Name (/Group Leader): Jayne Ngai Shi Yu

Student ID: 0353666

Module Code: BIO62204 Semester: Sem 2

Group Members:
Student ID Name

0352922 Akilram a/l Sivaram

0353614 Lye Elgene

Email (Individual/Group Leader): Contact No (Individual/Group Leader) :

[email protected] 60177468189
Title of experiment: Practical report 2 - Synthesis of Green Silver Nanoparticles using plant
extract from (Kaempferia galanga) optimised by temperature.
Module Lecturer: Dr. Nallammai Singaram

Student’s Statement:
I have read and understood the TU Regulations on cheating, plagiarism and
collusion and state that this piece of work is my own and does not contain any
unacknowledged work from any other sources. I authorise the University to test any
work submitted by me, using text comparison software, for instances of plagiarism. I
understand this will involve the University or its contractor copying my work and
storing it on a database to be used in future to test work submitted by others.
Note: The attachment of this statement on any electronically submitted assignments will be deemed to have the
same authority as a signed statement.

Date: 29/06/2022
Signed:
Received by: Date received from student:
1. Title and Summary (Author:Elgene)

Title: Synthesis of Green Silver Nanoparticles using plant extract from (Kaempferia
galanga) optimised by temperature.

Summary:

Materials having overall dimensions in the nanoscale, or under 100 nm, are referred
to as nanoparticles. These materials have become significant actors in contemporary
medicine in recent years, with uses ranging from contrast agents in medical imaging
to carriers for the transfer of genes to specific cells. The goal of this experiment is to
extract secondary metabolites from plants. With the extracted metabolites, we
synthesised silver nanoparticles using plant extract. Then we characterise
synthesised silver nanoparticles by UV-Spectrophotometry, FTIR and Zeta sizer.

2. Introduction (Author:Jayne)

The application of nanoscale materials and structures, usually ranging from 1 to 100
nanometers (nm), is an emerging area of nanoscience and nanotechnology (J.A.
Dahl et al., 2007, p. 2228). Nanomaterials may provide solutions to technological and
environmental challenges in the areas of solar energy conversion, catalysis,
medicine, and water treatment (J.E. Hutchison ACSNano, 2 2008, p. 395).
Nanomaterials often show unique and considerably changed physical, chemical, and
biological properties compared to their macro scaled counterparts. (L. Li et al., 2001,
p. 349). This is owing to their surface area to volume aspect ratio, allowing them to
interact with other particles better.

Synthesis of noble metal nanoparticles for applications such as catalysis, electronics,

optics, environmental, and biotechnology is an area of constant interest (T.
Masciangioli et al., 2003, pp. 102A-108A). Gold, silver, and copper have been used
mostly for the synthesis of stable dispersions of nanoparticles, which are useful in
areas such as photography, catalysis, biological labelling, photonics, optoelectronics
and surface-enhanced Raman scattering (SERS) detection (A.M. Smith et al.,2006,
p. 3895). Generally, metal nanoparticles can be prepared and stabilised by physical
and chemical methods; the chemical approach, such as chemical reduction,
electrochemical techniques, and photochemical reduction is most widely used (W.
Chen et al., 2001, p. 291). Studies have shown that the size, morphology, stability
and properties (chemical and physical) of the metal nanoparticles are strongly
influenced by the experimental conditions, the kinetics of interaction of metal ions
with reducing agents, and adsorption processes of stabilising agent with metal
nanoparticles (B. Knoll et al., 1999, p. 134).

The most common methods for preparing all these nanoparticles are wet-chemicals
(A. Tao et al., 2006, p. 4597). The chemical reduction method involves the reduction
of AgNO3 in aqueous solution by a reducing agent in the presence of a suitable
stabiliser, which is necessary in protecting the growth of the silver particles through
aggregation (Oliveira M et al., 2005, pp.429–435). In the formation of silver
nanoparticles by the chemical reduction method, the particle size and aggregation
state of the silver nanoparticles are affected by various parameters such as initial
AgNO3 concentration, reducing agent, AgNO3 molar ratios and stabiliser
concentration (Kim D et al., 2006, p.4019).

Microwave-assisted synthesis is a promising method for synthesis of silver NPs.

