Analytica Chimica Acta 795 (2013) 8–14

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Analytica Chimica Acta

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Polyaniline-iron oxide nanohybrid film as multi-functional label-free

electrochemical and biomagnetic sensor for catechol
Sudeshna Chandra a , Heinrich Lang b , Dhirendra Bahadur a,∗
a
Department of Metallurgical Engineering & Materials Science, Indian Institute of Technology Bombay, Powai, Mumbai 400076, Maharashtra, India
b
Lehrstuhl für Anorganische Chemie, Institut für Chemie, Technische Universität Chemnitz, Straße der Nationen 62, D-09111 Chemnitz, Germany

h i g h l i g h t s g r a p h i c a l a b s t r a c t

• Synthesis of mesoporous
polyaniline-iron oxide magnetic
(Fe3 O4 @PANI) nanohybrid with high
surface area of 94 m2 g−1 .
• Ligand-to-metal charge transfer is
seen between p␲ orbitals of catechol
and d␴* metal orbital of nanohybrid.
• Highly sensitive response
(312 ␮A ␮L−1 ) toward catechol
with low detection limit (0.2 nM)
was obtained by amperometry.
• Decrease in peak frequency of
AC susceptibility shows that the
Fe3 O4 @PANI nanohybrid be used as
biomagnetic sensor for catechol.

a r t i c l e i n f o a b s t r a c t

Article history: Polyaniline-iron oxide magnetic nanohybrid was synthesized and characterized using various spectro-
Received 8 April 2013 scopic, microstructural and electrochemical techniques. The smart integration of Fe3 O4 nanoparticles
Received in revised form 8 July 2013 within the polyaniline (PANI) matrix yielded a mesoporous nanohybrid (Fe3 O4 @PANI) with high surface
Accepted 16 July 2013
area (94 m2 g−1 ) and average pore width of 12.8 nm. Catechol is quasi-reversibly oxidized to o-quinone
Available online 31 July 2013
and reduced at the Fe3 O4 @PANI modified electrodes. The amperometric current response toward catechol
was evaluated using the nanohybrid and the sensitivity and detection limit were found to be 312 ␮A ␮L−1
Keywords:
and 0.2 nM, respectively. The results from electrochemical impedance spectroscopy (EIS) indicated that
PANI-iron oxide nanohybrid
Spectroelectrochemistry
the increased solution resistance (Rs ) was due to elevated adsorption of catechol on the modified elec-
Impedance trodes. Photoluminescence spectra showed ligand-to-metal charge transfer (LMCT) between p-␲ orbitals
Biomagnetic of the phenolate oxygen in catechol and the d-␴* metal orbital of Fe3 O4 @PANI nanohybrid. Potential
Catechol dependent spectroelectrochemical behavior of Fe3 O4 @PANI nanohybrid toward catechol was studied
using UV/vis/NIR spectroscopy. The binding activity of the biomagnetic particles to catechol through
Brownian relaxation was evident from AC susceptibility measurements. The proposed sensor was used
for successful recovery of catechol in tap water samples.
© 2013 Elsevier B.V. All rights reserved.

1. Introduction

Catecholamines are biogenic sympathomimetic hormones pro-

duced by adrenal glands in response to stress which play an
important role in the central nervous system. Catechols are typical
∗ Corresponding author. Tel.: +91 22 25767632. neurotransmitters that affect the regulation of blood pressure and
E-mail address: dhirenb@iitb.ac.in (D. Bahadur). metabolic processes, and are used as an index for several diseases,

