Assessment of the Efficacy of Silver Nanoparticle Synthesis Using Baobab (Adansonia

digitata) Seeds Crude Methanolic Extract from Hot Water Bath Method
Custodio, Jan Andrei F., Kalugdan, Luis Gabriel S., Villete, Dreanna Remz P.
Cavite National High
School – Senior High
Schools Division of Cavite City
Cavite, Philippines

ABSTRACT

Baobab seeds in Cavite City are currently overlooked and classified as waste due to their
abundance and lack of economic or practical applications. This study assesses the effectiveness
of Baobab (Adansonia digitata) seeds crude methanolic extract for synthesizing silver
nanoparticles. The extraction process involved sun-drying, pulverization, and a two-week soak in
methanol, followed by boiling off excess methanol using the hot water bath method. The
solution was created by combining the crude methanolic extract with a 1 mM silver nitrate
solution. Silver nanoparticle formation was assessed using two methods: color change tests and
UV-visible spectroscopy. The color change tests revealed an absence of silver nanoparticles,
with samples remaining white and turbid. UV-visible spectroscopy further confirmed this
absence by showing no absorbance peaks at 400 nm - 450 nm, diverging from successful
synthesis patterns. Turbidity in the extract, impeding light passage, was identified as the cause of
scattering and reflection, hindering the establishment of absorbance peaks. This study
contributes to understanding the constraints of Baobab seed extract in silver nanoparticle
synthesis, providing insights into potential avenues for waste utilization in Cavite City.
INTRODUCTION which causes them to be more reactive to other molecules,
In recent years, nanotechnology has been a modern making them an excellent catalyst.
field of science that plays a significant role in everyday life According to the Tropical Journal of Natural Product
(Tama, 2016). As Kamur, Kaur, and Sharma (2019) stated, Research, the formation of silver nanoparticles is
it is the application of science and technology to influenced by several key factors. With applications in
manipulate matter at atomic and molecular scales. Over the sensorics, energetics, nanomedicine, bioimaging,
past two decades, studies on synthesized nanomaterials photothermal effects, and optoelectronics, silver
and their characterization have been an emerging field of nanoparticles are a thoroughly investigated field. The
nanotechnology due to their vast applications in physics, optical, mechanical, thermal, electrical, magnetic,
chemistry, biology, and medicine (Panorulselvam et al., transport, reactive, and catalytic capabilities of
2017). In medical science, it is used as a tool in several nanoparticles are all significantly influenced by their size.
ways, like targeted drug delivery, gene delivery, artificial Longer periods of incubation yield higher concentrations
implants, cell repair, antibacterial treatment, and of silver nanoparticles; therefore, this is another critical
diagnostic techniques. These can reduce damage to healthy factor. Silver ion reduction to nanoparticles depends on the
cells in the body and allow for earlier disease detection plant extract utilized in the synthesis, which contains
(Seybon, 2019). Nanoparticles are described as phenolic chemicals such as flavonoids and tannins. This
nanotechnology's fundamental building blocks. They are extract aids in the production and stabilizes the produced
of great scientific interest as they are effectively a bridge silver nanoparticles by acting as reducing and capping
for atomic or molecular structures (Kamur et. al., 2019). agents. Furthermore, elements including composition and
Catalysts are substances that change the reaction rate the dielectric environment can have an impact on the
without being changed by the reaction itself. Reactions nanoparticles' size, shape, and morphology. As part of the
take place on the surface of the catalyst. The greater the synthesis process, color changes are visually observed to
catalyst's surface area compared with the catalyst's volume, provide a reaction indicator for the formation of
the more influential the catalyst will be in speeding up the nanoparticles. The synthesis of silver nanoparticles using
reaction. Due to their size, nanoparticles have a better UV-Visible spectroscopy is a cost-effective and relatively
surface area-to-volume ratio than particles of larger size, simple process compared to other methods, such as

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chemical synthesis or using green plants as reducing
agents. To synthesize silver nanoparticles, phytochemicals
derived from plant extracts are essential, acting as natural
reducing agents and stabilizers (Zuhrohtun et al., 2023).
Their inherent biocompatibility, along with the ability to
enhance antimicrobial properties, makes them crucial for
controlled, eco-friendly production and applications in
medicine and various fields.