Microwave heating is better than a conventional oil bath when it comes to
consistently yielding nanostructures with smaller sizes, narrower size distributions,
and a higher degree of crystallisation (Nadagouda MN et al., 2011, pp.469–478).
Microwave heating has shorter reaction times, reduced energy consumption, and
better product yields which prevents the agglomeration of the particles formed
(Nadagouda MN et al., 2011, pp.469–478). Moreover, microwave-assisted synthesis,
in conjunction with benign reaction media, can also drastically reduce chemical
wastes and reaction time in several organic syntheses and chemical transformations
(Polshettiwar V et al.,2009, pp.16–26).

The objectives of this experiment are to extract secondary metabolites from plant
(Kaempferia galanga) and synthesise silver nanoparticles using plant extract. Then,
characterise synthesised silver nanoparticles by UV-Spectrophotometry, FTIR and
Zeta sizer.

3. Materials and Methods (Author:Jayne)

3.1 Preparation of plant extract from (Kaempferia galanga):

Extract was prepared by weighing 3 g of biological material (Kaempferia galanga) to

300 ml of water to achieve 1% w/v solution. Then, place the mixture in a preheated
water-bath for 30 minutes at 80°C. After, pour the extract into a 50 ml centrifuge tube
and centrifuge at 3000 rpm for 5 minutes. Filter out the supernatant (plant extract)
using a funnel and filter paper. Lastly, collect the filtrate in a beaker.

3.2 Heating of mixtures to form silver nanoparticles:

To test the effect of microwave and hot-bath methods in forming silver nanoparticles.
According to table 1, two mixtures were prepared as a control with plant extract.
Control 1 is 5 mL extract to 45mL silver nitrate (AgNO3) while control 2 is 5 mL
extract to 45mL deionized water. Two samples are also prepared with sample 1
containing 5 mL extract in 45 mL silver nitrate microwaved at medium heat for 10
minutes whereas sample 2 with 5 mL extract in 45 mL silver nitrate being placed in
hot bath (80°C) for 10 minutes. Cool down the mixtures to room temperature. Then,
transfer 2 ml of each mixture into centrifuge tubes and centrifuge at 12,000-15,000
rpm including the control for ten minutes. Supernatant was discarded carefully
without shaking to ensure the pellet is not dislodged. 1 mL of deionized water was
added and centrifuge again at 12,000- 15,000 rpm for 10 minutes. Discard the
supernatant and add another 1 mL of deionized water and centrifuge again at 15,000
rpm for 10 minutes. The tubes are then dried by inverting them on a layer of paper
towels to drain the last drop of water. Lastly, tubes were stored at 4°C for future use.
Name of tube Tube contents

Control 1 5 mL extract to 45mL silver nitrate

Control 2 5 mL extract to 45mL deionized water

Sample 1 5 mL extract in 45 mL silver nitrate

Sample 2 5 mL extract in 45 mL silver nitrate

Table 1: Labelled tubes with different mixtures.

3.3 Characterization of silver nanoparticles:

a) UV-Visible absorption spectroscopy:

Set the UV-Spec to read between the range of 200 nm to 600 nm wavelength. 1 mL
of control 1 and 2, 1 mL of samples 1 and 2 were placed into individual cuvettes and
inserted into the spectrometer. A picture of the absorbance, ʎ reading and the
spectrum were taken. Formation of metal nano-particles to be verified by recording
the UV-Visible absorption spectrum by comparing the surface plasmon resonance
exists for the metal.

b) Zetasizer

By using a syringe 1 mL of control 1 and 2, 1 mL of samples 1 and 2 were taken and

injected into a special cuvette. Cuvette were placed in the analyzer and data were
analysed. The Zetasizer is used to measure three characteristics of particles or
molecules in a liquid medium. These three fundamental parameters are Particle size,
Zeta potential and Molecular weight.

c) Fourier Transform Infra-Red Spectroscopy (FTIR):

After cleaning the FTIR quartz surface with ethanol. Place 1-2 drops of the control 1
and 2, samples 1 and 2 onto the space and sealed for FTIR analysis. Spectrum
obtained were compared with the functional group table provided. Characterization of
nanoparticles by FTIR-UATR is to identify the functional groups contained in the
biological extract.
4. Results (Author:Akilram, Editor:Elgene)

Our Group Results:

Figure 1.1: Control 1, Zeta Potential Distribution

 Figure 1.2: Control 1, Size Distribution

Figure 1.3: Control 1, UV Spectrophotometer

Figure 2.1: Control 2, Zeta Potential Distribution
 Figure 2.2: Control 2, Size Distribution

Figure 2.3: Control 2, UV Spectrophotometer

Figure 3.1: Microwaved, Zeta Potential Distribution
 Figure 3.2: Microwaved, Size Distribution

