas

and Psidium guajava

5
 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 PHYTOCHEMISTRY

The discipline whose main objective is the study of the chemical constituents of plants is

Phytochemistry. The study of such compounds includes: their chemical structures, metabolism

(biosynthesis and degradation), natural distribution, biological function, extraction and

qualitative-quantitative evaluation. Before starting, any phytochemical analysis is important to

have an adequate preparation of the plant material. A practical and simple way of stabilization is

by heat treatment, applied, for example, in an oven at a reference temperature of 60 °C until the

samples reach constant weight; this way, one would make sure that the compounds will be in the

optimal conditions to be analyzed (Nadia and Eleazar, 2018).

Phytochemicals (from the Greek word phyto, meaning plant) are biologically active, naturally

occurring chemical compounds found in plants, which provide health benefits for humans further

than those attributed to macronutrients and micronutrients. They protect plants from disease and

damage and contribute to the plant’s color, aroma and flavor. In general, the plant chemicals that

protect plant cells from environmental hazards such as pollution, stress, drought, UV exposure

and pathogenic attack are called as phytochemicals. Recently, it is clearly known that they have

roles in the protection of human health, when their dietary intake is significant. More than 4,000

phytochemicals have been cataloged and are classified by protective function, physical

characteristics and chemical characteristics and about 150 phytochemicals have been studied in

detail (Mamta et al., 2013).

6
Phytochemical research of a plant includes several aspects:

- Extraction of the compounds to be analyzed from a sample or specimen.

- Separation and isolation of them.

- Identification and/or characterization of the isolated compounds.

- Investigation of the biosynthetic routes of a certain molecule.

- Determination or quantitative assessment.

The extraction and purification of organic compounds through the use of solvents, usually

follows certain rules based on structural analogies between the substance to be extracted and the

solvent that will be used for that purpose (Nadia and Eleazar, 2018).

The polarity of the compounds is another element to be taken into account, when considering the

solubility of a solute in a given solvent. Thus, strongly polar solvents dissolve ionic or highly

polar solutes, while low-polar solvents do not efficiently dissolve ionic solutes but do dissolve

low-polarity solutes (Nadia and Eleazar, 2018). The extraction of the vegetal material is done

consecutively using solvents, from a low polarity until reaching the water, which is the most

polar solvent. The obtained extracts can be clarified by filtration through celite with a vacuum

pump and then concentrated under reduced pressure. This is generally carried out in a rotary

evaporator, in which the solutions are concentrated until a volume reduction is achieved, at

temperatures between 30 and 50°C. The concentrated extracts must be stored refrigerated. In the

separation and identification of natural products, different techniques for isolation and

identification have been developed; this table is a summary of the main techniques

7
 Table 2.1 Types of separation and identification techniques

Chromatography Thin-layer chromatography (TLC)

Gas chromatography (GC)
High-resolution liquid chromatography (HPLC)
Capillary liquid chromatography (u-LC)
Electrophoresis Thin-layer electrophoresis (TLE)
Isotachophoresis (ITP) (electrophoresis at uniform speed)
Capillary electrophoresis (CE)
Spectroscopic UV spectroscopy
techniques Infrared spectroscopy (IR)
Near infrared spectroscopy (NIR)
Nuclear magnetic resonance spectroscopy (NMR)
Mass spectroscopy (MS)
Source: (Nadia and Eleazar, 2018).

2.2 TYPES OF PHYTOCHEMICALS

Phytochemicals have great antioxidant potential and are of great interest due to their beneficial

effects on health of human beings, and they give immense health benefits to the consumers.

Epidemiological and animal trials suggest that the regular consumption of fruits, and vegetables,

and whole grains reduces the risk of various diseases linked with oxidative damage (Cieslik

et al., 2006; Scalbert et al., 2005; Kris-Etherton et al., 2002).

Among the phytochemicals mentioned as potentially providing health benefits are polyphenols,

flavonoids, isoflavonoids, anthocyanidins, phytoestrogens, terpenoids, carotenoids, limonoids,

phytosterols, glucosinolates, and fibers.

The remaining organic chemicals, such as alkaloids, terpenes, flavonoids, lignans, plant steroids,

curcumines, saponins, phenolics, flavonoids, and glycosides, is considered as secondary

8
components. According to an analysis of relevant literature, phenolics are the most various and

chemically diverse plant phytochemicals (Saxena et al., 2013). Whereas phytochemicals is

categorized based on their function, a single compound may serve as both an antioxidant and an

antibacterial agent.

Phenolics

Since the late nineteenth century, when the French paradox was attributed with the high

consumption of phenolic content observed in red wine, researchers have been focused in plant

phenolics as anti carcinogenic and curative against chronic and degenerative diseases. Ever

since, studies investigated the biosynthesis, bioactivities, detoxification, and chemical

identification of phenolic content in different plants. Furthermore, analysis on the durability of

phenolic content in food industry and collection has become a primary concern (Jacob-velazquez

et al., 2017). Phenolic are the most abundant phytochemicals and are found throughout the

kingdom Plantae. Phenolics are a group of chemical compounds that contain hydroxyl groups,

where the (OH) group is directly bonded to an aromatic hydrocarbon group. Flavonoids,

phenolic acids, and polyphenols are the three most important types of dietary phenolic (Koche et

al., 2016). Many agronomic, pharmacological, chemical, and medical investigations have been

performed on phenolics compounds and functions (Cseke et al., 2016).

9
 Chemical structure of Phenolics.

Flavonoids

Flavonoids are derived from flavones and have two benzene rings separated by a propane unit. In

general, they are water- soluble compounds. The more complexed the compound, the more

vibrant it is. They are widely obtained from plants as glycosides, which can consider formation

persistence more nearly impossible (Hassanpour et al., 2016). Flavonoids have gained popularity

in recent years due to its wide pharmacological activities, anticancer activities to exert multiple

biological properties such as antimicrobial, cytotoxicity, anti-inflammatory.

Chemical structure of flavonoids

Tannins

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Tannins are phenolic compounds with high specificity ranging from 500 Da to more than 3000

Da identified in plants' leaves, bark, fruit, wood, and roots, mainly in the lysosomes. Plant

defense mechanisms against mammalian herbivores, birds, and insects have been linked to them.

Tannins are classified into two types based on their chemical characteristics and composition:

hydrolysable tannins and condensed tannins (Hassanpoour et al., 2016). Tannin-containing

phytoconstituents is used as exfoliates, diuretics, antitumor of the gastrointestinal system, as well

as anti-inflammatory properties, antibacterial, free radical scavenging, and haemostatic therapies.

Tannins are used in the food products to clarify wine, beer, and fruit juices.

Chemical structure of

Terpenoids

Terpenoids, already identified as isoprenoids, are the most diverse and abundant natural remedies

in terms of structure.2- methylbuta-1, 3-diene is the chemical formula for isoprene, the "building

block" of terpenoids (C5H8). Several terpenoids are industrially interesting due to their use as

flavours and fragrances in edible products and cosmetics, including such menthol and sclareol, as

well as because they are essential for agro-based quality product, such as the taste of fruits and

11
the aroma of flowers, such as linalool (Saxena et al., 2013). Terpenoids classification is based on

the number of isoprene units present. Monoterpenoids, sesquiterpenes, diterpenes, triterpenes,

and tetraterpenoids. According to preliminary research, terpenes play an important role in plant

signaling and growth regulation. Terpenoids may also have medicinal benefits such as anti-

mutagenic, anti-ulcer, hepaticidal, antimicrobial, or diuretic activity, as well as the

sesquiterpenoid antimalarial drug artimisinin and the diterpenoid anticancer drug taxol (Saxena

et al., 2013).

Chemical structure of terpernoids.

Saponins

Saponins have been found in a variety of plants as well as in a little under marine sources such as

starfish sea cucumber and fish (Guclu et al., 2007). Saponins are divided into two categories

based on the nature of their aglycone skeleton. The first category contains of steroidal saponins,

which are mostly primarily found in monocotyledonous angiosperms. The second group consists

of triterpenoid saponins, which are the most abundant and are found primarily in dicotyledonous

angiosperms (Sparg et al., 2004). Saponins are easily identified in phytoconstituents by their

haemolytic activity and ability to generate stable foams in aqueous medium, but for unequivocal

recognition, thin layer chromatography with a wide range of spray reagents is required. Thin

layer chromatography is also the preferred method for quantitative saponin analysis. Older

12
approaches, such as those based on haemolytic activity are not as accurate as they once were

(Oakenfull et al., 1981).

Chemical structure of saponins.

Table 2.2 Phytochemicals as well as their food sources and health benefits.

S/N Phytochemicals Sources Health benefits

13
1 Carotenoids Carrots, tomatoes, Antioxidants protect against
parsley, orange and uterine, prostate, colorectal,
green leafy lung, and digestive tract
vegetables, cancers
chenopods,
fenugreek, spinach,
cabbage, radish,
turnips

2 Phytosterols Vegetables, nuts, Suppress the growth of diverse

fruits, seeds tumors cell lines via initiation
of apoptosis and concomitant
arrest of cells in the G1 phase
of the cell cycle

3 Polyphenols Fruits, vegetables, Action against free radicals,

➢Flavonoids cereals, beverages, free radicals mediated cellular
➢Isoflavonoids legumes, signaling, inflammation,
➢Anthocyanidins chocolates, oilseeds allergies, platelet aggregation,
and hepatotoxins

4 Phytoestrogen Legumes, berries, Protection against bone loss

whole grains, and heart disease,
cereals, red wine, cardiovascular diseases, breast
peanuts, red grapes and uterine cancers
5 Terpenoids Mosses, liverworts, Antimicrobial, anti-parasitic,
(Isoprenoids) algae, lichens, antiviral, anti-allergic, anti-
mushrooms inflammatory,
chemotherapeutic, anti-
hyperglycemic, antispasmodic

6 Polysaccharides Fruits and Antimicrobial, antiparasitic,

vegetables antiviral, antiallergic,
antiinflammatory, lowering
serum, enhances defense
mechanisms

7 Saponins Oats, leaves, Protection against pathogens,

flowers, and green antimicrobial,
fruits of tomato antiinflammatory, antiulcer
agent
Source: (Monika Thakur et al., 2020).