0003-2670/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.aca.2013.07.043
 S. Chandra et al. / Analytica Chimica Acta 795 (2013) 8–14
such as hypertension, phaeochromocytoma, neuroblastoma septic
shock, Parkinson’s disease and pharmacological vasodilatation [1].
Any abnormality in the concentration of catecholamines in biologi-
cal fluids, such as urine, plasma, or serum sends a warning signal of
diseases. The pronounced significance of catecholamines in clinical
diagnosis and medical treatment therefore, makes its quantitative,
rapid and sensitive determination important. Though there are var-
ious chromatographic and spectroscopic methods for determining
catecholamines, unfortunately their high cost, low sensitivity and
selectivity, requirement of extraction and derivation steps make
the system less operative [2].
We have sought to address this problem through spectroelec-
trochemical, impedimetric and biomagnetic sensor based on an
electroactive porous nanohybrid with the combined benefits of
magnetic iron oxide [3], capable of oxidizing the catechol, and
an electrically conducting polyaniline. Electrochemical oxidation
of catechol in the mixed organic solvents is well reported which
showed one anodic and a cathodic peak that correspond to the
transformation of catechol to benzoquinone and vice versa within
a quasi-reversible two-electron process [4].
Porous materials with high surface area have a wide range
of applications in adsorption and separation processes, catalysis,
sensors and electrodes, biotechnology, etc. Commonly repre-
sented porous materials are carbon, glass, silicates (zeolites),
oxides, metals, and polymers. Usually, nanoporous materials can
be obtained by direct copolymerization of crosslinking monomers
using porogenic solvents or by the hypercrosslinking method.
The latter involves dissolution of non-crosslinked or marginally
crosslinked polymers followed by immobilization of the pores
by a secondary material. Germain et al. [5] followed the hyper-
crosslinking approach for preparing nanoporous PANI with high
hydrogen storage capacity. So far the synthetic approach involved
grafting of PANI on porous materials so as to produce porous micro-
spheres using interfacial polymerization.
Of late, efforts have been put to synthesize porous nanohybrids
with an aim to synergistically combine the merits of each compo-
nent in the hybrid material [6,7]. Smart intercalation of respective
components can form nanohybrids with much more enhanced
functions and capability. The expected advancement in the nanohy-
brids depends on the polymer-nanoparticle interfacial chemical
and electronic interactions arising due to polymer wrapping, cova-
lent/noncovalent interaction of molecules, etc. This may give rise
to different surface chemistries and electronic structures to the
nanohybrids [8]. It is indeed a great challenge to develop optimal
porous nanohybrids with a high specific surface area favorable for
catalytic sensing and adsorption. Recently, Chen et al. [6] designed
a Pt/C@PANI core–shell-structured catalyst for fuel cell in which
the carbon support is decorated by a layer of PANI and Pt nanopar-
ticles are anchored to the carbon support. Qiao and co-workers [7]
reported a mesoporous nanostructured PANI/TiO2 composite with
optimized biocatalytic and electrocatalytic properties for applica-
tion in microbial fuel cell.
We herein report tailoring of a porous nanohybrid
(Fe3 O4 @PANI) fabricated from iron oxide nanoparticles and
soluble PANI, wherein the Fe3 O4 nanoparticles are looked upon
as biocompatible, stable, and environmental friendly nanoparticle
with attractive electronic and magnetic properties which has
been recently explored for biosensing application [9]. On the
other hand, PANI with a ␲-conjugated structure provides high
electrical and proton conductivity in acidic environments and
possesses unique redox properties. However, its application is
limited by its intractable nature of being insoluble if synthesized
chemically; and brittle and thin if obtained electrochemically [10].
Proposed biosensing technology in this work exploits Fe3 O4 @PANI
nanohybrids and their optical, magnetic, electrochemical and spec-
troelectrochemical properties for enhancing sensing efficiency.
9
Morphological characterization of the nanohybrid plays an
important role since their properties affects the performance of