Figure 3.3: Microwaved, UV Spectrophotometer

Figure 4.1: Waterbath, Zeta Potential Distribution
 Figure 4.2: Waterbath, Size Distribution

Figure 4.3: Waterbath, UV Spectrophotometer

 Figure 5.1: Blank UV Spectrophotometer

Figure 5.2: Zinc Nitrate, UV Spectrophotometer

 Average Zeta
Sample Data Diameter Size Potential
(nm) (mV)

Wavelengt 259 277 300

h
(nm)
Zinc Nitrate
Absorban 0.249 0.166 0.062
ce
(A)

Wavelengt
h 268 290
Control 1 (nm)
561.4 -27.5
Absorban
ce 0.170 0.182
(A)

Wavelengt 229 249

Control 2 h
(nm) 403.2 -28.1

Absorban 0.146 0.086

ce
(A)

Wavelengt 287 296

Microwaved h
(nm) 288.5 -25.5

Absorban 0.390 0.389

ce
(A)

Wavelengt 242 268

Water Bath h
(nm) 454.2 -36.0

Absorban -0.063 0.090

ce
(A)
Table 2: Simplified version of all data
Characterization of nano-particles:

The nanoparticles were characterised into their respective heating processes. They
are grouped into microwaves and heated through a water bath. We also prepared 2
controls to act as a point of comparison.

UV- vis spectrum:

Figure 1.3 shows that the peaks are at 268nm and 290 nm and the absorbance is
-0.063 and 0.090for our control 1. Figure 2.3 shows us that the peaks are at 229nm
and 249nm and the absorbance is from 0.146 and 0.086 for our control 2. Figure 3.3
shows us that the peak for heating using microwave is at 287 nm and 296nm and the
absorbance 0.390 and 0.389 while figure 4.3 shows us the peak are at 242nm to 268
nm and the absorbance -0.063 and 0.090 for using the water bath. Figure 5.2 shows
the 3 different peaks at 259 nm,277nm, and 300 nm for Zinc Nitrate

FTIR:

For the FITR, our peaks we recorded for control 1 were 3338.09cm-1,
2120.24cm-1,1635.23cm-1,655.37 cm-1 and 597.79 cm-1 were observed. Peaks for
control 2 were 3337.37 cm-1, 2107.43 cm-1,1638.61 cm-1,666.79 cm-1 and 597.79 cm-1
were observed. Peaks for S1(microwaved) were 3343.95 cm-1,2120.11 cm-1, 1638.56
cm-1,666.85 cm-1 and 597.75 cm-1were observed. Peaks for S2(water-bath) were
2122.40 cm-1,1638.57 cm-1,666.76 cm-1 and 597.79 cm-1.

Zeta sizer:

For the Zeta sizer, three main things were recorded being size, zeta potential and the
molecular weight of the nanoparticles. For our control 1, the size of the particles in
the mixture was 561.4d.nm with a polydispersity index(Pdl) was 0.702 and the zeta
potential was -27.5mV with an area of 100% and a Zeta deviation of 5.43mV. For
control 2, the size of the particles in the mixture was 403.2d.nm with a polydispersity
index(Pdl) of 0.658 and the zeta potential was -28.1mVwith an area of 100% and a
Zeta deviation of 4.33mV.For our sample 1 which we microwaved, the size of the
particles in the mixture is 288.5d.nm with a polydispersity index(Pdl) of 0.603 and
the zeta potential was -25.5mVwith an area of 100% and a Zeta deviation of 4.59mV.
For our sample 2 which we placed in a water-bath, the size of the particles in the
mixture was 454.2d.nm with a polydispersity index(Pdl) of 0.577 and the zeta
potential was -36.0mVwith an area of 100% and a Zeta deviation of 4.33mV.
5. Discussion, Future Directions and Conclusion (Author:Akilram,
Editor:Elgene)

In this day and age people all over the world seem to be preoccupied with their busy
schedules, thus the popularity of instant foods that take less than 5 minutes to
prepare. When it comes to the packaging of these foods however, we aren’t so
trustworthy of how it is packaged. We worry if the packaging might react undesirably
with our food and cause more problems. Scientists have started looking at silver
nanoparticles as a solution due to its optical and antibacterial techniques. The
applications of silver nanoparticles also stretch beyond being anti-bacterial agents,
they are also good sensors, optical probes as well as good catalysts. (CD
Bioparticles, 2022).