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2.3 Jatropha curcas

The genus Jatropha belongs to the family Euphorbiaceae and has a great variety of species,

among them: J. multifida, J. curcas, J.molissima, J. gossypifolia that are currently the source of

studies for the production of biodiesel and also for the medicinal character that they have. They

are used in traditional folklore medicine to cure various ailments in Africa, Asia and Latin

America (Devappa et al., 2010). Their usage as traditional health remedies is the most popular

for 80% of the world population in Asia, Latin America and Africa and is reported to have

minimal side effects (Cowan, 1999). In this genus, Jatropha curcas have played major role in the

treatment of various diseases, including bacterial and fungal infections. All parts of Jatropha

(seeds, leaves, bark, etc) have been used in traditional medicine and for veterinary purposes for a

long time (Prasad et al., 2012).

It has many names in different regions like the physic nut, goat nuts, (pinhão-manso in Brazil),

Barbados nut, purging nut, nettle spurge, or just Jatropha (Wurdack et al., 2005) previous

researches showed that Jatropha is native to Central and South America (Pecina-Quintero et al.,

2014).

Jatropha curcas is a valuable multi-purpose crop; historically it was used as medicine for

wounds and leaves used as drinks against malaria. Jatropha plants are used to control soil

degradation, alleviate erosion, desertification and increase soil fertility, however, in last decades

there is more attention to use Jatropha oil for produce biodiesel, Jatropha curcas is easily

propagated by seeds or stem cutting, it is tolerant for drought for longtime, it grows well with

treated wastewater, also, it can be grown on marginal land. Jatropha curcas seeds have about 32-

40% valuable oil used to produce biofuel, therefore, it could be the source for biodiesel

production particularly in arid and semiarid regions.

15
2.3.1 Botanical description of Jatropha curcas

Jatropha curcas is a drought-resistant species which is widely cultivated in the tropics as a living

fence. Many parts of the plants are used in traditional medicine. The seeds, however, are toxic

to humans and many animals. Jatropha curcas by definition is a small tree or large shrub which

can reach a height of up to 5 m. The plant shows articulated growth, with a morphological

discontinuity at each increment. Dormancy is induced by fluctuations in rainfall and

temperature/light. The branches contain latex. Normally, five roots are formed from seedlings,

one central and four peripherals. A tap root is not usually formed by vegetative propagated plants

(Kobilke 1989). Jatropha curcas has 5 to 7 shallow lobed leaves with a length and width of 6 to

15 cm, which are arranged alternately. Inflorescences are formed terminally on branches and are

complex, possessing main and co-florescences with paracladia. Botanically, it can be described

as a cyme. The plant is monoecious and flowers are unisexual; occasionally hermaphrodite

flowers occur (Dehgan, 1979). Ten stamens are arranged in two distinct whorls of five each in a

single column in the androecium, and in close proximity to each other. In the gynoecium, the

three slender styles are connate to about two-thirds of their length, dilating to massive bifurcate

stigmata (Dehgan and Webster, 1979). Pollination of Jatropha curcas is by insects. Dehgan and

Webster, (1979) believe that it is pollinated by moths because of “its sweet, heavy perfume at

night, greenish white flowers, versatile anthers and protruding sexual organs, copious nectar, and

absence of visible nectar guides”. When insects are excluded from the greenhouse, seed set does

not occur without hand-pollination. The rare hermaphrodite flowers can be self-pollinating.

After pollination, a trilocular ellipsoidal fruit is formed. The exocarp remains fleshy until the

seeds are mature. The seeds are black, 2 cm long and 1 cm thick. The caruncle is rather small.

Wiehr (1930) and Droit (1932) described the microscopical anatomy of the seeds in detail, while

16
Singh (1970) described that of fruits. Gupta (1985) investigated the anatomy of other plant parts.

Jatropha curcasis a diploid species with 2n = 22 chromosomes.

Jatropha curcas

III.3.2 Taxonomic Classification of Jatropha curcas

Kingdom: Plantae

Order: Malpighiales

Family: Euphorbiaceae

Genus: Jatropha

Species: J. curcas

Source: (Kaushik, 2003)

17
2.3.3 Medicinal Benefit of Jatropha curcas

J. curcas is a multiple purposes plant and various parts of the plant are used in folk and

traditional medicine worldwide. All parts of J. curcas have been widely used in west and central

Africa (Neuwinger, 1996). The dried plant sap rubbed to a powder between the hands and

applied to wounds is regarded as “penicillin” in Congo. In Senegal, Nigeria, Congo and East

Africa, the leaf, stem sap or the dried powdered plant is spread on flesh wounds as a haemostatic.

In Ivory Coast grilled leaves are crushed together with saliva and the paste is applied to

abscesses and wounds. A few drops of diluted water solution of twig sap are given by mouth to

new-born babies affected by tetanus. The leaf has been used as haemostatic agent when applied

to cuts and bleeding wounds (Neuwinger, 1996; Staubmann et al., 1999). In Southeast Asia and

in some regions of Africa, the leaves are used as purgative while in Cape Verde and Cameroon,

the decoction of the leave is used internally and externally against fever. In Cameroon, the leaves

are also in use as the remedy against rheumatism and in Nigeria against jaundice (Staubmann et

al., 1999). In India, the juice from leaves is used to cure diseases such as dysentery and colic and

are also applied to the breast to promote lactation (Parveen et al., 2007).In many part of the

world, the seeds are used to ascites, gout, paralysis, skin diseases and as a purgative,

anthelminthic, abortifacient and as a laxative (Wole, 2009). The seed oil has been used as

ingredient in the treatment of rheumatism (Heller, 1996; Iwu, 1993). In Benin Republic, the

decoction of the leaves together with the roots and the fruits of Xylopiaethiopica is used as drink

to treat drepanocytosis while the fresh leaves mixed with kaolin pounded in water is also used as

a drink for the treatment of haemorrhoids (Neuwinger, 1996). In Nigeria, the latex is used as a

mouth rinse to treat bleeding gums and to sooth a baby’s inflamed tongue. In the Philippines,

Indonesia and in Benin Republic, a little latex on absorbent cotton is used to cure a toothache.

18
In the island of Tonga, in Oceania, the leaves of J. curcas have been used in folk medicine to

treat vaginal bleeding (Singh et al., 1984). Fagbenro-Beyioku (1998) investigated and reported

the anti-parasitic activity of the sap and crushed leaves of J. curcas. In Mali, the leaves are used

as treatment for malaria (Henning, 1997). The leaves are utilized extensively in West Africa

ethnomedical practice in different forms to cure various ailments like fever, mouth infections,

jaundice, guinea worm sores and joint rheumatism (Oliver-Bever, 1986).

Table 2.3 The uses of leaves of Jatropha curcas in folk and traditional medicine

Plant Treated disease Medical practice References

part

Leaves Malaria Decoction with Azadirachta Asase et al., (2005)

indica and Carica papaya
Leaves Wound healing Leaves Staubmann et al.,
applied to wounds (1999)
Leaves Fever Decoction is used internally Staubmann et al.,
and externally (1999)
Leaves Rheumatism Leaf decoction is applied Staubmann et al.,
externally (1999)
Leaves Jaundice Application of the leaves Staubmann et al.,
(1999)
Leaves Drink for diabetes Decoction of boiled leaves Gbolade (2009)
(J. curcas +
Syzygiumguineese) + palm
oil
Leaves Arthritis; against abscess in Raw leaves Sandberg et al.,
the stomach (2005)

Leaves Oedemas and cough (orally Leaf decoction Neuwinger, (1996)

and external use)
leaves Drepanocytosis (drink) Decoction of leaves + roots Neuwinger, (1996)
+ roots + fruits of Xylopiaethiopica

Leaves Haemorrhoids Fresh leaves + kaolin Neuwinger, (1996)

pounded in water (drink 150
ml once a day)
Source: (Bossou et al., 2019).

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2.3.5 Phytochemical compounds isolated from leaf extract of J. curcas

The review of the literature shows that J. curcas is a plant with many bioactive compounds,

especially from the family of diterpene, sesquiterpenoids and triterpenes. In fact among 76

compounds identified by Abdelgadir, (2013), 42 compounds belong to the family of diterpene.

Few phenolic compounds have been identified in this plant.

Compounds that have been isolated from J. curcas leaves include the flavonoid apigenin and its

glycosides vitexin and isovitexin, the sterols stigmasterol, β-D-sitosterol and its β-D-glucoside

(Chhabra et al., 1990). Furthermore, J. curcas leaves were reported to contain steroid

sapogenins, alkaloids, the triterpenae alcohol, 1-triacontanol and a dimer of a tripene alcohol

(Neuwinger, 1994; Staubmann et al., 1999). Staubmann et al., (1999) had isolated a complex of

5- hydroxypyrrolidin-2-one and pyrimidine-2, 4-dione from the leaves of J. curcas by extraction

with ethyl acetate. From the leaves gave α-amyrin, isovitexin, N-1-triacontanol, steroids,

campesterol, stigmasterol, β-sitosterol, apigenin, vitexin and isovitexin where isolated. The plant

also yielded tetradecyl-(E)-ferulate, 3-O- (Z)-coumaroyl oleanolic acid, heudelotinone,

epiisojatro -grossidiones, 2-methyanthraquinon, curcusones, coumaric acids, phydroxybenzoic

acid, protocatechuic acid, resorsilic acid, saponins and tannins (Najda et al., 2013; Ribeiro et al.,

2012). Zhang et al. (2009) isolated some phenolic compound from the areal part of J. curcas

which were identified as tomentin, 5-hydroxy-6,7 dimethoxycoumarin, 6-methoxy-7-

hydroxycoumarin and 2,3,7-trimethoxy-8-O-β-D- glucoside ellagic acid. Isolation of one

phytosterol compound named 5α-stigmasta-3,6-diene was also reported.

The leaves of Jatropha curcas contain apigenin, vitexin and isovitexin. α-amyrin, stigmasterol,

stigmastenes along with two new flavonoids founds inleaves and twigs.

