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Detection Limits of DLS and UV-Vis Spectroscopy in

Characterization of Polydisperse Nanoparticles Colloids.
Received 28th October 2019,
Synthesis and Characterization of Gold Nanoparticles
Riley Fink, Victoria Sanchez, Gabe Rodkey

Abstract 
Accepted 15th November 2019 In this experiment, gold nanoparticles (AuNPs) were synthesized and then
characterized using both UV-Vis spectroscopy and DLS characterization. The nanoparticle’s
behavior was observed when mixing chloroauric acid(HAuCl4) with glucose (C6H12O6) and salt
(NaCl) in order to determine the average size of the particles. The results illustrated the
formation of gold nanoparticles inside the cuvettes in which the experiment was conducted in.
Moreover, the size distribution of prepared gold nanoparticles varied with the different
concentrations of glucose and salt.

Introduction
Nanotechnology is one of the most important new targets of
science and technology. Nanotechnology is the formation of
1-100 nm sized particles and has become important in
modern technology systems due to their high surface area,
different optic and chemical properties. Nanoparticles have
been produced for various applications like polymeric PS,
PMMA, inorganic PbS, Ag2S, CdSe, TiO2, and Au, Ag, etc. Gold
nanoparticles have abundant use in the field of
biotechnology and biomedicine because they have large
surface bio conjugation with molecular probes and, as
previously mentioned, they also have many optical
properties. Within the experiment, the gold nanoparticles
will exhibit a particle size ranging from1 nm to 8 µm. To
understand why such a large range of NP sizes was observed,
it is import to understand the process of aggregation of NPs.
The initial stability of NPs can be attributed to the citrate
ion coating of the NPs that are present post-synthesis. When
ions are added to a solution of ion-coated NPs, however, the
negative charge of citrate is screened and the NPs are
allowed to then aggregate.3 This experiment with show the
characterization of NPs of various sizes and solute additions
using both DLS and UV-Vis spectroscopy.  synthesized gold nanoparticles. The peak wavelengths and
Materials and Methods
In this work, gold nanoparticle synthesis was
achieved. The materials used were gold salt concentration
(HAuCl4) mixing with both sodium chloride (NaCl) and glucose
(C6H12O6) concentrations to determine nanoparticle size
using UV/Vis spectra and DLS technology.  1.
Prepared was a 1.0 mM solution of hydrogen
tetrachloroaurate by diluting 5 mL of 10.0 mM stock solution
of hydrogen tetrachloroaurate to 50 mL volume. Transferred
to a vial was 10 mL of 1.0 mM hydrogen tetrachloroaurate.
The solution in the vial was then put on a hot plate that was
turned up to 6, and a magnetic stir bar was added with a stir
level of 4. After the solution came to a rolling boil, 1 mL of
38.8 mM citrate stock solution was pipetted into the solution
and left to continue to boil for 10 minutes after the color
change. Observations were recorded during this time. Then,
the solution was removed from the hot plate and left to cool
to room temperature. 
In order to characterize the gold nanoparticles, a
DLS characterization method was used. Two cuvettes were
rinsed 4 times with deionized water. One cuvette was then
filled with 2 mL of filtered DI water, and the other cuvette
was filled with 50 µl of the synthesized gold nanoparticle
solution and then filled the rest of the way to 2 mL with
filtered DI water. The blank cuvette was then placed in the
DLS machine and the data were recorded on the computer.
The same was repeated for the cuvette with the gold
nanoparticles. 
We then attempted to characterize the gold
nanoparticles using UV-Vis spectroscopy. Logger Pro software
and a spectrophotometer were used to read the absorbances
of a blank control (3 mL of DI water added to a cuvette) and
of a cuvette with 2.95 mL of DI water and 50 µl of
absorbances were recorded for each. Calculated were the
half peak height and the peak full width at half the
absorption maximum (FWHM). 
With each stock solution of 1 M sodium chloride
(NaCl) and 1 M glucose (FIX THIS c6h12o6), three cuvettes
were prepared: 
Blank Control: 3.0 mL DI water 

2. Low Solute Sample: 2.91 mL of DI water, 50 µl of
synthesized AgNPs, and 36 µl of one solute solution
(added last) 


3. High Solute Sample: 2.59 mL of DI water, 50 µl of

a.Santa Clara University 500 El Camino Real, Santa Clara, CA 95053
efink@scu.edu
b. Santa Clara University 500 El Camino Real, Santa Clara, CA 95053
vsanchezmonroy@scu.edu
c.Santa Clara University 500 El Camino Real, Santa Clara, CA 95053
grodkey@scu.edu
synthesized AgNPs, and 360 µl of solute solution
(same solution as 2) (added last)

All cuvettes were placed one at a time into the
spectrophotometer and the data were observed and
recorded in the computer. Graphs of the wavelength versus
absorbance were printed out for the blank cuvettes, glucose
solutions, and salt solutions. 

