CHAPTER ONE

1.0 INTRODUCTION

1.1 Background of the Study

Silver (Ag), a precious metal, has been used for centuries due to its unique properties,

including high electrical conductivity, reflectivity, and antimicrobial effects. Silver,

represented by the chemical symbol "Ag" and atomic number 47, is widely utilized in

jewelry, electronics, and industrial applications (Thirumurugan et al., 2016). With the rise of

nanotechnology, silver has gained even greater prominence through its use in synthesizing

silver nanoparticles (AgNPs), which have broad applications, especially in the industrial and

medical fields, due to their exceptional antimicrobial properties (Ahmad, A. et al., 2024).

Biosynthesis refers to the natural process by which living organisms produce complex

compounds from simple substances. In nanotechnology, biosynthesis involves using

biological agents such as plants, bacteria, or fungi to synthesize nanoparticles. This eco-

friendly method contrasts with conventional chemical synthesis, which often requires toxic

chemicals and harsh conditions (Mittal et al., 2017).

Biosynthesis of silver nanoparticles (AgNPs) specifically refers to the green synthesis of

silver nanoparticles through biological means. In this context, plant extracts are often used as

reducing and stabilizing agents in converting silver ions (Ag) into silver nanoparticles (Ag).

This method of biosynthesis is gaining popularity as it offers a sustainable alternative to

chemical synthesis, minimizing the environmental and health risks traditionally associated

with nanoparticle production (Khan et al., 2018).

Silver nanoparticles (AgNPs) are particles of silver with dimensions typically ranging from 1

to 100 nanometers. Due to their small size and large surface area, AgNPs exhibit unique

optical, electrical, and biological properties that differ significantly from bulk silver. These
1
nanoparticles have become central to various applications, particularly in antimicrobial

coatings, water treatment, and medical devices, due to their ability to disrupt microbial cells

and prevent infection (Rai et al., 2016).

Characterization is the process of analyzing and identifying the structure, properties, and

behavior of materials. In nanotechnology, characterization is essential to understand the

physical, chemical, and biological properties of nanoparticles. Various analytical techniques,

such as UV-Vis spectroscopy, Fourier-transform infrared spectroscopy (FTIR), and electron

microscopy, are used to characterize nanoparticles, determining their size, shape, surface

charge, and stability (Shanmugasundaram et al., 2017).

Characterization of silver nanoparticles involves the use of these techniques to analyze and

confirm the formation, structure, and functionality of AgNPs. Understanding the size, shape,

and surface properties of silver nanoparticles is crucial for optimizing their use in industrial

and medical applications. Techniques like UV-Vis spectroscopy provide information about

nanoparticle size and concentration, while FTIR helps identify the chemical bonds and

functional groups involved in the stabilization of the nanoparticles (Ibrahim et al., 2019).

Mahogany refers to the tropical hardwood species from the Meliaceae family, specifically the

Khaya senegalensis tree, commonly known as African mahogany. Mahogany is highly

valued for its strong, durable timber, but the tree also holds medicinal significance due to its

rich phytochemical content. Various parts of the tree, particularly its bark and stem, have

been used in traditional medicine for treating ailments such as fever, malaria, and bacterial

infections (Ali et al., 2020).

Mahogany stem extract is derived from the stem of the Khaya senegalensis tree and is rich in

bioactive compounds such as tannins, flavonoids, alkaloids, and phenolic acids. These

phytochemicals make mahogany stem extract an effective reducing and stabilizing agent for

2
the biosynthesis of silver nanoparticles. The bioactive compounds not only facilitate the

reduction of silver ions but also stabilize the formed nanoparticles, preventing aggregation

and ensuring long-term stability (Nguyen et al., 2018).

The industrial and biomedical applications of silver nanoparticles synthesized using

mahogany stem extract are vast. In the medical field, AgNPs are used in antimicrobial

coatings for wound dressings, surgical tools, and implantable devices due to their ability to

prevent bacterial infections. Their antiviral and antifungal properties make them useful in

pharmaceuticals and healthcare products (Kuppusamy et al., 2016). In industrial applications,

AgNPs are employed in water purification systems, food packaging, and electronics for their

antibacterial properties and excellent electrical conductivity (Javed et al., 2017). By utilizing

green synthesis methods, such as those involving mahogany stem extract, the environmental

impact of these applications is significantly reduced, offering a sustainable alternative to

conventional nanoparticle production.