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2.4 Psidium guajava

Psidium guajava. Known as Guava is a medicinal plant belonging to the family Myrtaceae. P.

guajava is a well-known traditional medicinal plant used in various indigenous systems of

medicine. It is a native of Central America but is now widely cultivated, distributed and the fruits

enrich the diets of millions of people in the tropics of the world. It is a genus of about 133 genera

and more than 3,800 species of tropical shrubs and a small tree of about 10 m high with

spreading branches that thrives on all kinds of soils. It is one of the most gregarious fruit trees

and is widely known by its common English name (guava). In Nigeria, it is called guava

(Hausa), gurfa (Yoruba) and Gwaibwa (Igbo). P. guajava also known as the ‘poor man’s apple’

of the tropics has a long history of traditional use, and a good proportion of which have been

validated by scientific research. Nutritional value of guavas are often included among super

fruits, being rich in dietary fiber, vitamins A and C, folic acid; and the dietary minerals,

potassium, copper and manganese. Having a generally broad, low calorie profile of essential

nutrients, a single common guava fruit contains about four times the amount of vitamin C as an

orange. (Shruthi et al., 2011)

2.4.1 Botanical description

Psidium guajava is an evergreen shrub like tree which reaches to the height of about 6 to 25 ft’s.

Plate 7 displays various parts of the plant i.e., leaves, flowers, fruit, seeds and bark. The plant has

a wide spreading network of branches. Mostly its branches are curved which display opposite

leaves with the small petioles of about 3 to 16 cm. The leaves are wide and clear green in color

and have clear and prominent veins (Arima et al., 2002; Rouseff et al., 2008). The plant

produces white flowers with incurved petals having a nice fragrant. Flowers have four to six

petals and yellow colored anthers and pollination occurs by the insects. Guava fruit ranges from

21
small to medium sized with 3 to 6 cm length. It has pear like shape and yellow color in ripen

condition (Das et al., 2011) It has a musky special odor when ripened which is strong but

pleasant (Morton, 2004). Its pulp is slightly darker in color which contains slightly yellowish

seeds. The size of seeds is very small and they are easily chewable. They are arranged in regular

patterns; their number ranges from 112 to 535 (Morton, 2004; Kumar et al., 2011). The guava

bark is thin and has green colored spots. It is very easy to remove it in long straps. It has a huge

content of antimicrobial and antibacterial compounds (Rahim et al., 2010).

22
Various parts of guava (a) Leaves (b) Flowers (c) Fruit (d) Seeds in the fruit (e) Bark

Source: (Naseer et al., 2018).

23
2.4.2 Uses of Psidium guajava in folklore medicine

There are quite number of species of guava but the most common is the lemon guava which is

Psidium guajava. In folk medicine, different parts of the guava plant are used for the treatment of

various human ailments such as wounds, ulcers, bowels, cholera (Begum et al., 2002) across

different parts of the world. The inflorescence axillary 1-3 flowered trees are used for treatment

of various diseases especially in developing countries. In Nigeria and its neighboring countries,

guava leaves are used in treatment of malaria, typhoid and yellow fever.

Guava is a well-known traditional medicinal plant and is used in various indigenous system of

medicine. The fruits are often included among super fruits, being rich in dietary fiber, vitamins A

and C, folic acid and dietary minerals such as potassium, copper and manganese. Having a

generally broad, low calorie profile of essential nutrients, a single common guava (P. guajava)

fruit contains about four times the amount of vitamin C as an orange ( Hassimotto et al., 2005).

These constituent has made it possible to be used traditionally for treatment of ailments since a

long time in history. More recent ethno pharmacological studies showed that guava is used in

many parts of the world for the treatment of number of diseases such as inflammatory for

diabetes, hypertension, carries wounds, analgesic and antipyretic effects (Gutierrez et al., 2008).

The barks and root are also used for medicinal purposes. In the form of decoction and poultice,

the bark is used as an astringent in the treatment of ulcers, wounds, diarrhea, dysentery and skin

ailments (Conway, 2001). It is also used as anti-amoebic as an infusion or decoction. In the form

of decoction and poultice, it is used to expel the placenta after childbirth and in infections of the

skin, carries, vaginal hemorrhage wounds, fever, dehydration and respiratory disturbance. The

leaf is the mostly and widely used part of guava plant for medicinal purposes across the world. In

Nigeria and other African countries, the leaves are used in treatment of conditions such as

24
malaria, gastroenteritis, vomiting, diarrhea, dysentery wounds, ulcers, toothache, coughs, sore

throat, inflamed gums and a number of other conditions (Jiarj et al., 1999; Abdelrahim et al.,

2012; Lutterodt, 1989). The decoction or infusion of the leaves is used as febrifuge,

antispasmodic and for rheumatism (Hernandez, 1971). It is also used as an antibiotic and in

management of diabetic and hypertension in America, Central and West Africa and South East

Asia. In some parts of the world, boiled leaf extract is used for the treatment of rashes caused by

scabies.

2.4.3 Taxonomic classification of Psidium guajava

Kingdom: Plantae

Clade: Tracheophytes

Clade: Angiosperms

Clade: Eudicots

Clade: Rosids

Order: Myrtales

Family: Myrtaceae

Genus: Psidium

Species: P. guajava

Source: (Vikrant Arya et al, 2012).

2.4.4 Some phytochemical composition of the leaf extracts of Psidium guajava.

Psidium guajava has many medicinal properties. The bioactive components of Psidium guajava

leaves have been evaluated. The chemical compositions of many bioactive compounds are

responsible for therapeutic applications. The qualitative analysis of ethanolic and aqueous extract

of Psidium guajava leaves show that tannin, phlobatannins, saponin, flavonoids, steroids,

25
terpenoids, triterpenoids, polyphenol and glycoside are present in extracts. The quantitative

determination of Psidium guajava methanolic leaves contain Vitamin C, Phenol, Tannin,

Flavonoids and Saponin. Psidium guajava leaves show anti-microbial activity against selected

bacterial and fungal pathogens (Anbuselvi and Jeyanthi, 2017).

A phytochemical screening of guava leaves extract reveals the presence of sponins,

alkaloid,volatile oil, steroid, balsmas, saponin glycosides, flavonoids, tannins and anthraquinone,

this indicates that leaves of guava plant contain some major bioactive compounds that can inhibit

the growth of microorganism, thereby proving it as an effective potentials source of antibiotic.

(Abdullahi Adamu 2021).

The phytochemicals detected in the leaves of P. guajava have been reported to offer countless

benefits of medicinal importance. For examples, phenolics and flavonoids demonstrate many

biological activities, such as antioxidant, antimicrobial, anticancer, anti-inflammatory and wound

healing properties. On the other hand, steroids are known to have antibacterial, insecticidal and

cardiotonic properties, while saponins can treat diabetes. Similarly, terpenoids have been used to

alleviate human diseases such as cancer, malaria, inflammation and various infectious diseases.

(Abdullahi Adamu 2021).

The phytochemical composition of leaf extracts of Psidium guajava comprises of various

beneficial phytochemicals such as flavonoids, steroids, saponins, phenols and terpenoids

alkaloids, tannins and carbodydrates (Vikrant Arya et al, 2012).

Guava is rich in tannins, phenols, triterpenes, flavonoids, essential oils, saponins, carotenoids,

lectins, vitamins, fiber and fatty acids. Guava fruit is higher in vitamin C than citrus (80 mg of

vitamin C in 100 g of fruit) and contains appreciable amounts of vitamin A as well (Baby et al.,

26
2010). Guava fruits are also a good source of pectin - a dietary fiber. The leaves of guava are rich

in flavonoids, in particular, quercetin. Much of guava's therapeutic activity is attributed to these

flavonoids. The flavonoids have demonstrated antibacterial activity (Nwinyi et al., 2008).

Quercetin is thought to contribute to the anti-diarrhea effect of guava; it is able to relax intestinal

smooth muscle and inhibit bowel contractions. In addition, other flavonoids and triterpenes in

guava leaves show antispasmodic activity.

Psidium guajava L. chemical composition includes compounds such as tannins, phenols,

flavonoids, saponins, carbohydrates, alkaloids, sterols, and terpenoids (Millones-Gómez et al.,

2020). It is important to consider that the type and abundance of phytochemicals can vary

depending on the microclimate and soil conditions of the habitat (Lavola et al., 2017), but also

depending on the plant tissue and seasonal changes (Hardege 2009).

Guava leaves are also a rich source of vitamins and minerals, such as calcium, potassium,

sodium, magnesium, iron, sulfur, vitamin B, and C (Andrian et al., 2015). Even, Thomas et al.,

(2017) mentioned that the leaves have a higher concentration of vitamin B (14.80 mg/100 g),

calcium (1,660 mg/100 g), magnesium (440 mg/100 g), phosphorus (360 mg/100 g), and iron

(13.50 mg/100 g) compared to the fruits, however, the fruit is richer in vitamin C (228.3 mg/100

g) and potassium (417 mg/100 g).

Naseer et al. (2018) also reported that the guava fruit contains vitamin A, C, iron, phosphorus

and calcium. It has more vitamin C than the orange. The fruit contains saponin, oleanolic acid,

lyxopyranoside, ara-bopyranoside, guaijavarin, quercetin and flavonoids (Arima et al., 2002,

Das, 2011). Ascorbic acid and citric acid are the major ingredients of guava that play important

role in anti-mutagenic activity (Grover and Bala, 1993). The skin of fruit contains ascorbic acid

27
in very high amount; however, it may be destroyed by heat. The strong pleasant smell of fruit is

credited to the carbonyl compounds (Dweck 1987). Guava fruit contains terpenes, caryo-

phyllene oxide and p-selinene in large quantity which produce relaxation effects (Meckes et al.,

1996). The flavonoid content is higher in the methanolic extract of the guava (Sanches et al,

2005). There are 41 hydrocarbons 25 esters, 13 alcohols and 9 aromatic compounds in guava

(Vernim et al., 1991). Titratable acidity and the total soluble solids are present in fruit (Reyes et

al., 1995). Guajadial is also present in guava (Yang et al., 2007). Essential oil is present in leaves

which contain α-pinene, limonene, β-pinene, isopropyl alcohol, men-thol, terpenyl acetate,

caryophyllene, longicyclene and β-bisabolene. Oleanolic acid is also found in the guava leaves

(Begum et al., 2004). Leaves have high content of limonene about 42.1% and caryophyllene

about 21.3% (Ogunwande et al., 2003). Leaves of guava have a lot of volatile compounds

(Taylor et al., 2001; Fu et al., 2010). The bark includes 12–30% of tannin and one

sourcedeclares that it includes tannin 27.4%, or polyphenols, resin and the crystals of calcium

oxalate. Tannin is also present in roots. Leukocyanidins, gallic acid and sterols are also present in

roots. Carbohydrates with salts are present in abundance.

