CHAPTER ONE

INTRODUCTION

Medicinal plants had played a pivotal role in the healthcare practices of various

civilizations throughout history. In many ancient cultures, such as those in Asia,

Africa, and South America, natural remedies had served as the primary source of

treatment for numerous ailments. Traditional healers and herbalists had accumulated

extensive knowledge about the therapeutic properties of local flora through

generations of empirical observation and practice (Kumar et al., 2021; Li et al., 2023).

This rich repository of indigenous knowledge had laid the groundwork for modern

pharmacology, as many contemporary drugs had been developed based on

compounds isolated from these plants (Brown & Green, 2023).

Historical records had shown that medicinal plants had been integral not only for

curing diseases but also for preventing illnesses. Ancient manuscripts and

ethnobotanical surveys had documented the use of specific plant species for their anti-

inflammatory, analgesic, and antimicrobial properties. For instance, herbal

preparations had been used to treat infections, alleviate pain, and boost the immune

system in traditional healing systems (Garcia et al., 2021). Moreover, the holistic

approach practiced in traditional medicine had integrated the use of these plants with

spiritual and cultural beliefs, thereby fostering a comprehensive view of health and

well-being that encompassed both physical and mental aspects (Sharma et al., 2022).

The significance of medicinal plants in traditional medicine had extended beyond

their immediate therapeutic effects. They had also been instrumental in guiding

scientific inquiry into the nature and activity of bioactive compounds. Researchers

had often turned to traditional medicinal practices to identify plants with promising

pharmacological properties, which then had been subjected to rigorous scientific

1
investigations. This approach had led to the isolation of many key compounds that

had been further developed into modern drugs, such as aspirin from willow bark and

quinine from cinchona (Kumar et al., 2021). The validation of these traditional

remedies through scientific methods had underscored the potential of natural products

as alternative sources for new therapeutic agents, especially in the context of rising

antibiotic resistance and emerging diseases (Li et al., 2023).

Furthermore, the sustainable use of medicinal plants had been an important

consideration within traditional medicine. Indigenous communities had often

practiced conservation and sustainable harvesting techniques to ensure that these

valuable resources were available for future generations. This traditional ecological

knowledge had contributed to the development of conservation strategies and had

informed modern practices in natural resource management (Garcia et al., 2021). The

integration of traditional wisdom with modern scientific approaches had not only

enriched the understanding of plant-based therapies but had also promoted

biodiversity conservation and sustainable healthcare practices.

2.1.1 Historical Perspectives on Medicinal Plants

Historically, medicinal plants had been the primary source of treatment before the

advent of synthetic drugs. Ancient civilizations such as those in Egypt, China, India,

and Greece had recorded the use of plant-based remedies in manuscripts and oral

traditions. For example, the Ebers Papyrus in ancient Egypt and the Ayurvedic texts

in India had documented the medicinal use of numerous plants for treating a wide

range of ailments (Garcia et al., 2021). These early records had underscored the

importance of plant-derived compounds and had laid the groundwork for the

systematic study of phytochemistry.

2
2.1.2 Ethnobotanical Contributions and Traditional Practices

Indigenous communities had developed extensive knowledge of local flora through

generations of observation and experimentation. Ethnobotanical studies had revealed

that traditional healers had employed specific plant species based on their observed

therapeutic benefits. These practices had not only contributed to the health and well-

being of local populations but had also provided valuable leads for researchers.

Detailed field studies had illustrated that plants were selected based on empirical

evidence and that their preparations were often tailored to address specific health

conditions (Sharma et al., 2022). This body of traditional knowledge had been crucial

in identifying plants with potential medicinal properties.

2.1.3 Transition from Traditional Knowledge to Modern Medicine

The wealth of traditional medicinal knowledge had gradually influenced modern

pharmacological research. Researchers had systematically investigated plants known

from traditional medicine to isolate and characterize their bioactive compounds. Many

modern drugs had been derived from these early studies—for instance, the discovery

of aspirin from willow bark and quinine from cinchona had directly stemmed from

traditional remedies (Brown & Green, 2023). This transition had demonstrated that

traditional practices, when scientifically validated, could lead to the development of

novel therapeutic agents and had bridged the gap between ancient wisdom and

modern science.

3
 2.1.4 Pharmacological Insights and Drug Discovery

Phytochemical investigations had revealed that medicinal plants contained a diverse

array of secondary metabolites, such as alkaloids, flavonoids, tannins, saponins, and

terpenoids. These compounds had been found to possess significant pharmacological

activities including anti-inflammatory, analgesic, and antimicrobial effects (Kumar et

al., 2021). Laboratory studies had confirmed that many of these compounds were

responsible for the therapeutic actions observed in traditional medicine.