1.2 Statement of the Problem

The disposal of agricultural waste, including mahogany (Swietenia spp.) stems, represents a

significant environmental challenge, often resulting in underutilization of these by-products.

At the same time, the high cost and environmental hazards associated with chemical methods

for synthesizing silver nanoparticles (AgNPs) hinder their widespread application,

particularly in fields such as agriculture, healthcare, and environmental remediation (Zahoor,

M. et al., 2021).

Chemical synthesis of AgNPs often involves toxic reducing agents and stabilizers, posing

risks to both human health and the environment. Additionally, these methods are resource-

intensive and costly, making them less feasible for large-scale applications in developing

3
economies. There is an urgent need for cost-effective, eco-friendly, and sustainable

alternatives to overcome these challenges (Al-Gethami, W. et al., 2024)

1.3 Aims and Objectives

The aim of this study is to extract, synthesize, and characterize silver nanoparticles using

mahogany stem methanol extract.

The specific objectives of the study are to:

1. To carry out solvent extraction using methanol

2. Phytochemical screening of the methanoic extraction of the mahogany stem.

3. Synthesis and characterization of AgNP

1.4 Significance of the Study

This study is significant as it promotes the development of eco-friendly and sustainable

methods for synthesizing silver nanoparticles, reducing reliance on toxic chemicals typically

used in nanoparticle production. The use of mahogany stem extract in the biosynthesis of

AgNPs contributes to green chemistry, aligning with global efforts toward environmentally

sustainable scientific practices.

1.5 Scope of the Study

The scope of this study is limited to the biosynthesis of silver nanoparticles using mahogany

stem extract. It focuses on the synthesis, characterization, and Analytical techniques such as

UV-Vis spectroscopy and FTIR will be employed for nanoparticle characterization.

4
 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Nanotechnology

Nanotechnology encompasses the design, synthesis and manipulation of materials at

nanoscale level to achieve exclusive properties, which can be suitably employed for the

required applications (Nasrollahzadeh et al., 2019). Recently, a wide range of applications

and numerous new disciplines in several fields, including optics, mechanics, chemical

industries, space industries, electronics, energy science, single electron transistors, light

emitters, nonlinear optical devices, photo-electrochemical, catalysis, biomedical, cosmetics,

drug or gene delivery and food and feed, have made extensive use of nanotechnology. Small

particles between 1 and 100 nm in with at least one dimension are defined as nanoparticles

(NPs). NPs have gained prominence in technological advancements due to their

physicochemical properties such as catalytic activity, electrical and thermal conductivity,

light absorption, melting point, wettability, quantum confinement, dispersed without

agglomeration and scattering resulting in enhanced performance over their bulk counterparts.

In recent years, Metallic nanoparticles (MNPs) have attracted considerable interest due to

their physicochemical properties, small size and high monodispersity (Ali et al., 2021).

Owing to unique characteristics, MNPs have found potential applications such as targeted

drug delivery, cancer therapy, bio sensing, optical device, electronic, magnetic, catalysis,

water treatment and antimicrobial applications. MNPs, in particular silver, copper and zinc

have attracted great attention and have emerged as a novel class of NPs. (Alavi et al., 2019).

2.2 Silver Nanoparticles (AgNPs)

Traditional chemical methods for synthesizing AgNPs involve the use of toxic chemicals

such as sodium borohydride, which can pose environmental and health risks. In contrast,

green synthesis using plant extracts eliminates the need for harmful chemicals, making it a

5
more sustainable and eco-friendly method (Dikshit et al., 2021). The plant extract contains

bioactive compounds (e.g., phenols, flavonoids, terpenoids) that not only act as reducing

agents but also help stabilize the nanoparticles, preventing agglomeration (Shafey, 2020).

2.3 Characterization of Silver Nanoparticles

Silver nanoparticles (AgNPs) are increasingly utilized in various fields due to their unique

physicochemical, optical, and biological properties (Almatroudi, 2020). Characterizing these

nanoparticles is essential for understanding their behavior, optimizing their applications, and

ensuring their stability and safety. The process involves determining key attributes such as

size, shape, surface properties, chemical composition, and optical behavior, using a variety of

analytical techniques (Pryshchepa et al., 2020)). One critical aspect of characterization is

assessing the size and shape of AgNPs. These parameters significantly influence their

properties, including their optical and antimicrobial behaviors. Transmission Electron

Microscopy (TEM) and Scanning Electron Microscopy (SEM) are commonly employed to

visualize nanoparticles and measure their dimensions. TEM provides high-resolution images,

allowing precise determination of size and morphology, while SEM gives insights into

surface features. For colloidal systems, Dynamic Light Scattering (DLS) is frequently used to

measure the hydrodynamic diameter and size distribution in a liquid medium.