the sensor. Besides size and structure, other parameters which
influence the properties of hybrid material are surface charge,
porosity, type and density of reactive surface groups. The stability
of the nanomaterials under varied temperature and pH conditions
is correctly oriented by choosing blocking agents and carrying
out post-immobilization modification. In the present case, the
electroactive PANI has been used as matrix materials in in situ
synthesis of Fe3 O4 nanoparticles which not only provided stability
to the nanohybrid but also provided efficient electron transfer for
catechol oxidation. Unlike conventional co-precipitation method,
the proposed modified method ensures control over porous
nature and matrix-natured nanostructures with enhanced elec-
tronic properties. The Fe3 O4 @PANI nanohybrids were found to be
extremely suitable mesoporous matrices for catechol sensing due
to increased internal porosity, high charge densities, super elec-
trochemical behavior and rapid charge transfer kinetics. Another
innovative aspect of the proposed technology is the biomagnetic
sensing using the nanostructures wherein Fe3 O4 @PANI can be
used for detecting the presence of small amounts of catechol. The
resulting nanohybrid has been studied in detail with respect to its
electronic, magnetic and optical properties, surface morphology,
porosity, and applicability in electrochemical, impedimetric and
biomagnetic sensing of catechols (CA).
2. Materials and methods
2.1. Materials and equipment
Ferric chloride hexahydrate (FeCl3 ·6H2 O), and ferrous chloride
tetrahydrate (FeCl2 ·4H2 O) were purchased from Sigma Aldrich
Chemical Co. All other chemicals were of analytical grade and used
as received. The phase purity and identification of the magnetic
nanoparticles were done by X-ray diffraction (XRD), and the pat-
tern of each sample was recorded with a PanAnalytical X-Pert
diffractometer using a monochromatized X-ray beam with nickel-
filtered Cu-K␣ radiation with 4◦ min−1 scan rate. A continuous scan
mode was used to collect 2 data from 5◦ to 70◦ . Fourier transform
infrared (FT-IR) spectra were obtained using a Jasco, FT-IR 300E
spectrometer with a resolution of 4 cm−1 . Scanning electron micro-
scope (SEM) with Energy Dispersive Spectroscope (EDS) S-3400N
Hitachi was used to image PANI and Fe3 O4 @PANI nanohybrids at
an acceleration voltage of 3 kV. Transmission electron microscopy
along with Electron energy loss spectroscopy (EELS) was used to
establish the distribution of iron oxide nanoparticles on PANI. EELS
were performed with a TEM 200 kV that incorporates an elec-
tron beam JEOL JEM 2010 F and an energy loss spectrometer. The
specific surface area, pore volume and pore size distribution of
the hybrids were measured by ASAP 2020 Micromeritics instru-
ment. Specific surface areas were determined by the multipoint
Brunauer–Emmet–Teller (BET) method. The corresponding pore
size distribution and total pore volume were determined by the
Brunauer–Joyner–Hallenda (BJH) method applied to the desorp-
tion branch. Prior to measurements, the samples were outgassed
at 40 ◦ C with a heating rate of 10 ◦ C min−1 for 1 h and then the
temperature was raised up to 50 ◦ C and maintained overnight. Pho-
toluminescence of all the samples was studied by Cary Eclipse
Fluorescence Spectrometer in the range of 300–700 nm.
AC magnetic susceptibility under different frequencies, and the
dependence of magnetization with the applied magnetic field were
measured by using a Physical Properties Measurement System
(PPMS from Quantum Design).
The electrochemical measurements (Cyclic Voltammetry and
Amperometry) were conducted in a 3-electrode single-cell
10 S. Chandra et al. / Analytica Chimica Acta 795 (2013) 8–14
system in phosphate buffer along with 0.1 M KCl as electrolyte.
Glassy carbon electrode (GCE, diameter ϕ = 2 mm), Pt-wire and
Ag/AgCl electrodes were used as working, counter and reference
electrodes, respectively with CHI1140A electrochemical worksta-
tion (CHI110, Austin, TX). All electrochemical measurements were
carried out at room temperature. To eliminate the effect of dis-
solved oxygen, the electrolyte was purged with nitrogen gas for
half an hour. The Electrochemical impedance measurements (EIS)
of the fabricated electrodes were measured by the (Gamry EIS 300)
instrument in a single compartment 3-electrode cell and analyzed