Nanotechnology is the usage and management of incredibly tiny structures, ranging

in size from 1 to 100 nanometers. Because of their tiny size, nanoparticles have the
potential to enter the body. This could have a positive impact on medicine, but it also
poses a risk to public health. There are potential concerns associated with
nanotechnology for both the environment and human health. Many of these dangers
are still poorly understood. Investments have been made in the study of nanoparticle
applications. To thoroughly investigate the concerns, particularly how nanoparticles
affect the human body, will need more time and money.

In this day and age people all over the world seem to be preoccupied with their busy
schedules, thus the popularity of instant foods that take less than 5 minutes to
prepare. When it comes to the packaging of these foods however, we aren’t so
trustworthy of how it is packaged. We worry if the packaging might react undesirably
with our food and cause more problems. Scientists have started looking at silver
nanoparticles as a solution due to its optical and antibacterial techniques. The
applications of silver nanoparticles also stretch beyond being anti-bacterial agents,
they are also good sensors, optical probes as well as good catalysts. (CD
Bioparticles, 2022).

This discussion is to bring closure on and further elaborate on my results and also
talk about my future plans and ways to improve this overall experiment. Firstly, let's
talk about our results of the UV-Vis Spectrophotometer. Figure 3.3 shows us that the
peak for heating using microwave is at 287 nm and 296 nm and the absorbance
0.390 and 0.389 while figure 4.3 shows us the peak are at 242 nm to 268 nm and the
absorbance -0.063 and 0.090 for using the water bath. This shows us that
temperature do play a role in the UV- spectroscopy. One example of this result can
be dated back to an experiment done by (Xin-Xia, 2013). They did an experiment on
how temperature affects Cimetidine tablets (400 mg2). Firstly the researchers,
Cimetidine was first weighed accurately then it was dissolved in hydrochloric
solutions to make stock solutions. A set of standard stock solutions with Ph 1.2 at
concentrations of 89.48, 178.24, 266.96 and 356.68 μg/m and the concentrations
were identical with the dissolution rate of the Cimetidine tablets being at 20.13%,
40.10%, 60.07% and 80.25%. The absorbance was then carried out at different
temperatures being 19,24,30 and 37degreees. The detecting wavelength was then
set at 244 nm and the reference wavelength was set at 300 nm. The results showed
that the same concentration had a higher absorbance at a higher temperature. We
can then say that the standard curves were different at different temperatures. This
proves my hypothesis that molecular movement increases at a higher temperature
therefore more energy will be needed which will therefore increase the absorbance.

Next, we shall proceed with the FTIR. Fourier- transform infrared spectroscopy or
FTIR is an analytical method used to understand structures of individual molecules
and also the composition of molecular mixtures. (METTLER TOLEDO, 2022). For
control 1 our peaks were 3338.09cm-1, 2120.24cm-1,1635.23cm-1,655.37 cm-1 and
597.79 cm-1. Based on the FTIR chart, we can find out that there are carbon triple
bond Functional groups (stretching), Hydrogen-bonds OH stretches (stretching).
Peaks for control 2 were 3337.37 cm-1, 2107.43 cm-1, 1638.61 cm-1, 666.79 cm-1 and
597.79 cm-1..Based on the FTIR chart, we can find out that, there are
hydrogen-bonded O-H stretch functional groups (stretching),carbon triple bond
functional group (stretching), carbon-bromine functional group (stretching) and a
carbon- Iodine functional group (stretching). Peaks for S1 (microwaved) were
3343.95 cm-1, 2120.11 cm-1, 1638.56 cm-1, 666.85 cm-1 and 597.75 cm-1. Based on
the FTIR chart, we can find out that there are hydrogen-bonded O-H stretch
functional groups (stretching),carbon triple bond functional group (stretching),
carbon-bromine functional group (stretching) and a carbon- Iodine functional group
(stretching). Peaks for S2 (water-bath) were 2122.40 cm-1,1638.57 cm-1,666.76 cm-1
and 597.79 cm-1. Based on the FTIR chart, we can find out that there are carbon
triple bond functional groups (stretching), carbon-bromine functional group
(stretching) and a carbon- Iodine functional group (stretching).