The structural properties of silver nanoparticles are equally important. X-Ray Diffraction

(XRD) is widely used to confirm the crystalline nature of AgNPs and identify specific

phases. This technique also allows the estimation of particle size using Scherrer’s equation.

Fourier Transform Infrared Spectroscopy (FTIR) complements XRD by identifying surface

functional groups, providing insights into capping agents and stabilizers used in the synthesis

process.

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Optical properties are another hallmark of silver nanoparticles, primarily due to their unique

localized surface plasmon resonance (LSPR). This phenomenon causes AgNPs to exhibit

strong light absorption and scattering, which is typically monitored using UV-Visible

Spectroscopy. The LSPR peak, generally observed between 400 and 450 nm, is sensitive to

size, shape, and the surrounding medium. Photoluminescence (PL) Spectroscopy can further

reveal electronic transitions and fluorescence behavior, useful in applications such as imaging

and sensing.

Surface charge and stability are crucial for the long-term usability of silver nanoparticles,

particularly in colloidal systems. Zeta Potential Analysis measures the electrostatic potential

of nanoparticle surfaces, which is indicative of their stability. A high absolute zeta potential

value suggests strong repulsive forces between particles, minimizing aggregation and

ensuring colloidal stability.

Chemical composition analysis is vital for confirming the purity of AgNPs and detecting

impurities or oxidation states. Energy-Dispersive X-Ray Spectroscopy (EDX) and X-Ray

Photoelectron Spectroscopy (XPS) are often used for elemental and chemical state analysis,

respectively. These techniques help identify the presence of silver in its metallic or ionic

forms, as well as other elements or compounds associated with the particles.

Lastly, thermal stability of AgNPs can be assessed using Thermogravimetric Analysis

(TGA), which examines weight loss as a function of temperature. This analysis helps

determine the robustness of surface coatings and the thermal degradation profile of the

nanoparticles.

By integrating these characterization techniques, researchers gain a comprehensive

understanding of silver nanoparticles, enabling their tailored application in fields such as

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medicine, environmental remediation, and industrial processes. Proper characterization not

only enhances their functionality but also addresses potential safety and regulatory concerns,

ensuring their effective and responsible use.

2.4 Applications of Silver Nanoparticles (AgNPs)

Silver nanoparticles (AgNPs) are one of the most widely researched and applied

nanomaterials due to their unique properties, such as a high surface area-to-volume ratio,

optical properties, and antimicrobial effects (Beyene et al., 2017). These properties make

AgNPs suitable for a variety of applications across several fields, including medicine,

electronics, agriculture, and environmental management. (Burdușel et al., 2018)

2.5 Synthesis of silver nanoparticles

2.5.1 Conventional and green methods

There are three fundamental methods through which nanoparticles may be synthesised. These

include physical, chemical and biological processes (Jamkhande et al., 2019). Chemical

methods are the most common yet have few benefits. Their major shortfall is that they are not

green methods of synthesis. Physical methods show promise in terms of greenness, but

biological methods seem to uphold the principles of green chemistry almost absolutely

(Andraos et al., 2022).

In the case of physical methods, evaporation-condensation and laser ablation are the most

common and principal methods. In the evaporation-condensation method, synthesis is done

with the help of a tube furnace at atmospheric pressure. Despite giving a sizeable yield of

nanoparticles, this method has a range of disadvantages. The tube furnace occupies lots of

space, the process consumes enormous amounts of energy, and this increases the temperature

of the surroundings. Lastly, the reaction is slow as thermal equilibrium is not achieved easily,

and it is difficult to arrive at the optimum temperature (Ayuk et al., 2017).

