Sensors and Actuators B 199 (2014) 330–338

Contents lists available at ScienceDirect

Sensors and Actuators B: Chemical

journal homepage: www.elsevier.com/locate/snb

Development of electrochemical biosensor with ceria–PANI

core–shell nano-interface for the detection of histamine
Manju Bhargavi Gumpu a,b , Noel Nesakumar a,c , Swaminathan Sethuraman a,b ,
Uma Maheswari Krishnan a,b , John Bosco Balaguru Rayappan a,c,∗
a
Centre for Nanotechnology & Advanced Biomaterials (CeNTAB), SASTRA University, Thanjavur 613 401, Tamil Nadu, India
b
School of Chemical and Biotechnology, SASTRA University, Thanjavur 613 401, Tamil Nadu, India
c
School of Electrical & Electronics Engineering, SASTRA University, Thanjavur 613 401, Tamil Nadu, India

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

Article history: A mediator-free electrochemical biosensor with CeO2 –PANI nano-interface for sensing histamine using
Received 27 January 2014 diamine oxidase (DAO) has been developed. CeO2 –PANI core–shell nanoparticles were prepared by
Received in revised form 19 March 2014 hydrothermal method. The field emission scanning electron microscopy (FE-SEM) revealed the aggre-
Accepted 3 April 2014
gated spherical morphology of CeO2 . The core–shell formation of CeO2 –PANI was confirmed with field
Available online 15 April 2014
emission transmission microscopy (FE-TEM). The polycrystallinity of CeO2 and CeO2 –PANI was confirmed
using X-ray diffraction (XRD). Immobilization of DAO with CeO2 –PANI was confirmed with Fourier trans-
Keywords:
form infrared spectroscopy (FT-IR). Electrochemical studies were carried out through cyclic voltammetry
Histamine
CeO2 –PANI
and amperometry using modified GCE/CeO2 –PANI/DAO as a working electrode, Ag/AgCl saturated with
Diamine oxidase 0.1 M KCl as a reference electrode and platinum (Pt) wire as a counter electrode. The linear range was
Cyclic volatammetry observed from 0.45 to 1.05 mM with a sensitivity of 724.94 ␮A cm−2 mM−1 . Michaelis–Menten constant
Amperometry was calculated as 0.798 mM. It exhibited limit of detection of 48.7 ␮M, limit of quantification of 132.4 ␮M
Tiger prawn with a response time of <1 s and good shelf life of 86% till 18 days. Also the developed biosensor was
applied on tiger prawn to estimate the histamine content.
© 2014 Published by Elsevier B.V.

1. Introduction became a key determinant to avoid adverse effects. As a part of food

Histamine is a heterocyclic low molecular weight organic com-
pound formed by enzymatic removal of carboxyl group from
histidine, found in various foods such as meat, fish, alcoholic bever-
ages, vinegar, fermented foods etc [1,2]. Improper handling, storage
conditions, availability of free amino acids and optimum tempera-
tures are the main reasons for increase in histamine concentration
thereby leading to the toxification of fish related food items [3,4].
Consumption of such fish leads to “Histaminosis” or “Scombroid
fish poisoning” causing both physiological and toxicological effects
based on dosage [5].
According to World review of fisheries and aquaculture in 2011,
production of fish is estimated to be 154 million tonnes all over
the world which signifies that the rate of consumption is being
increased. As a result, safety evaluation of fish and its products
∗ Corresponding author at: Centre for Nanotechnology & Advanced Biomaterials
(CeNTAB) & School of Electrical & Electronics Engineering (SEEE) SASTRA University,
Thanjavur 613 401, India. Tel.: +91 4362 264 101x255; fax: +91 4362 264120.
E-mail address: rjbosco@ece.sastra.edu (J.B.B. Rayappan).
safety evaluation, levels of histamine in fish are limited to 0.45 mM
by Food and Drug Administration (FDA) [3,5].
To detect histamine levels in fish and its related products,
many enzymes such as methyl amine dehydrogenase, monoamine
oxidase (MAO), diamine oxidase (DAO) etc., have been used.
Among these, DAO is of particular interest as it shows high speci-
ficity towards histamine and also it can oxidatively deaminate
histamine [6]. To estimate histamine levels for fish freshness deter-
mination various analytical methods such as high performance
liquid chromatography [7,8], Spectrofluorimetry [9], thin layer
chromatography [10], ELISA, capillary electrophoresis, impedance
spectroscopy [11], microdialysis [12] etc., have been used. But all
these methods require sample preparation and time consuming
[13].
In a step of advancements in biosensors, Telsing et al. developed
a voltammetric pea seedling amine oxidase modified biosensor
which was able to detect cadaverine, putrescine with a linear range
of 30–88 ␮g mL−1 and 24–67 ␮g mL−1 respectively, but it failed
to detect histamine [14]. Lomillo et al. developed a disposable
biosensor which showed poor precision in terms of repeata-
bility [15]. Also, various electrochemical based biosensors were

