INDIAN INSTITUTE OF TECHNOLOGY ROORKEE

NPTEL

NPTEL ONLINE CERTIFICATION COURSE

Biomedical Nanotechnology

Lec-03
Synthesis of Nanomaterials by Physical and Chemical Methods

Dr. P. Gopinath
Department of Biotechnology
Indian Institute of technology Roorkee

Hello everyone, today we are going to see the third lecture of this course. The third lecture is on
the synthesis of nanomaterials by physical and chemical methods.

So in this lecture, we are going to learn how to synthesize nanomaterials by physical methods
and chemical methods. And we are also going to learn what is nucleation and growth, and how to
synthesis metal nanoparticles and metal nanorods. And we are also going to see a small simple
experiment to understand how we can synthesis silver nanoparticles in the lab with the available
chemicals.

So let us see the physical methods. Under the physical methods, there are several types. In this
lecture, we are going to mainly focus on mechanical milling and laser ablation methods.

First, we will see what mechanical milling is. Mechanical milling is a top-down process. Top-
down process means, you are converting the bulk material to nanomaterials. Here, the
nanoparticles are formed in a mechanical device, generally referred to as a mill. So here we are
going to use the metallic ball which will break the bulk material into nano-sized particles.

Let us see the principle of milling. So the fundamental principle of the size reduction in a
mechanical device is the energy imparted to the sample, during the impacts between the milling
media. So this model represents the moment of collision. Here, you can see that the particles are
trapped between the two colliding balls, within the space occupied by the mass of powder
particles.

You can see here rotating metallic balls. So the metallic drum is rotating and the metallic balls
are falling on the powder, which is going to break the micro sized powder into nano size
particles. This is called as the mechanical milling or ball milling. We can also call it like a ball
milling, because we are using metallic balls. And this is an electron microscopic picture of the
particles. As I told you earlier, for seeing these nanoparticles, you need an electron microscope.
So this is the scanning electron microscopic picture. You can see here, figure A is after 3 hour of
milling, figure B is after 9 hour of milling, C is after 15 hour and D is after 20 hour of milling.
We can see here with respect to time, the particle size is going down. Whenever you take the
picture you have to insert the scale bar. The scale bar is very important which will be useful for
calculating the size of the particles.

So let us see the next method that is, laser ablation. Here you will be using the laser light to
vaporize your material. The generation of nanoparticles is by laser ablating a solid target. We can
get the nanoparticles in the gaseous or in the liquid environment. As it depends on the
environment, the particles which we get can be in the nano powdered form, or it can be in a
colloidal solution form. Laser ablation is one of the easiest and fastest methods. When you
compare with other methods, it does not involve long reaction times or multiple steps. Further, it
does not require any toxic chemicals for its synthesis. So it is also called as environment friendly
or green method.

In this method, the resulting nanoparticles which we are obtaining in the colloidal solutions are
ultra pure. These particles do not contain any counter ions or any reaction by products, and so we
can use these particles as such for biological applications. And again, we can also add ligands to
these particles after the synthesis process or in the suitable solvent itself to coat the surface of the
nanoparticles.

How to synthesis nanoparticles by laser ablation? If you want to synthesis silver nanoparticles by
laser ablation you need a high purity silver slice. And if you want to make the copper
nanoparticles you need a high purity copper slice. The slice should be at the bottom of the glass
vessel containing 20ml of water. We have to apply the laser light, like 1064 nm pulsed Nd:YAG
laser and operating at fixed energy of 30 minutes. This will produce yellow colored solution in
the case of silver nanoparticles or light green colored solution in the case of copper
nanoparticles.

So this is the experimental setup of laser ablation. You can see that the synthesis of nanoparticles
by laser ablation liquids is one of the simplest experimental configuration. If you want to make
the silver nanoparticles, we are going to use the silver slice, and if you want to make the copper
nanoparticles, you can keep the copper slice. When you apply the laser light, it will vaporize the
material and the nanoparticles can be obtained in the colloidal solution form. And here, we can
use the X, Y and Z stage to move the target and unitize the complete material for making the
nanoparticless.