Baobab seeds in Cavite City are considered a waste
primarily due to their abundance in the small city, with no
significant utilization or value attributed to them. Due to
their abundance, baobab seeds are sometimes seen as
wasted or underutilized resources, particularly in areas like
Samonte Park and walking trails. In this context, the seeds
are not being utilized for any meaningful purpose, whether
it be for consumption, commercial applications, or local
products. The lack of awareness, demand, or established
practices for incorporating baobab seeds into daily life or
economic activities contributes to their classification as
waste in Cavite City. In the absence of a specified purpose,
these seeds wind up being overlooked and scattered
throughout public areas.
The Baobab tree and its related species belong to the
Malvaceae family and the Adansonia genus. The baobab is
known for its distinctive appearance, broad trunk, unusual
root-like branches, and huge, velvety fruit. Due to the
many distinct uses of its many sections, the tree is steeped
in tradition, and locals commonly refer to it as the "tree of
life”. The name "Baobab" is derived from the Arabic “bu
hobab”, which means "fruit with many seeds" (Jackson,
2015). The Baobab tree serves as both a source of
protection and raw materials for numerous valuable items,
such as food, clothing, and medicine. Baobab fruit pulp,
seeds, leaves, flowers, roots, and bark are all edible, and
scientists have looked at their potential purposes.
Numerous biological characteristics of baobab include
antibacterial, anti-malarial, antiviral, antioxidant, and anti-
inflammatory activity, among others (Rahul et al., 2015).
Baobab pulp and leaf extract synthesized with methanolic
extract had similar antibacterial and inhibitory effects to
different antibiotics against Enterococcus faecalis. (El
Bakkali et al., 2022).
Studies on the antioxidant potential of baobab seeds
have focused mainly on their phenolic components,
especially flavonoids. Findings reveal that baobab seeds
contain considerable levels of flavonoids, as suggested by
a study on their phenolic and fatty acid contents. One type
of flavonoid that has been detected in notable
concentrations is procyanidin B2, which is as high as
1488.90± 7.43 mg/100 g fresh weight. Additionally,
preliminary research indicates that baobab seeds have
about four times more flavonoid content overall than
phenolic content overall. Baobab seeds have a high
flavonoid concentration, making them a viable source for
the extraction of important flavonoids with potential uses
in the food, pharmaceutical, and nutraceutical industries
(Guo et. al., 2019). The qualitative phytochemical
screening assay of powdered baobab fruit seeds was
carried out using an aqueous extract. The results of the
assay indicated the presence of five out of the eight
targeted phytochemical groups, specifically flavonoids,
phenols, saponins, tannins, and terpenoids in the aqueous
extract (Wasihun et al., 2023). The presence of these
phytochemical groups in baobab seeds will directly affect
the outcome in synthesizing silver nanoparticles.
Through the analysis of spectral data, UV-Visible
spectroscopy can be utilized to ascertain the composition
of a material. Using this method, light absorption in the UV
and visible spectrum is measured, and the resulting spectra
are interpreted. The chemical substances included in the
material can be identified by examining the spectra's peaks
and patterns. The shift in wavelength of the surface
plasmon resonance (SPR) peak in the UV-Vis spectra,
which is caused by the size-dependent dielectric functions
of the nanoparticles, can be used by UV-Visible
spectrophotometers to identify silver nanoparticles (Car &
Krstulović, 2022). UV-Visible spectroscopy is a powerful
tool for studying catalysts in liquid and solid states,
offering insights into their electronic transitions. It applies
to various catalyst types, including heterogeneous,
homogeneous, electrocatalysts, and photocatalysts.
However, interpreting UV-Visible absorption bands can be
challenging due to their broad nature, requiring advanced
theoretical calculations and chemometrics. This technique
measures the absorption of ultraviolet and visible light by
a sample, creating an absorption spectrum that identifies
compounds or elements (Bert & Weckhuysen, 2023). The
scattering of suspended matter in water causes the overall
spectral curve to rise nonlinearly, which can interfere with
the accuracy of experimental results in UV-Visible
spectroscopy. Spectral turbidity can lead to inaccurate
measurements of water quality parameters when using
UV-Visible spectroscopy. The interference of spectral
turbidity can affect the detection accuracy of organic
matter content in water when using UV-Visible
spectroscopy (Huang & Wang, 2022). The visibility of
nanoparticles in UV-Visible spectrophotometers can be
impacted by turbidity. The transmission of light through
the sample can be reduced by the scattering and absorption
of UV light brought on by suspended particles in water. As
a result, measurements may become less accurate, and the
UV irradiation in a UV reactor may be underestimated.