by Echem Analyst, Gamry software.
2.2. Synthesis of Fe3 O4 @PANI nanohybrid
The most common method to prepare Fe3 O4 @PANI nanohybrid
is chemical/electrochemical oxidation polymerization, wherein,
the aniline monomer is polymerized in situ in the presence of
Fe3 O4 nanoparticles and surfactant [11] or by the in situ emul-
sion polymerization/reverse micelle method [12]. However, for
these methods challenges like preventing aggregation of the Fe3 O4
nanoparticles, solubility of the nanohybrids, and processability
issue which arise due to polymerization had to be managed. We
sought to address the problem by developing a simple yet highly
efficient and easy method for preparing the Fe3 O4 @PANI nanohy-
brid which retained the unique behavior of both the components.
The nanohybrid were synthesized by a modified co-precipitation
method in which FeCl2 ·4H2 O (0.86 g) and FeCl3 ·6H2 O (2.35 g)
were dissolved in toluene–isopropanol (2:1 mixture) containing
an already synthesized PANI [10,13]. As the reaction solution was
heated to 80 ◦ C, 2 mL hydrazine hydrate was added slowly and the
reaction was allowed to proceed for 1 h with constant and vigorous
stirring to produce a black suspension. The reaction mixture was
cooled to ambient temperature, and purified by repeated washing
sequentially with ethanol and acetone using a permanent mag-
net.
2.3. Fabrication of Fe3 O4 @PANI electrode
Glassy carbon working electrodes were cleaned by polishing
on aqueous slurries of 0.3 ␮m and 0.05 ␮m alumina powder. The
electrode was then rinsed with deionized water and was kept in
dilute sulphuric acid for 5 min followed by repeated ultrasonica-
tion in methanol and water for 20 min each. After final sonication
in ethanol, the electrode was left to dry in ambient air for a few
minutes. Fe3 O4 @PANI nanohybrids were dispersed in Milli-Q water
and drop-casted onto the surface of the working electrode and
air-dried to obtain a thin layer of coating. The electrode was then
gently washed with deionized water to remove unbound particles
and used fresh in electrochemical experiments. The electrochemi-
cal behavior of the modified electrode was investigated by cyclic
voltammetry (CV) and electrochemical impedance spectroscopy
(EIS). Amperometric i − t measurements were used to detect cat-
echol quantitatively at a fixed operating potential of 800 mV vs.
Ag/AgCl in 0.1 M PBS/0.1 M KCl electrolyte.
2.4. Real sample analysis
Modified Fe3 O4 @PANI electrodes were tested for their applica-
bility for analysis of catechol in tap water samples. The samples
were spiked with known concentration of catechol and recovery
experiments were performed by measuring the CV responses to
the samples in which the known concentration of catechol were
added. The results were also compared with high performance liq-
uid chromatography (HPLC) using water:methanol (ratio 90:10)
mobile phase with C18 column and UV detector.
3. Results and discussion
3.1. Structural analysis of the nanohybrid
The X-ray diffraction pattern of Fe3 O4 @PANI nanohybrid exhib-
ited well resolved peaks of crystalline iron oxide (Figure S1 in SI).
The crystallite size as calculated from the most intense peak was
found to be 16.7 nm. The FTIR spectrum of Fe3 O4 @PANI nanohy-
brid exhibited similar IR bands as that of PANI (Figure S2 in SI). The
decrease in the intensity of the NH stretching bands (1170 cm−1 )
in Fe3 O4 @PANI nanohybrid is attributed to NH· · ·Fe3 O4 inter-
actions [14]. The strong band at 574 cm−1 is assigned to Fe O
stretching vibrations [15].
Surface morphology of PANI and Fe3 O4 @PANI nanohybrids was
determined by Scanning electron microscopy (SEM) which shows
porous and irregular PANI (Figure S3a). Images of Fe3 O4 @PANI
nanohybrids (Figure S3b) show the presence of Fe3 O4 nanoparticles
as white balls and patches on the PANI matrix. The energy disper-
sive X-ray (EDX) shows that the white spots (marked as # in Figure
S3b) are Fe3 O4 with weight % as C (19.02), N (7.64), O (27.30) and
Fe (44.68). In contrast to the above, these white spots are absent
in PANI (Figure S3a) and the EDX gives an weight % as C (56.64), N
(31.67), and O (11.58).
Transmission electron microscopy (TEM) along with electron
energy loss spectroscopy (EELS) was used to establish the dis-
tribution of iron oxide nanoparticles on PANI which showed
monodispersed Fe3 O4 nanoparticles within the PANI matrix (Figure
