World Journal of Nano Science and Engineering, 2018, 8, 1-31

http://www.scirp.org/journal/wjnse
ISSN Online: 2161-4962
ISSN Print: 2161-4954

Characterization of Polyvinyl Pyrollidone

Coated Sodium Borohydride Stabilized Particle
Colloidal Silver Fresh None Filtered and Nano
Filtered Solution Made up of Magnetic Stirring
and Cooling Method

Jyoti Prakash Pani1*, Royana Singh2, Sanjay Singh3

1
Department of Anatomy, Sir Nandkumar Y. Tasgaonkar Institute of Medical Science and Research, Mumbai, India
2
Department of Anatomy, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India
3
Department of Pharmaceutical Division, Indian Institute of Technology, Banaras Hindu University, Varanasi, India

How to cite this paper: Pani, J.P., Singh, Abstract

R. and Singh, S. (2018) Characterization of
Polyvinyl Pyrollidone Coated Sodium Bo- Characterization is absolutely necessary and is a must in order to understand
rohydride Stabilized Particle Colloidal and estimate different silver nanoparticle (nm) size in specific group wise
Silver Fresh None Filtered and Nano Fil- manner which corresponds to group wise in number & sizes, and their im-
tered Solution Made up of Magnetic Stir-
ring and Cooling Method. World Journal
portance and effect on biological tissue and organs with agglomeration for
of Nano Science and Engineering, 8, 1-31. nano toxicological studies in environments, the acute toxicity of colloidal sil-
https://doi.org/10.4236/wjnse.2018.81001 ver nano particles (AgNps) were studied in fresh dissected tissues of Swiss Al-
bino mice and their fetuses. In this manuscript, an attempt is made to demon-
Received: March 16, 2018
Accepted: March 28, 2018
strate the synthesis and characterization of silver nano particles with a wide
Published: March 31, 2018 range of sizes (from 2.75 nm up to 1908.2 nm in radius) by reducing silver ni-
trate powder with polyvinyl pyrollidone in aqueous solutions in the presence
Copyright © 2018 by authors and of a sodium borohydride stabilizer. The resulting particles were found spheri-
Scientific Research Publishing Inc.
cal aggregates with a rough surface and poly dispersity index below 18.26%
This work is licensed under the Creative
Commons Attribution International (>0.783 PDI). The particle optical, cumulative, diluents and electrical conduc-
License (CC BY 4.0). tivity properties were examined by dynamic light scattering and zeta potential
http://creativecommons.org/licenses/by/4.0/ but morphology was evaluated after examination by transmission electron
Open Access microscopy & image-j. Silver nanoparticles were directly coated with poly-
vinyl pyrollidone with a sodium borohydride stabilizer. Optical properties on
a single-particle level were studied by means of auto correlation function
measurements. The effective poly dispersity index of the charged silver nano-
particles was low enough to form a colloidal crystal at low ionic strength.
Colloidal form is found more toxic than suspended particles in 1.5 molar so-
dium chloride solution; this shows increase of silver nanoparticles size due to

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J. P. Pani et al.

agglomeration, will reduce the toxicity but increase teratogenicity.

Keywords
Intensity Distribution, Auto Correlation Function, DelsaTM Nano Beckman
Coulter Machine, Poly Dispersity Index, Refractive Index, Viscosity

1. Introduction
Colloidal metallic nano silver solution exhibits significant importance on fun-
damental, textile and industrial sector, and also on the metallodielectric silver
nanoparticle which also expresses great interest in electrical conductivity in cir-
cuit. Colloidal nanosilver particles are vividly used as agents for breakable
chemical reaction & optics [1] [2] relevant sensor for biological and chemical
products [3]. It also represents the study of lights with no parallel alignment [4]
[5] [6] spectroscopy discovered by scientist Raman C V with extended floor
(SERS) [7] [8] and the field of electronics and electrical [9]. Photonic crystal
study is absolutely a recently created field in universe of nano science which di-
rectly has a link and relation with successful characterization of particle colloidal
silver. In the recent time, silver nano metals of smallest, smaller, small, bigger
and biggest sizes and metallodielectric spheres found new applications in field of
photonic crystals [10] [11] [12] [13] [14] to synthesize NaBH4 stabilized and
PVP coated colloidal silver nano metal particles by magnetic stirring and cooling
method more than one protocols have been developed, but in this manuscript,
only specific protocol articulated and others mentioned as material and method
section in characterization articles [15] [16] [17]. Sodium borohydrate (NaBH4)
and polyvinyl pyrollidone (PVP) [18] are utilized as chemical stabilizer and coa-
ter to nanosilver because of the high electronegative characteristic of silver nano
metal (+0.483 PDI; −17.52 mV zp). The nanosilver may be coated with reducing
agents like citrate which is also called capping agent [19]. It also includes free
radicals [19]. Many synthesis methods are famous for the preparation of silver
nano metal spheres which are ultimately developed to produce small silver na-
noparticles (radius less than 2.75 nm). In this article, we strongly emphasize the
characterization of silver nanoparticles after magnetic stirring and cooling syn-
thesis of silver particles in colloidal form with a wide range of small and big sizes
and narrow with broad size distribution by reducing silver nitrate with poly
vinyl pyrollidone in an aqueous medium and 1.5 molar sodium chloride sus-
pender with sodium borohydried stabilizer. The important properties of silver
nanoparticle are shape, size, aggregation state, external surface modification
properties, solubility in solution, nano size structure and comprise of chemical
constituent with make-up certainly affected silver nanoparticle behavior and
toxicity. For silver nanoparticle summarization of synthesis and characteriza-
tion, a wide range of spectroscopic and biological methods are also available in

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 J. P. Pani et al.