28
 CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Area of study

This study was conducted in Charis Rhema Research Laboratory at high level, Benue State.

Further studies were conducted at Joseph Sarwuan Tarkaa University, Makurdi. Nigeria.

Makurdi is a city located in Central Nigeria on Latitude 7 o33o50oN and 8o32o10oE along river

Benue. Population of about 438,000 people. The city shares boundaries with Guma local

government area to the north-east, Gwer to the south and Gwer west to the west (Amy Mckenna,

2020).

29
 Map of Makurdi.

Source: Benue State Ministry of Lands, Survey and Solid Minerals. (2015)

30
3.2 Plant samples collection

Fresh leaves of Jatropha curcas and Psidium guajava were collected in the month of February

from Joseph Sarwuan Tarkaa Universtiy, Benue State Makurdi. They were identified by a plant

scientist in the Department of Biological sciences, Benue State Universtiy. The samples were air-

dried. The leaves were pounded into powdered form using a mortar and pestle and were stored in

an air- tight container for further use (Oseni and Alphonse 2011).

3.2.1 Preparation of Extracts

Exactly 100g each of the powdered leaves were extracted separately in cold using absolute

ethanol and methanol (750 mL) for 96 hours (4days) with occasional shaking. The extract was

then separately filtered through Whatman’s No. 1 filter paper and the filtrates were concentrated

to dryness in vacuo using a rotary evaporator to remove the solvents.

3.3 Qualitative analysis of leaf extract of Jatropha curcas and Psidium guajava

Qualitative screening was performed for biochemicals like phenols, flavonoids, saponins,

tannins, alkaloids, terpenoids, coumarins, anthocyanin and anthraquinone,

3.3.1 Assessment for alkaloids

Mayer’s test: To 2 mL of each fraction, 2 mL of HCl was added and then few drops of Mayer’s

reagent were mixed to it. Formation of white precipitate or green color indicated the presence of

alkaloids (Archana et al., 2012)

31
3.3.2 Assessment for anthraquinones

1. To 1 mL of extract, 1 mL benzene was included; addition of 1mL of ammonia solution (10%]

was followed. A red color appearance upon ammonia solution addition was indicative of

anthraquinones presence (Abubakar et al., 2017)

2. 2% HCl drop wise added in plant extract. Formation of Red precipitate was indicative of

anthraquinones (Shah et al., 2014).

3.3.3 Assessment of flavonoids

Alkaline reagent test: To the 1 mL plant extract 2N NaOH of 1 mL was added. Flavonoids

presence indicated through the appearance of yellow color (Sajid et al., 2016).

FeCl3 test: FeCl3 solution, only few drops were added to each extract of 1 mL. The blackish red

precipitate formation showed the occurrence of flavonoids.

3.3.4 Assessment of glycosides

Conc. H2SO4 test: One micro litter of each extract was treated by 1 mL of concentrated H 2SO4.

After that solution was kept for 2 minutes. Glycosides were confirmed by analyzing the

precipitation of red color (Khan et al., 2012).

3.3.5 Assessment of tannins

1 Alkaline Reagent test: 2 ml NaOH was added in 2 ml of extract. Presence of tannins was

confirmed if color changes from yellow to red.

2. FeCl3 test: A volume of 2 mL of 5% FeCl 3 was treated to 1 mL of plant extracts. Presence of

tannins was represented by greenish black or dark blue color (Khan et al., 2012).

32
3.3.6 Assessment of steroids

Salkowski test: 10 mL of chloroform was added in 1 mL of each extract in a test tube. After that

10 mL concentrated sulphuric acid was dissolved in a test tube. Two layers were formed; lower

layer expressed yellow color along green fluorescence while upper layer showed red. The

formation of these layers indicates steroids were present (Khan et al., 2012).

3.3.7 Assessment of saponins

Froth formation with distilled water: 2 mL of distilled water was added to 2 mL of extract.

Then shake in a graduated cylinder lengthwise for 15 min. A layer of foam produced that is of 1

cm, this layer was indicative of saponins (Okerulu et al., 2017).

3.3.8 Assessment of Phenols

To the 1 mL of extract of plant, 2 mL distilled water was added and then 10 % FeCl 3, only few

drops were added. Formation of blue green color was showing phenol presence (Ali et al., 2017).

3.3.9 Assessment of Terpenoids

Salkowski test: Each crude extract (1 mg) was mixed with chloroform (2 mL) and concentrated

sulphuric acid (1 mL). The formation of reddish-brown colour at the interface indicates the

presence of terpenoids (Iqbal et al., 2015).

3.4 Quantitative analysis of leaf extract of Jatropha curcas and Psidium guajava

Quantitative screening was performed for biochemicals like phenols, flavonoids, saponins,

tannins, alkaloids, terpenoids, anthraquinone, steroids and glycosides.

3.4.1 Quantification of total Cardiac glycosides

Cardiac glycosides of each generation of suspension culture were quantitatively determined

according to Solich et al., (1992) with some modifications. For determination of cardiac

33
glycosides, 10 mL each of extract was mixed with 10 mL of freshly prepared Baljet's reagent (95

mL of 1% picric acid + 5 mL of 10% NaOH). After an hour, the mixture was diluted with 20 mL

distilled water and the absorbance was measured at 495 nm.

3.4.2 Quantification of anthraquinone

The anthraquinone compounds were identified by the Borntrager reaction. The determination of

anthraquinone UV-Visible spectrophotometry at 325nm was used.

3.4.3 Quantification of flavonoid

Total flavonoid content was determined by aluminum chloride method using catechin as a

standard. 1 mL of test sample and 4 mL of water was added to a volumetric flask (10 mL

volume). After 5 min 0.3 mL of 5 % Sodium nitrite, 0.3 mL of 10% aluminium chloride was

added, incubation at room temperature was done after 6 minutes. 2 ml of 1 M Sodium hydroxide

was added to the reaction mixture. Immediately the final volume was made up to 10 mL with

distilled water. The absorbance of the reaction mixture was measured at 510 nm against a blank

spectrophotometer.

3.4.4 Quantification of Terpenoids

Powdered form of 10 g of each extract was soaked in alcohol for a day. Later on, it was filtered

and petroleum ether was use for purpose of extraction. The extracted material was calculated and

considered as terpenoids.

3.4.5 Quantification of Saponin

Quantitative Estimation of Saponins Test extract were dissolved in 80% methanol, 2 mL of

vanilin in ethanol was added and mixed well. 2 mL of 72% sulphuric acid solution was added,

34
mixed well and heated on a water bath at 600 c for 10 minutes; absorbance was measured at

544nm against reagent blank.

3.4.6 Quantification of tannins

Buren and Robinson, (1969) approach was followed in order to assessed quantity of tannin with

little advancement. 500 mg extract was measured in (50 mL) plastic bottles. Addition of 50 mL

distilled water was done and shaken for one hour in mechanical shaker. Later on, filtration was

performed and volume was raised up to 50 mL in a volumetric flask. 5 mL of filtered solution

was taken in test tube, 2 mL of FeCl 3 (0.1 M), HCl (0.1 N) and potassium ferrocyanide (0.008

M) was added. 120 nm absorbance was measured using a spectrophotometer. Results were

depicted in the form of GAE (mg of Gallic Acid Equivalent) per gram of dried extracts (Sharma

et al., 2015).

3.4.7 Quantification of Alkaloids

To 1 mL of test extract, 5 mL pH 4.7 phosphate Buffer was added and 5 mL BCG solution and

shake a mixture 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 was measured at 470

nm against blank prepared as above but without extract. Atropine is used as a standard material

and compared the assay with Atropine equivalents.

3.4.8 Quantification of Steroids

Quantitative Estimation of Steroids 1 mL of test extract of steroid solution was transferred into

10 mL volumetric flasks. Sulphuric acid (4N, 2 mL) 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 ± 20 C for 30 minutes with occasional

35
shaking and diluted to the mark with distilled water. The absorbance was measured at 780 nm

against the reagent blank.

3.4.9 Quantification of total phenolics

The total phenolic content in different solvent extracts was determined with the Folin-

Ciocalteu’s reagent (FCR). In the procedure, different concentrations of the extracts were mixed

with 0.4 mL FCR (diluted 1:10 v/v). After 5 min, 4 mL of sodium carbonate solution was added.

The final volume of the tubes was made up to 10 ml with distilled water and allowed to stand for

90 min at room temperature. Absorbance of sample was measured against the blank at 750 nm

using a spectrophotometer.

3.5 Data analysis

The statistical analysis used for this data was ANOVA and t-test. Mean values were separated

using Fisher’s Least Significant Difference where differences exist at 5% level of significance.

36
 CHAPTER FOUR

4.0 RESULTS

The qualitative and quantitative phytochemical composition of the ethanolic and methanolic leaf

extracts of Jatropha curcas and Psidium guajava were determined in this study.

4.1 Qualitative analysis of the phytochemical composition of Jatropha curcas and Psidium
guajava
The qualitative analysis of the phytochemical composition of Jatropha curcas and Psidium

guajava in table 4.1 shows the presence of Alkaloids, Phenols, Flavonoids, Tannins, Glycosides,

Anthraquinone, Steriods, Saponins and Terpenoids.

37
Table 4.1 The qualitative analysis of the phytochemical composition of Jatropha curcas and
Psidium guajava

Phytochemicals Jatropha Jatropha curcas Psidium Psidium guajava

curcas methanolic guajava methanolic
ethanolic extract ethanolic extract
extract extract
Alkaloids +++ +++ +++ +++

Tannins ++ ++ + +

Saponins + + + +

Steroids + + + +

Terpenoids ++ ++ +++ +++

Anthraquinone +++ +++ + +

Phenolics + ++ +++ +++

Flavonoids +++ +++ + ++

Glycosides + + + +

Key = strong ++ + moderate ++ weak +

38
 4.2 Quantitative analysis of the ethanolic and methanolic leaf extracts of Jatropha curcas

The quantitative analysis of the ethanolic and methanolic leaf extracts of Jatropha curcas in

table 4.2 shows Alkaloids, Phenolics, Tannins, Flavonoids, Saponins, Steriods, Glycosides,

Terpernoids and anthraquinone were observed and quantified.