Consequently, the isolation and characterization of these bioactive molecules had

become a cornerstone of drug discovery, highlighting the continued relevance of

medicinal plants in modern therapeutic development (Li et al., 2023).

2.1.5 Conservation and Sustainable Harvesting Practices

Traditional medicinal practices had often incorporated principles of conservation and

sustainability. Indigenous communities had practiced selective harvesting and had

implemented measures to ensure that plant resources were not depleted. These

sustainable practices had been critical in preserving biodiversity and had provided a

model for modern conservation strategies. Researchers had acknowledged that the

sustainable use of medicinal plants was essential not only for the continuation of

traditional medicine but also for ensuring that future generations could benefit from

these natural resources (Garcia et al., 2021).

1.2 STATEMENT OF THE PROBLEM

Despite the long-standing traditional use of Ochna schweinfurthiana for medicinal

purposes, its phytochemical constituents and antibacterial properties had not been

comprehensively investigated in modern scientific studies. The emergence and rapid

spread of antibiotic-resistant bacteria had created a significant public health challenge,

4
 and conventional antibiotics had been losing their efficacy (Garcia et al., 2021). This

development had underscored the need to explore alternative sources of antibacterial

agents derived from natural products.

SIGNIFICANCE OF THE STUDY

The study had been significant in addressing the growing global challenge of

antibiotic-resistant bacteria by exploring alternative sources of antibacterial agents.

By investigating the phytochemical composition and antibacterial activities of Ochna

schweinfurthiana, the research had provided valuable scientific insights that supported

the traditional use of the plant and contributed to the field of natural product research

(Kumar et al., 2021; Li et al., 2023).

1.3 AIM AND OBJECTIVES

1.3.1 Aim

The aim of the study is to investigate the phytochemical composition and antibacterial

activities of Ochna schweinfurthiana.

1.3.2 Specific objectives

i. To detect the presence of bioactive compounds such as flavonoids, tannins,

alkaloids, saponins, and terpenoids e.t.c

ii. To carry out antimicrobial activity of the plant extracts against test microbes such

as methicillin resist staphylococcus aureus, E. coli, S. typhi e.t.c

5
 CHAPTER TWO

LITERATURE REVIEW

Ochna schweinfurthiana, a plant belonging to the Ochnaceae family, has been widely used in

African traditional medicine for treating a variety of ailments, including bacterial

infections, fever, wounds, and inflammatory diseases (Odeyemi et al., 2022).

Ethnobotanical records and traditional medicinal practices had long recognized Ochna

schweinfurthiana for its therapeutic properties. Indigenous communities had

traditionally used various parts of the plant to treat ailments such as infections,

inflammation, and wounds (Jones, 2020; Li et al., 2023). These traditional

applications had provided the impetus for scientific investigations into the plant’s

phytochemical composition and pharmacological potential. The longstanding use of

Ochna schweinfurthiana in folk medicine had suggested that it contained bioactive

compounds that might be responsible for its observed medicinal effects (Kumar et al.,

2021). By integrating traditional knowledge with modern scientific methods,

researchers had aimed to validate the ethnobotanical relevance of the plant and

explore its potential as a source of novel antibacterial agents.

Phytochemical screening is a fundamental step in the study of medicinal plants as it

helps identify bioactive compounds responsible for pharmacological activities. Ochna

schweinfurthiana is believed to contain various secondary metabolites that contribute

to its medicinal properties. Previous studies have identified essential phytochemicals

such as flavonoids, alkaloids, saponins, and phenolics in other species of the Ochna

genus, demonstrating significant antimicrobial and antioxidant activities (Eze et al.,

2023). Despite its traditional uses, limited scientific research has been conducted to

validate its medicinal properties, particularly its antibacterial potential. The

6
Importance of Medicinal Plants in Drug Discovery with the rising global concern over

antimicrobial resistance (AMR), the search for novel antibiotics from natural sources

has become imperative. The World Health Organization (WHO) has classified AMR

as one of the top public health threats, with bacterial pathogens such as

Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa developing

resistance to commonly used antibiotics (WHO, 2023). As a result, researchers are

turning to medicinal plants as alternative sources of antimicrobial agents. Studies have

shown that plant-derived secondary metabolites, including alkaloids, flavonoids,

tannins, and terpenoids, exhibit significant antibacterial properties by targeting

microbial cell walls, inhibiting essential enzymes, and disrupting bacterial replication

(Aliyu et al., 2022).

