GOLD NANOPARTICLE IN DOPAMINE

BIO-NANOSENSORS

SUBMITTED BY

A.Nakkiran

CECRI
ABSTRACT

Sensor technology is one of the most important key technologies of the future with
a constantly increasing number of applications, both in the industrial and in the private
sectors. More and more bio sensors are used for the control of processes in environment
monitoring, healthcare, and Military (against bioweapons). Consequently, the
development of fast and sensitive bio sensors is the subject of intense research, propelled
by strategies based on nanoscience and nanotechnology. This paper highlights the recent
developments and reflects the impact of nanoscience on electrochemical sensor
technology by elucidates size dependent sensitivity of gold electrode to sense dopamine,
an important neurotransmitter.
Introduction

According to the modern definition, biosensors are analytical devices comprising

a biological or biologically-derived sensing element either integrated within or intimately
associated with a physicochemical transducer1 (Fig1). The transducer is an important
component in a biosensor through which the measurement of the target analyte(s) is
achieved by selective transformation of a biomolecule-analyte interaction into a
quantifiable electrical or optical signal. A wide range of optical and electrochemical
instruments have been employed in conjunction with biological sensing.

Figure 1 Schematic diagram showing the main components of a biosensor

(a) A bio-component, (b) a transducer which converts the biochemical reaction into a
physical signal, (c) an amplifier which converts a physical signal into an electrical signal,
which is processed and displayed by a recorder or PC (d).

Electrochemical transducers detect an electrochemical signal that is generated by the

interaction between the analyte and the receptor. It could be a change of a redox potential
(Potentiometric), the conductivity of the solution (conductometric) or the production of
redox active molecules that generate a current (voltammetric, amperometric,
coulometric).
Dopamine and Electrochemistry

Dopamine is an important neurotransmitter because it is involved in physical and

cognitive functions. To understand the challenges associated with measuring dopamine it
is important to understand the environment in which dopaminergic neurons function.

Neuron Function

• Nerve cells, or neurons, are the basic building blocks of the nervous system. The
neuron is responsible for sending and receiving nerve impulses or messages. A neuron
that is excited will transmit its energy to neurons that are next to it (Fig.2).

Figure 2: Transformations message through neurons

•Neurons have a central cell body attached to slender, branching arms. There are
two types of “arms”: dendrites are like antennae and carry messages, or impulses, to the
cell body, while axons carry messages away from the cell body.

• Impulses travel from neuron to neuron from the axon of one cell to the dendrites
of another by crossing over a tiny gap between the two nerve cells called a synapse.
 • Incoming messages from the dendrites are passed to the end of the axon where
sacs containing neurotransmitters (dopamine) open into the synapse.

• The dopamine molecules cross the synapse and fit into special receptors on the
receiving cell.

• That cell is stimulated to pass the message on

• After the message is passed on, the receptors release the dopamine particles
back into the synapse where the excess dopamine is “taken up” or recycled within the
releasing neuron

So inadequately stimulation of dopamine (DA) can cause fatal disease such as

Parkinson’s and schizophrenia. Recent clinical studies have demonstrated that the
content of dopamine in biological fluids can be used to assess the amount of oxidation
stress in human metabolism and excessive oxidative stress has been linked to cancer,
diabetes mellitus, and hepatic disease.

Electrochemical Detection

Dopamine (DA), the most important among the class of neurotransmitters, plays
an important role in the function of the central nervous system. The development of
methods for dopamine quantification in nerves and biological fluids is the subject of
intense current investigation in neurochemical studies.Electrochemical method is an ideal
choice for the quantitative determination of dopamine, because

•Dopamine is easily oxidizable

•R. MARK WIGHTMAN Research group in University of North Carolina has

employed in-vivo voltammetry to measure the dopamine release and uptake in freely
moving animals and found that a behavioral stimulus can evoke a transient increase in
dopamine, providing how a neurotransmitter controls behavior on second and sub second
timescales and revealing how critical rapid, selective, and sensitive measurements for
real-time detection of chemical changes in the brain(Fig 3)
 Figure 3 X-Ray image of microelectrode in brain