plenty manner. Dynamic light scattering, zeta potential UV-Vis and infrared
spectroscopy and image-j estimation offer the best possibility to characterize sil-
ver nanoparticles which are chosen in this work for characterization of size. Es-
pecially quantum dots and organic based nanoparticles like fullerenes and car-
bon nanotubes needs characterization Fourier transformation infrared (FTIR)
and UV-Vis spectroscopy which have been used to compare aqueous colloidal
suspensions of C60 and this is typically showing similarity to characterization
method adopted for various size nanosilver metal [20]. Pesika the main scientist
with team also used UV spectroscopy to study the relationship between absor-
bance spectra and particle size distributions for quantum-sized nanocrystals
[21]. Particularly in the case of silver nanoparticles, the UV-Vis absorption
spectra have proved to be quite sensitive as the Plasmon peak and full width at
half maximum (FWHM) depends on the extent of colloid aggregation [22].
X-ray diffraction (XRD) is a non-destructive technique and can reveal informa-
tion about the crystallographic structure, size and elemental composition of nat-
ural and manufactured materials. It has been widely applied in the characteriza-
tion of silver and other metal nanoparticles including those synthesized using
biological agents. Microscopic techniques can be applied not only to visualize
silver nanoparticles but also to generate useful data on the size, shape distribu-
tion and other measurable properties [23]. Electron microscopy is one of the
techniques that have played a major role on studying silver nanoparticles. Since
the early conventional bright field images and the intermediate resolution dark
field techniques, to the high-resolution atomic images of nanoparticles the re-
sults have shown that indeed the nanoparticles, in the range of a few nanome-
ters, can have well-defined crystal structures [24]. The relationship between the
biological activity and size/shape of silver nanoparticles has been investigated.
As a general trend, it has been found that smaller particles generally present an
enhanced antibacterial power if compared with their larger counterparts. Al-
though little is known about how the biological activity is affected by the silver
nanoparticles shape, some scientists have shown that truncated triangular silver
nanoparticles present stronger biocidal effect than those presenting spherical or
rod-shaped morphologies. However, all studies performed until now have used
distinct reactions to synthesize silver nanoparticles with different sizes or shapes.
Although, similar trends have been obtained for different groups, the fact of
comparing the biological activity of nanoparticles obtained from distinct synthe-
sis was always a point of discussion since the final result could be masked by ar-
tifacts coming from different syntheses procedure. This study attempts to made
verification of the silver nanoparticle characterization process which was done
by the techniques of UV-Vis absorption spectroscopy, transmission electron
microscopy (TEM) and dynamic light scattering (DLS) with zeta potential.
1) Material & Methods
All the reagents and chemicals for this silver nanoparticle colloidal solution
synthesis and characterization experiment were purchased in raw or powdered

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J. P. Pani et al.

form, from Trimurthy Scientific Chemical Organization Pvt. Ltd. Varanasi Ut-
tarpradesh, India.
2) Synthesis of Silver Nano Particle Colloidal Solution
Procedure
Step 1. 30 ml of 0.002 M Sodium Borohydride (NaBH4) added to an Erlen-
meyer flask. Conformity achieved that solution used must be made fresh right
before the experiment. A 1 inch magnetic stir bar was added and place the flask
in an ice bath on a stir plate and stirred. Keeping the Sodium Borohydride
(NaBH4) on ice reduce the rate of decomposition during the experiment.
Step 2. 2 ml of 0.001 M silver nitrate (AgNO3) dripped into the stirring fresh
NaBH4 solution at approximately 1 drop per second from a graduated bottle
fixed with infusion set. Stirring was stopped as soon as all of the AgNO3 is add-
ed.
Step 3. The presence of a colloidal suspension can be detected by the reflection
of a laser beam from the particles.
Step 4. A small portion of the solution transferred to a test tube. The addition
of a few drops of 1.5 M sodium chloride (NaCl2) solution causes the suspension
to turn darker yellow, then grey as the nanoparticles aggregate.
Step 5. A small portion of the solution transferred to a test tube. A drop of
0.3% polyvinyl pyrollidone (PVP) added to it. PVP prevents aggregation. Addi-
tion of NaCl2 solution then has no effect on the color of the suspension. Slowly
PVP solution added to the stirred Silver colloid solution.
Step 6. Silver nanoparticles color the yellow stained glass in medieval
churches. To make “stained glass” the mixture decanted into a mold leaving air
bubbles [25].
3) Materials required
Stock Solutions for 8 batches.
• 0.001 M AgNO3: 0.017 g of AgNO3 was dissolved into 100 ml distilled water.
This solution can be kept for later usage.
• 0.002 M NaBH4: 0.0189 g of NaBH4 was dissolved into 250 ml distilled water.
This solution must be made fresh before the experiment.
• 0.3% PVP solution: 0.1 g of PVP was dissolved into 33 ml distilled water.
4) Equipment (Figure 1)
5) Instrument required
• Small Erlenmeyer flask.
• Large dish of ice.
• Stirrer hotplate.
• 1" magnetic stir bar.
• Droppers.
• Laser pointer.
• Mold.
6) Characterization of colloidal silver solution done by Figure 2
7) Characterization of silver nano particles

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 J. P. Pani et al.

Figure 1. Synthesis of AgNps colloidal solution by magnetic stirring and cooling method
upon a magnetic stirrer.

Figure 2. (A-D): Methods adopted for characterization of silver nano particles colloidal
solution (A), (B). Dynamic light scattering and zeta potential (C). Image-j (D). Transmis-
sion Electron Microscopy.

Further, the Silver nanoparticle colloidal solution was observed by TEM

(Transmission electron microscopy), DLS (Dynamic light scattering) and Zeta
potential for characterization to know particle size and mean, standard devia-
tion, standard error of mean are evaluated from number of particles. Image-J
software was also used for this purpose [26] [27] [28].
8) Procedures for segregating bigger and smaller size silver nano particle of 20
nm mean size (Figure 3)
The procedure adopted for segregating bigger size particles from smaller size
particles was to filter the bigger size silver particles colloidal silver with the help
of a nano filter device. The device comprises of 2 knobs made up of hard plastic
one washer and a fine filter with less than 100 nm pore size. (2 - 20 nm size
range) The raw colloidal silver which was freshly prepared and collected inside a
20 ml big syringe with silver foil wrapped and light protected. The shining yel-
low colored none filtered colloidal silver after filtration becomes amber color the
shinning yellow color cause harm to eye ball. The amber colored solution was

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J. P. Pani et al.

Figure 3. (A-D): (A), (B)-Nano filter device with (C). Falcon tube in which colloidal sil-
ver is kept by wrapping silver foil (D). Filtered and non filtered AgNps colloidal solution
None filtered AgNps colloidal solution shinning yellow colored Filtered AgNps colloidal
solution amber colored.

collected in a properly leveled falcon tube after filtration with silver foil wrapped
and light and temperature protected as it was highly sensitive. Both non filtered
and filtered colloidal silver bearing falcon tube was properly leveled to differen-
tiate between the two.
9) Transmission electron microscopy
To obtain the size and morphology of the Silver nanoparticles and to get the
TEM image of the tissue which is 1, 1.5 and 10 lakh times magnified, TEM
processing with viewing performed. TEM analysis of the PVP coated Sodium
Borohydried stabilized Silver nanoparticles colloidal solution performed at an
accelerating voltage of 100 KV. Silver nanoparticles was examined in the form of
colloidal solution suspended in NaCl2 (Ag-1.8 mg/ml) and subsequently depo-
sited on the form var/carbon coated TEM grid. Digital TEM camera was cali-
brated for size and measurement of the silver nanoparticular mass. Mean, Stan-
dard deviation, Individual differences from mean were calculated from procured
information measuring over 100 Silver nanoparticles in the random fields of
view with addition to it images procured by specific compacted cassettes which
is viewed by special software named soft imaging viewer or SIS image viewer
that showed general morphology of the scattered silver nanoparticles in the solu-
tion.