These compounds are known to interact with microbial cells, leading to growth

inhibition and cell death (Okafor & Igwe, 2023). The antibacterial potential of

medicinal plants is typically assessed using in-vitro techniques which including: Agar

well diffusion method – Measuring the zone of inhibition against test bacteria,

Minimum Inhibitory Concentration (MIC) – Determining the lowest concentration at

which bacterial growth is inhibited and Minimum Bactericidal Concentration (MBC)

– Identifying the concentration at which bacterial cells are completely killed. This

study was employed these methods to determine the antibacterial efficacy of Ochna

schweinfurthiana against selected pathogenic bacteria. The results will provide insight

into its potential as a natural antibiotic and support its integration into pharmaceutical

research for new drug development. Given the widespread bacterial resistance to

synthetic antibiotics, exploring plant-based antimicrobials is an urgent need. If Ochna

schweinfurthiana is found to possess strong antibacterial properties, it could serve as a

basis for developing alternative antimicrobial agents. This study aims to contribute to

7
the growing body of scientific knowledge by: Identifying the phytochemical

constituents present in Ochna schweinfurthiana, evaluating its antibacterial activities

against common bacterial pathogens and Assessing its potential for pharmaceutical

and therapeutic applications. By conducting phytochemical screening and

antibacterial evaluation, this research provided valuable data on the medicinal

potential of Ochna schweinfurthiana, paving the way for further clinical studies and

drug formulation efforts. Previous studies on medicinal plants had demonstrated that

bioactive compounds such as flavonoids, tannins, alkaloids, saponins, and terpenoids

played crucial roles in exerting antibacterial effects (Kumar et al., 2021; Li et al.,

2023). However, limited research had been conducted on Ochna schweinfurthiana,

leaving a critical gap in the literature regarding its potential to serve as a source of

novel antibacterial agents. Moreover, existing reports on the phytochemical profile of

the plant had been inconsistent, and the antibacterial activities of its extracts had not

been evaluated against a broad spectrum of pathogenic bacteria (Sharma et al., 2022).

Consequently, the study had been initiated to address these issues by conducting a

detailed phytochemical screening of Ochna schweinfurthiana and evaluating its

antibacterial activities against selected bacterial and fungal strains. This research had

aimed to validate the traditional claims regarding the medicinal properties of the plant

and to determine whether its bioactive compounds could contribute to the

development of alternative antibacterial therapies.

2.2.1 Overview of Phytochemicals

Phytochemicals had been recognized for their roles in plant defense and as agents of

therapeutic benefit in human medicine. These naturally occurring compounds had

been responsible for many of the medicinal properties observed in plants, and their

complex mixtures often had led to synergistic effects in biological systems (Kumar et

8
al., 2021; Li et al., 2023). Their study had provided valuable insights into alternative

treatments, particularly in the context of rising antibiotic resistance.

2.2.2 Alkaloids

Alkaloids, which were nitrogen-containing compounds, had been known for their

potent pharmacological activities. Historically, they had been associated with a wide

range of therapeutic effects including analgesic, antimalarial, and anticancer

properties. Their complex structures allowed them to interact with various biological

targets, contributing significantly to their antibacterial and cytotoxic activities (Garcia

et al., 2021).

2.2.3 Flavonoids

Flavonoids, a major class of polyphenolic compounds, had been widely distributed

among medicinal plants. They had exhibited strong antioxidant, anti-inflammatory,

and antibacterial effects. Their ability to neutralize free radicals and modulate cell

signalling pathways had been well documented, making them key contributors to the

overall therapeutic profile of many plant extracts (Brown & Green, 2023).

2.2.4 Tannins

Tannins had been high molecular weight polyphenolic compounds known for their

astringent properties. They had demonstrated antimicrobial, anti-inflammatory, and

wound-healing activities. By precipitating proteins and interfering with microbial

enzymes, tannins had contributed to the inhibition of bacterial growth, a feature that

had been exploited in traditional remedies (Sharma et al., 2022).

9
2.2.5 Saponins

Saponins, characterized by their glycosidic structures and foaming properties, had

been investigated for their immunomodulatory, anticancer, and antimicrobial effects.

Their amphiphilic nature allowed them to interact with cell membranes, which had

often resulted in increased cell permeability and subsequent biological activity. These

interactions had rendered saponins useful in both pharmaceutical formulations and

traditional medicine (Kumar et al., 2021).