http://dx.doi.org/10.1016/j.snb.2014.04.009
0925-4005/© 2014 Published by Elsevier B.V.
 M.B. Gumpu et al. / Sensors and Actuators B 199 (2014) 330–338
developed to detect biogenic amines in fish using mediators such
as ferrocene derivatives, 2,6-dichlorophenolindophenol, potassium
ferricyanide and 4-amino diphenylamine [5], manganese dioxide
[14], hydroxy methyl ferrocene [15]. However, use of such media-
tors leads to leaching of untethered mediator from enzyme layer,
causes toxicity in biological tissues and suffers from redox interfer-
ence [16]. Besides, anodic iridium oxides were used to determine
pH and histamine levels in guinea pigs [17]. Also, a different
approach namely heat-transfer method was chosen by Peeters et al.
for the detection of histamine in saliva, blood [11].
Usage of nano-interface enhances direct electron transfer
between enzyme and electrode thereby enhancing bio sensing
properties [18] showing high electron transfer rates and sensi-
tivity of a biosensor. In this work, ceria–polyaniline (CeO2 –PANI)
core–shell nanoparticles were considered as a nano-interface
because CeO2 exhibits nano-morphological, functional, biocompat-
ible and catalytic properties with faster electron transfer kinetics
[19]. PANI in the core–shell structure helps further to increase the
sensitivity; also it acts as a localization layer for electronic transi-
tion between DAO and CeO2 . In this present work, both physical,
optical properties of CeO2 –PANI core–shell nanoparticles and elec-
trochemical properties of mediator-free DAO based biosensor were
studied for the determination of histamine levels in tiger prawn.
2. Materials and methods
2.1. Reagents
Cerium (III) nitrate hexahydrate (Ce(NO3 )3 ·6H2 O), sodium
hydroxide (NaOH), ammonium per sulfate (APS), and aniline
hydrochloride (C6 H8 ClN) were purchased from Sigma Aldrich,
India. 10 mg ␮L−1 solutions of diamine oxidase (DAO) (E.C 1.4.3.6.,
activity: 0.18 U mg−1 , Sigma Aldrich, India) in 0.1 M phosphate
buffered saline (PBS), pH 7.4 (mono basic sodium phosphate, diba-
sic sodium phosphate from Merck, India), 0.1 M potassium chloride
(7.45 mg mL−1 ), hydrochloric acid (HCl), acetone were procured
from Merck, India. Histamine (66 mg mL−1 ) was reconstituted in
5 mL of 0.1 M PBS, pH 7.4 was used as stock and stored at 278 K.
Other solutions used were 1.7 ␮g mL−1 of ascorbic acid reconsti-
tuted in 100 mL of 0.1 M PBS (pH 7.4), 0.34 mg mL−1 of sucrose,
0.06 mg mL−1 of urea reconstituted in 10 mL of 0.1 M PBS, (pH 7.4)
each. All these were purchased from Sigma Aldrich, India. Nafion
(50 mg mL−1 ) was procured from Sigma Aldrich, India. Live fresh
tiger prawn was purchased from local market and stored at room
temperature for 6 days after its death.
2.2. Sample preparation
Histamine was extracted from fish muscle Penaeus monodon
(tiger prawn) according to the following procedure. 60 mg of mus-
cle tissue was homogenized with 30 mL of 0.1 M PBS, (pH 7.4). This
mixture was centrifuged for 30 min at 12,000 rpm and the super-
natant solution was transferred and made up to 30 mL with water,
which was stored at 277 K.
2.3. Synthesis of ceria–polyaniline (CeO2 –PANI) core–shell
nanoparticles
CeO2 nanoparticles were prepared by a simple hydroxide
method [20]. Briefly, 0.3 M NaOH solution (12 mg mL−1 ) was added
slowly to 0.1 M Ce(NO3 )3 ·6H2 O (42 mg mL−1 ), stirred for 24 h when
a yellow coloured precipitate was formed at room temperature