2. Photography
Analysis and photography of scattered 100 silver nanoparticles in the random
fields of view in the solution was done under FEI Tecnai G2 spirit twin trans-
mission electron microscope equipped with Gatan digital CCD camera (Nether-
land) at 60 or 80 KV.

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1) Statistical analysis
Percentage of outputs evaluated from each groups and finally the percentages
were compared between and within various size group nanosilver. Continuous
data were compared by the application of one way ANOVA, Manova, post hoc,
multiple comparison Dunnett test, F, t test, Newmann Keul test and descriptive
statistics. Mean, standard deviation, standard error of mean were evaluated. Dif-
ferences were considered significant when evaluated P value shows less than
0.005 (P < 0.005).
2) Result
3) Characterization of silver nano particles colloidal solution
With an aim to know the exact size and diameter of all the silver nano par-
ticles in nano meter (nm) scale the present study was undertaken. AgNps col-
loidal solution was used to estimate mean and standard deviation of the particle
size. Successful characterization of the fresh prepared AgNps colloidal solution
by the process of Dynamic light scattering, zeta potential, Transmission electron
microscopy and Image-J estimation was done which led to proper segregation of
bigger and smaller size silver nanoparticles from none filtered (a(fresh),
a1(preserved)) and nano filtered (b(fresh), b1(preserved)) AgNps colloidal solu-
tion.
4) Dynamic light scattering by DelsaTM Nano Common Beckman Coulter
machine showing intensity distribution and auto correlation function
5) Overall none filtered colloidal AgNps solution analysis (a (fresh), a1 (pre-
served))
The Dynamic light scattering (DLS) yielded 2 types of result such as distribu-
tion result and cumulative result. The DLS analysis (distribution result) of none
filtered colloidal silvernano solution (a) resulted in form of 5 peak values its
Diameters, standard deviation analyzed from intensity distribution and auto
correlation function (ACF) graph. The analysis is shown in Table 1 given below.
The average diameter liberated by Beckman Coulter from distribution result
was 6151.2 nm with standard deviation 10,181.3 (6151 ± 10,181.3). The cumula-
tive result part from analysis liberated by DelsaTM Nano Beckman Coulter pur-
chased from United Kingdom was shown to be 1340.3 nm as average diameter of
all bigger silver nanoparticles. The poly dispersity index value of the same

Table 1. The DLS (distribution result) analysis of none filtered colloidal silver solution
(a) resulted in form of 5 peak values its diameters, standard deviation analyzed from in-
tensity distribution and ACF graph shown in Table 1.

Peak Diameter Standard deviation

1 272.6 69.1
2 22,601.5 5138.13
3 0.0 0.0
4 0.0 0.0
5 0.0 0.0

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J. P. Pani et al.

colloidal silver (a) liberated by the same machine was 0.538 at a measurement
condition when temperature is maintained 25.1˚C inside the water diluents of
the colloidal silver with refractive index 1.3328 and viscosity 0.8858 and scatter-
ing intensity was found to be 11101 (CPS) (Figure 4).
The distribution result part of DLS analysis of none filtered colloidal silver
solution (a1) also yielded by Beckman Coulter in form of 5 peak values its di-
ameters, standard deviation analyzed from intensity distribution and ACF graph
shown in Table 2 given below.
The average diameter of colloidal silver (a1) liberated by Beckman Coulter
was 6750.8 nm with standard deviation 10,979.6 (6750.8 ± 10,979.6) which is
seen moderate higher in comparison to colloidal silver (a). The cumulative

Figure 4. Intensity distribution and ACF graph after DLS analysis of none filtered colloidal silver solution (a) showing distribu-
tion result in the form of peak, diameter (nm), std.dev., average with poly dispersity index and diffusion constant.

Table 2. The DLS analysis of none filtered colloidal silver solution (a1) also resulted in form of 5 peak values its diameters, stan-
dard deviation analyzed from intensity distribution and ACF graph table.

Peak Diameter Standard deviation

1 33.9 7.1
2 627.7 371.3
3 21,330.5 10,096.9
4 0.0 0.0
5 0.0 0.0

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 J. P. Pani et al.

analysis of the same (a1) liberated by Beckman Coulter was 1944.9 nm as aver-
age diameter of all bigger silver nanoparticles scattered in matrix of solution
which showed significant increase in average size of silver nanoparticles present
in colloidal silver (a1) in comparison to colloidal silver (a) as the concentration
and agglomeration of silver particle in colloidal solution increase. The poly dis-
persity index of the same colloidal silver (a1) liberated by Beckman Coulter was
0.722 which also showed increase PDI in comparison to colloidal silver (a) at a
measurement condition when temperature is maintained 25.1˚C inside the water
diluents of the colloidal silver with refractive index 1.3328 and viscosity 0.8858
and scattering intensity having 11101 (CPS). The refractive index, viscosity and
scattering intensity of colloidal solution (a1) is found similar to colloidal solu-
tion (a). The increments of all those parameters of colloidal silver (a1) found
significant (P < 0.001) (Figure 5).
The overlay graph of normalized intensity distribution analysis of colloidal
silver (a) liberated by Beckman Coulter machine showed, 2 phase analysis of PI
with values 0.483 & 0.517 with average calculated 0.483 + 0.517/2 = 0.500 which
more or less matches with PI liberated in Cumulant analysis (0.538) by intensity
distribution & ACF graph of colloidal silver (a). 2 phase analysis of Diameter of
10% silver nanoparticles of colloidal silver (a) showed 200 to 373.2 nm ranges

Figure 5. Intensity distribution and ACF graph of DLS analysis of none filtered colloidal silver (a1) showing distribution result
part in the form of peak, diameter (nm), std.dev., average with poly dispersity index, and diffusion constant.

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J. P. Pani et al.

and 200 to 324.6 nm ranges, Diameter of 50% silver nanoparticles showed 200 to
1011.1 nm ranges and 200 to 896.6 nm ranges, Diameter of 90% silver nanopar-
ticles showed 200 to 3147.1 nm ranges and 200 to 3199.0 nm ranges. The overall
average size of bigger silver nanoparticles of colloidal silver (a) resulted 200 to
1378.8 nm ranges (Figure 6).
The over lay graph of normalized intensity distribution analysis of colloidal
silver (a1) yielded 2 phase analysis of PI by Beckman Coulter machine with val-
ues 0.722 & 0.538 with average calculated 0.722 + 0.538/2 = 0.630 which in-
creased significantly (P < 0.001) from the PI value of normalized intensity dis-
tribution analysis by overlay graph of colloidal silver (a). 2 phase analysis of di-
ameter of 10% silver nanoparticles of colloidal silver (a1) scattered in matrix of
solution showed 200 to 204.2 nm ranges and 200 to 189.9 nm ranges, Diameter
of 50% silver nanoparticles showed 200 to 685.4 nm ranges and 200 to 288.5 nm
ranges, Diameter of 90% silver nanoparticles showed 200 to 23,286.7 nm ranges
and 200 to 23,080.1 nm ranges. The overall average size of bigger silver nanopar-
ticles of colloidal silver (a1) calculated 200 to 1642.6 nm ranges. The overall in-
crements of values found significant (P < 0.001) in comparison to bigger silver
nanoparticles of colloidal silver (a) (Figure 7).