2.2.6 Terpenoids

Terpenoids, which were derivatives of isoprene units, had been recognized for their

structural diversity and extensive pharmacological properties. They had demonstrated

anti-inflammatory, antimicrobial, and anticancer activities, largely due to their ability

to interact with multiple molecular targets. The structural variations among terpenoids

had contributed to a wide range of biological effects, validating their use in traditional

medicinal systems (Li et al., 2023).

.2.7 Glycosides

Glycosides, composed of a sugar moiety linked to a non-sugar component (the

aglycone), had exhibited various therapeutic properties including cardiotonic and

antimicrobial activities. Their biological effects had often been attributed to the

release of the active aglycone upon hydrolysis, which then had interacted with cellular

receptors and enzymes (Garcia et al., 2021).

2.2.8 Biological Activities of Phytochemicals

Collectively, these phytochemicals had contributed to the remarkable biological

activities observed in medicinal plants. Their antibacterial activities, in particular, had

10
 been of significant interest given the increasing challenge of antibiotic resistance. The

synergistic interactions among these diverse compounds had frequently enhanced

their therapeutic efficacy, supporting the use of plant extracts as potential alternatives

or complements to conventional antibiotics (Brown & Green, 2023; Sharma et al.,

2022).

2.3 Phytochemical screening methods

Phytochemical screening had been an essential step in identifying the bioactive

constituents present in medicinal plants. Various qualitative methods had been

employed to detect the presence of key phytochemical groups. The following sub-

sections detailed the procedures and techniques that had been used in the screening of

Ochna schweinfurthiana extracts.

2.3.1 Sample Preparation and Extraction

Plant materials had been collected, cleaned, and dried before being ground into a fine

powder. The powdered samples had then been subjected to solvent extraction using

solvents such as methanol, ethanol, or aqueous solutions, depending on the target

compounds. The choice of solvent had been critical in ensuring maximum yield of the

bioactive constituents (Kumar et al., 2021; Li et al., 2023). The extracts had been

concentrated using a rotary evaporator, and the dried residues had been stored for

further phytochemical analysis.

2.4 Antibacterial activities of plant extracts

Several studies had investigated the antibacterial activities of plant extracts using

various methodologies such as agar diffusion, disc diffusion, and determination of

minimum inhibitory concentrations. Researchers had found that extracts from

medicinal plants exhibited significant antibacterial effects against both Gram-positive

11
 and Gram-negative bacteria. This antibacterial activity was largely attributed to the

presence of bioactive compounds—such as flavonoids, tannins, and alkaloids—that

had been documented to inhibit microbial growth (Kumar et al., 2021; Li et al.,

2023). Moreover, the effectiveness of these extracts had been influenced by the

extraction methods and solvents used, which had affected the yield and potency of the

bioactive constituents (Garcia et al., 2021). These findings had provided a solid

foundation for considering plant extracts as potential alternatives or complementary

agents to conventional antibiotics.

2.5 Antibiotic resistance and the need for alternative therapies

Over the past decades, the rapid emergence of antibiotic-resistant bacteria had become

a major public health concern. Numerous investigations had revealed that many

bacterial strains had developed resistance to conventional antibiotics, leading to

treatment failures and escalating healthcare challenges (Garcia et al., 2021). In light of

this global crisis, researchers had increasingly turned their attention to alternative

therapies, including plant-derived compounds, which had shown promising

antibacterial properties (Sharma et al., 2022; Brown & Green, 2023). These natural

products had demonstrated the ability to inhibit resistant bacterial strains, thereby

offering a potential solution to overcome the limitations of existing antibiotic

regimens. Consequently, the urgency to develop novel antibacterial agents from

natural sources had been recognized in both clinical and research settings,

emphasizing the need for alternative therapeutic approaches.

2.6

12
 CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Sample Collection and Identification

The stem bark of Ochna schweinfurthiana was collected from phase II ABU Zaria.

The Plant was identified by a botanist from the department of Biological science

faculty of life sciences Ahmadu bello university Zaria with voucher No. ABU002898.

3.1.2 Sample Preparation

The plant sample was thoroughly washed with distilled water to remove any dirt and

impurities. They will then be air-dried in the shade at room temperature for 10-14

days to avoid the degradation of bioactive compounds. Once completely dried, the

plant material was grounded into a fine powder using an electric grinder and stored in

airtight containers for further analysis.