S4). EELS were performed with a TEM 200 kV that incorporates an
electron beam JEOL JEM 2010 F and an energy loss spectrometer. In
the EELS spectrum, K-edges of C, N and O can be clearly identified.
The sharply defined ␲* and ␴* of the C K and N K-edges are char-
acteristic of sp2 bonding of PANI network. The O-K edges of Fe3 O4
showed two peaks: one at 533 eV derived from O 1s to 2p core level
hybridized with Fe 3d orbital and the other at 548 eV which origi-
nates from O 2p states hybridized with the Fe 4s and 4p states. The
peaks at 562 and 568 eV are due to the scattering of the third and
the first oxygen coordination shells by outgoing or backscattering
electrons [16,17]. The EELS analysis also confirms the presence of
the Fe L2 and L3 edge signals at 715 and 727 eV which correspond to
excitations from the spin-orbit level transitions 2p3/2 → 3d3/2 3d5/2
and 2p1/2 → 3d3/2 , respectively. Thus, from the above observations
of TEM-EELS, it may be inferred that the Fe3 O4 particles interacted
with the PANI matrices through non-covalent bonds, such as ␲–␲
stacking and hydrophobic wrappings [18,19].
The BET surface area of the Fe3 O4 @PANI nanohybrids was cal-
culated based on adsorption data in the partial pressure (P/Po )
range 0.01–0.99. The N2 adsorption–desorption isotherm has a
hysteretic loop in 0.7–0.9 range of relative pressure, which is a char-
acteristic of the adsorption–desorption of a mesoporous material
(Fig. 1). The hysteresis loop remains unsaturated even at high rel-
ative pressure (H3 type) which suggests slit shaped pores (as also
evidenced from the hump in the pore distribution) that may be
due to the deposition of Fe3 O4 nanoparticles on the PANI-matrix.
The external and the BET surface area of the nanohybrid were
experimentally found out to be 133 and 94 m2 g−1 , respectively
with the adsorption average pore width of 12.8 nm. The high sur-
face area and the narrow pore size distribution could render the
nanohybrid very useful for adsorption of any analyte. It can also
be anticipated that the hydrogen acceptors (NH ) of the PANI
may act as stabilizers for the Fe3 O4 nanoparticles and can pro-
vide a bed matrix for its deposition. As expected, the wealth of
a simple synthetic protocol of mesoporous nanohybrids with high
surface area might lead to many prospective studies on biosens-
ing, which prompted us to examine its sensing capability toward
catechol, a neurotransmitter and a prognostic marker for several
diseases.
 S. Chandra et al. / Analytica Chimica Acta 795 (2013) 8–14
Fig. 1. Nitrogen adsorption/desorption isotherm and pore size distribution (inset)
of the Fe3 O4 @PANI nanohybrids.
Photoluminescence (PL) spectroscopy was used to study the
optical and electronic interaction of Fe3 O4 @PANI nanohybrids
with catechol and for elucidation of the probable mecha-
nism of the sensing system (Fig. 2). The spectrum of PANI in
toluene–isopropanol (2:1 mixture) shows emission peaks at 322,
409 and 426 nm and a band around 450–500 nm which are caused
by the reduced benzenoid group and localized exciton associated
with the quinoid ring, respectively [20]. The room temperature
PL spectrum of Fe3 O4 @PANI nanohybrids shows emission bands
at ∼322 and 426 nm corresponding with that of PANI, however,
the emission intensity is higher than that of PANI which is proba-
bly due to the introduction of Fe3 O4 nanoparticles in the hybrid.
The Fe3 O4 nanoparticles increase the density of the carrier and
confine the non-radiative decay of PANI thereby improving the
emission efficiency of material [21]. The band at 409 nm disap-
pears and new bands appear at ∼360, 502, and 531 nm, which
may be due to the interfacial states of the Fe3 O4 and PANI [22].
There is also a significant increase in the intensity of the weak band
at 502 nm due to increased ␲-electron mobility in the compos-
ite favoring formation of singlet excitons which decay radiatively
to the ground state resulting in an enhanced photoluminescence
[23]. Interestingly, the PL intensity is enhanced on addition of
25 Fe3O4@PANI
426
PANI
Fe3O4@PANI-Catechol
20
409
502
Intensity /-
15
10
531
5 surface and PANI enhances the electron transfer and accelerates
322
360
0 favors the reaction at lower potential. It is quite possible that dur-
300 350 400 450 500 550 600
Wave length (nm)
Fig. 2. Photoluminescence spectra of PANI, Fe3 O4 @PANI nanohybrids and its