Drawbacks

But Interference due to the co-existence of ascorbic acid (AA) in the biological
fluids, which also undergoes oxidation more or less at the same potential, is the major
flaw in dopamine sensor. Also, the concentration of AA is relatively higher than that of
DA in these samples (103 times higher than DA), which results in poor selectivity and
sensitivity for DA detection.So the detection of DA in the presence of excess of AA is a
challenging task in electro analytical research.
EXPERIMENTAL SECTION

Chemicals

Hydrogen tetrachloroaurate, dopamine (DA) and ascorbate (AA) were obtained

from Aldrich and were used as received. All other chemicals used in this investigation
were of analytical grade and were used without further purification. The phosphate buffer
solution (PBS) was prepared from NaH2PO4 and Na2HPO4 (0.1 M).

Preparation of Au colloids

Au colloids were prepared according to the literature. Typically, 1 ml of 1%

HAuCl4 was added to 90ml of water at room temperature. After 1 min of stirring, 2 ml of
38.8 mM sodium citrate was added. Subsequently,1 ml of freshly prepared 0.075%
NaHB4 in 38.8mM sodium citrate was added and the colloidal solution was stirred for
another 5-10 min and stored in a dark bottle at 4 8C. The concentration of the Au
nanoparticles was estimated to be 0.32 mM.

Immobilization of Au colloidal particles

The Au electrodes of 1.6 mm diameter were polished with alumina powder (1.0
and 0.06 mm) and sonicated in water for 5-10 min. The polished electrodes were then
electrochemically cleaned by potential cycling between /0.2 and 1.5 V at a scan rate of 10
V s-1 in 0.05 M H2SO4 for 10 min or until the cyclic voltammogram characteristic for a
clean Au electrode was obtained. The electrochemically cleaned Au electrode was
immersed into an aqueous solution of 10 mM of amine-terminated monolayer of
cystamine (CYST) for 1 h. The CYST monolayer-modified electrode was rinsed well
with water and kept in water for at least 30 min to remove the physically adsorbed CYST.
The CYST electrode was subsequently soaked in the Au colloidal solution for 12 h. The
resulting electrode was washed with copious amount of water and subjected to
electrochemical experiments. Here after the Au nanoparticle-immobilized electrodes will
be referred as the nano-Au electrode (fig.4).
Figure 4 Schematic representation of the fabrication of the nano-Au self-assembly (note
that this is a pictorial representation and is not on the correct scale).
1) Bare gold electrode 2) CYSTMINE modified –Au electrode 3) Au nanoparticle
immobilized electrode 4) TEM of Au nanoparticle immobilized electrode

RESULT AND DISSCUSSION

Fig. 5 shows the SW voltammograms obtained for DA at the bare and nano-Au
electrodes. The voltammetric response at the bare electrode is rather broad, whereas it is
sharp and well defined at the nano-Au electrode, suggesting that the electrochemical
behavior of the nano-Au electrode is quite different from the bulk Au electrode.
Furthermore, as can be readily seen from this figure, the peak current at the nano-Au
electrode is significantly larger compared to that of the bare electrode. For instance a
1.7fold enhancement in the peak current at the nano-Au electrode was observed, which
indicates that the nano-Au electrode possesses excellent sensitivity towards DA.
 Fig. 5 Square-wave voltammograms obtained for the oxidation of DA (50 mM) at
the bare Au (a) and nano-Au (b) electrodes in 0.1 M PBS (pH 7.2).

Since ascorbic acid (AA) is the major interferent in the voltammetric

measurement of DA, its voltammetric behavior at the Au nano-assembly was studied.
Fig. 6 shows the cyclic voltammograms obtained for the oxidation of AA at the nano-Au
electrode at different scan rates. At the nano-Au electrode the oxidation occurs at around
0.03 V and an enormous increase in the peak current compared to the bare electrode was
observed. These results indicate that the nano-Au electrode effectively catalyzes the
oxidation of AA. It has been reported quite recently that nanometer-sized Au particles
exhibit excellent electro catalytic activity.