3.2 Methods

3.2.1 Extraction (Maceration Method)

The powdered sample was subjected to maceration for the extraction of the powdered

plant materials. Approximately 500 grams of the powder was soaked in 1.5 L of n-

hexane and in a transparent glass container. The mixture was stirred occasionally and

left to stand for 7 days at room temperature. After maceration, the extract was filtered

using Whatman No. 1 filter paper to separate the marc from the filtrate. The filtrate

was allowed to stand for some days to remove excess solvent until a semi-solid

residue was obtained, which was stored for subsequent analysis (Azwanida, 2015).

The same procedure was carried out to obtain the methanol extract.

13
3.2.2 Phytochemical Screening

3.2.2.1 Alkaloid

The presence of alkaloids was determined using Mayer's and Wagner's reagents.

About 0.5 grams of the extract was dissolved in 5 ml of dilute hydrochloric acid and

filtered. A few drops of Mayer's reagent (potassium mercuric iodide) and Wagner's

reagent (iodine in potassium iodide) was added to the filtrate. The formation of a

yellowish-white precipitate with Mayer's reagent and a brownish precipitate with

Wagner's reagent indicates the presence of alkaloids (Trease & Evans, 2002).

3.2.2.2 Saponin

To detect saponins, the foam test was used. Approximately 0.5 grams of the extract

was shaken with 5 ml of distilled water in a test tube. The formation of stable foam,

persisting for at least 15 minutes, indicates the presence of saponins (Trease & Evans,

2002).

3.2.2.3 Phlobatannins

1ml of the concentrated extract is mixed with 1 % hydrochloric acid and then boiled.

The presence of phlobatannins is indicated by the formation of a red precipitate,

signifying the hydrolysis of tannin structures into insoluble phlobaphenes (Trease &

Evans, 2002).

3.2.2.4 Tannin

For tannins, the Ferric Chloride test was employed. Approximately 0.5 grams of the

extract was mixed with 5 ml of distilled water, boiled, and then filtered. A few drops

of 0.1 % Ferric chloride were added to the filtrate. The appearance of a brownish-

green or blue-black coloration indicates the presence of tannins (Trease & Evans,

2002).

14
3.2.2.5 Phenolic Compound

The presence of phenolic compounds was determined using the Folin-Ciocalteu

reagent. About 0.5 grams of the extract will be dissolved in 5 ml of distilled water,

and 0.5 ml of Folin-Ciocalteu reagent was added. After 3 minutes, 2 ml of 20 %

sodium carbonate solution was added, and the mixture was allowed to stand for 1

hour. A blue color indicates the presence of phenolic compounds (Trease & Evans,

2002)

3.2.2.6 Steroid

Steroids was detected using the Liebermann-Burchard test. Approximately 0.5 grams

of the extract was dissolved in 2 ml of chloroform. Acetic anhydride and concentrated

sulphuric acid were added. A blue-green color change indicates the presence of

steroids.

3.2.2.7 Flavonoids Test

To 1m of the concentrated extract, 5 ml of 1 % aluminium chloride in methanol is

added. The mixture is allowed to stand for 30 minutes at room temperature. The

formation of a yellow coloration indicates the presence of flavonoids (Trease &

Evans, 2002).

3.2.2.8 Anthraquinone Test

For the qualitative test, 3 ml of the extract is mixed with 10% potassium hydroxide

solution. The mixture is observed for the formation of a red, pink, or violet color,

indicating the presence of free anthraquinones (Trease & Evans, 2002).

15
3.2.2.9 Cardiac glycosides

To 3 ml of the extract, 3 drops of sulphuric acid and a vanillin reagent were added to

detect the presence of cardenolides. Subsequently, glacial acetic acid and

Concentrated sulphuric acid were added a characteristic of red-violet color upon

treatment shows the presence of cardiac glycosides (Trease & Evans, 2002).

3.3 Antimicrobial Screening

3.3.1 Zone of Inhibition

The antimicrobial activities of Ochna schweinfurthiana extracts was determined by

Agar diffusion method using some pathogenic microbes obtained from the

Department of Medical microbiology ABU teaching hospital Zaria. Mueller Hinton

agar was the medium used as the growth medium for the microbes which, was

prepared according to the manufacturer instructions, sterilized at 121ºC for

15minutes, poured into the sterile petri dishes and was allowed to cool and solidify.

0. 1 g of the extracts was weighed and dissolved in 10 mls of DMSO to obtain a

concentration of 10 mg/ml. This was the initial concentration of the extracts used to

determine its antimicrobial activities. The sterilized medium was seeded with 0.1ml of

the standard inoculum of the test microbe, the inoculum was spread evenly over the

surface of the medium by the used of sterile swab. Using a standard cork borer of 6

mm in diameter, a well was cut at the centre of each inoculated medium and 0.1ml of

solution of the extract of the concentration of 10 mg/ml was then introduced into the

well on the inoculated medium. This was incubated at 37 0C for 24 hours, after which

the plates of the medium were observed for the zone of inhibition of growth, the zone

was measured with a transparent ruler and the result recorded in millimetre (CLSI,

2018).