In the ethanolic extract, alkaloid was observed to have the highest concentration of 6.38 ± 0.18

(P<0.05). This was followed by anthraquinone (4.49 ± 0.06) and flavonoids (4.27 ± 0.06). The

phytochemical with the least concentration was the phenolics with the concentration of 1.48 ±

0.18 (P<0.05).

Alkaloid was also observed to have the highest concentration of 6.13 ± 0.18 in the methanolic

extract, followed by the flavonoids (4.83 ± 0.24) (P<0.05). The least concentration was observed

in glycosides (1.87 ± 0.08) (P<0.05).

A comparative analysis of the ethanolic and methanolic extracts showed a significantly higher

concentration of Phenolics (P= 0.010), Flavonoids (P=0.018), saponins, and steroids (P=0.026)

in the methanolic extracts compared to the ethanolic extract. Alkaloids was also slightly higher

in the ethanolic extract, however no significant difference was observed (P=0.293).

39
Table 4.2 The quantitative analysis of the ethanolic and methanolic leaf extracts of
Jatropha curcas

Phytochemicals Ethanolic extract Methanolic extract P-value

Alkaloids (%) 6.38 ± 0.18 6.13 ± 0.18 0.293

Phenolics 1.48 ± 0.18 3.12 ± 0.13 0.010
(mgGAE/100g)
Tannins 3.35 ± 0.35 3.36 ± 0.33 0.979
(mgTAE/100g)
Flavonoids 4.27 ± 0.06 4.83 ± 0.24 0.018
(mgQE/100g)
Saponins 1.57 ± 0.16 2.54 ± 0.13 0.022
(mgOAE/100g)
Sterioids 2.70 ± 0.01 2.88 ± 0.04 0.026
(mgTE/100g)
Glycosides 1.70 ± 0.03 1.87 ± 0.08 0.115
(mgDE/100g)
Terpernoids (%) 3.61 ± 0.02 3.11 ± 0.01 0.001
Anthraquinone 4.49 ± 0.06 4.13 ± 0.18 0.123
(m/m%)
0.350 0.358
FLSD (0.05)

40
 4.3 Quantitative analysis of the ethanolic and methanolic leaf extract of Psidium guajava.
The quantitative analysis of the ethanolic and methanolic leaf extract of Psidium guajava in table

4.3 shows the concentration of alkaloids was observed to be the highest in the ethanolic extract

(7.00 ± 0.35) (P<0.05), followed by phenolics (5.40 ± 0.13). The least concentration observed

was in steroids (1.03 ± 0.000) and saponins (1.08 ± 0.08) (P<0.05).

In the methanolic extract, alkaloids also had the highest concentration (7.50 ± 0.00), followed by

the phenolics (5.42 ± 0.08) (P<0.05).

The comparative analysis of the ethanolic and methanolic extracts showed significantly higher

flavonoids, (P=0.026) in the methanolic extract and Tannins in the ethanolic extract (P=0.008).

41
Table 4.3 The quantitative analysis of the ethanolic and methanolic leaf extract of Psidium
guajava.

Phytochemicals Ethanolic Methanolic P-value

extract extract

Alkaloids (%) 7.00 ± 0.35 7.50 ± 0.00 0.184

Phenolics 5.40 ± 0.13 5.42 ± 0.36 0.948
(mgGAE/100g)
Tannins 2.88 ± 0.01 2.49 ± 0.05 0.008
(mgTAE/100g)
Flavonoids 2.78 ± 0.05 3.01 ± 0.02 0.026
(mgQE/100g)
Saponins 1.08 ± 0.08 1.28 ± 0.01 0.081
(mgOAE/100g)
Sterioids 1.03 ± 0.00 1.12 ± 0.08 0.272
(mgTE/100g)
Glycosides 2.20 ± 0.20 1.63 ± 0.13 0.077
(mgDE/100g)
Terpernoids (%) 4.81 ± 0.09 5.13 ± 0.16 0.136
Anthraquinone 2.06 ± 0.06 2.00 ± 0.00 0.346
(m/m%)

FLSD (0.05) 0.343 0.328

42
4.4 Comparative analysis of the ethanolic extracts of Jatropha curcas and Psidium guajava
The comparative analysis of the ethanolic extracts of Jatropha curcas and Psidium guajava in

table 4.4 shows Jatropha curcas ethanolic extract was observed to have significantly higher

flavonoids (P=0.001), steroids (P=0.000) and anthraquinones (P=0.001) than the ethanolic

extract of Psidium guajava. The extract of Psidium guajava on the other hand had significantly

higher phenolics (P=0.002) and terpenoids (P=0.003). A slightly higher but insignificant

concentration of alkaloids (P=0.155) and glycosides (P=0.072).

43
Table 4.4 showing the comparative analysis of the ethanolic extracts of Jatropha curcas and
Psidium guajava

Phytochemicals Jatropha Psidium P-value

curcas guajava

Alkaloids (%) 6.38 ± 0.18 7.00 ± 0.35 0.155

Phenolics 1.48 ± 0.18 5.40 ± 0.13 0.002
(mgGAE/100g)
Tannins 3.35 ± 0.35 2.88 ± 0.01 0.198
(mgTAE/100g)
Flavonoids 4.27 ± 0.06 2.78 ± 0.05 0.001
(mgQE/100g)
Saponins 1.57 ± 0.16 1.08 ± 0.08 0.065
(mgOAE/100g)
Sterioids 2.70 ± 0.01 1.03 ± 0.00 0.000
(mgTE/100g)
Glycosides 1.70 ± 0.03 2.20 ± 0.20 0.072
(mgDE/100g)
Terpernoids (%) 3.61 ± 0.02 4.81 ± 0.09 0.003
Anthraquinone 4.49 ± 0.06 2.06 ± 0.06 0.001
(m/m%)

44
 4.5 Comparative analysis of the methanolic extracts of Jatropha curcas and Psidium
guajava
The comparative analysis of the methanolic extracts of Jatropha curcas and Psidium guajava in

table 4.5 shows the methanolic extract of Jatropha curcas had significantly higher concentration

of flavonoid (P = 0.001), saponins (P = 0.005), steroids (P = 0.001) and anthraquinones (P =

0.000). Psidium guajava on the other hand had higher alkaloids (P=0,008), phenolics (0.014) and

terpenoids (P=0.006).

45
Table 4.5 The comparative analysis of the methanolic extracts of Jatropha curcas and
Psidium guajava

Phytochemicals Jatropha Psidium P-value

curcas guajava

Alkaloids (%) 6.13 ± 0.18 7.50 ± 0.00 0.008

Phenolics 3.12 ± 0.13 5.42 ± 0.36 0.014
(mgGAE/100g)
Tannins 3.36 ± 0.33 2.49 ± 0.05 0.067
(mgTAE/100g)
Flavonoids 4.83 ± 0.24 3.01 ± 0.02 0.001
(mgQE/100g)
Saponins 2.54 ± 0.13 1.28 ± 0.01 0.005
(mgOAE/100g)
Sterioids 2.88 ± 0.04 1.12 ± 0.08 0.001
(mgTE/100g)
Glycosides 1.87 ± 0.08 1.63 ± 1.13 0.521
(mgDE/100g)
Terpernoids (%) 3.11 ± 0.01 5.13 ± 0.16 0.006
Anthraquinone 4.13 ± 0.18 2.00 ± 0.00 0.000
(m/m%)

46
 CHAPTER FIVE

5.0 DISCUSSION, CONCLUSION AND RECOMMENDATIONS

5.1 DISCUSSION

Throughout the world, plants are considered important source of raw material for the synthesis of

ancient as well as modern drugs, the research on medically important species of plants, play role

toward the confirmation of plant to treat disorders and it is probably a better solution to develop

low cost and effective medicines from available raw material. (Abdullahi Adamu, 2021)

Screening of phytochemicals provides detail description about the medicinal importance and

aptitude of plant. In the current study, bioactive constituents that report biological active nature

to the plant were analyzed and results confirmed the existence of anthraquinones, steroids,

terpernoids, alkaloids, flavonoid, glycosides saponins, tannins and phenolics. By using different

solvents with different polarities. This is similar to a study conducted by Abdullahi Adamu

(2021) who worked on the qualitative screening of the methanolic leaf extract of Psidium

guajava and observed higher amounts of alkaloids, flavonoids and tannins. Vikrant Arya (2012)

also conducted a similar study on Psidium guajava and observed significant amount of alkaloids,

tannins, steriods and flavonoids in the ethanolic extract. These results imply that a variety of

pharmacological actions, such as antioxidant, anti-inflammatory, antibacterial, and anticancer

capabilities, may be shown by the bioactive chemicals found in these plant extracts.

Compounds belonging to the respective groups have been reported to impart various medicinal

characteristics to the plants. Due to the presence of flavonoids, plants possess antioxidant

properties as flavonoids are a water-soluble antioxidant having free radical scavenging properties

as well as anticancer activities (Yadav et al., 2014). Alkaloids are known to possess analgesic as

47
well as antibacterial properties (Nassar et al., 2010) while terpenoids are well known to possess

antibacterial, anti-inflammatory, anticancer, and antiviral properties (Chung et al., 1998).

Tannins have anti-cancerous and antibacterial activities (Radhika et al., 2013) and have ability to

inter-fare the protein synthesis (Sabbah et al., 2017; Lira et al., 2017). Steroids are well known to

have cardio-tonic effect as well as insecticidal and antibacterial effect (Iqbal et al., 2015; Riviere

et al., 2009). Phenolic compounds present in plants responsible for the antimicrobial,

antiallergic, antidiabetic, antioxidant, anti-inflammatory, antimutagenic and anticarcinogenic

properties (Khan et al., 2015; Manjunatha et al., 2006). Presence of saponins in plant are very

important because of use in the treatment of hypo cholesterols and hyperglycaemia due to

anticancer, antifungal, antioxidant, antibacterial and weight loss (Iloki et al., 2015; Xiao 2017).

Glycosides play role as anticoagulant activity, antitumor activity, ant degranulating activity and

influenza virus inhibition (Ntie-Kang et al., 2014; Xiao, 2017). Anthraquinones present in plants

are responsible for the regulation of immunity and play therapeutic role in autoimmune diabetes

(Rastogi et al., 2015).