Fig. 6 Cyclic voltammograms obtained for the oxidation of AA (100mM) at the nano-Au
electrode in 0.1MPBS (pH 7.2). Scan rate: 25, 50, 75, 100, 125, 150 and 175 mV s_1.
 The important attribute of the nano-sized catalysts is the

• High surface area and

•Interface-dominated properties that differs from the atomic, molecular and bulk
counterpart.

In the present investigation the facilitated oxidation of ascorbic acid (AA) at the
nano-Au electrode is believed to be due to the excellent catalytic activity of nano-sized
Au particles. Because the main objective of the present investigation is the determination
of DA in the presence of AA, our attention is focused on the voltammetric detection of
DA in the presence of AA.

Figure 7 Cyclic voltammograms of a binary mixture solution of AA and DA at the bare

Au (a) and nano-Au (b) electrodes (pH 7.2). Scan rate: 100 mV s-1. (B) Corresponding
square-wave voltammograms obtained at bare Au (a) and Nano-Au (b) electrodes.

Fig. 7 shows the cyclic and SW voltammograms obtained for DA and AA coexisting at
bare and nano- Au electrodes. We can see that the bare electrode cannot separate the
voltammetric signals of AA and DA. Only one broad voltammetric signal was observed
for both analytes and the voltammetric peak decreased in the subsequent sweeps.
Therefore it is impossible to use the bare electrode for the voltammetric determination of
DA in the presence of AA. But, the nano-Au electrode resolved the mixed voltammetric
signals into two well-defined voltammetric peaks at 0.015 and 0.185 V corresponding to
the oxidations of AA and DA, respectively. The nano-Au electrode shows good
selectivity and excellent sensitivity in the detection of DA in the presence of AA.AA is
readily oxidized well before the oxidation potential of DA is reached. Thus the catalytic
oxidation of AA by the oxidized DA is completely eliminated and the precise
determination of DA in the presence of AA is possible at the nano-Au electrode. The
voltammetric signals of AA and DA remained unchanged in the subsequent sweeps,
indicating that the nano-Au electrode does not undergo surface fouling. Furthermore, the
separation between the voltammetric peaks of AA and DA is large (165 mV) and thus the
simultaneous determination of AA and DA or the selective determination of DA in the
presence of AA is feasible at the nano-Au electrode

Insights on Influence of Particle Size on Electrochemical bio-sensing

The result shows that nano-sized Au is largely different from the bulk counterpart
and it shows a surprisingly high electro catalytic activity. The nano-Au electrode
successfully distinguishes the voltammetric signals of AA and DA, which are
indistinguishable at the bare Au electrode. Because each incorporated nanoparticle
operate as an individual electrode (electrode of nanosize), and act as an active site for
interfacial electron transfer. Electron transfer at nanoscale electrodes is much different
from that of bulk, because reducing the electrode size increases the diffusion-controlled
transport rate in steady-state voltammetric measurements. Consider the simple electron-
transfer reaction

Ox + e–
Red

Occurring at a spherical electrode of radius a, Diffusion of Ox to the electrode

surface occurs before the electron-transfer step, and either step may be rate-limiting. At
steady state, the rate constant for diffusion (cm/s) of redox molecules to the spherical
electrode is simply D/a, in which D is the diffusion constant (cm2/s) of Ox. Since a is in
nanometer, D/a is comparable to or significantly larger than the standard electron-transfer
rate constant ket (cm/s).

D/a ≥ket

So the overall rate of the electrode reaction is solely controlled by the interfacial
electron-transfer step (i.e. transport rate of analyte from the bulk of the solution to the
interface is not accounted).Hence nanoelectrode open up possibilities for work in very
low concentration (nanomolar) of analyte.whatever can be done at a planar electrode can
be done at concentration of about 106 times lower by using nanoelectrode without
reaching the limiting current. But in bulk electrode the reaction is under diffusion control,
so low concentration analyte reactions are not easily distinguishable from the high
concentrated interference molecule. Hence reason for the high sensitivity of
nanoelectrode is well explained.

CONCLUSION

The present work reveals the fact that; size miniaturization is the principal
cause for high selectivity and high sensitivity of DA sensor. Also proposed reason
soundly corroborates the obtained result. Thus the potential application of nano-
sized Au for the fabrication of a voltammetric DA sensor is demonstrated.