Figure 6. Overlay graph of normalized intensity distribution analysis of none filtered colloidal silver (a) showing pH, average di-
ameter, PDI, segregated analysis of 10%, 50% and 90% silver nanoparticles size in nm range and its overall average.

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 J. P. Pani et al.

Figure 7. Overlay graph of normalized intensity distribution analysis of none filtered colloidal silver (a1) showing pH, average
diameter, PI, segregated analysis of 10%, 50% and 90% silver particles size in nm range and its overall average.

6) Overall nano filtered colloidal AgNps solution analysis (b (fresh), b1 (pre-

served))
The DLS analysis of nano filtered colloidal silver solution (b) yielded by
Beckman Coulter liberated in form of 5 peak values its diameters, standard devi-
ation analyzed from intensity distribution and ACF graph shown in Table 3
given below.
The average diameter in distribution result part yielded by Beckman Coulter
machine liberated 1500.1 nm with standard deviation 1317.7 (1500.1 ± 1317.7).
The cumulative analysis of the same yielded by Beckman Coulter machine libe-
rated 200.1 nm as average diameter of all smaller silver nanoparticles. The poly
dispersity index of the same colloidal silver (a) yielded by Beckman Coulter ma-
chine liberated 0.483 at a measurement condition when temperature is main-
tained 25.8˚C inside the water diluents of the colloidal silver with refractive in-
dex 1.3327 and viscosity 0.8719 and scattering intensity having 6453 (CPS)
(Figure 8).
The DLS analysis of nano filtered colloidal silver solution (b1) yielded by
Beckman Coulter machine liberated in form of 5 peak values its diameters,
standard deviation analyzed from intensity distribution and ACF graph shown
in Table 4 given below.

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J. P. Pani et al.

Table 3. The DLS analysis of nano filtered colloidal silver solution (b) yielded by Beck-
man Coulter liberated in form of 5 peak values its diameters, standard deviation analyzed
from intensity distribution and ACF graph table.

Peak Diameter Standard deviation

1 1500.1 1317.7
2 0.0 0.0
3 0.0 0.0
4 0.0 0.0
5 0.0 0.0

Table 4. The DLS analysis of nano filtered colloidal silver solution (b1) also yielded by
Beckman Coulter machine liberated in form of 5 peak values its diameters, standard dev-
iation analyzed from intensity distribution and ACF graph table.

Peak Diameter Standard deviation

1 1372.9 1415.9
2 0.0 0.0
3 0.0 0.0
4 0.0 0.0
5 0.0 0.0

Figure 8. Intensity distribution and ACF graph of DLS analysis of nano filtered colloidal silver (b) showing distribution result in
the form of peak, diameter (nm), std.dev., average with poly dispersity index and diffusion constant.

The average diameter yielded by Beckman Coulter machine liberated 1372.9

nm with standard deviation 1415.9 (1372.9 ± 1415.9). The shown value which

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 J. P. Pani et al.

significantly decrease from the value of smaller particle colloidal silver. (b) The
cumulative analysis of the same yielded by Beckman Coulter machine liberated
71 nm as average diameter of all smaller silver nanoparticles. The poly dispersity
index of the same colloidal silver (b1) yielded by Beckman Coulter machine li-
berated 0.461 which significantly decreased from PI value of colloidal silver (b)
at a measurement condition when temperature is maintained 25.8˚C inside the
water diluents of the colloidal silver with refractive index 1.3328 and viscosity
0.8878 and scattering intensity having 6292 (CPS) (Figure 9).
The overlay graph of normalized intensity distribution analysis of nano fil-
tered colloidal silver (b) yielded by Beckman Coulter machine liberated 2 phase
analysis of PI with values 0.679 & 0.243 with average calculated 0.679 + 0.243/2 =
0.461 which decrease significantly (P < 0.001) from the value of normalized in-
tensity distribution analysis by overlay graph of bigger particle colloidal silver
(a & a1). 2 phase analysis of diameter of 10% smaller size silver nanoparticles
colloidal silver yielded by Beckman Coulter machine liberated 2.3 to 5.2 nm
ranges with average 3.75 nm, diameter of 50% smaller size silver nanoparticles
yielded by Beckman Coulter machine liberated 71 to 717.5 nm ranges and 30.5
to 30.5 nm ranges with an average of 71 to 374 nm ranges, Diameter of 90%
smaller size silver nanoparticles yielded by Beckman Coulter machine liberated
71 to 3605.5 nm ranges and 71 to 198.1 nm ranges. The overall average size of
smaller silver nanoparticles of colloidal silver yielded by Beckman Coulter ma-
chine liberated 71 to 300.1 nm ranges. The overall decrease of values found

Figure 9. Intensity distribution and ACF graph of DLS analysis of nano filtered colloidal silver (b1) showing distribution result in
the form of peak, diameter (nm), std.dev., average with poly dispersity index and diffusion constant.

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J. P. Pani et al.

significant (P < 0.001) in comparison to bigger silver nanoparticles of colloidal

silver (a & a1) (Figure 10).
The overlay graph of normalized intensity distribution analysis of nano fil-
tered colloidal silver (b1) yielded by Beckman Coulter machine liberated 2 phase
analysis of PI with values 0.479 & 0.223 with average calculated 0.479 + 0.223/2 =
0.351 which decrease significantly (P < 0.001) from the value of normalized in-
tensity distribution analysis by overlay graph of bigger particle colloidal silver (a
& a1). 2 phase analysis of diameter of 10% smaller size silver nanoparticles col-
loidal silver (b1) yielded by Beckman Coulter machine liberated 2.3 to 5.2 nm
ranges with average 3.75 nm, diameter of 50% smaller size silver nanoparticles
(b1) yielded by Beckman Coulter machine liberated 21 to 117.5 nm ranges and
30.5 to 30.5 nm ranges with an average of 21 to 74 nm ranges, Diameter of 90%
smaller size silver nanoparticles yielded by Beckman Coulter machine liberated
21 to 360.5 nm ranges and 21 to 198.1 nm ranges. The overall average size of
smaller silver nanoparticles of colloidal silver yielded by Beckman Coulter ma-
chine liberated 21 to 71.1 nm ranges. The overall decrease of values found sig-
nificant (P < 0.001) in comparison to bigger silver nanoparticles of colloidal sil-
ver (a & a1) (Figure 11).
7) Zeta potential analysis of none and nano filtered fresh and preserved col-
loidal silver (Figure 12).