8
In the laser ablation method, no chemical reagent is used, and it has been deemed as efficient.

The absence of chemical solvents in this process makes it reasonably environmentally

friendly, and it adheres to the fifth principle of Green chemistry that speaks of solvent-less

reactions. Moreover, it is also possible to control the particle size of the nanoparticle by

adjusting the number of laser pulses. The purity of the resultant silver nanoparticles is also

exceptionally high when synthesised by this method. In fact, in the field of antimicrobials,

nanoparticles synthesised via laser ablation are deemed to showcase a higher reactivity and

antimicrobial action than their chemically synthesised counterparts (Alhajj et al., 2024).

Thus, these facts make it seem as if the laser ablation methodology is an excellent one and is

extremely environmentally friendly. While this is true for most of the part, one factor reduces

the overall greenness and efficiency of this process. In the laser ablation process, very high

temperatures are required, and it proceeds with the consumption of large amounts of energy.

Thus, it does not adhere to principle six of Green chemistry- "Energy requirements of

chemical processes should be recognised for their environmental and economic impact and

should be minimised. Synthetic methods should be conducted at ambient pressure and

temperatures." Furthermore, the application of laser ablation has not been scaled-up for

industrial use, primarily due to the low productivity of this method. Therefore, at present, this

process is not very economical (Vishwanath et al., 2021).

2.5.2 Chemical Synthesis of Nanoparticles

Chemical methods of synthesis of silver nanoparticles are plentiful. They are also extremely

common and make use of water or organic solvents for the manufacture of silver

nanoparticles. In chemical synthesis procedures, three reactant components are necessary for

the preparation of nanoparticles, and these include a metal precursor, reducing agents, and a

stabilising agent (Yaqoob et al., 2020). Out of the various chemical methods proposed, the

9
simplest one involves the reduction of silver nitrate in an aqueous solution, in the presence of

a reducing and a stabilising agent. A variety of reducing agents are employed, including

citrate, ascorbate, borohydride, and hydrogen gas. Stabilising agents used are surfactants, and

ligands or polymers with specific functional groups such as polyvinylpyrrolidone, and

polyethylene glycol. The size of the resultant nanoparticles largely depends on the initial

concentration of the silver nitrate solution, the concentration of the stabilising agent and the

molar ratio of silver nitrate to the reducing agent (Yaqoob et al., 2020). The polyol process is

a type of chemical method that has commonly been used to synthesis silver nanoparticles. In

this method, silver nitrate is reduced in the presence of ethylene glycol that performs a dual

role of a solvent and a stabilising agent. Particular reagents and reducing agents are

employed, keeping in mind the end size and shape of nanoparticle required. For instance,

chemical synthesis methods rely on inorganic reducing agents such as sodium citrate and

sodium borohydride. Sodium citrate, a weaker reducing agent, usually leads to larger sized

nanoparticles compared to those yielded by borohydride, a strong reducing agent. Similarly, a

standard chemical reagent used to prepare spherical nanoparticles is oleyl amine-liquid

paraffin mixture (Vishwanath et al., 2021). The biggest advantage of chemical synthesis

methods is their versatility, low cost and ability to produce defined nanoparticles of definite

size, dimension, and structure. However, chemical methods rank the lowest in terms of

greenness. The solvents and reducing agents used in the synthesis process are toxic to both

human health and the environment. (Joshi et al., 2019).

2.5.3 Mechanism of Green Synthesis of Nanoparticles

The biosynthesis of AgNPs using plant extracts involves the reduction of silver ions (Ag) to

form elemental silver (Ag). Bioactive compounds from the plant extract, such as phenolic

compounds, flavonoids, and alkaloids, play a dual role in this process: they act as both

10
reducing agents and stabilizers. These compounds interact with Ag⁺ ions, leading to the

formation of silver nanoparticles. The stabilization is essential to prevent aggregation and to

control the size and shape of the nanoparticles. Biological or green synthesis of nanoparticles

has emerged as a promising alternative to chemical methods. This process involves using

biological materials like plant extracts, bacteria, fungi, or enzymes to reduce metal ions into

nanoparticles (Kaviya et al., 2020).

2.6 Mahogany (Swietenia spp.) as a Source for Silver Nanoparticle Synthesis

Mahogany is a tropical hardwood tree that has medicinal, antibacterial, and anti-

inflammatory properties. The leaves, bark, and stems of mahogany are rich in flavonoids,

phenolic compounds, and tannins, which have been shown to possess reducing properties.

Swietenia macrophylla, the most common species of mahogany, is known to contain

compounds such as quercetin, kaempferol, and catechins. These bioactive compounds could

be involved in the reduction of silver ions to form silver nanoparticles (Martha et al., 2021).