confirming CeO2 formation. It was centrifuged at 8000 rpm for
20 min and heated at 424 K for 6 h. Later it was subjected to
annealing at 524 and 624 K. Then synthesis of PANI was carried
331
out by adding 0.25 M APS (110 mg mL−1 ) slowly to 0.2 M ani-
line hydrochloride (50 mg mL−1 ) and then stirred for 1 h. After
12 h, PANI which gets precipitated was filtered and then washed
with 0.2 M HCl (7.3 mg mL−1 ) followed by washing with 100 mL
of acetone (10 mg mL−1 ). Then it was dried at 334 K and for fur-
ther polymerization it was kept in ice at 274–276 K. Later it was
acidified by replacing 10 mL of deionized water with 10 mL HCl
(7.3 mg mL−1 ) [21]. Then, 1:1 ratio of CeO2 and PANI were ground
together and sonicated for 1 h. The solution mixture was stirred for
a period of 24 h and the precipitate of CeO2 –PANI was filtered and
dried.
2.4. Preparation of GCE/CeO2 –PANI/DAO/nafion thin film
80 mg of CeO2 –PANI nanoparticles in 100 ␮L of 0.1 M PBS, (pH
7.4) was subjected to ultrasonication for about 15 min. Later 50 ␮L
of DAO stock solution (10 mg ␮L−1 ) was added and further sub-
jected to ultrasonication for 1 min followed by drying. Finally DAO
tagged CeO2 –PANI was obtained and was coated onto the glassy
carbon working electrode (GCE) electrode along with 5 ␮L of nafion
(50 mg mL−1 ) and allowed to air dry.
3. Instrumentation
3.1. CeO2 –PANI characterization
Morphology of CeO2 nanoparticles was studied by subject-
ing to cold field emission scanning electron microscopy (FE-SEM)
(JSM 6701F, JEOL, Japan). Structural characteristics of CeO2 and
CeO2 –PANI crystallinity was studied using X-ray diffractometer
(XRD) with Cu K␣ radiation (Rigaku Ultima III, USA) from an angle
of 20–75◦ . Core–shell morphology of CeO2 –PANI was studied using
field emission transmission electron microscope (FE- TEM) (JSM
2100, JEOL, Japan). CeO2 and CeO2 –PANI tagged with DAO were
analysed by using Fourier transform infrared spectroscopy (FT-IR)
(Spectrum 100, Perkin Elmer, USA).
3.2. Analysis
Image J software (Image J 1.48q) was used to estimate the size of
CeO2 –PANI nanoparticles from FE-TEM micrographs. Electrochem-
ical analysis was carried out using an electrochemical analyzer
(CH600C, CH Instruments, USA). Electrochemical studies were car-
ried out using GCE/CeO2 –PANI/DAO as working electrode (3 mm
dia, CHI104, CH Instruments, USA), Ag/AgCl (CHI111, CH Instru-
ments, USA) saturated with 0.1 M KCl as a reference electrode
and a Pt wire electrode (CHI115, CH Instruments, USA) as counter
electrode. The cyclic voltammograms were recorded in 0.1 M PBS
(pH 7.4) at room temperature. The performance of the calibration
models was verified using statistical analysis (Supplementary. Eqs.
(1)–(3)).
4. Results and discussion
4.1. XRD data for CeO2 –PANI nanoparticles
The X-ray diffraction pattern of CeO2 synthesized at both 524
and 624 K are represented in Fig. 1(a). The characteristic diffraction
peaks corresponding to (1 1 1), (2 0 0), (2 1 0), (2 2 0), (3 1 1), (2 2 2)
and (4 0 0) planes confirmed the polycrystalline nature of CeO2 with
cubic fluorite structure and were in good agreement with [JCPDS
34–0394]. The XRD pattern of CeO2 –PANI at both 524 and 624 K
were analysed and represented in Fig. 1(b). CeO2 –PANI prepared at
524 and 624 K showed a decrease in the diffraction peak intensity
corresponding to (1 1 1), (2 0 0), (2 2 0), (3 1 1) and (4 0 0) planes. The
332 M.B. Gumpu et al. / Sensors and Actuators B 199 (2014) 330–338