16
3.3.2 Minimum Inhibitory Concentration (MIC)

The minimum inhibition concentration of the extract was determined using the broth

dilution method. Mueller Hinton broth was prepared, 10mls was dispensed into test

tubes and was sterilized at 1210C for 15mins, the broths were allowed to cool.

McFarland’s turbidity standard scale number 0.5 was prepared to give solution.

Normal saline was prepared, 10mls was dispensed into sterile test tube and the test

microbe was inoculated and incubated at 37 0C for 6hrs. Dilution of the microbe was

done in the normal Saline until the turbidity marched that of the McFarland’s scale by

visual comparison at this point the test microbe has a concentration of about 1.5x 108

c/ml. Two-fold serial dilution of the extract was done in the sterile broth to obtain the

concentrations of 10 mg/ml, 5 mg/ml, 2.5 mg/ml, 1.25 mg/ml and 0.63 mg/ml. The

initial concentration was obtained by dissolving 0. 1g of the extract in 10 mls of the

sterile broth. Having obtained the different concentrations of the extract in the sterile

broth, 0.1 ml of the test microbe in the normal saline was then inoculated into the

different concentrations, incubation was made at 37 0C for 24hours, after which the

test tubes of the broth were observed for turbidity (growth) the lowest concentration

of the extract in the sterile broth which shows no turbidity was recorded as the

minimum inhibition concentration. (CLSI, 2018)

3.3.3 Minimum Bactericidal/Fungicidal Concentration (MBC/MFC)

MBC/MFC were carried out to determine whether the test microbes were killed or

only their growth was inhibited. Mueller Hinton agar was prepared sterilized at 121 o C

for 15 minutes, poured into sterile petri dishes and was allowed to cool and solidly.

The contents of the MIC in the serial dilutions were then sub cultured onto the

prepared medium, incubation was made at 37 0C for 24 hours, after which the plates

17
of the medium were observed for colony growth, MBC/MFC were the plates with

lowest concentration of the extracts without colony growth. (CLSI, 2018)

18
 CHAPTER FOUR

RESULTS

Table 4.1 revealed the percentage yields of the various extracts obtained from the extraction
of the stem bark of Ochna schweinfurthiana.

Table 4.1 Results of percentage yield of the stem bark extracts of O. schweinfurthiana

Solvents weight of plant sample (g) % yields

Methanol 250 2.5
n- Hexane 250 1.5

Table 4. 2: Results of phytochemical screening of the stem bark extracts of O.

schweinfurthiana

The table 4.2 shows the phytochemicals present or absent in the methanol and hexane
extracts of the stem bark O. schweinfurthiana

Phytochemicals methanol extract n-Hexane extract

Alkaloids
1. Mayer’s test + -
2. Wagner’s test - -
Saponins - -
Glycosides - -
Phlobatannins + +
Tannins + +
Steroids - -
Flavonoids + -
C. glycosides - -
Phenolics + +
Anthraquinones + +
Key: + == Present, - == Absent

19
Table 4.3 below shows the results of the zones of inhibition of the methanol and hexanes
extract of O. schweinfurthiana against the test microbes and its comparison with those if
standard antibiotics ciprofloxacin, Sparfloxacin and Fluconazole

Table 4. 3: result of antimicrobial activity of the stem bark extracts of O. schweinfurthiana

Test organisms Methanol Hexane Cipro Spar Fluco

extract extract (10 mg/ml)

MRSA R R R S R

VRE S S S R R

S. aureus S R R S R

H. pylori S S S R R

E. coli S S S R R

S. typhi S R S R R

C. jejuni S R R S R

P. mirablis R R S R R

C. albicans S S R R S

C. krusei S R R R S

Key: S = Sensitive, R= Resistant, Cipro = ciprofloxacin, S = Sparfloxacin and Fluco =

Fluconazole

Table 4.4 above shows the results of the zones of inhibition of the extracts against the test
organisms and against standard drugs

20
Table 4. 4: Result of zones of inhibition of the stem bark extract of O. schweinfurthiana

Test organisms Methanol Hexane Cipro Spar Fluco

extract extract (5mg/ml)
MRSA 0 0 0 30 0

VRE 25 18 28 0 0

S. aureus 28 0 0 31 0

H. pylori 24 22 26 0 0

E. coli 23 20 35 0 0

S. typhi 20 0 40 0 0

C. jejuni 22 0 0 36 0

P. mirablis 0 0 30 0 0

C. albicans 25 18 0 0 32

C. krusei 20 0 0 0 30

Key: Cipro = ciprofloxacin, S = Sparfloxacin and Fluco = Fluconazole

The table 4.5 shows the results of the MIC of the methanol extract against the test organisms
at various concentrations of the extract
.

