IMMOBILIZING GOLD NANOPARTICLES IN MESOPOROUS SILICA COVERED

REDUCED GRAPHENE OXIDE: A HYBRID MATERIAL FOR CANCER CELL

DETECTION THROUGH HYDROGEN PEROXIDE SENSING
Raia Quitoriano || raiaquitoriano@yahoo.com
Department of Physical Sciences and Mathematics
University of the Philippines Manila
ABSTRACT
A novel 2-D hybrid material with a potential for nonenzymatic sensing of hydrogen peroxide (H 2O2) was developed by the immobilization of gold nanoparticles (AuNPs) on the surface of reduced graphene oxide (RGO) covered with periodic mesoporous silica (PMS). The synergistic effects of AuNPs, RGO,
and PMS were utilized and the resulting hybrid (RGO-PMS@AuNPs) was characterized through various instrumental techniques and electrochemical methods. The sensor exhibited wide linear detection range (0.5 M to 50 mM), low detection limit (60 nM), good reproducibility, selectivity, sensitivity, and stability for the determination of H2O2 concentrations in aqueous solutions. The sensor was also tested for the applicability in the analysis of H 2O2 in human urine
and in the distinction of cancer cells from normal cells, both of which were successful. Thus the sensor has an application potential for diagnosis of cancer.
The findings of this study open a new area of research regarding the actual applicability of the sensor for diagnosis of diseases associated with high levels
of H2O2 using urine and real cell samples from humans.

SIGNIFICANCE OF THE STUDY

Hydrogen peroxide has been labeled as a chemical threat agent primarily because its excess may lead to tremendous effects in both biotic
and abiotic systems. In biological systems, it is a by-product of many enzymatic reactions, particularly those of the oxidases in the mitochondria.
H2O2 in living cells results to the generation of reactive oxygen species
(ROS), excess of which may lead to oxidative stress which is related to
a variety of biological damages including aging, cancer, cardiovascular
diseases, and neuronal death. Moreover, tumor cells can generate
H2O2, which recruits leukocytes to the injured cells. Thus, the concentration of H2O2 in human cells may be an appropriate indicator of the biological activities of the cells.
Consequently, many studies have explored methods for the quantitative measurement of hydrogen peroxide concentration, including fluorometry, spectroscopy, cell imaging, and electrochemical methods, but
none of them were sensitive enough to detect H2O2 in nanomolar concentrations as produced by cellular activities.

OBJECTIVES
The general objective of this study is to develop a novel hybrid material
with a promising potential in biosensing and diagnostics of cancer. To be
able to achieve this, the study is aimed to develop a synthetic approach in
the preparation of a two-dimensional nanohybrid system consisting of
AuNPs over PMS coated RGO, to quantify its ability in electrochemical
sensing of H2O2 and glucose in human urine, and to utilize the developed
sensor in distinguishing cancer cells from normal cells.

RESULTS

Figure 1. RGO-PMS@AuNPs/GC (b) CV curves with H2-O2 and (d)

calibration plot
The cathodic peak of RGO-PMS@AuNPs/GC at 0.75 V and the
anodic peak at 0.65 V exhibited linear increase as H2O2 was added.
The calibration plot from chronoamperometric methods showed that
the linear detection range of the sensor is 0.5 M to 50 mM and the
limit of detection is 60 nM.

METHODOLOGY
Synthesis of
RGO-PMS

Cancer Cell
Detection
Electrochemical
Measure-

Synthesis of
RGO-PMS@
AuNPs

Characterizations

Figure 2. RGO-PMS@AuNPs/GC (b) amperometric response

curve with and without HeLa, (c) amperometric response curve
comparison of HEK 293, HeLa, and HepG2 cells
Amperometric response curves of HeLa cells at -0.75 V showed
increase in current signal at the addition of PMA, while after the addition of catalase, the signal decreased to its background level. The
H2O2 concentration released HeLa cells was found to be ~0.8 M. In
HEK 293 cells, the addition of PMA and catalase did not have significant effect in current signal while HepG2 cells exhibited response to
PMA and catalase addition similar to that of HeLa cells.

DISCUSSION
Morphological characterizations showed that the AuNPs had a good structure and strong interaction with the RGO-PMS hybrid. Using electrochemical
techniques, the sensor was found to have a very wide linear detection range and low detection limit compared to previously developed H2O2 sensors.
Analysis of urine samples showed that the calculated H2O2 concentrations using the sensor agree with a previous report. The sensor exhibited high reproducibility, stability, and selectivity which are all desirable properties of a nondisposable sensor. When the sensor was tested to analyze cancer cells in vitro, it
was found to be effective in distinguishing cancer cells from normal cells.

CONCLUSION AND RECOMMENDATION

A novel two-dimensional RGO-PMS@AuNP hybrid was developed and was characterized through electrochemical techniques. The hybrid was found to
be an excellent sensor of H2O2 in aqueous medium with wide linear range, low LOD, good reproducibility, stability, and selectivity. The sensor was also able
to analyze H2O2 in human urine samples and to distinguish cancer cells from normal cells through the concentration H 2O2 production. Therefore, the sensor
indeed has a promising potential for biosensing and diagnostics of diseases such as cancer.
While this study has done an intensive investigation of the sensing capabilities of RGO-PMS@AuNPs, study on the actual application of the hybrid in the
diagnosis of cancer or other diseases associated with high levels of H2O2 has not been done. Hence the use of real human samples such as urine and cell
samples for future study is recommended. It is also recommended that the diagnosis of other diseases associated with high levels of ROS and H2O2 such
as neurodegenerative disorders and cardiovascular diseases be studies since the sensor is capable of sensing H2O2 with high sensitivity.
REFERENCE
Maji, S. K., Sreejith, S., Mandal, A. K., Ma, X., & Zhao, Y. (2014). Immobilizing Gold Nanoparticles in Mesoporous Silica Covered Reduced Graphene Oxide:
A Hybrid Material for Cancer Cell Detection through Hydrogen Peroxide Sensing. ACS Applied Materials and Interfaces .