Furthermore, the accuracy of experimental data may be
impacted by the nonlinear rise in the overall spectral curve
due to the scattering of suspended particles.
Research Objectives
The main objective of this study is to determine if
Baobab (Adansonia digitata) seeds crude methanolic
extract can synthesize silver nanoparticles.
Specifically, it aims to:
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1. Obtain crude methanolic extract from Baobab Figure 1
(Adansonia digitata) seeds. Project Design of the Study
2. Determine if it is possible to synthesize silver
nanoparticles with Baobab (Adansonia digitata)
seeds crude methanolic extract.
3. Determine it is possible to obtain a crude
methanolic extract from Baobab (Adansonia
digitata) seeds using hot water bath method.

Research Questions
This study aims to determine if Baobab
(Adansonia digitata) seeds crude methanolic extract
are capable of synthesizing silver nanoparticles.
Specifically, it seeks to answer the following
questions:
1. Is it possible to obtain a crude methanolic extract
from Baobab (Adansonia digitata) seeds?
2. Can Baobab (Adansonia digitata) seeds crude
methanolic extract synthesize silver
nanoparticles?
3. Is it possible to obtain a crude methanolic extract
from Baobab (Adansonia digitata) seeds using
hot water bath method?
Figure 2
METHODOLOGY Project Development of the Study
This chapter intends to present the research
design, project development, research procedure,
research instrument, data analysis procedure, and
potential ethical issues of the study. This part will
precisely tackle all the processes and techniques
that will be accomplished to obtain reliable
results and information.
Research Design
This study employed an experimental research
design to investigate the interactions between
independent and dependent variables. The primary
objective was to determine if Baobab seed crude
methanolic extract could synthesize silver
nanoparticles. The extract underwent two tests: color
change, where the extract showed no change in color,
and a UV-VIS spectrophotometer, which showed no
absorbance peak due to the extract’s turbidity. Research Procedure
Correlating the results, the study aimed to determine Plant Identification
if Baobab seed crude methanolic extract can
synthesize silver nanoparticles. The methods of
A sample of the fruit used in this study was
making baobab seed crude methanolic extract and
data gathering from Adebolabola, Akintola, Azeez, brought to the University of the Philippines-
and Kehinde (2020) were followed with some Diliman, Institute of Biology, College of
modifications. Science at Diliman, Quezon City, for plant
identification. The result of the process was sent
via email.

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Preparation of Materials
The materials needed to conduct this study
are shown in Figure 3. The baobab fruits were
collected in local areas of Cavite City and
brought to the University of the Philippines
Biology to identify the plant specimen. To
obtain the methanolic extract of baobab seeds,
333 grams of washed, dried, and powdered
baobab seeds and 1L of methanol are needed. In
preparation for the 1mM Silver Nitrate
solution, a magnetic stirrer was used to dissolve
1 gram of Silver nitrate in 5.892 L of distilled
water.
Figure 4
Materials Used in the Study
Methanolic Extraction of Baobab Seeds
The extraction of the methanolic extract of
Baobab seeds will be based on the research
procedures of Adebolabola, Akintola, Azeez,
and Kehinde (2020), with some changes. Two
kilograms of Baobab seeds were collected,
washed, and sundried for two weeks. The
seeds were pulverized and then soaked in
methanol for two weeks. 333 grams of the
pulverized Baobab seeds were soaked in 1 L
of methanol. After obtaining the methanolic
extract, the excess solvent was removed from
the extract using the hot water bath method.
Using the Delta20 rule, 20°C was added to the
boiling point of methanol, which is 65°C, to
obtain the water temperature of the hot water
bath, which is 85°C
Figure 5
Methanolic Extraction of Baobab Seeds
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Preparation of Silver nitrate solution
Adapted from the method of Adebolabola,
Akintola, Azeez, and Kehinde (2020), 1 mM of
silver nitrate was prepared and used for the
synthesis. In preparing one milli molar AgNO 3
solution, 1 gram of AgNO was dissolved in
3
5892 mL distilled water. The AgNO and water
3
were mixed for 10 minutes using a magnetic
stirrer. Figure 6 shows the process of preparing
the AgNO solution.