Quantitative analysis was also done on major phytochemicals such as flavonoids, terpenoids,

saponins, tannins, anthraquinones, steroids, alkaloids, phenolics and glycosides. Different

extracts of Jatropha curcas and Psidium guajava expressed different quantity of phytochemicals.

In this study, higher amount of alkaloid contents was found in ethanolic and methanolic extracts

of both plants, higher concentration of alkaloids, phenolics, flavonoids and saponins in Jatropha

was observed in the methanolic extract, higher concentration of alkaloid, flavonoid and

anthraquinone was observed in the ethanolic extract of Jatropha, higher concentration of

alkaloids, phenols and terpernoids was observed in the ethanolic extract of Psidium guajava,

48
higher concentration of alkaloids, Phenolics, flavonoids and terpernoids were observed in the

methanolic extract of Psidium guajava

This is a similar finding of Anbuselvi and Jeyanthi, (2017) who worked on Phytochemical

biochemical and antimicrobial activity of Psidium guajava leaf extracts, the qualitative as well as

quantitative analysis of phytochemicals was performed. They observed highest amount of

alkaloids and phenols followed by tannins and flavonoids.

Nor et al. (2020) worked on Psidium guajava and reported the methanolic extract to contain

higher concentration of flavonoids, steroids, saponins, phenols and terpernoids.

As at the time of this study, not a lot of research articles about phytochemical composition of

Jatropha curcas were published. Zengin et al. (2021) however, worked on the chemical

composition and biological properties of two Jatropha species and reported the leaf extracts of

Jatropha curcas to show higher total phenol content (TPC) and total flavonoid content (TFC)

than the stem barks.

The qualitative and quantitative analysis of this research and others in cite further confirms the

presence of these phytochemicals in both plant. Differences in the total quantification would be

influenced by either time of research, season and even storage of extracts, variations in the

polarities of the solvents used for extraction as well as differences in the chemical composition

of the plant samples used.

49
5.2 Conclusion

Phytochemical screening of the leaf extracts of Jatropha curcas and Psidium guajava revealed

the presence of alkaloids, glycosides, flavonoids, tannins, saponins, phenols, steroids, terpenoids

and anthraquiones in variable quantities. These may be responsible for their numerous medicinal

properties.

5.3 Recommendations

I. Indigenous knowledge on the use of these medicinal plants should be recognized,

protected and promoted since they are found to have the major bioactive compounds that

can inhibit the growth or microorganisms known to cause disease.

II. The potential of these plants should be explored by pharmaceutical and therapeutic

industries for the products of antibiotics.

50
 REFERENCES
Abdelgadir, H.A. and Van, S. (2013). Ethnobotany, ethnopharmacology and toxicity of Jatropha
curcas L. (Euphorbiaceae): A review. South African Journal of Botany, 88: 204–218.

Abdelgadir, H.A., Jäger, A.K., Johnson, S.D. and Van Staden, J. (2010). Influence of Plant
Growth Regulators on Flowering, Fruiting, Seed Oil Content, and Oil Quality of
Jatropha curcas. South African Journal of Botany, 76: 440–446.

Abdullahi-Adamu. (2021). Phytochemical Screening of Guava Leaves Extract. International

Journal of Pure and Applied Science Research, 12: 2384-5918.

Abubakar, Z., Ogidi, O. C. and Oyetayo, V. O. (2017). Assessment of anti-staphylococcal

activity of ethanolic extract of Lenzitesquercina (L) and P. Karsten against clinical
Staphylococcus species Clinical Phytoscience, 2(1), 8.

Adrian, J. A. L., Arancon, N. Q., Mathews, B. W. and Carpenter, J. R. (2015) Mineral

composition and soil-plant relationships for common guava (Psidium guajava L.) and
yellow strawberry guava (Psidium cattleianum var. lucidum) tree parts and fruits.
Communications in Soil Science and Plant Analysis, 46:1960–79.
Ahirrao, R.A., Patel, M. R., Pokar, D.M., Patil, J. K. and Suryawanshi, H. P. (2020).
PhytochemicalScreening of Leaves of Jatropha curcas. International Journal of
Research in Ayurveda and Pharmacy, 2(4): 1324-1327.

Ali, S., Khan, M. R. and Sajid, M. (2017). Protective potential of Parrotiopsis jacquemontiana
on Carbon Tetrachloride Induced Hepatotoxicity in Experimental Rats. Biomedicine
and Pharmacotherapy, 95: 1853-1867.

Amy Mckenna. (2020). Makurdi: Location, Facts and Population. Encyclopedia Britannica,
Retrieved 2023-08-07.
Anbuselvi, S. and Jeyanthi, R. (2017). Phytochemical Biochemical and Antimicrobial Activity of
Psidium Guajava Leaf Extract.Pharmaceutical Scienceand Research, 9(12): 2431-
2433.
Anil, A., Patil, N.S., Kumar, R. and Mandal. D. (2017). Irrigation Scheduling and Fertilization
Improves Production Potential of Jatropha (Jatropha curcas L.) international Journal
of Current Microbiology and Applied Sciences, 6(5): 1703-1716.

Archana, P., Samatha, T., Mahitha, B. and Chamundeswari, N. R. (2012). Preliminary

phytochemical screening from leaf and seed extracts of Senna alata L. Roxb-an
ethnomedicinal plant. International Journal of Pure and Applied Science Research, 3:
82-9.

Arima, H. and Danno, G. (2002). Isolation of Antimicrobial Compounds From Guava (Psidium
guajava L.) and Their Structural Elucidation. Bioscience, Biotechnology, and
Biochemistry, 66:1727–30.

51
Asase, A., Oteng-Yeboah, A. A., Odamtten, G. T. and Simmonds, J. (2005). Ethnobotanical
Study of some Ghananian Anti-malarial Plants. Journal of Ethnopharmacology, 99,
273–279.

Begum, S., Hassan, S. I., Ali, S. N. and Siddiqui, B. S. (2005). Chemical Constituents from the
Leaves of Psidium guajava. Natural Product Research, 18(2):135–40.

Bossou, A. F. A. D., Olayé T., Bogninou, G. S. R., Koudoro, Y. A., Agbangnan D. C. P.,
Bothon, F. T. D., Alitonou, G. A., Avlessi, F. and Sohounhloue, C. K. D. (2020).
Medical Benefit, Pharmacology and Toxicity of Jatropha Curcas L. (Euphorbiaceae)
International Journal of Advance Research, 8(01): 856-864.

Cavalcante, N. B., Santos, A. D. and Almeida, J. R. G., (2020). The Genus Jatropha
(Euphorbiaceae): A Review on Secondary Chemical Metabolites and Biological
Aspects. Chemical Interactions, 318, 108976.

Chang, L. W., Chang, W. J., Yen, S. C. and Huang, P. D. (2002). Antioxidant Activity of Sesame
Coat. Food Chemistry, 78 :(3) 347-354.

Chhabra, S. C., Mahunnah, R. L. A. and Mshiu, E. N. (1990). Plants Used in Traditional

Medicine in Eastern Tanzania. III Angiosperms (Euophorbiaceae to
Menispermaceae). Journal of Ethnopharmacology, 28: 255.

Chung, K. T., Wong, T. Y., Wei, C. I., Huang, Y. W. and Lin, Y. (1998). Tannins and Human
Health: A Review: Critical Reviews in Food Science and Nutrition, 38(6), 421-464.

Cseke, L. J., Kirakosya, A. and Kaufman P. B. (2016). Antioxidant Activity of Guava. CRC
Press, 28: 255.

Dahake, R.., Roy, S., Patil, D., Rajopadhye, S. and Chowdhary, A. (2013). Potential anti-HIV
activity of Jatropha curcas Linn. Leaf extracts. Journal of Antivirals and
Antiretrovirals, 5 160–165.

Das, Adams. (2011). Review on Nutritional, Medicinal and Pharmacological Properties of

Centella asiatica (Indian pennywort). (2011). Journal of Biologically Active Products
from Nature, 1(4): 216–28.

Deepak, P., Soumen, R. and Abhay, C. (2019). Evaluation of Jatropha curcas Linn. Leaf
Extracts for Its Cytotoxicity and Potential to Inhibit Hemagglutinin Protein of
Influenza Virus. Indian Journal of Virology, 24(2): 220–226.

Dehgan, B. (1984). Phylogenetic significance of interspecific hybridization in Jatropha

(Euphorbiaceae). Systematic Botany, 9(4):467-478.
Dehgan, B. and Schutzman, B. (1994). Contributions Toward a Monograph of Neotropical
Jatropha: Phenetic and Phylogenetic Analyses. Anatomy of Botanic Garden, 81:349-
367.
52
Dehgan, B. and Webster. G. L. (1979). Morphology and Infrageneric Relationships of the genus
Jatropha (Euphorbiaceae). University of California Publications in Botany,
94(4):467-478.

Devappa, R, K., Makkar, H. P. S. and Becker, K. (2010). Jatropha Diterpenes: A Review.

Journal of the American Oil Chemists' Society, 2010, 88, 301–322.

Devika, M. (2021). Guava (Psidium guajava L.) Leaves: Medicinal Uses and Phytochemical
Profile. Medicinal and Aromatic Plants, (Los Angeles) 10: 383.
Divakara, B. N., Upadhyaya, H. D., Wani, S. P. and Laxmipathi, G. C. L. (2010). Biology and
Genetic Improvement of Jatropha curcas L.: A Review. Applied Energy, 87(3): 732-
742.

Dreyfuss, M. M. and Chapela, I. H. (2010). Potential of Fungi in the Discovery of Novel, Low-
Molecular Weight Pharmaceuticals, Natural Products with Therapeutic Potential,
80(3): 732-845.

Droit, S. (1932). Research on the Seed and Oil of Purghère or Indian PINE nuts (Jatropha curcas
L.). Dissertation. Université de Paris, Faculté de Pharmacie, 10: 383.

Dweck, A. C. (1987). A Review of Guava (Psidium guajava). Journal of Biologically Active

Products from Nature, 1(4): 216–28.