Figure 10. Overlay graph of normalized intensity distribution analysis of nano filtered colloidal silver (b) showing pH, average
diameter, PI, segregated analysis of 10%, 50% and 90% silver particles size in nm range and its overall average.

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 J. P. Pani et al.

Figure 11. Overlay graph of normalized intensity distribution analysis of nano filtered colloidal silver (b1) showing pH, average
diameter, PI, segregated analysis of 10%, 50% and 90% silver particles size in nm range and its overall average.

The zeta potential analysis of the none and nano filtered fresh and preserved
particle colloidal silver yielded by Beckman Coulter liberated −17.52 mV zeta
potential value in common, same for all type of colloidal solution. Mobility
yielded by Beckman Coulter liberated −1.369e−004 (cm2/Vs) in common, same
for all type of colloidal solution which indicates that colloidal solution is thin in
nature. Conductivity rate of the current in all types colloidal solution showed
0.4356 (mS/cm). Zeta-potential analysis of individual flow cells of all types of
colloidal silver yielded by Beckman Coulter liberated in the following form that
is upper surface −52.08 (mV) and lower surface 9.63 (mV). The Doppler shift of
all types of solution showed 10.7 Hz .The base frequency of all types of solution
showed 124.8 Hz. The refractive index of all types of solution showed 1.3328
with dielectric constant 78.3. The viscosity of all types of colloidal silver showed
0.8858 which proofs that the colloidal solution is thin and it contains mixture of
smaller particles both which easily allows the current to pass fast through the
solution. The average electric field for all types of colloidal solution showed
−16.18 V/cm. The average amount of current pass through all types of colloidal
solution showed −0.36 mA which also indicates all types of colloidal silver solu-
tions are thin in consistency and it allow the current to pass easily through the

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J. P. Pani et al.

Figure 12. Zeta potential analysis of none & nano filtered fresh and preserved particles colloidal silver showing zeta potential val-
ue, mobility, conductivity value of the current passing through the solution. Zeta potential value of the cell upper and lower sur-
face. Its Doppler shift, refractive index, viscosity of the colloidal. It also shows Avg. electric field and Avg. current rate.

field. Agglomeration of solution which causes inflammation and increase in size

of the silver nanoparticles has no relation to rate of conductivity of electric cur-
rent in matrix of the solution.
8) Spectroscopic evaluation of silver nanoparticles size from nano filtered col-
loidal silver
The primary size of the silver nanoparticles evaluated by spectroscopic analy-
sis from original nano filtered colloidal silver. The spectroscopy found, highest
absorbance (almost 0.35) at liberation of 400 nm wave length inside the colloidal
silver solution field due to highest excitement whereas lowest absorbance is
achieved (almost 0.07) when 600 nm wave length was liberated inside the origi-
nal colloidal field due to lowest excitement. So this analysis indicates the solution
must be thin in consistency and low in density and it must contains smaller size
silver nanoparticles (2 to 5.2 nm range) because at lowest wave length liberation
from spectroscope colloidal silver showed highest absorbance and highest exci-
tation (Figure 13).
9) Result of Image-J analysis of none filtered colloidal silver (Figure 14)

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 J. P. Pani et al.

Figure 13. Spectroscopic analysis of nano filtered smaller particles colloidal silver show-
ing rate of emission wave length and its respective absorbance value inside colloidal silver.

Figure 14. (A-B): Image-J analysis of none filtered bigger particles colloidal silver photo
showing mean, standard deviation, minimum & maximum size of particles and area of
photograph (Image 5 of bigger particles).

8 transmission electron microscopy photograph of bigger particles none fil-

tered colloidal silver obtained and edited by Image-J software. They resulted in
the following form:
Image 1 Bigger Particle mean size-: 220.040 nm
Image 2 Bigger Particle mean size-: 208.010 nm
Image 3 Bigger Particle mean size-: 232.076 nm
Image 4 Bigger Particle mean size-: 200.111 nm
Image 5 Bigger Particle mean size-: 238.069 nm
Image 6 Bigger Particle mean size-: 222.010 nm
Image 7 Bigger Particle mean size-: 196.077 nm

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J. P. Pani et al.

Image 8 Bigger Particle mean size-: 201.055 nm

Over all mean of the bigger particle size = 214.618 nm
The maximum particle size of bigger particles calculated 255 nm and mini-
mum size evaluated by Image-J software 2.3 to 0 nm. Respective mean, standard
deviation, minimum, maximum and total area that was scanned and evaluated
by Image-J of image 1to 8 of TEM plot of bigger particle colloidal silver which
calculated, the overall mean of bigger particles is 214.618 nm with over all stan-
dard deviation is 124.628 (214.618 ± 124.628) (Graph 1), showing here only
Image-J analysis of Image 5 of bigger particle.
10) Image-J analysis of smaller particles nano filtered colloidal silver (Figure
15)
6 transmission electron microscopy photograph of smaller particles nano fil-
tered colloidal silver obtained and edited by Image-J software. They resulted in
the following below form:

Graph 1. Mean and standard deviation overall size of all bigger particles.

Figure 15. (A-B): Image-J analysis of nano filtered smaller particles colloidal silver photo
showing mean, standard deviation, mode, minimum & maximum size of particles and
area of photograph (image 4 of smaller particle).

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 J. P. Pani et al.

Image 1 smaller Particle mean size-: 8.503 nm

Image 2 smaller Particle mean size-: 10.909 nm
Image 3 smaller Particle mean size-: 8.604 nm
Image 4 smaller Particle mean size-: 71.56 nm
Image 5 smaller Particle mean size-: 10.808 nm
Image 6 smaller Particle mean size-: 9.706 nm
Over all mean of the smaller particle size = 20.015 nm
The maximum particle size of smaller particles calculated 255 nm and mini-
mum size evaluated by Image-J soft ware 0 nm. Respective mean, standard devi-
ation, minimum, maximum and total area that was scanned and evaluated by
Image-J of image 1to 6 of TEM plot of smaller particle colloidal silver resulted,
the overall mean of smaller particles is 20.015 nm with over all standard devia-
tion is 12.628 (20.015 ± 12.628) (Graph 2) showing here only Image-J analysis of
Image 4 of smaller particle.
11) Summary and histogram comparison of bigger and smaller silver nano-
particles
The summary of mean of image 5 of bigger particle calculated 238.069 nm.
Over all mean out of 8 bigger particles TEM image under gone Image-J analysis
and calculated 214.618 nm and Mean ± S.D. of 8 bigger particles images calcu-
lated 214.618 ± 124.628 where as the histogram of image 5 of bigger particle
generated Mean ± S.D. value 233.132 ± 66.921. This more or less matches when
going for comparison between the two (Figure 16).
At the same time, the summary of mean of image 4 of smaller particle calcu-
lated 71.56 nm. Over all mean out of 6 smaller particles TEM image under gone
Image-J analysis calculated 20.015 nm and Mean ± S.D. of 6 smaller particles
images calculated 20.015 ± 12.628 where as the histogram of image 4 of smaller
particle generated Mean ± S.D. value 143.298 ± 126.518. Here the analysis mere-
ly differentiated (Figure 16).
The overall comparison between bigger and smaller silver nanoparticles from
the angle of summary and histogram of Image-J says present study used silver
nanoparticles size ranged between 20.015 to 71.1 nm in maximum percentage

Graph 2. Mean and standard deviation overall size of all smaller particles.