response toward catechol.
11
Fig. 3. Cyclic voltammograms of modified electrodes in the presence of 10−6 M cate-
chol; scan rate 50 mV s−1 , inset shows calibration curve obtained from amperometry
with varying [catechol].
catechol to the solution of Fe3 O4 @PANI nanohybrids, which may
be due to the combined effect of ligand-to-metal charge transfer
(LMCT) between the p␲ orbitals of the phenolate oxygen of the
catechol and the d␴* metal orbital of the Fe3 O4 @PANI nanohybrids
and reduction of non-radiative pathways.
3.2. Electrochemical performance of the nanohybrids
Cyclic voltammograms (CVs) of PANI and Fe3 O4 @PANI nanohy-
brids in 0.1 M PBS/0.1 M KCl are given in Figure S5. CVs were then
recorded to examine the electrochemical response of neat PANI
and Fe3 O4 @PANI nanohybrids toward catechol in 0.1 M PBS/0.1 M
KCl at a scan rate of 50 mV s−1 . Buffer mixtures are of particular
significance with respect to chemical processes in physiology, and
since we propose to use the developed biosensor for detection of
catechol in living systems, we used PBS as electrolyte. In our pro-
posed application, buffer system is adjusted to a physiological pH
of 7.4, which means while, on one hand, PBS can act as an elec-
trolyte, and on the other hand a definite pH is maintained by it
which is of particular importance in living system. Unlike bare GCE,
on all the modified electrodes, the oxidation and the reduction
peak of catechol appeared which indicates that the catechol can
be oxidized to o-quinone and reduced at the electrodes (Fig. 3).
However, there is a remarkable increase in the peak oxidation cur-
rent of catechol using Fe3 O4 @PANI nanohybrid electrodes which
shows that Fe3 O4 @PANI nanohybrid have a good electrocatalytic
activity toward analyte detection. Catechol exhibited one-electron
redox couple at 0.23 and 0.5 V and an irreversible oxidation peak
at 0.78 V, with a peak-to-peak separation potential (Ep ) of 60 mV.
The first redox peak corresponds to the quasi-reversible oxidation
of catechol to semiquinone radical, while the second reduction
peak is attributed to the reduction of the semiquinone radical
followed by proton abstraction to give catechol. Probably, Fe3 O4
helps in adsorbing and facilitating catechol to reach the electrode
the reaction rate. Thus, the positive shift in oxidation potential is
attributed to the binding of catechol with the nanohybrids which
650 700 ing oxidation of catechol, electron transfer takes place through the
bonds between PANI–NH· · ·Fe3 O4 and the porous structure of the
nanohybrids which further augments the interface between them,
thus resulting in an improved electrocatalytic performance [24]. It
may be assumed that the external surface area of the nanohybrids
12 S. Chandra et al. / Analytica Chimica Acta 795 (2013) 8–14
-4 Catechol
1.5x10
ascorbic acid
-4 dopamine
1.2x10
-5
9.0x10
Current (A)
-5
6.0x10
-5
3.0x10
0.0
-5
-3.0x10
-0.2 0.0 0.2 0.4 0.6 0.8
Potential (V)
Fig. 4. Cyclic voltammograms of 1.0 × 10−6 M catechol in the presence of
1.0 × 10−4 M dopamine and 1.0 × 10−4 M ascorbic acid. The voltammograms rep-
resents selective determination of catechol in the presence of higher amounts of
other interferents.
afforded by the pores increases the active site on the electrode
surface thereby playing a vital role in the catalytic reaction.
Cyclic voltammetry for oxidation–reduction of catechol at
Fe3 O4 @PANI nanohybrid modified GCE was also carried out in
the potential range −0.1 to 1.0 V vs Ag/AgCl at scan rates of
5–200 mV s−1 (Figure S6 in SI). With increase in the scan rate,
the anodic (Ipa ) and cathodic (Ipc ) peak currents increased. Also,
the Ipa moved to more positive potential while the Ipc shifted to
more negative values. The separation of the peak potential of cate-
chol increased with increasing scan rates. The peak currents were
proportional to the square root of scan rate () in the range of
5–200 mV s−1 with R2 value of 0.99591 and 0.99709 for Ipa and
Ipc , respectively, suggesting an adsorption-controlled process. It is
also seen that proportional to the augmentation of potential sweep
rate, there is an increase in the height of cathodic peak indicat-
ing that the overall mechanism involved a chemical step after the
electron transfer [25]. An EC mechanism is proposed in which the
quasi-reversible chemical and electrochemical reactions take place