Going forward I believe we as a group can improve our skills in testing and preparing
the sample. Firstly, I will talk about the different ways to improve when testing using a
UV-Vis Spectrophotometer. We can improve the accuracy of the UV-Vis
spectrophotometer. A study done by (News-Medical.net, 2017) using extremely short
path lengths, can measure across a wide range. This therefore resulted in improved
accuracy of measurements and saves time and reduces the waste of samples. Next,
we believe we could have improved our skills when preparing and delivering our
spectrophotometric sample. The same study done by (News-Medical.net, 2017)
states that we should always check if the samples are purified prior to the
measurement. The same study also mentions that, we should thoroughly clean the
bottom and top of the measurement surfaces to remove any contaminants as a dirty
or contaminated measurement surface can lead to anomalous results like a negative-
spectra.

Next, we can improve our way of measuring FTIR in the future. We believe that to
improve, we can reduce the size of the diameter of the FTIR. A study done by
(McGreivy, Nick ORCiD, Jaworski, 2018) states that this design allows for scannable,
high spatial resolution absorbance of complex molecular gas mixtures. In that
experiment to achieve the 2nm lateral spatial resolution, the researchers sent the IR
beam through a 2nm to 2nm slits to reduce the beam area. It was then sent through
the plasma discharge and reflected back before being sent to a detector. The results
show that there is an increase in my resolution when the diameter is reduced.

In conclusion, throughout this experiment We as a group have learnt so many

important things like the importance of nanoparticles and the impact it holds. We are
not professionals therefore we have made many errors throughout this experiment.
There are many ways to improve our experiment to reach a much desired outcome
as our results for the most part were not on par with the results we have found online.
Most of our errors were from lack of knowledge and awareness when handling the
samples which could’ve led to contamination and made us not reach our desired
outcome.This report was a good learning experience as I feel it better trains us to
produce scientific reports of great quality.

6. Safety and Precaution (Author:Jayne)

To ensure safety of an individual in the laboratory, some precaution steps are

required when conducting the experiment. Wearing proper PPE is required such as
latex gloves, mask, lab coats, covered shoes, full pants and washing hands before
leaving the lab. Dry nanomaterials need to be transferred in closed containers. Mask
is important for spills that might result in airborne nanoparticles. Avoid brushing or
sweeping spilled or dried nanoparticles instead use a vacuum. While removing
materials from the hot water bath or microwave, do not use bare hands instead use a
clipper or cloth. Lastly, disposing waste in appropriate containers is important for
proper waste segregation.
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A feedback proforma needs to be included with each lab report. Please complete all details clearly.

Student ID: 0353666

Module code: BIO62204

Email: [email protected] Contact No: 60177468189

Title of assignment/report: Synthesis of Green Silver Nanoparticles using plant extract from
(Kaempferia galanga) optimised by temperature.

Module Lecturer: Dr. Nallammai Singaram

B. This section will be completed by the lecturer/tutor assessing your report:

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underlying the lab. made on post-lab questions
lab. Most post-lab answering post-lab are incomplete
Post-lab questions questions. and inaccurate.
questions answered
answered accurately.
accurately.
 Conclusion (5) Conclusion Conclusion Conclusion No conclusion
ncludes whether ncludes whether includes what was was included in
the findings the findings learned from the the report OR
supported the supported the experiment. shows little effort
hypothesis, hypothesis and and reflection.
possible sourceswhat was learned
of error, and what from the
was learned from experiment.
the experiment.

Bibliography (5) Several reputable A few reputable A few references Only one
references were references are are used and cited eference is given;
used and cited used and cited correctly. errors in citation
correctly. correctly. of references.

Format Lab report is Lab report is Lab report is neatly Lab report is
Layout/ Spelling/ typed and uses neatly written or typed, handwritten and
Punctuation/ headings and handwritten and but formatting does looks sloppy with
Grammar (5) subheadings to uses headings not help visually cross-outs,
visually organize and subheadings organize the multiple erasures
the material. to visually material. and/or tears and
One or fewer organize the Detectable (< five creases.
errors in spelling, material. errors) in spelling, More than five
punctuation and Minor (two or punctuation and errors in spelling,
grammar in the three errors) in grammar in the punctuation and
report. spelling, report. grammar in the
punctuation and report.
grammar in the
report.
Safety/ Lab is carried outLab is carried out Lab is carried out Safety procedures
experimental competently with with attention to with some were ignored
initiative full attention to relevant safety uncertainties. and/or some
(5) relevant safety procedures, but Attention to aspect of the
procedures. one safety relevant safety experiment posed
The set-up and procedure needs procedures, but a threat to the
experiment, to be reviewed. several safety safety of the
posed no safety procedures need tostudent or others.
threat to any be reviewed.
individual.

Any additional comments:

Assessed by: Date:

Sample Moderated by (if any): Date: