Hydrodynamic sizing of citrate-reduced gold colloid using dynamic light

scattering analysis

Muhammad Anas Yuzairi Mohd Yusri, Mohd Hafiz Abu Hassan, Syaza Azhari
Faculty of Science & Technology, Universiti Sains Islam Malaysia
anas.yuzairi00@gmail.com

Abstract : Colloidal gold remarkably known with a set with intriguing characteristics that can
be used in various applications, including detecting biochemical compounds with proper
sensing moiety. The colloidal gold in this study was produced using varying the temperature
and concentration of each reagent used in the synthesis and primarily characterized using
dynamic light scattering analysis as a facile and economical method of analysis. Dynamic light
scattering analysis showed the produced colloidal gold with nanoparticles measured between
18 – 80 nm with decent polydispersity (PdI 0.331–0.695). It was shown varied synthesis
temperature affected minimally on the resulting hydrodynamic size of the nanoparticles,
meanwhile concentration of both reagents, gold (III) chloride and sodium citrate, impacted the
hydrodynamic size notably.
Keyword: Gold nanoparticles, dynamic light scattering, hydrodynamic size

Introduction
Gold nanoparticles or colloidal gold are defined by Kuppusamy et al (2014), Louis and
Pluchery (2012), and Shah et al (2014) as a suspension of dispersed nanoscaled gold particles
in a liquid matrix, commonly water. Due to its nanoscaled dimension, colloidal gold acquired
various advantageous properties such as high surface-to-volume ratio, low toxicity, and size
and geometry-dependent optical properties. Smaller size nanoparticles exhibit surface plasmon
resonance significantly as observed its peak maxima can be observed around 500 – 550 nm in
UV-vis spectrum. Furthermore, numerous researchers reported larger nanoparticles redshifted
the peak maxima and altered the colour of the colloid, from brilliant red to pale purple or
colourless.
In terms of measuring the dimensions of nanoparticles, typical and exemplary method
to acquire the measurements is electron microscopy techniques, namely scanning electron
microscopy and tunneling electron microscopy. Electron microscopy techniques visualized the
physical structure of the inquired nanoparticles via digital imaging as used by de Melo et al
(2019) and Gubicza et al (2013). Apart from commonly used electron microscopy techniques
to measure the dimension of colloidal gold, spectroscopy method especially dynamic light
scattering (DLS) recently popularized to be a facile and non-destructive analytical method for
most nanotechnology research applications. DLS analysis, as discussed by Ramos (2017), is
used to determine the size of particles along with its size distributions, ranged from 1 nm to 10
μm. Zheng et al (2016) highlighted DLS as an economical and simple operation analysis
method that renders out direct output data from raw measurement. In contrary to the
aforementioned advantages of DLS analysis, particle size measurement of colloidal gold using
DLS does not translate directly to other measurement methods such as UV-vis spectroscopy
method and electron microscopy method as polydispersed nanoparticles samples were not able
to reproduce the results that agreed with other methods. Furthermore, the poor reproducibility
of the results also caused by the variations in the aggregation states of the colloid.
This study was one of several efforts to produce a suitable colloidal gold for
biochemical compounds detections as well as determine the optimal synthesis condition to
produce the aforementioned colloidal gold. As mentioned in several reports, colloidal gold
suited for chemical sensor development were characterized with small particle size that within
range of DLS analysis measurement. Therefore, it is desirable to produce colloidal gold using
a reliable method with modified parameters and DLS as the nanoparticle measuring method.

Experimental procedure
Colloidal gold in this study was produced using citrate reduction method, established
by Turkevich et al (1951). Varied concentrations of sodium citrate (1, 2, 3 %) were used to
reduce the gold (III) chloride solution with different concentrations (1, 2, 3 mM) to form the
colloidal gold. In addition to the synthesis parameters, the temperature used (100, 80, 60˚C)
also varied to study the effects of varied temperature onto the hydrodynamic size of the
colloidal gold.
The reaction was done in a 50-ml beaker on a hotplate equipped with magnetic stirrer.
All laboratory glassware used were cleaned and rinsed thoroughly with aqua regia to remove
any contaminants and prevent early nucleation and aggregation of AuNPs. Gold (III) chloride
solution (20 ml) heated up to required temperature along with stirring on constant speed, and
then mixed with sodium citrate solution (2 ml). Upon introducing sodium citrate solution into
the reaction solution, the pale yellow colour of gold (III) solution turned bright red after several
minutes of stirring and heating at the specified temperature. The reaction stopped after several
seconds of constant colour observed as the heating and stirring ceased. The products then stored
in a chilling 4˚C storage. For DLS analysis, the colloidal gold samples were diluted beforehand
with 1:10 dilution factor in distilled water.
DLS measurements of the colloidal gold were done using Micromeritics Nanoplus 3
zetasizer, equipped with solid-state green laser (532 nm, 4 mW), complimented with Particulate
System Nanoplus HD software for raw data analysis. The particle size measurements were
performed with 60 seconds runtime, 90˚ scattering angle and operating temperature at 25˚C.

Results and findings

DLS measurements rendered out primary data in form of intensity distribution of the
size of the produced colloidal gold (Figure 1), showing most of the variable yielded unimodal
distribution except colloidal gold synthesized using 3% sodium citrate solution that yielded
bimodal distribution. The peak of each distribution indicated the average hydrodynamic size
of each produced colloidal gold along with its modified synthesis parameter. Meanwhile, high
deviations of the sizes produced wide distribution curve, which explained by the polydispersity
index of each synthesized colloidal gold.
Increasing size of colloidal gold was explained by the occurrence of Ostwald ripening,
a thermodynamic-driven formation of a large particle from a copious amount of smaller
particles. However, in this study, synthesis temperature affected the change of hydrodynamic
size of colloidal gold at a minuscule level. Tran et al (2016) further explained on delicate
changes on the particle size in colloidal gold resulting from lowering the synthesis temperature.
On the other hand, increasing gold (III) chloride solution significantly heightened the
hydrodynamic size of the produced colloidal gold as increasing the amount of gold (III)
complexes in reaction solution also heightened the growth and formation rates of individual
particles. In addition, the hydrodynamic size of colloidal gold produced using varied sodium
citrate concentration was increased at 2% sodium citrate level, yet decreased at 3% sodium
citrate and produced two average hydrodynamic sizes. Fluctuations of the sizes were caused
by high pH level and large amount of stabilizing and capping agents available, explained by
Tran et al (2016) and supported by Wang et al (2009). Sodium citrate is reported as an alkaline
solution and increasing its concentration heightened the pH level of reaction solution. At high
pH level, gold (III) complexes became more stable and impervious to citrate reduction,
prolonging the synthesis process. On contrary, high amount of citrate ions provided more
capping agent to prevent further aggregation and produce smaller sized nanoparticles, hence
the intensity peaked on the average size of colloidal gold.
The broadness of the distribution showed the polydispersity of the colloid, further
quantified and stated in the table along with the average sizes of the colloidal gold (Table 1).
Polydispersity of the colloidal gold ranged from 0.331 to 0.695, indicating the distribution of
hydrodynamic sizes of every individual nanoparticle was decently dispersed in the colloid.
However, highest polydispersity index (PdI > 0.5) was observed in the intensity distribution of
adjusted concentration of sodium citrate (Figure 1B). High polydispersity index was due to the
average hydrodynamic size was weighted on the larger sizes compared to smaller sizes,
reflected by the positive skewness of the size distribution of the colloidal gold. Despite the
broad distribution of nanoparticle sizes, all variables involved in the synthesis process affected
significantly on the polydispersity index as discussed by Cavalli et al (2009) and required
further analysis and investigation.

A B C

Figure 1: Intensity distributions of colloidal gold produced using different synthesis

parameters; A: synthesis temperature, B: sodium citrate concentration, C: gold (III) chloride
concentration

Table 1: Average size and its polydispersity index of colloidal gold synthesized using
different synthesis parameters

Gold (III)
Sodium citrate
Synthesis chloride Average size Polydispersity
concentration
temperature (˚C) concentration (nm) index
(%)
(mM)
100 1 1 33 0.346
80 1 1 33 0.331
 60 1 1 50 0.369
100 2 1 30 0.350
100 3 1 80 0.377
100 1 2 46 0.695
100 1 3 18, 30 0.615

Conclusion
Summarily, in this study, DLS analysis was used to measure the hydrodynamic size of
each colloidal gold produced using varied temperature and concentration of reactants. Intensity
distribution of the sizes of the nanoparticles from DLS analysis showed the increase in
hydrodynamic sizes due to increase in concentration of gold (III) chloride and sodium citrate,
and lower synthesis temperature. However, increase in sodium citrate also reduced the
hydrodynamic size as the amount of capping agent available to stabilize the nanoparticles was
heightened. Medium polydispersity index shown from DLS analysis indicated decently
distributed sizes of produced nanoparticles.

Acknowledgements
The authors gratefully appreciated the financial support for this research from USIM’s
internal research funding under research grant code PPPI/FST/0217/051000/12118.

References
Cavalli, R., Donalisio, M., Civra, A., Ferruti, P., Ranucci, E., Trotta, F., & Lembo, D. (2009).
Enhanced antiviral activity of Acyclovir loaded into β-cyclodextrin-poly(4-acryloylmorpholine)
conjugate nanoparticles. Journal of Controlled Release.
https://doi.org/10.1016/j.jconrel.2009.04.004
de Melo, F. M., Klimuk Uchiyama, M., Nakamura, M., Kawassaki, R. K., Pelleschi Taborda, C., &
Toma, H. E. (2019). Unravelling the nature of the spongy dark material in aged Turkevich gold
nanoparticles colloidal solutions by CytoViva® dark-field imaging and HRTEM analysis.
Micron, 121, 21–25. https://doi.org/10.1016/j.micron.2019.03.001
Gubicza, J., Lábár, J. L., Quynh, L. M., Nam, N. H., & Luong, N. H. (2013). Evolution of size and
shape of gold nanoparticles during long-time aging. Materials Chemistry and Physics, 138(2–3),
449–453. https://doi.org/10.1016/j.matchemphys.2013.01.012
Kuppusamy, P., Mashitah, M. Y., Maniam, G. P., & Govindan, N. (2014). Biosynthesized gold
nanoparticle developed as a tool for detection of HCG hormone in pregnant women urine
sample. Asian Pacific Journal of Tropical Disease, 4(3), 237. https://doi.org/10.1016/S2222-
1808(14)60538-7
Louis, C., & Pluchery, O. (2012). Gold Nanoparticles for Physics, Chemistry and Biology. Imperial
College Press.
Ramos, A. P. (2017). Dynamic Light Scattering Applied to Nanoparticle Characterization.
Nanocharacterization Techniques. Elsevier Inc. https://doi.org/10.1016/B978-0-323-49778-
7/00004-7
Shah, M., Badwaik, V. D., Agrisciences, C., Kherde, Y., & Waghwani, H. (2014). Gold
nanoparticles : various methods of synthesis and antibacterial applications. Frontiers in
Bioscience, (19), 1320–1344.
Tran, M., DePenning, R., Turner, M., & Padalkar, S. (2016). Effect of citrate ratio and temperature on
gold nanoparticle size and morphology. Materials Research Express, 3.
https://doi.org/10.1088/2053-1591/3/10/105027
Turkevich, J., Stevenson, P. C., & Hillier, J. (1951). A study of the nucleation and growth process in
the synthesis of colloidal gold, 55–75.
Wang, S., Qian, K., Bi, X., & Huang, W. (2009). Influence of speciation of aqueous HAuCl4 on the
synthesis, structure, and property of Au colloids. Journal of Physical Chemistry C, 113(16),
6505–6510. https://doi.org/10.1021/jp811296m
Zheng, T., Bott, S., & Huo, Q. (2016). Techniques for Accurate Sizing of Gold Nanoparticles Using
Dynamic Light Scattering with Particular Application to Chemical and Biological Sensing
Based on Aggregate Formation. ACS Applied Materials and Interfaces, 8(33), 21585–21594.
https://doi.org/10.1021/acsami.6b06903