Fig. 1. The XRD pattern of CeO2 prepared at (a) 524 and 624 K, (b) CeO2 –PANI at 524 and 624 K. The FE-SEM image of CeO2 prepared at (c) 524 K, (d) 624 K. The FE-TEM
micrograph of CeO2 –PANI core–shell nanoparticles prepared at (e) 524 K (inset(i)), (f) 624 K (inset (ii)). The FT-IR spectra of (g) polyaniline (PANI) (h) CeO2 –PANI prepared
at 524 and 624 K and CeO2 –PANI prepared at 524 and 624 K tagged with DAO.
 M.B. Gumpu et al. / Sensors and Actuators B 199 (2014) 330–338
grain size of CeO2 and CeO2 –PANI nanoparticles was calculated by
using Debye’s Scherrer formula [22]. Crystallite size of CeO2 at 524,
624 K and CeO2 –PANI at 524, 624 K was estimated to be 9.8, 9.2 and
5.6, 5.3 nm respectively.
4.2. FE-SEM and FE-TEM analysis
Fig. 1(c) and (d) represents the FE-SEM images of CeO2 at 524
and 624 K. It showed spherical and aggregated morphology with a
mean size of 63 ± 4 and 60 ± 12 nm respectively. Fig. 1(e) and (f)
shows the FE-TEM micrograph of CeO2 –PANI with the darker core
of CeO2 embedded in the lighter PANI matrix indicating core–shell
nanoparticle formation with a crystallite size of 9 ± 2 and 10 ± 1 nm
for CeO2 –PANI at 524 and 624 K respectively.
4.3. FT-IR spectral analysis
In the preparation of polyaniline, initially emeraldine base
was formed which is blue in colour. This reaction is exother-
mic in nature, a raise in temperature was observed which helps
emeraldine base to be converted to emeraldine salt form i.e., the
conducting form of polyaniline (PANI) [23]. Additionally, the pre-
pared polyaniline was treated with 0.2 M HCl to provide uniform
protonation to polyaniline with chloride counter ions. Also, the
treatment of PANI with concentration HCl helps in the enhance-
ment of the conductivity. This conductive form of PANI was
characterized using FT-IR.
Fig. 1(g) showed a peak at 3385 cm−1 indicating N H stretching
of benzoid ring, 1585 cm−1 showing the quinoid ring, 1502 cm−1
benzoid ring, 1395 cm−1 C N stretching, 1309 cm−1 aromatic
amine for the combination modes of benzoid and quionoid com-
pounds, 1134 cm−1 corresponds to high electrical conductivity
and high degree of localization, 825 cm−1 a C H stretching was
observed indicating polyaniline formation. [24]. The FT-IR spec-
tra of CeO2 –PANI and CeO2 –PANI tagged with DAO are presented
in Fig. 1(h). The absorption band of CeO2 –PANI at 3439 cm−1 cor-
responds to free N H stretching, band at 1642 cm−1 corresponds
to C O stretching, 1587 cm−1 and 1494 cm−1 corresponds to C C
stretching in aromatic ring of emeraldine base confirming the pres-
ence of CeO2 along with PANI. Bands at 1309 cm−1 represents C N
stretching of aromatic secondary amine in PANI, band at 1133 cm−1
indicates high electrical conductivity. Band at 818 cm−1 is due to the
Ce O metal oxygen bond and all bands are in good agreement with
other reported data [24–26]. CeO2 –PANI tagged with DAO was con-
firmed from FT-IR with absorption bands observed at 3445 cm−1
corresponding to N H stretching of secondary amine, 1634 cm−1
corresponding to N H bending of primary amine. The band at
2925 cm−1 indicates C H stretch of methyl group indicating the
presence of DAO. Also the remaining bands of CeO2 –PANI tagged
with DAO showed a shift in corresponding peaks of CeO2 –PANI [27]
suggesting association of the enzyme with the CeO2 –PANI matrix.
4.4. Electrochemical studies
Electrochemical studies were carried out in the absence of his-
tamine in 0.1 M PBS (pH 7.4) by measuring current 5 times from the
bare GCE working electrode. It was observed that instrument noise
was low and was equal to 73 nA with a relative standard deviation
(RSD) of 0.05. In order to optimise the experimental conditions,
control experiments were carried out using bare GCE, GCE/DAO,