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J. P. Pani et al.

Figure 16. Image-J analyses of none & nano filtered bigger & smaller particles colloidal
silver histogram and summary and comparison between the two.

from total whereas rest of all measured 71.56 to 200.1 nm size ranged.
12) Comparison of line graph, fractal box count and surface plot between big-
ger and smaller silver particle
The upper mention Figure 17 generated by Image-J which compares between
bigger and smaller particles of study used none and nano filtered colloidal silver
says line graph of image 5 of bigger particles shows bigger size column graphs
where as image 4 of smaller particles shows significant smaller column graphs
and less amount in comparison to the bigger particles. The fractal box count of
image 5 of bigger particles calculated D = 1.9823 whereas the fractal box count of
image 4 of smaller particles calculated D = 1.8504 which is less value in compar-
ison to image 5. The surface plot generated by image 5 also indicates the sizes of
bigger particles are significantly bigger in comparison to image 4 of smaller par-
ticles. The status of surface plot of image 4 of smaller particles indicates the
same.
13) Particle characterization by transmission electron microscopy (Figure 18)
8 plots of bigger particles TEM images resulted average size of bigger silver
nanoparticles ranged from 1300 nm to 1900.8 nm whereas 6 plots of smaller par-
ticles TEM images resulted average size of smaller silver nanoparticles ranged
from 2.75 to 71 nm. The overall size of all silver nanoparticles ranged from 71 to
200 nm (Figure 19).
There is close approximation observed in dynamic light scattering, zeta po-
tential, Image-j estimation and transmission electron microscopy process after
making a comparison of a with a1 and b with b1 type of AgNps colloidal solu-
tion after characterization and segregated size estimation.

3. Discussion
The characterization of 4 types of silver nano particles colloidal solution is a vital
step done in the beginning phase of this present study which reveals primary size
of the study used silver nano particles by dynamic light Scattering, zeta potential,
image-j estimation and transmission electron microscopy (TEM) and for their
agglomeration in cell medium by photon cross correlation spectroscopy (PCCS)

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 J. P. Pani et al.

Figure 17. (A-F): Image-J analyses of none & nano filtered AgNps colloidal solution (A-C) bigger (5); (D-F) smaller (4) particles
colloidal silver showing comparison of line graph, fractal box count and surface plot between the two.

and ultraviolet-visible (UV-vis) spectroscopy, which is ultimately resulted 20 to

71.1 nm mean ranges though close approximation in analysis found in between
all procedures . Representative of TEM pictures of the bigger and smaller AgNps
were showed. The TEM images of the poly vinyl pyrollidone (PVP) coated
AgNps dispersed and suspended in 1.5 molar sodium chloride (NaCl2) con-
firmed the primary silver nanoparticle size stated by the manufacturers while
buying it from market in raw powdered form called silver nitrate (AgNO3) and
also in all types of colloidal AgNps solution. Among these, silver precipitation in
aqueous colloidal media which is most commonly used in experiment because it
is easy, cheap, and versatile. Silver nanoparticle nature is guided by precipitation
technique which offers best possibilities by changing the experimental parame-
ters, such as reactant concentrations, temperature, pH, reducing agents, and sta-
bilizers [15] [29] [30]. The uncoated bigger silver nano particles found hetero-
geneous distribution within the average ranges of 71.1 to 300.1 nm, most of the

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J. P. Pani et al.

(A) (B)

(C) (D)

(E)

Figure 18. (A-E): Images of TEM of none & nano filtered colloidal silver (A-C) bigger
particles & (D-E) smaller particles.

smaller being around 20 to 71.1 nm ranges. Changes in hydrodynamic size dis-

tribution and state of agglomeration of all size types of AgNps monitored di-
rectly after dispersion and suspended after 0 h and after 8 h as well as 24 h in
NaCl2 medium are presented as by volume, density distribution and size together

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 J. P. Pani et al.

Figure 19. (A-B): TEM mode comparison of Mean ± S.D. of

overall sizes of bigger and smaller nanoparticles of none and na-
no filtered colloidal silver.

with the changes in scattered light intensities for the corresponding time points.
The silver nanoparticle size distribution by volume (D10, D50 and D90) is in-
cluded as Table 5.
1) 2 phase analysis of bigger silver nanoparticles (Table 6)
2) 2 phase analysis of smaller silver nanoparticles
When interpreting the data it was noted that the intensity of the scattered
light increases non-linearly with increasing nanoparticle size due to agglomera-
tion, and that two processes hence occur simultaneously, agglomeration (in-
creased intensity) and sedimentation (reduced intensity). Furthermore, there is a
risk of overestimation of the proportion of small silver particles in the size dis-
tributions curves due to rotational diffusion that can take place for non-spherical
particles and give rise to a peak at small particle sizes (<3 nm). Furthermore, it is
also possible to synthesize and characterize composite bigger and smaller silver
nanoparticle and silver ions (Ag+) [31] [32] [33]. Because of the high electropo-
sitive character of silver (+0.799/−16.18 v/cm2 V36), agents which reduces in
various reaction [15] which also includes free radicals and peroxidases, [34] so-
dium borohydrate (NaBH4), [35] can be used as stabilizer for colloidal solution
for prolong duration preservation. However, most protocols in synthesis and
characterization is famous for the preparations of spherical silver nanometals
were designed to produce small particles (radius 2 to100 nm ranges). Coating of
colloidal particles with a poly vinyl pyrollidone layer has many advantages, be-
cause it allows for surface modification and deaggregation, decreases the poly

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J. P. Pani et al.

Table 5. S1 (None filtered).

Poly
Diameter D (10%) D (50%) D (90%)
Number Repeat Ph Avg dispersity
(nm) nm nm nm
index

1 1 NA 200 - 1356.6 0.483 200 - 373.2 200 - 1011.1 200 - 3147.1

2 2 NA 200 - 1400.9 0.517 200 - 324.6 200 - 896.6 200 - 3099.0

200 - 1378.8 0.500 200 - 348.9 200 - 953.85 200 - 3173.5

Table 6. S2 (None filtered).