on the modified electrodes in the presence of catechol. The decrease
in current ratio (Ipa /Ipc ) with increase in scan rate indicates good
electroactivity of the material toward catechol.
3.3. Interference studies of dopamine (DA) and ascorbic acid (AA)
Both AA and DA coexist with catechol in physiological fluids
and possess oxidation potentials close to each other at electrode,
resulting in the electrochemical response of catechol being almost
overlapped by AA or DA. In this study, it was found that well
resolved redox peak of catechol in the presence of higher concen-
tration of DA and AA could be obtained by using the Fe3 O4 @PANI
modified GCE. In order to confirm the sensing of catechol in the
presence of AA and DA, we scanned a mixture solution contain-
ing 1.0 × 10−6 M catechol, 1.0 × 10−4 M AA, and 5.0 × 10−4 M DA by
cyclic voltammetry. As is shown in Fig. 4, the cyclic voltammograms
demonstrated that the coexistence of AA and DA with catechol
(pH 7.0; PBS) produced no obvious change in peak potentials of
catechol, though slight lowering of peak current was observed. Fur-
ther, in the presence of only dopamine, the Fe3 O4 @PANI-modified
electrode showed a cathodic peak separation of about 90 mV for
catechol-DA which indicates that DA could produce no substan-
tial effect on the assay of catechol. Similarly, there was no overlap
of peaks of catechol with that of ascorbic acid suggesting the fea-
sibility of reliable determination of catechol. Based on the above
discussion, it may be concluded that neither DA nor AA will
1.0
Fig. 5. Nyquist plots of Fe3 O4 @PANI modified electrodes with varying concentration
of catechol; the inset shows equivalent circuit diagram used to model impedance
data.
interfere in the sensing of catechol and the proposed system can
be well used for selective determination of catechol in the mixture
solution containing other possible interferents.
A typical steady state current response of the Fe3 O4 @PANI
nanohybrids modified GCE toward successive addition of catechol
in a stirred solution of in 0.1 M PBS/0.1 M KCl electrolyte is shown
in Figure S7 in SI. For this, 20 ␮L of catechol was added to the elec-
trolyte solution and amperometric detection was carried out at a
stirring of 250 rpm for a period of 600 s. A fast and stable current
response was observed and the sensitivity of catechol was found
to be about 312 ␮A ␮L−1 with a detection limit of 0.2 nM (inset of
Fig. 3).
3.4. Impedimetric and spectroelectrochemical characteristics of
the nanohybrids
Electrochemical impedance spectroscopy (EIS) was used to
monitor the impedance changes of Fe3 O4 @PANI modified electrode
surface and its interfacial properties in the presence of different
concentrations of catechol. The frequency was varied from 0.1
to 105 Hz with 20 point impedance readings per decade. Nyquist
diagram (Fig. 5) displays that the modified electrode showed an
increase in Rs values (from 31.43 to 182.7 ) with increase in con-
centration of catechol in solution where the analyte tends to adsorb
faster on the electrodes giving rise to more Rs indicating that the
electrochemical impedance signal is directly related to the amount
of catechol adsorbed onto the electrode. A good linear relationship
was obtained with a correlation coefficient of 0.9894, the limit of
detection (LOD) was calculated to be 16.5 nM (S N−1 = 3).
To compliment the findings and to gain better insight into the
mechanism of the interaction between Fe3 O4 @PANI nanohybrids
and catechol, spectroelectrochemical studies were performed.
Potential dependent optical absorption spectra of Fe3 O4 @PANI
nanohybrids and Fe3 O4 @PANI nanohybrids-catechol in 0.1 M
dichloromethane solutions of [NBu4 ][B(C6 F5 )4 ] as supporting elec-
trolyte was studied using UV/vis/NIR spectroscopy. For this, a
stepwise increase (100 mV) of the potential from −100 mV to
700 mV vs. Ag/AgCl in an optically transparent thin-layer electro-
chemistry (OTTLE) cell has been applied (Fig. 6).
Fe3 O4 @PANI nanohybrids showed the soret band at 380 nm
which increased in intensity on increasing the potential from −100
to 600 mV. Fe3 O4 @PANI nanohybrids-catechol showed strong
absorption bands at 275 nm in the absence of any potential. On
applying 100 mV, a new broad band appears at ∼390 nm. The
 S. Chandra et al. / Analytica Chimica Acta 795 (2013) 8–14 13