3
Figure 6
Preparation of Silver nitrate solution
Synthesis of Silver Nanoparticles using
Baobab seeds methanolic extract
Following the method of Adebolabola,
Akintola, Azeez, and Kehinde (2020), two set-
ups were prepared for the Baobab seeds
methanolic extract; set up A was 60.0% Baobab
seeds methanolic extract mixed with 40.0%
AgNO3 solution; set up B was 40.0% Baobab
seeds methanolic extract mixed 60.0% AgNO3
solution. Setups A and B were mixed using a
magnetic stirrer for 5 minutes. The color of the
samples will then be assessed before subjecting
the samples to a UV-visible spectroscopy test to
validate the formation of silver nanoparticles.
Figure 7
Synthesis of silver nanoparticles with Baobab
seeds crude methanolic extract.
Data Gathering Procedure
This section of the study indicates the
procedure for data collection and measurement.
To acquire information regarding the efficacy of
Baobab (Adansonia digitata) seeds crude
methanolic extracts in synthesizing silver
nanoparticles, the color and UV-visible results
of the samples were monitored and analyzed.
Color Change
When silver nitrate is mixed into the plant
extract, a change in color can be observed. This
can be an indicator. The formation of silver
nanoparticles can be determined by observing
the changing of color of the obtained extract
from colorless to brown. According to the study
of Muthusamy in 2014, the color brown is an
indicator that nanoparticles are formed.
Ultraviolet–visible spectroscopy
UV-visible spectroscopy, the absorbance
test, was used to validate the formation of silver
nanoparticles. The process was conducted at De
La Salle University - Manila: Chemistry
Department Laboratory. Approximately 3mL of
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the liquid samples were used in each cuvette.
3mL of distilled water was also tested to prove
that the scattering of lights did not cause the test
results. Both the samples will be undergo the
testing twice to obtain accurate readings. The
wavelength range was set to 200 nm-500 nm.
The absorption peak determines the light
absorbed by the particles in the sample.
Figure 8
UV-visible spectroscopy test
Data Analysis
The change in color of the extract will be
analyzed to determine if silver nanoparticles
have formed. A solution that synthesized silver
nanoparticles will change its color from yellow
to dark yellow, then green, and finally to brown.
For further verification, the graph obtained
from the UV-Vis spectrophotometer will be
analyzed to determine the formation of silver
nanoparticles. Based on the study of
Adebolabola, Akintola, Azeez, and Kehinde
(2020), an absorbance peak between 400 - 450
nm should appear in the UV-Vis Absorption
spectrum to confirm the synthesis of silver
nanoparticles. If no absorbance peak appears in
the graph, the silver nanoparticles have failed to
synthesize. The figure below shows the result of
a silver nanoparticle sample with an absorbance
peak.
Figure 9
Ultraviolet–visible spectroscopy with an
absorbance peak
Potential Ethical Issues
Equipment handling, such as the UV-Vis
spectrophotometer, should be performed
cautiously to prevent unwanted accidents and
injuries. To avoid injuries, the researchers should
cautiously handle and obtain the baobab seeds
crude methanolic extract. When mixing, the
Baobab (Adansonia digitata) seeds crude
methanolic extract to 1mM silver nitrate,
ensuring not inhaling any hazardous fumes that
may be produced during the extraction process is
a priority. Safety goggles, face masks, and gloves
should be worn throughout the experiment.
RESULTS AND DISCUSSIONS
This part of the paper illustrates all of the data
gathered from the methodology conducted. Each
of the tests done was explained along with the
results gathered. The objectives and the research
questions were all given the application and
answers.
CONCLUSION
The following conclusions were derived as
the data were analyzed and interpreted:
a. Baobab (Adansonia digitata) seeds crude
methanolic extract cannot synthesize silver
nanoparticles.
b. Modifications to the extraction method are
needed to synthesize silver nanoparticles.
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 for water quality monitoring based on UV-Vis
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