Poly
Diameter D (10%) D (50%) D (90%)
Number Repeat Ph Avg dispersity
(nm) nm nm nm
index
1 1 NA 20 - 520.6 0.679 2.3 20 - 717.5 20 - 3605.5

2 2 NA 20 - 20 0.243 5.2 20.5 - 30.5 20 - 198.1

20 - 300.15 0.461 3.75 20 - 374 20 - 1901.8

dispersity index of the particles and makes the colloidal solution thinner. It
boosts colloidal solution stability by depleting Van Der Wall force on wall. Poly
vinyl pyrollidone acts as successful coating agent for many metal nanoparticle
especially silver nanoparticle which helps in deaggrgation and agglomeration ul-
timately causes for increase size of the particle due to inflammation [36]-[42].
poly vinyl pyrollidone directly over surfactant stabilized silver particles was re-
ported recently in an experimental work by one of the eminent scientist [43]. In
this article, we describe the characterization of silver particles of a wide range of
bigger sizes and narrow size distribution by reducing silver nitrate with sodium
borohydried in an aqueous medium with poly vinyl pyrollidone mixed as deag-
gregator agent. The various sizes silver nanoparticle morphology was studied by
dynamic light scattering, zeta potential, image-j estimation and transmission
electron microscopy. Electrical conductivity response on multiple silver nano-
particle level and colloidal solution matrix was studied by means of zeta poten-
tial and simulated to scattering theory. The results showed that particle agglo-
meration as well as sedimentation, indicated by a reduction of the scattered light,
and adding a drop of poly vinyl pyrollidone into it as a deaggregator agent was
evident for all AgNps with time but there was a clear difference between the ci-
trate coating and PVP coatings though citrate coating is not the part of this
study. By use of the procedure described above, we obtained spherical but irre-
gular and spiny silver nanoparticle with a radius in the ranges from 71.1 to 300.1
nm as bigger size range and 20 to 71.1 nm as smaller size range. Particle poly
dispersity index was more difficult to control and was varied according to size
range. (Bigger PI = 0.722 and Smaller PI = 0.483) However, poly dispersity
found as low as 14.6% was achieved in several samples (Table 7).
The particles are spherical aggregates with a rough surface where the rough-
ness is on the order of a few nanometers. The particle upper and lower surface

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 J. P. Pani et al.

Table 7. Radius (R) and Polydispersity (δ) of silver particles obtained at different experi-
mental conditions as determined by TEM.

Sample [AgNO3] δ (mol/L) R (nm) δ (%)

Nonefiltered(a)F 0.538 1340.3/Avg.-: 6151.2 13.5

Nonefiltered(a1)P 0.722 1944.9/Avg.-: 6750.8 14.6

Nanofiltered(b)F 0.483 1356.6/Avg.-: 1500.1 10.8

Nanofiltered(b1)P 0.461 71/Avg.-: 1372.9 6.7

charge is determined by the polyvinyl pyrollidone layer. The surface charge plays
an important role in the particle self-organization after sedimentation in
aqueous suspensions. In poly vinyl pyrollidone coated silver nanoparticle the
presence of an inert dielectric layer around metal nanoparticle allows tailoring
their properties and interaction potential. For instance, a thin (∼16 nm) poly
vinyl pyrollidone layer is sufficient to screen the Van Der Waals attraction hold
responsible for increase of the silver nanoparticle stability. The most important
factor observed while conducting coating process is the stability of the initial
colloidal suspension at the reaction conditions (high pH and temperature). The
71.1 nm PVP coated AgNps initially showed a trimodal size distribution, with
peaks centered approximately at 71, 300.15 and 1901.8 nm. The two larger mod-
es refer to agglomerates, also seen from the TEM and DLS investigation, and are
expected due to the high ionic strength in the NaCl2 medium. The peak at 1901.8
nm refers both to particles of this size and to the rotational diffusion effect, as
described above. After 8 h, the two larger modes were shifted towards larger siz-
es indicating further agglomeration. However, after 24 h the size distribution
was similar as the initial observations with smaller size particle/agglomerate dis-
tributions. An evident reduction in scattered light intensity with time indicated
fewer particles in solution and thus, the discrepancy between 8 h and 24 h is
predominantly explained by sedimentation of the largest particles from which
follows a reduced intensity and reduced size distribution of particles still in solu-
tion. The 20 nm PVP coated NaBH4 ( Sodium Borohydried) stabilized AgNps
agglomerated directly after dispersion (0 h), were less stable with time in NaCl2
medium, and sedimented to a larger extent when compared with the 2.31 nm
PVP coated AgNps. The latter after nano filtration procedure the silver nano
particles showed mostly small particles (<3 nm) even after 24 h, and only a low
amount of agglomerates of larger sizes. Also the scattered light intensity was rel-
atively stable with time, indicating a higher stability due to NaBH4 (Sodium Bo-
rohydried). The observed differences in agglomeration and sedimentation beha-
vior of the none filtered and nano filtered PVP coated 20 nm particles were fur-
ther confirmed by UV-Vis measurements, showing a reduced absorbance with
time for the no filtered PVP coated particles due to sedimentation. The rate of
sedimentation was higher for the no filtered PVP coated particles as compared
to the nano filtered PVP coated AgNps, in agreement with the PCCS findings.
Also there was a slight broadening of the peaks with time, explained by the for-

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J. P. Pani et al.

mation of larger agglomerates. The freshly prepared 20 nm PVP coated AgNps

had a trimodal size distribution, with the peaks broadening out with time up to 8
h. The proportion of the peak of the largest agglomerates (1908.1 - 3173.5 nm)
was reduced and vanished after 24 h. Similar to findings for the 20 nm PVP
coated particles, the intensity of the scattered light was reduced at the same time
as the size distribution became bimodal and more narrow again due to further
agglomeration of the smallest particles and sedimentation of the larger agglo-
merates. The 20 nm PVP coated AgNps initially showed a trimodal distribution
and an increased agglomeration with time. After 24 h the larger agglomerates
sedimented and the smaller particles became more agglomerated. The uncoated
AgNps also agglomerated with time but, after 24 h there were no large agglome-
rates (>200 nm) in solution. This might be explained by a higher rate of agglo-
meration for the uncoated particles, resulting in large agglomerates that due to
sedimentation were not detected. The observed presence of silver particles sized
less than <71 nm has been verified for the same batch of AgNps elsewhere. The
zeta potential analysis was done by the same machine. The DelsaTM Nano C,
Beckman Coulter machine purchased from United Kingdom. None filtered and
nano filtered AgNps colloidal solutions were used in this analysis. Both type par-
ticles showed EOS plot −17.52 mV (zeta potential) and mobility distribution
−1.369c−004 (cm2/Vs). The machine measured conductivity of 0.4356 (mS/cm).
The calculated Doppler shift was found 10.90 Hz for larger and smaller sized
particle with a base frequency of 124.8 Hz. Dielectric constant showed 78.3 for
both fresh made solution. Base on the results of zeta sizer instrument, hydrody-
namic size distribution of AgNps in the suspended powder stock solution,
ranged from about 20 to over 200 nm; with a mean average size (ZAve) of 71.2
µm (198.1 nm). In the colloidal solution, hydrodynamic size distribution of
AgNps ranged from 71.2 to 1908.1 nm and the zeta average (mean particles size)
was 20.0 - 200 nm; also in this case, totally tree classes of particles were distin-
guishable: 200 - 348.9 nm (10), 200 - 953.85 nm (50%), and 200 - 3173.5 nm
(90%). Also according to zeta sizer instrument % information, zeta potential of
colloidal and suspended powder AgNps had an average of EOS plot −17.52 mV
and +41.03 ± 0.13 mV respectively. A zeta potential range from ±40 to ±60 mV
is a sign of good stability for colloids. (ASTM, 1985) The colloidal AgNps ob-
served by TEM were spherical in shape, with a maximum diameter of 200nm:
90% of the particles had diameters between 200 and 3173.5 nm, just 10% of the
particles had diameters more than 100 nm, and the CMD (count median diame-
ter) for the particles was 71 nm. Also the geometric mean diameter (GMD) and
geometric standard deviation (GSD) of colloidal silver nano particles were 33.9
nm and 7.1 nm respectively. The AgNps colloidal solution dried to powdered
form stalk on tungsten grid observed by TEM were spherical in shape, with a
maximum diameter of 20 - 25 nm: 85.97% of the particles had diameters be-
tween 11 and 24 nm, just 10% of the particles had diameters more than 100nm,
and the CMD for the particles was 374 nm. Also the GMD and GSD nano fil-
tered silver nanoparticles were 2.7 nm and 1.1 In the case of suspended powder