The use of plant-based materials for the synthesis of silver nanoparticles (AgNPs) has gained

significant attention due to its eco-friendly and cost-effective nature. Among the diverse plant

sources explored, mahogany (Swietenia spp.), a genus known for its medicinal and

phytochemical properties, has emerged as a promising candidate for green synthesis of

AgNPs. The rich bioactive compounds present in mahogany, including flavonoids, phenols,

and tannins, play a pivotal role in reducing silver ions and stabilizing the synthesized

nanoparticles (Sen et al., 2023)

2.6.1 Mechanism of Synthesis

The synthesis of silver nanoparticles using mahogany extracts involves a straightforward

process. Typically, aqueous or ethanol extracts of mahogany leaves, seeds, or bark are

prepared and mixed with a silver nitrate solution. The phytochemicals in the extract serve as

11
reducing agents, converting silver ions (Ag) to metallic silver (Ag), while also acting as

capping agents to stabilize the nanoparticles. This bioreduction process is marked by a visible

color change in the reaction mixture, indicating the formation of AgNPs (Malik et al., 2021).

2.6.2 Characterization of AgNPs Synthesized from Mahogany

Silver nanoparticles synthesized using mahogany extracts exhibit distinctive characteristics

influenced by the plant’s phytochemical profile. UV-Visible Spectroscopy typically shows a

characteristic surface plasmon resonance (SPR) peak between 400 and 450 nm, confirming

the formation of AgNPs (Sivalingam et al., 2024). Transmission Electron Microscopy (TEM)

reveals their morphology, often showing spherical nanoparticles with sizes ranging from 10

to 50 nm. X-Ray Diffraction (XRD) analysis confirms their crystalline nature, while Fourier

Transform Infrared Spectroscopy (FTIR) identifies functional groups responsible for

reduction and stabilization, such as hydroxyl (-OH) and carbonyl (-C=O) groups (Salame et

al., 2018).

2.6.3 Advantages of Using Mahogany for AgNP Synthesis

Eco-Friendly Process: The use of mahogany extracts eliminates the need for hazardous

chemicals commonly used in traditional nanoparticle synthesis methods, making it

environmentally sustainable (Pindiga et al., 2022).

Abundance of Biomolecules: Mahogany is rich in secondary metabolites like flavonoids,

alkaloids, and tannins, which efficiently drive the reduction and stabilization of nanoparticles.

Cost-Effectiveness: Mahogany trees are widely available, and their extracts can be prepared

using simple methods, reducing the overall cost of AgNP production.

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Enhanced Antimicrobial Activity: The bio-functionalized AgNPs exhibit enhanced

antimicrobial properties due to the synergistic effects of silver and the phytochemicals from

mahogany.

2.6.4 Applications of Mahogany-Derived AgNPs

The silver nanoparticles synthesized using mahogany extracts have diverse applications. In

biomedicine, they demonstrate potent antimicrobial, antifungal, and antiviral activities,

making them suitable for use in wound dressings, coatings, and drug delivery systems. In

environmental science, they are effective in water purification and pollutant removal. Their

catalytic and optical properties are also harnessed in industrial processes and sensor

development (Yin et al., 2018)

Mahogany (Swietenia spp.) provides an efficient and sustainable platform for the synthesis of

silver nanoparticles. Leveraging its phytochemical richness not only ensures eco-friendly

synthesis but also enhances the functional properties of the resulting nanoparticles.

2.7 Synthesis of Nanoparticles from Plant Extracts

Biomolecules from plant extracts are commonly employed for the production of silver

nanoparticles. The method of using plant extracts to synthesise nanoparticles is termed as

phytosynthesis (Rajeshkumar et al., 2017). Usage of plants is often preferred over microbes, as

phytosynthesis is more economical and can be easily scaled for industrial purposes. When

plants are used to enable silver nanoparticle synthesis, an aqueous extract is prepared from

their leaves, roots, or stems, after which an aqueous solution of a silver salt is added

(Rajeshkumar et al., 2017). Usually, an aqueous solution of silver nitrate is taken, and

reduction occurs with the help of biomolecules that are present in the pant extract. Thus, the

overall process resembles the chemical methods very closely, but the reagents used are non-

toxic and derived from plants. The total number of reagents used in this type of synthesis is