Table 4. 5: Results of minimum inhibitory concentration of the methanol extract of O.

Schweinfurthiana against the test organisms
Test organisms Concentration in mg/ml

10 5 2.5 1.25 0.63

MRSA

VRE - - 0* + ++

S. aureus - - - 0* +

21
H. Pylori - - 0* + ++

E. coli - - 0* + ++

S. typhi - - 0* + ++

C. jejuni - - 0* + ++

P. mirabilis

C. albicans - - 0* + ++

C. krusei - - 0* + ++

Key: - = No turbidity (no growth), 0* = MIC, + = Turbid (light growth),

++ = moderate turbidity and +++ = high turbidity

The table 4.6 shows the results of the MIC of the hexane extract against the test organisms at
various concentrations of the extract.

Table 4.6: Results of minimum inhibitory concentration of the hexane extract of O.

Schweinfurthiana against the test organisms
Test organisms Concentration in mg/ml

10 5 2.5 1.25 0.63

MRSA
VRE - 0* + ++ +++ -

S. aureus

H. Pylori - 0* + ++ +++

E. coli - 0* + ++ +++

22
S. typhi

C. jejuni

P. mirabilis

C. albicans - 0* + ++ +++

C. krusei

Key: - = No turbidity (no growth), 0* = MIC, + = Turbid (light growth),

++ = moderate turbidity and +++ = high turbidity

The table 4.7 shows the results of the MBC/MFC of the methanol extract against the test
organisms at various concentrations of the extract.

Table 4.7: Results of minimum Bactericidal minimum fungicidal concentration of the

methanol extract of O. schweinfurthiana against the test organisms
Test organisms Concentration in mg/ml

10 5 2.5 1.25 0.63

MRSA

VRE 0* + ++ +++ +++

S. aureus 0* + ++ +++ +++

H. Pylori 0* + ++ +++

E. coli 0* + ++ +++ +++

23
S. typhi 0* + ++ +++ +++

C. jejuni 0* + ++ +++ +++

P. mirabilis

C. albicans 0* + ++ +++ +++

C. krusei 0* + ++ +++ +++

Key: - = No turbidity (no growth), 0* = MBC/MFC, + = Turbid (light growth),

++ = moderate turbidity and +++ = high turbidity

The table 4.8 shows the results of the MBC/MFC of the hexane extract against the test
organisms at various concentrations of the extract.

Table 4.8: Results of minimum Bactericidal / minimum fungicidal concentration of the

hexane extract of O. schweinfurthiana against the test organisms

Test organisms Concentration in mg/ml

10 5 2.5 1.25 0.63

MRSA

VRE 0* + ++ +++ +++

S. aureus

H. Pylori 0* + ++ +++ +++

E. coli 0* + ++ +++ +++

S. typhi

C. jejuni

P. mirabilis

C. albicans 0* + ++ +++ +++

C. krusei

24
Key: - = No turbidity (no growth), 0* = MBC/MFC, + = Turbid (light growth),
++ = moderate turbidity and +++ = high turbidity

CHAPTER FIVE

5.0 DISCUSSION, CONCLUSION AND RECOMMENDATIONS

5.1 Discussions

The extraction method employed in this study was maceration method categorized

under solvent extraction method. This involve soaking 429.642kg of the powdered

stem bark of Ochna schweinfurthiana using two different solvents namely methanol

and n-hexane for specific period of time. The percentage yield of crude extract is as

presented in table 4.1. The result revealed that highest yield of crude extract was

recorded in methanol with 2.4% followed by n-hexane with 1.4% yield. This highest

percentage yield recorded from methanol could be attributed to its high polarity index

compared to n-hexane a non-polar solvent. The choice of solvent, therefore, plays a

significant role in determining the efficiency and completeness of the extraction

process (Azwanida, 2015).