So this animation will give a clear idea how the laser ablation works. We can see that YAG laser
is applied to the metal target, for example, if you want to make the silver nanoparticles you will
be keeping the sliver metal here and if you want to make the copper nanoparticles you will be
keeping the copper metal here. When you apply the laser light, it will vaporize the metal and this
nanoparticles can be collected in the aqueous media. Using the X, Y stage we can move this
target to the X, Y and Z direction. So you understood how we to make nanoparticles with
physical methods like mechanical milling as well as laser ablation. Now we will see how we can
make nanoparticles by chemical method.

For synthesizing nanoparticles by chemical methods, we need three important components. First,
is the metal salt, second one is reducing agent, and third is capping agent. Some of the examples
for reducing agents are sodium citrate, sodium borohydrate and ascorbic acid. These are some of
the examples of capping agents-Thiols, Citrate, and Polymers like polyvinyl alcohol and PVP.

So what happens here is, the reducing agent will reduce your metal salt into metal nanoparticles
but these metal nanoparticles are highly unstable so it will try to join together and form
agglomerations or aggregations. So to make the nanoparticles stable, we have to add a capping
agent or stabilizing agent which will prevent the aggregation or agglomeration.

Synthesis of any nano materials involves two important steps- one is nucleation and another one
is growth. What is nucleation? Nucleation is the phenomena of initiation of formation of the first
nano crystals in the solution. It involves the appearance of very small particles or nuclei of the
new phase which are capable of growing. How nucleus is important for a living cell, similarly for
nanoparticles this nuclei is very important. There are two type of nucleation. First one is
homogenous nuclei. It forms simultaneously and uniformly throughout the solution. Next one is
heterogeneous nuclei which gets nucleated at different time points.

Let us see this picture of LaMer’s diagram which shows the time verses solute concentration. So
once the nucleus is formed, growth will start. So if the nucleation is homogeneous, all nuclei
forms simultaneously. Then, you will get mono dispersed particles. Mono dispersed means
uniformly sized particles. If the nucleus is heterogeneous, it means it is nucleated at different
times. This produces poly dispersed particles, which means you do not get uniformly sized
particles. So, when you see the nanoparticles under a transmission electron microscope, you can
see uniformly sized particles in mono dispersed particles. In the case of poly dispersed
nanoparticles, you get randomly sized particles.

So nucleation plays a very important role in controlling the properties of final product like size
distribution as well as the nature of the phase. For making very small size nanoparticless, you
need strong nucleation and a slow growth. This means, you can get highly uniform sized
particles, if the process of nucleation and growth are successfully separated.

This is the typical precipitation reaction. Here, we are going to use the metal salt and reducing
agent. So the reducing is going to reduce your metal salt into metal atoms. These metal atoms
combine to form the clusters, these clusters combine to form the crystal size. So this crystallite
will combine and form the primary particles. This is the nucleation step and this is the growth
step. So these primary particles combine and start growing. We have to stop the growth at this
stage or else all the particles will combine and grow into a big particle which can be
agglomerated particles. Agglomerated particles are not useful. We need small particles for our
applications. We can achieve that by arrested precipitation method. What is arrested
precipitation? Here a large number of nucleation centers are formed by vigorous mixing of the
reactant solution. So when you make your nanoparticles, we have to mix the solution vigorously.
When you mix it vigorously, you will have more number of nucleation. This I will show in the
last, with a simple experiment how to mix it vigorously and how to make these small particles.

And here if you keep the concentrations small, then the nuclei growth will be stopped due to lack
of the material. This called as arrested precipitation. We have to stop the particles from the
Oswald ripening. What is Oswald ripening? It is a growth mechanism, where smaller particles
dissolve and are considered by the larger particles. What happens as a result of this is, the
average nanoparticle size will be increased with time and so the particle concentration will
decrease. So if the particle size is increased, it decreases at the solubility. This is called Oswald
ripening. By using a stabilizing agent, we can protect it from the Oswald ripening.