1.50 9.9x10
-5
Fe3O4@PANI
without potential
1.25 -100 mV -5
1.2 9.6x10
0 mV
100 mV
1.00 0.9 9.3x10
-5
Absorption /-

200 mV

Absorption /-

χ " (μB/Oe)
300 mV 0.6
0.75 400 mV 9.0x10
-5

500 mV 0.3
600 mV 8.7x10
-5
0.50 700 mV 0.0
Fe3O4@PANI-
200 300 400 500 600 700 -5
Wave length (nm) 8.4x10 catechol
0.25
-5
8.1x10
0.00

0 2000 4000 6000 8000 10000

500 1000 1500 2000 2500 Frequency (Hz)
Wave length (nm)
Fig. 7. Imaginary part of the AC magnetic susceptibility as a function of frequency
Fig. 6. UV/vis/near-IR spectroelectrochemical measurements of modified elec- for Fe3 O4 @PANI (solid symbols) and Fe3 O4 @PANI-catechol (open symbols).
trodes in the presence of catechol, recorded in an OTTLE cell. Inset shows the
expanded region from 200–700 nm.

which diminished the peak frequency. Since the peak frequency of

intensity of the band at 275 nm started decreasing on applying a
potential of 200 mV with a rise in intensity of the band at 390 nm.
The change in the intensity of the absorption continued till a poten-
tial of 400 mV, after which the band at 275 nm started diminishing.
This transition indicates a reduction in the concentration of the
reduced species and their conversion into the oxidized species.
Polarization of the electrode takes place from 0.2 to 0.7 V which
corresponds to the oxidation state of the catechol resulting in a
steady growth of the band at 390 nm which grows at the expense
of the band at 275 nm. The intervalence charge transfer band due to
the semiquinone-catecholate transition was observed at 1900 nm
which is shifted to lower energy with increase in potential which
could be attributed to various substituents in the catecholate moi-
ety [26].
We further explored this sensing through Brownian relaxation
upon binding of the nanohybrids with the catechol. Fig. 7 shows
the imaginary part of AC magnetic susceptibility as a function of
frequency in the presence of an AC magnetic field. On interac-
tion with the catechols, the peak frequency of the nanoparticles
decreased considerably along with a frequency shift toward higher
side [27]. The interaction led to an increase in the hydrodynamic
radii of the nanoparticles and their rotational motion is blocked
the AC susceptibility is inversely proportional to the particle vol-
ume, this scheme can be used to monitor the change in the particle
volume upon binding of catechol to Fe3 O4 @PANI. The use of Brown-
ian relaxation time measurements in frequency domain provides a
platform for developing a biomagnetic sensor.
3.5. Real sample analysis
The proposed sensing method was used to determine catechol in
tap water collected from two different sources and a known amount
of catechol was added. Recovery experiments were performed and
the quantitative recoveries were evaluated from the calibration
curve obtained by cyclic voltammetry. The results were also com-
pared using HPLC and are tabulated in Table 1. The results show that
the two methods are in good agreement and the Fe3 O4 @PANI mod-
ified electrodes can be used for effective determination of catechol
in the water samples.
Table 2 gives an overview of various materials and techniques
like amperometry, square wave voltammetry (SWV), differential
pulse voltammetry (DPV) and cyclic voltammetry (CV) used for
sensing and detection of catechol. It also provides a comparative
performance of various catechol sensors in terms of detection limit,

Table 1
Recoveries of catechol concentration in tap water samples by the proposed electrode.

Sample By HPLC (g g−1 ) By cyclic voltammetry using Fe3 O4 @PANI modified GCE Recovery (%)

Added (␮M) Found (␮M) Added (␮M) Found (␮M)

Tap water 1 100 105 50 56 112

Tap water 2 120 123 100 95 95

Table 2
Comparison of various catechol biosensors based on different electrode materials in terms of performance.

Sl. No Electrode material Technique Limit of detection Sensitivity Interference Reference

1 Graphene DPV 1.8 × 10−8 M NM None with hydroquinone [28]

2 Ferrocene Amperometry 10.8 ␮M 1.14 ␮A mM−1 None with UA [29]
3 Poly(malachite green)coated MCNT film DPV 5.8 ␮M 1.3 mA cm−2 mM−1 None with quinol [30]
4 DeniLite Laccase Amperometry 70 nM 70 nA ␮M−1 None with AA [31]
5 Graphite SWV 2.9 × 10−7 M NM None with AA [32]
6 Au/TiO2 Amperometry 1 × 10−6 M 51.58 ␮A ␮M−1 NM [33]
7 PANI Amperometry 0.05 ␮M NM None with AA, DA [2]
8 Mesoporous Pt DPV NM 1.20 mA cm−2 mM−1 NM [34]
9 Fe3 O4 @PANI/GCE Amperometry 0.2 nM 312 ␮A ␮L−1 None with AA, DA Present work
EIS 16.5 nM
14 S. Chandra et al. / Analytica Chimica Acta 795 (2013) 8–14
sensitivity and interference which clearly shows that the present
system is much better than the previously reported ones.
4. Conclusion
In summary, we present a synthetic strategy for easy evolu-
tion of a mesoporous Fe3 O4 @PANI nanohybrid and demonstrate
an ultra-sensitive electro- and spectroelectrochemical platform
for label-free sensing of catechol. With the unique distribution of
Fe3 O4 on the conducting PANI matrix, the catechol detection was
achieved at nanomolar levels (the limit of detection was 0.2 nM).
The decrease in the peak frequency of the imaginary part of the AC
magnetic susceptibility demonstrates the feasibility of using the
nanohybrid as biomagnetic sensor based on Brownian relaxation.
Besides this, we provide a thorough understanding of the inter-
action of catechols with the nanohybrid system, thereby opening
new horizons in the area of spectroelectrochemical analysis and
biosensing.
Acknowledgements
Authors are grateful to the Nanomission of Department of Sci-
ence and Technology, Govt. of India, and Humboldt Foundation,
Germany for providing fellowships.
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.aca.2013.07.043.
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