Ekeleme, K., Paul, T., Istifanus, N. and Ufomadu, U. (2017). Phytochemical Analysis and
Antibacterial Activity of Psidium guajava L. Leaf Extracts. Global Scholarly
Communication Biological and Pharmaceutical Sciences, 3(5): 2581-3250.
Elsevier, B. V. (2023). Science Direct, A Review on Phytochemicals. Retrieved From
URL: https://www.sciencedirect.com/science/article/pii/B9780128185933000117 (6-
3-2023)

Fagbenro-Beyioku, A. F, Oyibo, W. A. and Anuforom, B. C. (1998). Disinfectant/Antiparasitic

Activities of Jatropha curcas. East African Medical Journal, 75: 508-511.

Fazel, S., Hamidreza, M., Rouhollah, G. and Mohammadreza, V. (2008). Spectrophotometric

determination of total alkaloids in some Iranian medicinal plants, Thai Journal of
Pharmaceutical Science, 32 (2008) 17-20.

Félix-Silva, J., Giordani, R. B., Silva, A. A. D., Zucolotto, S. M., Fernandes, G. and Pedrosa, M.
D. F. (2014). Jatropha gossypiifolia L. (Euphorbiaceae): A Review of Traditional
Uses, Phytochemistry, Pharmacology, and Toxicology of This Medicinal Plant.
Evidence-Based Complementary and Alternative Medicine, 369-204.

Fu H. Z., Luo, Y. M., Li, C. J., Yang, J. Z. and Zhang, D. M. (2010). Three Unusual
Meroterpenoids from the Leaves of Psidium guajava L. Organic Letters, 12(5):5135–
8.

53
Gbolade, A. (2009). Inventory of Antidiabetic Plants in Selected District of Lagos State, Nigeria.
Journal of Ethnopharmacology, 121, 135–139.

Gokhan, Z., Mohamad, F. M., Kouadio, I. S., Gunes, A., Ouattara, K. E., Jugreet, B. S., Luigi,
B., Sheila, L., Simonetta, C., Lucia, R., Annalisa, C., Luigi, M., Giustino, O., József,
J., Zoltán, C. and Claudio, F. (2021). Chemical Composition and Biological
Properties of Two JatrophaS pecies: Different Parts and Different Extraction
Methods. Antioxidants, 10: 792.

Grover, I. S. and Bala, S. (1993). Studies on Antimutagenic Effect of Guava (Psidium guajava)
in Salmonella typhimurium. Mutual Reserve, 300:1–3.

Güçlü-Üstündağ, Ö. and Mazza, G. (2007). Critical Review on Food Science and Nutrition,
Journal of Ethnopharmacology, 47(3):231-258.

Gupta, R. C. (1985). Pharmacognostic studies on ‘Dravanti’. Part I: Jatropha curcas Linn.

Indian Academy of Sciences (Plant Science.), 94(1):65-82.
Hardege, J. (2009). Chemical ecology. In: Encyclopedia of Life Support Systems; Earth and
Atmospheric, 47(3):231-258.

Hassanpour, S., Maherisis N. and Eshratkhah, B. (2016). International Journal of Forest, Soil
and Erosion, 908-654.

Heller, J., (1996). Physic nut Jatropha curcas L., Promoting the Conservation and use of
Underutilized and NEGLECTED crops, 1st edition. International Plant Genetics and
Crop Plant Research Institute, (Rome, Italy). 765-887.

Henning, K. and Gubitz G. M., (1997). Fuel Production Improves Food Production: The
Jatropha project in Mali. In: Biofuels and Industrial products from Jatropha curcas,
Journal of Ethnopharmacology, 92-97.

Iloki-Assanga, S. B., Lewis-Luján, L. M., Lara-Espinoza, C. L., GilSalido, A. A., Fernandez-

Angulo, D., Rubio-Pino, J. L. and Haines, D. D. (2015). Solvent effects on
Phytochemical Constituent Profiles and Antioxidant Activities, Using Four Different
Extraction Formulations for Analysis of Bucidabuceras L. BMC research notes, 8(1),
396.

Iqbal, E., Salim, K. A. and Lim, L. B. L. (2015). Phytochemical screening, total phenolics and
antioxidant activities of bark and leaf extracts of Goniothalamusvelutinus (airy shaw)
from Brunei Darussalam. Journal of King Saud University – Science, 27: 224-232.

Iwu, M. M. (1993). Handbook of African Medicinal Plants. CRC Press, Florida, 24–33.

Jacobo-Velázquez, D. A. and Cisneros-Zevallos, L. (2017) Methanolic and Ethanolic Extracts of

leaf Extracts of Psidium guajava. Molecule, 22(8):1249.

54
Kaushik, N. (2003). Effect of capsule maturity on germination and seedling vigour in Jatropha
curcas. Seed Science and Technology, 31:449-454.

Khan-Pathan, R., Gali, P. R., Pathan, P., Gowtham, T. and Pasupuleti, S. (2012). In vitro
Antimicrobial Activity of Citrus aurantifolia and its Phytochemical Screening. Asian
Pacific Journal of Tropical Disease, 2, S328-S331.

Kobilke, H. (1989). Understanding the plant Jatropha (Jatropha curcas L.). Diploma thesis.
University Hohenheim, Stuttg. 45-86

Koche, D. E., Shirsat, R. U., Kawale, M. A. and Hislopia, J. (2016). Jatropha curcas Linn.
Indian Academy of Sciences. (Plant Science.) 9:1-1.

Kumar, A. and Sharma, S. (2008). An Evaluation of Multipurpose Oil Seed Crop for Industrial
Uses (Jatropha curcas L.): A Review. Indian Crop Production, 28, 1–10.

Kumar, K. P. V., Pillai, M. S. N. and Thusnavis, G. R. (2011). Seed extract of Psidium guajava
as Eco-friendly Corrosion Inhibitor for Carbon Steel in Hydrochloric Acid Medium.
Journal of Material Science and Technology. 27(12):1143–9.

Lavola, A., Salonen, A., Virjamo, V. and Julkunen-Tiitto, R. (2017). Phytochemical Variation in
the Plant-part Specific Phenols of Wild Crowberry (Empetrumhermaphroditum
Hagerup) Populations. Phytochemical Letters. 21:11–20.

Ludwiczuk, A., Skalicka-Woźniak, K. and Georgiev, M. I. (2017). Effect of Guava (Psidium

guajava) in Salmonella typhimurium. Journal of Pharmacognosy, 987-65.

Mamta, S., Jyoti, S., Rajeev, N., Dharmendra, S. and Abhishek, G. (2013). Phytochemistry of
Medicinal Plants. Journal of Pharma-cognosy and Phytochemistry of Medicinal
Plants, 1(6): 2278- 4136.

Manjunatha, B. (2006). Antibacterial Activity of Pterocarpus santalinus. Indian journal of

pharmaceutical sciences, 68:115.

Manoj, K., Maharishi, T. and Varsha, S. (2021). Guava (Psidium guajava L.) Leaves: Nutritional
Composition, Phytochemical Profile and Health-Promoting Bioactivities.
Foods,10(4): 752.

Meckes, M., Calzada, F., Tortoriello, J., Gonzalez, J. L. and Martínez, M. (1996). Terpenoids
Isolated from Psidium guajava hexane extract with depressant activity on central
nervous system. Phyther Research, 10(7):600–3.

Millones-Gómez, P. A, Maurtua-Torres, D., Bacilio-Amaranto, R., Calla-Poma, R. D.,

RequenaMendizabal, M. F. and Valderrama-Negron, A. C. (2020). Antimicrobial
Activity and Antiadherent Effect of Peruvian Psidium guajava (guava) leaves on a
Cariogenic Biofilm model. Journal of Contemporary Dent Practice, 21:733–40.

55
Morton, J. F. (2004). Fruits of Warm Climates, Journal of Ethnobotany, 425–8.

Nadia, M. and Eleazar, M. E. S. (2018). Introduction to Phytochemicals: Secondary Metabolites

from Plants with Active Principles for Pharmacological Importance. Physical
Sciences, Engineering and Technology,10: 5772-78226.

Najda, A., Almehemdi, A. F. and Zabar, A. F. (2013). Chemical Composition and Nutritional
Value of Jatropha curcas L. Leaves. Journal of Genetic and Environmental
Resources Conservation, 1(3): 221-226.

Naseer, S., Hussain, S. and Naeem, N. (2018). The phytochemistry and Medicinal Value of
Psidium guajava (Guava). Clinical Phytoscience, 4, 32.

Nassar, Z. D., Aisha, A. A. and Majid, A. M. S. A. (2010). The pharmacological properties of

terpenoids from Sandoricum koetjape. Academic research of medicinal plants, 345-
987

Neuwinger, H. D. (1994). African Traditional Medicine. Biomedical and life science. 450. 35.

Neuwinger, H. D. (1996). African Ethnobotany: Poisons and Drugs: Chemistry, Pharmacology,

Toxicology. Chapman and Hall, New York 36:500–509.

Nisha, S. T., Malini, S. and Agarwal, R. M. (2015) Phytochemical Analysis of Jatropha curcas
L. During Different Seasons and Developmental Stages and Seedling Growth of
Wheat (Triticumaestivum L) as Affected by Extracts/Leachates of Jatropha curcas.
Physiological Molecular Biology of some Plants, 21(1): 83–92.
Nor, A. J., Nurul, A. A. A. and Nurul, I. A. (2020). Phytochemical Screening and Antioxidant
Activity of Psidium guajava. Malaysian Journal of Analytical Sciences, 24(2): 173 -
178

Ntie-Kang, F., Onguéné, P. A., Lifongo, L. L., Ndom, J. C., Sippl, W. and Mbaze, L. M. (2014).
The Potential of Anti-malarial COMPOUNDS derived from African Medicinal
Plants, Part II: A Pharmacological Evaluation of Non-alkaloids and Non-Terpenoids.
Malaria journal, 13- 81.

Oakenfull, D. (1981). Saponins in Food. Food Chemisty, 7(1):19-40.

Oduola, T., Adeosun, G. O., Oduola, T. A., Avwioro, G. O. and Oyeniyi, M. A. (2005)
Mechanism of Action of Jatropha Gossypifolia Stem Latex as a Haemostatic Agent.
Electronic Journal of General Medicine, 2:140–143.

Ogunwande, I. A., Olawore, N. O., Adeleke, K. A., Ekundayo, O. and Koenig, W. A. (2003).
Chemical Composition of the Leaf Volatile Oil of Psidium guajava L. Growing in
Nigeria. Flavour Fragrance Journal, 18:136–8.