How can we tune the size of the particles? Tuning of the size of the particles can be achieved by
controlling the nucleation as well as the growth rate. For example, fast nucleation will provide
high concentration of nuclei, and it will yield smaller nano crystals. But when you use this slow
nucleation, it provides low concentration of seeds and the same concentration of precursors. This
results in larger particles. The simple example is, consider, we have 5 person and we have 50
rupees. Then each will get 10 rupees. But if we have only 2 persons to share, then each will get
25 rupees. Similarly here, when we have more number of nuclei and same amount of precursors,
we will get small sized particles. A slow nucleation process with only two nucleus but same
quantity of precursors will produce bigger sized particles.

What is the role of a stabilizing agent? It is preventing the uncontrollable growth of particles. It
prevents the particle aggregation and control the growth rate. It will also control the particle size
and allow the particles to dissolve in various solvents.

Let us see how the stabilization agent works. There are two types of stabilization. One is
electrostatic stabilization and the other one is steric stabilization. What is electrostatic
stabilization? It means the adsorption of irons to the surface. It creates an electrical double layer
which results in a Coulombic repulsion force between the individual particles. The uniform
particle’s charge will repel each other. This will give an electrostatic stabilization. The next one
is steric stabilization. This means the metal is covered by a layer of material which is sterically
bulky. For example, the polymers and surfactants could be used.

So these are the parameters which is going to affect the particle growth, shape and structure. The
kind of capping agent you are going to use, the kind of reducing agent you are going to use, and
what is the concentration of reactants determine the nucleation process. Moreover, the pH value
of the solution is important and duration of heat treatment in some cases is going to favor the fast
nucleation.

So this is a typical picture of a metal nanoparticle synthesis. You can see here, if you want to
make gold nanoparticles, you can use the gold metal precursor. You are going to reduce using
reduce agent which will reduce the metal salt into metal nanoparticle.

Let us see how we can make gold colloids. For synthesizing gold colloids in water, we have to
use the metal precursor hydrogen tetrachloroaurate. A solution of trisodium citrate is your
reducing agent. Here we can get gold nanoparticles of the size 10 to 20 nm. In this reaction, the
citrate acts like a weak reducing agent and also it acts like a stabilizer. So a layer citrate anions
adsorbs around each nanoparticle and it prevents them from aggregating because the citrate ions
give a uniform charge to nanoparticles. By electrostatic repulsion, the nanoparticles will be kept
separated out. Here the citrate act like a reducing agent as well as a stabilizing agent. So finally
you will get a ruby-red color solution which will confirm the presence of gold nanoparticles.

Why these metal nanoparticles show different kinds of colors based on the size and shape? This
is the silver and it is showing different kind of colors with respect to size and shape. These are
the gold spheres of different size and gold nano rods, showing different colors. So what is the
reason for formation of different colors? The reason is the surface plasmon resonance. The metal
nanoparticles show the surface plasmon which means the incident light creates oscillation in the
conduction electrons on the surface of the nanoparticles and the electromagnetic radiation is
absorbed.

So let us see what surface plasmon resonance is. When a nanoparticle is much smaller than the
wave length of light, coherent oscillation of the conduction electron bands is induced by the
interaction with an electromagnetic field which creates a resonance called surface plasmon
resonance.

And these are the factors which are going to control the size and the shape of the nanoparticles.
You can see here, that the concentration of the reducing agent and the string rate is going to
control the size of the particles. You always have to use a fresh filter solution to make the
nanoparticles. The kind of stabilizing agent you are going to use is going to decide the size and
shape of your nanoparticles.

Now, let us see how to synthesize gold nano rods. It is a two step process. If you want to make
silver nano rods, you have to use a silver salt and if you want to make gold nano rods, you have
to use a gold salt. Also we are going to use the NaBH4, that is sodium borohydride, which is a
very good strong reducing agent. When you use sodium borohydride, it will reduce the gold into
very small sized particles and it will get the seed formation. . The citrate will act like a capping
agent, stabilizing agent and it will inhibit the particle growth. So you will get the seed and in next
step, you will use the seed and add the gold or silver salt.