The results of phytochemical screening carried out on the methanol and n-hexane

stem bark extracts of Ochna schweinfurthiana is presented in Table 4.2. The results

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revealed the presence of phlobatannins, tannins, and phenolics in both methanol and

n-hexane extracts, while anthraquinones, flavonoids and alkaloids were only present

in the methanol extract. Notably, saponins, glycosides, steroids, and cardiac

glycosides were absent in both extracts. These findings are in agreement with Oladeji,

et al. (2020), who reported the presence of alkaloids, tannins, and phenolics and

flavonoids, which were detected in this study. Conversely, Mohammed et al., (2022)

reported the presence of both flavonoids and saponins. The presence of these

phytochemicals supports the traditional use of Ochna schweinfurthiana in treating

various ailments. The results of antimicrobial screening of the methanol and hexane

stem bark extracts of Ochna schweinfurthiana was carried against ten pathogenic

bacteria and fungi namely; Methicillin resistant staphylococcus aureus, Vancomycin

resistant enterococci Staphylococcus aureus, Helicobacter pylori, Escherichia coli,

Salmonella typhi, Confylobacter jejuni, C. mirabilis, Candida albicans and Candida

krusei. The findings indicated that both extracts were effective against VRE, H.

pylori, Escherichia coli and C. albicans. MRSA and P. mirabilis were resistant to

both methanol and hexane extracts respectively. This demonstrates that the methanol

and hexane extracts of Ochna schweinfurthiana exhibit broad-spectrum antimicrobial

activity. The sensitivity of both extracts against H. pylori indicates the traditional use

of the plant in treating ulcers. Nwachukwu et al. (2018) reported sensitivity against

Streptococcus pyogenes, Klebsiella, and Aspergillus fumigatus using methanol extract

of Ochna ovata.

The results of the zones of inhibition carried revealed that the methanol extracts

exhibited inhibition zones ranging from 20-28 mm, while hexane extract had zones of

inhibition of between 18-20 mm. S. aureus showed the highest inhibition zone for

methanol extract (28 mm), while S. typhi recorded the least zone of inhibition (20

26
mm). For hexane extract, E. coli had the highest zone (20 mm), and Candida species

recorded the lowest the lowest zone of inhibition (18 mm). These results align with

Okigbo and Ajalie (2015), who reported inhibition zones of 18-25 mm for methanol

extract against S. aureus, and Jafari-Sales and Shadi-Dizaji (2019), found zones of

inhibition of between 15-22 mm against E. coli for the methanol extracts of Allium

sativum. Jagessar et al. (2018) noted lower efficacy for hexane extracts against the

fungi (10-15 mm). Findings from this work shows that the methanol extract had

comparable zones of inhibition (20-28 mm) for the bacteria species with those of

control drugs Ciprofloxacin and Sparfloxacin which showed inhibition zones of (31 -

40 mm) against the bacteria while the fungi species had comparable zone of 20-25

mm with fluconazole which showed zone inhibition of (30- 32 mm). The results of

minimum inhibitory concentration of the methanol extract is as presented in table 4.3.

The results showed minimum inhibitory concentration (MIC) of 2.5 mg/ml against

VRE, H. pylori, E. coli, C. jejuni, C. albicans, and C. krusei while S. aureus and S.

typhi had MIC. of 1.25 mg/ml and 5 mg/ml respectively. The result of the MIC of the

hexane extract revealed that VRE, H. pylori, E. coli and C. albicans had MIC of 5

mg/ml respectively. The results of minimum bactericidal concentration / minimum

fungicidal concentration (MBC/ MFC) revealed that the methanol extract had

MBC/MFC of 10 mg/ml against all the organism except MRSA and P. mirabilis

whereas, the hexane extract had MBC/MFC of 10 mg/ml against VRE, H. pylori, E.

coli and C. albicans. These results indicate a significant antimicrobial potential of

Ochna schweinfurthiana at low concentrations. Muema et al. (2020) also reported

high activity of methanol extract against Gram-positive bacteria, especially S. aureus.

Studies have attributed the antibacterial activity of plant extracts to the presence of

alkaloids, tannins, and flavonoids that possess antibacterial properties (Cowan, 1999).

27
The observed antimicrobial properties in Ochna schweinfurthiana may be attributed

to this phytochemical constituents of the extracts which justified the use of the plant

in traditional medicine

CONCLUSION

This project work aims to analyze the phytochemical composition and antimicrobial

activity of methanol and n-hexane extracts of the stem bark of Ochna

schweinfurthiana. Phytochemical analysis identified several bioactive compounds

including alkaloids, tannins, flavonoid, phlobatannins, phenolics, and anthraquinones.

Antimicrobial screening against selected pathogenic microorganisms demonstrated

high activity. These findings support the traditional medicinal use of Ochna

schweinfurthiana for the treatment of various ailments

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