13
also lower, as plant extracts play a double role as reducing and stabilising agents. The

reaction often proceeds via a single step; it is highly efficient and quick. In fact, according to

(Husen et al., 2018). The process of generating silver nanoparticles is so fast that 90% of the

initial concentration of Ag+ ions was converted to Ag in approximately 11 min, at a

temperature of 95 °C. It was also found that the size of the resultant silver nanoparticles could

be effectively regulated by varying the temperature, concentration of silver nitrate, the

concentration of the plant extract. The ease of regulating various properties such as shape and

size of the resultant nanoparticles is a huge benefit to generate nanoparticles with high

antimicrobial activity. Usually, production of silver nanoparticles is facilitated by leaf

extracts, but as mentioned above other plant extracts, such as the garlic clove, may also be

utilised for this purpose (Nasir, S. et al., 2022).

2.8 Factors Affecting the Synthesis of Silver Nanoparticles

The synthesis of silver nanoparticles (AgNPs) is influenced by various factors that affect their

size, shape, distribution, and stability. Understanding these factors is critical for optimizing

the synthesis process to achieve nanoparticles with desired properties for specific

applications. These factors can broadly be categorized into chemical, physical, and

environmental parameters. (Mukherji et al., 2019)

2.8.1 Concentration of Silver Nitrate

The concentration of silver nitrate in the reaction mixture plays a critical role in determining

the final size of the nanoparticles. A higher concentration of silver nitrate typically leads to

larger nanoparticles, whereas a lower concentration may result in smaller particles ( Mukherji

et al., 2019)

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2.8.2 Temperature

Temperature can affect the rate of reduction and the size of the nanoparticles. Generally,

higher temperatures promote faster synthesis, but can also lead to the formation of larger

nanoparticles. Low temperatures may result in slower reduction, leading to smaller

nanoparticles.

2.8.3 pH of the Solution

The pH of the solution can influence the stability and morphology of the nanoparticles. A

slightly alkaline pH (around 7–9) is generally favorable for the synthesis of silver

nanoparticles.

2.8.4 Reaction Time

The length of time the reaction mixture is allowed to react can also impact the size and

uniformity of the nanoparticles. Longer reaction times generally lead to the formation of

larger nanoparticles. (Panariello et al., 2018).

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 CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Apparatus and Reagents

The reagents used for this research were of analytical grade. All apparatus used were

thoroughly cleaned in other to avoid any contamination. The apparatus and reagent used are

as follows;

3.1.1 Apparatus

i. Beakers

ii. Test tubes

iii. Dropper pipette

iv. Erlenmeyer flask

v. Thermometer

vi. Volumetric flask

vii. Filter paper (Whatman no. 1)

viii. Measuring cylinder

ix. Aluminum paper

x. Magnetic stirrer

xi. Analytical weighing balance

xii. Electrical weighing

xiii. Funnel

xiv. Incubator

xv. Heating mantle

xvi. Oven

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xvii. Centrifuge

xviii. UV-visible spectroscopy

xix. FT-IR spectroscopy

3.1.2 Reagents

i. Methanol

ii. Ferric chloride

iii. Silver Nitrate

iv. HCI

v. H2SO4

vi. NaOH

vii. HCCl3

viii. NH3

ix. Fehling reagents (a and b)

x. Meyer reagent

xi. Wagner reagent

xii. Double distilled water

3.2 Sample Collection and Preparation

The Mahogamy stem sample were collected in the botanical garden of Joseph Sarwuan Tarka

University Makurdi. The sample was cult into pieces, cleaned, and air dried to a constant

weight at room temperature until fully dried with a reliable weight record. A mortal and

pastle was used to pulverize the dried sample into fine powder for solvent extraction .

3.3 Extraction

17
About 1 kg of dried powdered of Mahogamy stem sample was taken in separate 250 mL

conical flasks with screw cap, and then 700 mL of methanol was added to the flask. The flask

was kept at room temperature with 60 mins. stirring using a magnetic stirrer after each 24 hrs

For 3 days. The extract was then filtered using Whatman filter paper (No. 1) under vacuum

and dried at room temperature in watch glass dish. Weight of each dish was noted prior to

drying of the extract.