And when you add a weak reducing like ascorbic acid, what happens is, it will favor the growth.
Here you are using a different kind of stabilizer that is CTAB. This CTAB stabilizer will provide
a template to get rod shaped nano rods in this step. So, when you use a strong reducing agent,
you will get the seeds and when you use a weak reducing agent you will get the nano rods.
Further, the CTAB stabilizer plays a major role in formation of nano rods.

So, this is the synthesis of gold nano rods. Here, in the first step, you are going to use the gold
solution and sodium citrate. You are going to mix them with sodium borohydride and you will
get nanoparticles of 4 nm in diameter. So this is your seed. The second step is stock solution.
Here we are going to use the gold solution plus you are adding the CTAB stabilizer. Also you are
going to add the ascorbic acid.

When you add the ascorbic acid, what happens is, it is going to reduce the gold Au3+ to Au 1+.
This results in the disappearance of color. It is a 3 step protocol for nano rod synthesis. The first
step is you take 1 ml of seed and mix with 9 ml of your stock solution. From this solution, you
take 1 ml and mix 9 ml of your stock solution and finally, you take 10 ml of this solution + 90ml
of this stock solution to get the nano rods.

This size of nano rod synthesized depends on the seed size as well as the length of tails of the
CTAB. If we have more CTAB, it is going to play a major role in the nano rod size and
dimension.

What is the growth mechanism of gold nano rods? The first step is nucleation, followed by the
growth, which will be followed by the development of facet. Once the facet is formed, the CTAB
stabilizer will go and bind to this facet. It will continue to grow until all the reagents are
exhausted. This CTAB is a positively charged bilayer and it is going to stabilize the nano rods.

By changing the concentration of reducing agent and stabilizing agent, we can get different kinds
of shapes like cubes, hexagon, and triangular nanoparticles.

In a simple example, we are going to use ascorbic acid. We are going to increase the
concentration of ascorbic acid from A to C. So when you are increasing the concentration of
ascorbic acid from A to C, we are increasing the seed concentration from C to D. When you play
around with the concentration of ascorbic acid, we will get different kinds of size and shapes of
the nanoparticles.

So if I ask you to select a nanoparticle based on size for a catalytic reaction, which one will you
select? We will select the smallest size, because the smallest nanoparticles have the highest
surface area to volume ratio and so it will have more catalytic activity.
So now, let us see how to synthesize silver nanoparticles. Here we will use silver nitrate, AgNO3
and the sodium borohydride as the reducing agent. This reducing agent will also act like a
stabilizer. You can see here, from Ag+ it became Ag0 and the borohydride is protecting these
nanoparticles from aggregation or agglomeration.

So in the case of silver nanoparticles, you will get yellow colored solution. We can see here that
these silver nanoparticles are of different sizes. If the particles are aggregated it is not useful for
the further applications. And how do you control and how do you stabilize these nanoparticles?
The ratio between the metal salt and the reducing agent is very very important. If the ratio is 2, it
is highly stable, if the ratio is 2.1 it is stable for 30 minutes and if the ratio is 1.9, it is stable for
only 20 minutes, and if the ratio is 1.8 it is stable only for 5 minutes. So for any nanoparticle
synthesis protocol, you have to think and optimize the ratio properly. This is very important for
making stable nanoparticles.

After you have made the nanoparticles, the first step of the characterization is UV visible
spectroscopy. I guess most of you know what UV visible spectroscopy is. Most of the labs will
be having this UV visible spectroscopy. So what you have to do is, you have to transfer 1ml of
the synthesized solution to a quartz cuvette and scan the solution in the UV visible spectrometer.
So when you start the scanning from 200 to 800 nm, you will get the λmax at 400 nm wave length.
Usually, silver nanoparticles give a λmax at 400 nm range. If we get this kind of peak, you can be
sure that you got silver nanoparticles. From the silver nanoparticles peak, you can approximately
calculate the size. For example, if you are getting the λ max at 395 nm, the particle size is
approximately 10 to 14 nm. If we get the peak at 420 nm, then the size of the particle is 35 to 50
nm, and if we get the peak at 438 nm, then the size of the particle is 60 to 80 nm.