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 J. P. Pani et al.

AgNps, TEM images showed that in an aqueous environment the silver nano
particles clumping and form large aggregates, many of which are larger than
200nm. This result is consistent with the results of hydrodynamic size distribu-
tion which was obtained from the dynamic light scattering method (zeta sizer).
Electron dispersive X-Ray study (EDX) analyses were confirmed that only ele-
mental silver was presented in both colloidal and dry powdered AgNps. Ac-
cording to the ICP-OES results, the concentrations of Ag ions in the acid di-
gested stocks of colloidal and suspended powder AgNps were 1908.1 and 447.2
mg·l−1, respectively. Also results of the XRD pattern confirms the crystallinity of
powdered type AgNps and presence of elemental crystalline silver, also other
phases, except metallic silver, were not present in the sample. In addition the
XRF results showed that purity of powdered type AgNps was 97.86%.The spec-
troscopic examination and size evaluation of the silver nanoparticles resulted al-
so 20 to 200 nm range with highest absorbance (0.35) at 400 nm wave length and
lowest absorbance (0.075) at 600 nm wavelength. The image-j estimation of the
TEM images also yielded same result as DLS, zeta potential, TEM and spectros-
copy that is the present study used particle size ranged between 20 to 200 nm
sizes.

4. Conclusion
The characterization of silver nano particle colloidal solution indicates the mean
of silver nano particles size varies between 20 - 71.1 nm ranges (smaller) and
71.1 to 300.1 nm ranges (bigger) which more or less matches with Image-j, DLS
and TEM analysis. The smallest particles mean size measured 2.3 to 20 nm. In
conclusion, AgNps of 20 nm mean size in hydrodynamic diameter and with a
zeta potential of −17.52 mV expresses significant cytotoxicity. In conclusion, we
developed a unique magnetic stirring and cooling method to synthesize large
and small silver colloidal particles with multi radius larger than 100 nm and
smaller than 20 nm via controlled aggregation by poly vinyl pyrollidone coated
silver nanoparticles after. After agglomeration, smaller particles flow in medium
of matrix of colloidal solution and bigger particles settle at bottom of the solu-
tion as sediment after sedimentation process. The remaining smaller silver na-
noparticles are spherical but irregular and spiny aggregates with a low polydis-
persity (<14.6%) and surface roughness on the order of 2.3 nm, as determined by
transmission electron microscopy. The particles were coated with a protective
poly vinyl pyrollidone layer. The overall zeta potential analysis of all types of ex-
periment used AgNps colloidal solution found −17.52 mV which is nothing but
the capacity of electrical conductance inside matrix of solution. More or less
there is close approximation found between dynamic light scattering, zeta po-
tential, Image-j estimation and transmission electron microscopy analysis for all
fresh and preserved types of bigger and smaller nanosilver colloidal solution re-
garding characterization and segregated size estimation. The overall estimation
shows there is very kin difference between a with a1 and b with b1 type of

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J. P. Pani et al.

AgNps colloidal solution which is coated with protective poly vinyl pyrollidone
and stabilizer NaBH4.

Acknowledgements
We thank Dr Royana Singh (Proctor Banaras Hindu University) for inspiring
this research and Dr Sanjay Singh (IIT, BHU Pharmaceutical division) for cha-
racterization process which includes DLS and zeta potential analysis. We also
thank Dr Chatterji and Mr. L.K. Chauhan for transmission electron microscopy
and himself did image-j estimation and analysis. This work is part of the re-
search program of Department of Anatomy Institute of Medical Sciences, Bana-
ras Hindu University, which is financially supported by the University Grant
Commission, India.

Conflict of Interest
Panels of author and co authors declare no conflict of interest.

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Abbreviation
SERS: Surface Enhanced Raman Spectroscopy, NaBH4: Sodium Borohydried,
PVP: Poly Vinyl Pyrollidone, PDI: Poly Dispersity Index, DLS: Dynamic Light
Scattering, ZP: Zeta Potential, FTIR: Fourier Transformation Infra Red, UVVis:
Ultra Violate Visa Vis, TEM: Transmission Electron Microscopy, mV: milli Volt,
zp: zita potential, AgNO3: Silver nitrate, NaBH4: Sodium Borohydride, NaCl2:
Sodium chloride, SIS: Soft Imaging System viewer, FWHM: Full Width at Half
Maximum, XRD: XRay Diffraction, ACF: Auto Correlation Function, cps: cycle
per second, cp: cycle per, MG: Milli Gram, CCD: Computerized Captured De-
vice, FEI: Fourier Electron Image, ANOVA: Analysis of Variance, MANOVA:
Multi Analysis of Variance, AgNps: Silver nanoparticles, nm: nanometer, NID:
Normalized Intensity Distribution, PCCS: Photon Cross Correlation Spectros-
copy, GSD: Geometric Standard Deviation, CMD: Count Median Diameter,
GMD: Geometric Mean Diameter, I4SP: Image 4 Smaller Particle, I5BP: Image 5
Bigger Particle, zs: zita sizer, Ag+: Silver ion, R: Radius, δ: Poly Dispersity Index,
EDX: Electron dispersive XRay study, EOS: Equation of State, ICP OES: Induc-
tively Coupled PlasmaOptical Emission Spectroscopy, XRF: X-Ray Fluorescence.

DOI: 10.4236/wjnse.2018.81001 31 World Journal of Nano Science and Engineering