3.4 Phytochemical Screening

Qualitative phytochemical screening of crude methanolic extract Chemical tests was carried

out using the extract from the plant, using standard procedures to identify the constituents as

described by Harborne (1998). The tests for the presence or absence of various secondary

metabolites are summarized below:

3.4.1 Test for alkaloids.

 Meyer reagent

Few drops of Meyer's were added to 3ml of filtrate. A cream color was observed which

indicated the presence of alkaloids.

 Wagner's reagent

Few drops of Wagner were added to 3ml of filtrate. A reddish-brown color was observed

which indicated the presence of alkaloids.

3.4.2 Flavonoids

 Ferric chloride test

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Few drops of neutral ferric chloride were added to a small quantity of alcoholic solution of

extract. A blackish red coloration indicated the presence of flavonoids.

3.4.3 Test for tannins

About 0.5 mg of crude extract will be boiled in 20 ml of water in test tubes then filtered.

Addition of few drops of 0.1% ferric chloride will produce brownish green or blue black

colouration indicating presence of tannins.

3.4.4 Test for phytosterols (Salkowski test)

5 mL of the extract will be mixed with 2 ml of chloroform and concentrated sulphuric acid to

form a layer. A cherry red coloration of the interface showed the presence of phytosterols.

3.4.5 Test for phlobatannins

Sample will be boiled with 1% aqueous hydrochloric acid to produce red precipitate

indicating the presence of phlobatannins.

3.4.6 Test for Carbohydrates and Glycosides

 Fehlings test

1 mL of crude extract was boiled on water bath with 1 mL each of Fehling solutions A and B.

A red precipitate indicated the presence of sugar.

3.4.7 Test for carboxylic acid

Extract was dissolved in water and treated with sodium bicarbonate. Brisk effervescence was

observed indicating the presence of carboxylic acid.

3.5 Silver nanoparticle synthesis using methanolic extract

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90 ml of double distilled water/methanol (90/10%v/v) solution of the crude methanolic

extract was prepared and introduced into a conical flask. 1.0 mM AgNO 3, solution was

prepared and then added in drop wise manner while been stirred continuously using a

magnetic stirrer. Stirring was allowed for 60 minutes till when a color changed was observed

from light to intensely dark green. The reaction process was stopped and the mixture was

kept to stand for 24hr. The resulting solution was centrifuged for 60 minutes at 3400 rpm and

filtered. The filtrate (Ag-NPs) was then subjected for further analysis.

3.6 Characterization

All synthesized samples were characterized using 'UV spectrophotometer (7415 Jenway) at

the central instrument laboratory of the department of chemistry, Joseph Sarwuan Tarka

University, Makurdi. FTIR was carried out Using FTIR spectrophotometer (Happ Genzel) at

National Research Institute Chemical Technology, Ahmadu Bello University Zaira.

20
 CHAPTER FOUR

4.0 RESULTS AND DISCUSSION

4.1 Results

Table 1: Phytochemical Screening Result

TEST Alkaloids Observation Inference

i. Wagner +_ Presences of
ii. Hager + Alkaloid
iii. Picric +
iv. Iodine +
Reducing Sugar
i. Felling Test + Prescence of
reducing sugar
Cardiac Glycoside
i. Keller-Killant + Prescence of
Test Cardiac Glycoside
ii. Badjet Test +
Flavonoids
i. Alkaline + Presence of
ii. Conc. H2SO4 + Flavonoids
iii. Ferriec Chloride +
iv. Potassium +
Dichromate
v. Ellagic Acid +
Tannis
i. Brugers Test + Presence of Tannis
ii. Nacht Test +
Phlobatannis
i. Hydrochloric + Prescence of
Test Phlobatannis
Phytosterols
i. Salkowski Test + Prescence of
ii. Acetate + Phytosterols
Anhydride

21
22
23
24
4.2 Discussion of Finding

UV-Vis Absorption Spectrum

Figure 1 displays the UV-Vis absorbance spectrum for sample, measured across a

wavelength range of 200 to 700 nm. The spectrum highlights two key absorption peaks: a

prominent one at 423 nm and a smaller peak at 675 nm. The absorbance, presented in

arbitrary units (a.u.), is plotted on the y-axis against wavelength in nanometers (nm) on the x-

axis. These peaks provide insight into the electronic transitions within the sample. The 423

nm peak suggests the presence of a chromophore, with transitions likely associated with

π→π* or n→π* processes, characteristic of conjugated systems or unsaturated molecular

structures (Smith and Clark, 2021). The peak at 675 nm may indicate another electronic

transition, possibly linked to metal-ligand complexes or extended conjugation, as suggested

by Liu et al. (2020).