Next step is the analysis by using a transmission electron microscope. When you see under a
transmission electron microscope, you can see the particles size. When you take any microscopic
picture, you have to include a scale bar. By comparing with the scale bar, the size of the particles
can be found. So based on the scale bar, we get easily estimate and calculate the particle size.
Now, let us see the nano rod formation. When you use a silver salt and make nano rods, two
peaks will appear. When you see the nano rods under UV visible spectroscopy it will give two
peaks. One peak corresponds to the seed and remaining all are nano rods of different size. You
can see that the nanoparticles are giving a single peak but the nano rods are giving two peaks.
What is the reason? Nano rods are giving two peaks because it has two dimensions- longitudinal
and transverse.

Let us see a simple experiment. I will demonstrate you, how to make nanoparticles in your lab
with simple chemicals available in your lab. So for this, we have to use 1ml of deionized water in
1.5 ml micro centrifuge tube. We will be adding 1.67 µl of silver nitrate solution that is of 0.1M,
to the above tube. We will immediately add 1.33 µl of freshly prepared 2mg /ml NaBH4 solution
to it. The sodium borohydride solution should be freshly prepared. Then, you add 90 µl of SDS
solution to the above solution. This SDS solution is the stabilizer and it is going to stabilize your
silver nanoparticles. Then you have to mix the solutions vigorously. If you see yellow color, that
means you are sure that the silver nanoparticles is formed.

In another 1.5 ml tube, where I did not add any reducing agent, will remain transparent.
Nanoparticles are not formed in this tube. From this tube, I am going to remove 93 µl of water
because in the subsequent step, we are going to add 90 µl of SDS and 1.67 µl of silver nitrate
solution and 1.33 µl of sodium borohydride solution. The silver nitrate solution is light sensitive
and so we have to cover it with an aluminum foil. Always wear gloves and also lab coat when
you make silver nanoparticles.

So I am adding the silver nitrate solution to both the tubes. Now I am going to add 1ml of water
to the sodium borohydride. As I told you earlier, the sodium borohydride should be prepared
fresh. We will take 2 mg of sodium borohydride in a 1.5 ml tube and add 1ml of water to it to
make a fresh solution of sodium borohydride. Mix it thoroughly, so that the sodium borohydride
can dissolve completely. Add 1.33 µl of this freshly prepared sodium borohydride to only one of
the tubes and not to both the tubes. Now I am adding 90 µl of SDS solution to both the tubes. I
am going to mix it vigorously. Now you can observe the formation of yellow color, which means
the nanoparticles are formed. You can see clearly that the tube to which reducing agent was
added became yellow in color indicating the formation of silver nanoparticles and the tube to
which the reducing agent was not added remains transparent.

That means, the silver nanoparticles are formed in the solution where I added the reducing agent.
The other tube, where there is no reducing agent, there is no formation of silver nanoparticles.
Hope you understood this experiment to make nanoparticles in the lab.

So finally to summarize, in this lecture, we have learnt what mechanical milling is and what laser
absorption is. So these are two physical methods and using these physical methods, we learnt
how to make nanoparticles. You learnt that the mechanical milling is a top down approach. The
laser absorption is a green method and ecofriendly method because it does not involve the use of
toxic chemicals. We also learnt how to make nanoparticles by chemical reaction method.

For chemical reaction method, you need three important components. What are these important
components? You need metal salt, a reducing agent as well as a capping agent or stabilizing
agent. We also learned what nucleation is and what growth is. So nucleation is the formation of
first nano crystals in the solution which will be followed by the growth.

And we have also learnt how to make metal nanoparticles and metal rods. By a simple
experiment, we also understood how to make nanoparticles in the lab with the available
chemicals. I will end my lecture here. Thank you all for listening. I will see you in another
interesting lecture.

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