1 reference: Gold Nanoparticle Properties

2 reference: Salt Stability of Nanoparticles

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Detection Limits of DLS and UV-Vis Spectroscopy in Characterization of Polydisperse Nanoparticles Colloids

other particles getting in the solution and causing the light

to be absorbed or refracted differently than it should. The
two methods are both fairly quick to use and relatively
inexpensive. The DLS has the issue that larger particles can
completely block the presence of smaller particles, and the
smaller particles will neglect to show up in the histogram of
particle size.1 The downside of UV-Vis is that one cannot see
the separate peaks for the particles of different sizes within
each sample. All of the solutes affect the color of the overall
solution because the solution was diluted with a clear
substance and therefore the dark red color was diluted to a
lighter pink. However, the high salt concentration sample
was a lighter pink than any of the other solute samples.
Adding the solutes increased λmax  by 2.9-4.9 nm in the cases
of all solutions except for the high salt concentration, which
had its λmax 14.7 nm higher than the original gold
nanoparticle solution. FWHM decreased from the original 100
nm in both of glucose solutions, but increased in the low salt
solution. The high salt solution peak was so flat that the
FWHM was not applicable to the graph. 
When the nanoparticles are exposed to salt as they
were in the salt samples, agglomeration occurs because of
the ionic properties of salt. This causes the particles to act
more like large particles.2  This is why the estimated
nanoparticle size increases with the addition of salt. There
was no difference in the λmax for the high glucose and low
glucose solutions, and only a small difference between their
absorbances at λmax. This shows that glucose has less of an
effect on the size of the gold nanoparticles than salt does,
which makes sense, because glucose is not ionic. 
Our DLS results for the gold nanoparticle solution
indicated the majority of particles around the 13-15 nm
range. Our estimation based on the UV-Vis results was 5-10
nm, so the UV-Vis gave us a smaller result than the DLS. We
did not collect information from the DLS for any of the
solutions containing glucose or salt, so the results from the
DLS and UV-Vis for those solutions cannot be compared. The
DLS and UV-Vis might give different results because, as
previously mentioned, the DLS has the possibility of blocking
smaller particles from being detected.1 This would cause the
size of the particles to be slightly over estimated on the DLS,
which agrees with out data. 
The estimations for the size of the gold
nanoparticles were based on two things. The peak
wavelength (λmax) was used in conjunction with information
increase in λmax and the estimated particle size, as well as
the elongated peak on the UV-Vis graph of the high salt
solution. tables from Cytodiagnostics3 to obtain the
estimation of the diameters of nanoparticles based on the
assumption that each solution did not contain salt or
glucose. Because the addition of salt creates larger particles
do to agglomeration,2 we estimated that the size of the
Top to Bottom:
nanoparticles in both of the salt solutions would actually be
Graph 1: Blank solution (HAuCl4)
Graph 2: Salt Solution (top two curves, high concentration, lower two, low
larger than the estimation determined from treating the
concentration) data as having no salt. 
Graph 3: Glucose solution (top curve, high, second second, low, lower curve, The color change from light pink to an even lighter pink in
blank) the high salt concentration corresponds to the significant

Discussion

The DLS results from the water control indicate

that there is still a possibility of particles found in the
solution in addition to gold nanoparticles. Thus, we cannot
conclude that all data points on the DLS were necessarily all
from nanoparticles as opposed to other particles in the
solution. Both UV-Vis and DLS results could be skewed by

2 |1-3 This journal is © The Royal Society of Chemistry 2019

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Detection Limits of DLS and UV-Vis Spectroscopy in Characterization of Polydisperse Nanoparticles Colloids

3. Pamies, R.; Cifre, J. G. H.; Espín, V. F.; Collado-González,

M.; Baños, F. G. D.; Torre, J. G. D.L. Aggregation Behaviour
of Gold Nanoparticles in Saline Aqueous Media. Journal of
Nanoparticle Research 2014, 16 (4).

4. Tomaszewska, E.; Soliwoda, K.; Kadziola, K.; Tkacz-

Szczęsna, B.; Celichowski, G.; Cichomski, M.; Szmaja, W.;
Grobelny, J. Detection Limits of DLS and UV-Vis Spectroscopy
in Characterization of Polydisperse Nanoparticles Colloids.
Journal of Nanomaterials 2013, 2013, 1–10.

Conclusions
In the present work, we reported a simple and novel method
to synthesize AuNPs through light penetration of aqueous
HAuCl4 ions using UV/Vis and Dynamic Light Scattering (DLS)
technology. The developed method resulted in the formation
of AuNPs with an average size of 31.0 ± 0.5 nm.
Spectroscopic characterization of the synthesized AuNP’s by
DLS analysis further confirmed the distribution and stability
of the synthesized AuNPs. The overall results showed that
DLS synthesis method is comparable to the UV/Vis method
and can be considered as a more accurate method by
relating the intensity of light penetration to the size of
particles in order to determine the frequency of size in
which the particles were being formed.

Conflicts of interest
There are no conflicts to declare.

Acknowledgements

We would like to acknowledge Dr. Steven Suljak for

supervising our lab procedure and providing the necessary
equipment for us to carry out our experiment. We are also
acknowledging the institution of Santa Clara University for
the facilities in which we performed the lab.

References

1. Cytodiagnostics. Gold Nanoparticle Properties. http://

www.cytodiagnostics.com/store/pc/viewcontent.asp?
idpage=2 (accessed Nov 8, 2019).

2. nanoComposix. Salt Stability of Nanoparticles. https://

nanocomposix.com/pages/salt-stability-of-nanoparticles
(accessed Nov 8, 2019).

This journal is © The Royal Society of Chemistry 2019 1-3 | 3

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