FTIR Spectrum

Figure 2 presents the FTIR spectrum of sample, which highlights the functional groups

present in the material. A broad absorption band around 3205 cm⁻¹ corresponds to O-H or N-

H stretching vibrations, indicative of hydroxyl or amine groups. Peaks observed between

2922 cm⁻¹ and 2851 cm⁻¹ are associated with C-H stretching vibrations, typically from

aliphatic CH₂ or CH₃ groups. A strong absorption band at 1707 cm ⁻¹ suggests the presence

of carbonyl (C=O) functional groups, such as those found in ketones, aldehydes, or

carboxylic acids (Ibrahim et al., 2020).

In the region near 1500–1600 cm⁻¹, the spectrum reveals peaks indicative of aromatic C=C

stretching vibrations, further supporting the presence of aromatic rings. Peaks below 1000

cm⁻¹ are characteristic of C-H bending or out-of-plane vibrations, which are also associated

25
with aromatic compounds (Rahman and Khan, 2019). Collectively, these FTIR results

confirm the presence of hydroxyl, carbonyl, and aromatic functional groups, suggesting that

sample S32 has an organic molecular structure.

The UV-Vis and FTIR results provide complementary insights into the molecular

composition of sample S32. The presence of π-conjugated systems and aromatic rings, as

indicated by UV-Vis peaks at 423–427 nm and 675–674 nm, is consistent with the FTIR

identification of aromatic C=C vibrations and carbonyl functionalities. The broad O-H

stretching band observed in the FTIR spectrum further supports the possibility of hydrogen

bonding or polar interactions, which can influence electronic transitions in the UV-Vis range.

These findings align with studies by Smith and Clark (2021) and Liu et al. (2020), which

demonstrate the interplay between functional groups and optical properties in conjugated

systems. Additionally, the material’s absorbance profile and functional group composition

suggest potential applications in light-absorbing materials, as highlighted by Johnson et al.

(2019) and Zhang et al. (2018), particularly in dyes, pigments, or energy-harvesting systems.

X-Ray Diffraction (XRD)

X-rays through the atomic planes of a sample. In the graph, the horizontal axis (2θ) shows

the diffraction angles, while the vertical axis indicates the relative intensity of the diffracted

X-rays. The pattern shows several distinct peaks at specific 2θ angles, with a particularly

intense peak around 38o-39°, indicating a dominant crystal plane. The sharpness of the peaks

reflects the material's high degree of crystallinity, suggesting that the sample is

predominantly crystalline. The presence of multiple smaller peaks corresponds to other

crystallographic planes, indicating additional structural orientations within the material. The

XRD pattern can be analyzed further to identify the material's phases by comparing the

observed peak positions with reference databases. If desired, additional calculations, such as
26
determining crystal size using the Scherrer equation, can be performed to extract more

structural information. The results are consistent with a material that is highly ordered at the

atomic level.

27
 CHAPTER FIVE

5.0 CONCLUSION AND RECOMMENDATION

5.1 Conclusion

The study successfully demonstrated the biosynthesis and characterization of silver

nanoparticles (AgNPs) using extract from the mahogany stem (Khaya senegalensis). The

phytochemical analysis revealed the presence of several bioactive compounds, including

flavonoids, alkaloids, and tannins, which played crucial roles as reducing and stabilizing

agents in the synthesis of AgNPs. Characterization techniques, such as UV-Vis spectroscopy

and FTIR, confirmed the formation of silver nanoparticles with desirable properties,

including size and stability. The synthesized AgNPs exhibited significant antimicrobial

activity against selected bacterial strains, highlighting their potential for application in both

industrial and biomedical fields. This eco-friendly approach not only reduces reliance on

toxic chemical methods but also aligns with sustainable practices in nanotechnology.

5.2 Recommendations

Based on the results of the study the following recommendation are made:

1. It is recommended to optimize extraction methods to enhance the yield of bioactive

compounds for more efficient nanoparticle synthesis.

2. Future studies should explore varying conditions to improve the synthesis efficiency

and quality of silver nanoparticles.

3. It is recommended to utilize additional characterization techniques to gain a

comprehensive understanding of the nanoparticles’ properties.

28
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