56
Okerulu, I. O., Onyema, C. T., Onwukeme, V. I. and Ezeh, C. M. (2017). Assessment of
Phytochemicals, Proximate and Elemental Composition of Pterocarpus soyauxii
(Oha) Leaves. American Journal of Analytical Chemistry, 8(06), 406.

Oliver-Bever, B. (1986). Medicinal Plants in Tropical West Africa, Cambridge University Press,
London. 34-98.

Oseni, L. A., Alphonse and Prince, K. (2011). Comparison of Antibacterial Properties of

Solvent. Extracts of Different Parts of Jatropha curcas. International Journal of
Pharmacy and Phytopharmacology,1(3): 117-123

Parveen, B. U., Shikha, R. and Ashawani, K. (2007). Traditional Uses of Medicinal Plants
Among the Rural Communities of Churu District in the Thar Desert, Indian Journal
of Ethnopharmacology, 113: 387-399.

Pecina-Quintero, V., Anaya-López, J. L., Zamarripa-Colmenero, A. and Núñez-Colín, C.A.

(2014). Genetic structure of Jatropha curcas L. in Mexico and Probable centre of
Origin. Biomass Energy, 60: 147-155

Radhika, B., Murthy, J. V. V. S. N. and Grace, D. N. (2013). Preliminary Phytochemical

Analysis and Antibacterial Activity Against Clinical Pathogens of Medicinally
Important Orchid cymbidium aloifolium (l.). International Journal of Pharmaceutical
Sciences and Research, 4(10), 3925.

Rahim, N., Gomes, D. J., Watanabe, H., Rahman, S.R., Chomvarin, C., Endtz, H.P. and Alam,
M. (2010) Antibacterial Activity of Psidium guajava Leaf and Bark Against Multi
Drug-resistant Vibrio cholerae. Japanese Journal of Infectious Disease, 63:271–4.

Rastogi, S., Pandey, M. M. and Rawat, A. K. S. (2015). Medicinal plants of the genus Betula—
Traditional uses and a phytochemical– pharmacological review. Journal of
ethnopharmacology, 159, 62-83.

Reyes, M. U. and Paul, R. E. (1995). Effect of Storage Temperature and Ethylene Treatment on
Guava (Psidium guajava L.) fruit ripening. Postharvest Biology Technology,
5214(95):357–65.

Ribeiro, S.S., da Silva, T.B., de Souza., Moraes, V.R., de Lima, Nogueira, P.C., Costa, E.V.,
Bernardo, A. R., Matos, A. P., Fernandes, J. B., das Graças, M. F., da Silva, F., dos
Santos., Pessoa, A.M. and Silva-Mann, R. (2012). Chemical Constituents of
Methanolic Extracts of Jatropha curcas L and Effects on Spodoptera frugiperda)
Quím Nova, 35 (11): 2218 – 2221.

Rivière, C., Pieters, L., Dejaegher, B., Vander Heyden, Y., Van, M. C. and Quetin-Leclercq, J.
(2009). Polyphenols isolated from antiradical extracts of Psidium guajava.
Phytochemistry, 70(1), 86-94. 36

57
Rouseff, R. L., Onagbola, E. O., Smoot, J. M. and Stelinski, L. L. (2008). Sulfur volatiles in
Guava ( Psidium guajava L .) Leaves: Possible defense mechanism. Journal of
Agriculture and Food Chemistry, 56:8905–10.

Sabandar, C.W., Ahmat, N., Jaafar, F.M. and Sahidin, I. (2013). Medicinal property,
phytochemistry and pharmacology of several Jatropha species (Euphorbiaceae): A
review. Phytochemistry, 85, 7–29.

Sabbah, A., Nasser, M., As-sadi, F., Hijazi, A., Rammal, H., and Nasser, G. (2017). Chemical
composition and antioxidant activity of lebanesepunica granatum peels. International
Journal of Pharma Research and Health Services, 5(1), 1552-7.

Sajid, M., Khan, M. R., Shah, N. A., Shah, S. A., Ismail, H., Younis, T. and Zahra, Z. (2016).
Phytochemical, antioxidant and hepatoprotective effects of Alnus nitida bark in
carbon tetrachloride challenged Sprague Dawley rats. BMC complementary and
alternative medicine, 16(1), 268.

Sanches, N. R., Aparício, D., Cortez, G., Schiavini, M. S., Nakamura, C. V. and Prado B, (2005).
An Evaluation of Antibacterial Activities of Psidium guajava (L.). Brazilian Archives
of Biology Technology, 48:429–36.

Sandberg, F., Perera-Ivarsson, P. and El-Seedi, H.R. (2005). A Swedish Collection of Medicinal
Plants from Cameroon. Journal of Ethnopharmacology, 102, 336–343.

Saxena, M., Saxena, J. and Nema, R. (2013). Phytochemical of Medicinal Plant. Journal of
Pharmacognosy and Phytochemistry, 09-76.

Sharma, A., Goyal, R. and Sharma, L. (2015). Potential Biological Efficacy of Pinus plant
Species Against Oxidative, Inflammatory and MICROBIAL disorders. BMC
complementary and alternative medicine, 16(1), 35.

Siddiqui, A. and Hira, M. (2022). An Introduction to Phytochemical Analysis and their Types.
Scholars Research Library, Der Pharmacia Lettre,14(2): 01-05
Singh, I. D. (1983). New leaf Spot Diseases of Two Medicinal plants. Madras Agricultural
Journal, 70(7):490.

Singh, Y. N., Ikahihifo, T., Panuve, M. and Slatter, C (1984). Folk Medicine in Tongga. A study
on the Use of Herbal Medicines for Obstetric and Gynecological Conditions and
Disorders. Journal of Ethnopharmacology, 12: 305-98.

Solich, P., Sedliakova, V. and Karlicek, (1992). Spectrophotometric Determination of Cardiac

Glycosides by Flow-Injection Analysis. Analytica Chemica Acta, 269(2): 199-203.
Soliman, W. M. and He, X.R. (2015). The Potentials of Jatropha Plantations in Egypt: A
Review. Modern Economy, 6, 190-200.

58
Sparg, S., Light, M. E. and Van, S. (2004). The potential of Psidium guajava phytochemical
composition. Journal of Ethnopharmacology, 2004, 94 (3):219-243.

Staubmann, R., Ncube, I., Gubitz, G.M., Steiner, W. and Read, J.S., (1999). Esterase and lipase
activity in Jatropha curcas L. seeds. Journal of Biotechnology, 75, 117–126. 56.

Staubmann, R., Schubert-Zsilavecz, M., Hiermann, A. and Karting, T. (1999). The

antiinflammatory effect of Jatropha curcas. Journal of Biotechnology, 45-43.

Sukohar, A., Herawati, H. and Puspasari, G. (2017). Anticancer Activity of Jatrophone an

isolated compound from Jatropha gossypifolia Plant Against Hepatocellular Cancer
Cell hep G2 1886. Biomedical Pharmacology, 10, 667–673.

Taylor, P., Pino, J. A., Agüero, J., Marbot, R. and Fuentes, V. (2001). Leaf oil of Psidium
guajava L. from Cuba. Journal of Essential Oil Research, 13:61–2.

Thomas, L. A. T., Anitha, T., Lasyaja, A. B., Suganya, M., Gayathri, P. and Chithra, S. (2017)
Biochemical and mineral analysis of the undervalued leaves–Psidium guajava L.
International Journal of Advance Science Research. 2:16–21.

Threats Reports. (2019) Antibiotic Resistance. Centre for Disease Control and Prevention.

Vernin, G., Vernin, E., Vernin, C. and Metzger, J. (1991). Extraction and GC-MS-SPECMA data
bank analysis of the aroma of Psidium guajaua L. fruit from Egypt. Flavour and
fragrance Journal, 6:143–8.

Vijayta, S., Dr. Renu, M., Sikha, M. and Nishi, Y. (2015). Analysis of the Phytochemical
Content of Jatropha gossypifolia L. Chemical and Process Engineering Research, 35:
2225-0913.
Vikrant, A., Narender, T. and Kashyap, C. P. (2012). Preliminary Phytochemical Analysis of the
Extracts of Psidium Leaves. Journal of Pharmacognosy and Phytochemistry,1(1):
4563-0094.
Waleed F. A. (2019) Jatropha curcas: An Overview. Journal of Advances in Agriculture,10:
2349-0837.

Warra, A. (2019). Cosmetic potentials of physic nut (Jatropha curcas Linn.) seed oil: A review.
American Journal of Science and Industrial Research, 3, 358–366.

Wiehr, E. (1930). An Overview of Jatropha Phytochemical Compostion. Mathematical-Natural

Scientific Faculty, 908-564

Wole, O. M. and Ayanbode, O. F. (2009). Use of Indigenous Knowledge by women in a

Nigerian RURAL community. Indian Journal of Traditional Knowledge 8, 287–295.

59
Wurdack, K. J., Hoffmann, P. and Chase, M. W. (2005). Molecular phylogenetic analysis of
uniovulate euphorbiaceae (euphorbiaceae) using plastid RBCL and TRNL-F DNA
sequences. American Journal of Botany, 92(8):1397-1420.

Xiao, J. (2017). Dietary flavonoid aglycones and their glycosides: Which show better biological
significance? Critical Reviews in food science and nutrition, 57(9), 1874-1905.

Yadav, R. N. S. and Agarwala, M. (2017). Phytochemical Analysis of Some Medicinal Plants.

Journal of phytology, 3.

Yang, X., Hsieh, K., Liu, J. and Guajadial. (2007) An Unusual Meroterpenoid from Guava
Leaves Psidium guajava. Organic Letters, 9:5135–8.

Zengin, G., Mahomoodally, M. F., Sinan, K. I., Ak, G., Etienne, O. K., Sharmeen, J. B., Brunetti,
L., Leone, S., Di Simone, S. C. and Recinella, L. (2021) Chemical Composition and
Biological Properties of Two Jatropha Species: Different Parts and Different
Extraction Methods. Antioxidants, 10, 792.

Zhang, X., Zhang, M., Su, X., Huo, C., Gu, Y. and Shi, Q. (2009). Chemical constituents of the
plants from genus Jatropha. Chemistry and Biodiversity, 6, 2166–2183.

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