GCE/CeO2 /DAO, GCE/CeO2 –PANI/DAO with both CeO2 synthesized
at 524 and 624 K. The cyclic voltammograms of different modified
electrodes were recorded in the presence of 0.45 mM of histamine
(66 mg mL−1 ) in 0.1 M PBS (pH 7.4) with a scan rate of 0.1 V s−1 and
are presented in Fig. 2.
333
Fig. 2. Cyclic voltammograms of modified electrode in 0.1 M PBS (pH 7.4) in pres-
ence of 0.45 mM histamine.
In the presence of oxygen, DAO catalyzes histamine
to Imidazoleacetaldehyde and peroxide. i.e., Histamine +
DAO
Oxygen−→Imidazoleacetaldehyde + peroxide [28]. Generally,
the standard reduction potential (E0 ) of Ag/AgCl is 222.3 mV.
And, the E0 of peroxide to oxygen (H2 O2 → O2 + 2H+ + 2e− ) is
−682 mV (Vs Standard Hydrogen electrode (SHE) at 298 K). If
the electrochemical reaction is performed with Ag/AgCl as ref-
erence electrode, the E0 will shift to −904.3 mV [29]. GCE/DAO,
GCE/CeO2 at 524 K/DAO, GCE/CeO2 at 624 K/DAO, GCE/CeO2 at
524 K–PANI/DAO and GCE/CeO2 at 624 K–PANI/DAO electrodes
showed irreversible cyclic voltammetric current with the Epc of
−1039 mV, −1036 mV, −990 mV, −985 mV and −1037 mV respec-
tively. This trend confirmed the occurrence of reduction reaction
of peroxide to oxygen in the cathodic process. It also showed
that the DAO biomolecule retains its catalytic activity even after
the adsorption on to different biomaterials. In order to study the
effects of the PANI and CeO2 in CeO2 –PANI composites on the
histamine determination, Epc of different modified electrode were
compared. When the Epc of GCE/CeO2 at 524 K/DAO and GCE/CeO2
at 624 K/DAO is compared with GCE and GCE/DAO bioelectrode,
only the CeO2 nanoparticles prepared at 624 K allows the GCE/CeO2
at 624 K/DAO to operate at low reduction potential than in the
presence of CeO2 nanoparticles prepared at 524 K. Further, the
effects of CeO2 –PANI composites on the histamine determina-
tion were studied. In the case of GCE/CeO2 at 524 K–PANI/DAO
bioelectrode, PANI allows the bioelectrode to operate at low
potential (Epc (GCE/CeO2 at 524 K/DAO) − Epc (GCE/CeO2 at 524
K–PANI/DAO) = −51 mV) where as in the case of GCE/CeO2 at
624 K–PANI/DAO bioelectrode, PANI allows the bioelectrode to
operate at high potential (Epc (GCE/CeO2 at 624 K/DAO) − Epc
(GCE/CeO2 at 624 K–PANI/DAO) = 47 mV). It also showed that PANI
in the GCE/CeO2 at 524 K–PANI/DAO bioelectrode can reduces the
interfering reactions and inaccuracy.
It is evident that GCE/CeO2 at 524 K–PANI/DAO in presence
of 0.45 mM histamine showed higher sigmoidal cathodic wave
current (IW = 8.95 ␮A) which is 1.05 times greater than that of
GCE/CeO2 at 624 K–PANI/DAO (IW = 8.54 ␮A). In case of GCE/CeO2
at 624 K–PANI/DAO, IW got lowered than in absence of PANI
(IW = 8.7 ␮A) which might be due to electron hindrance property
of PANI (Supplementary Fig. S1). The electron hindrance property
of PANI in GCE/CeO2 at 624 K–PANI/DAO was verified by compar-
ing RSD of current response for various modified GCE electrodes
with respect to bare GCE in the presence of 0.45 mM histamine.
Generally RSD of current response for various modified GCE elec-
trodes with respect to bare GCE in the presence of 0.45 mM
histamine should be higher. This statement was in good agreement
334 M.B. Gumpu et al. / Sensors and Actuators B 199 (2014) 330–338
Table 1
Comparison of various parameters of GCE/CeO2 at 524 K–PANI/DAO and GCE/CeO2 at 624 K–PANI/DAO.
Modified electrode FWHM (mV)
GCE/CeO2 at 524 K–PANI/DAO 538
GCE/CeO2 at 624 K–PANI/DAO 637
between GCE/CeO2 at 524 K/DAO (RSD = 0.144) and GCE/CeO2
at 524 K–PANI/DAO (RSD = 0.169). But in case of GCE/CeO2
at 624 K/DAO (RSD = 0.141) and GCE/CeO2 at 624 K–PANI/DAO
(RSD = 0.119) electrode, RSD of current response in presence of
PANI got decreased. This might be due to the low surface cover-
age of DAO at GCE/CeO2 at 624 K–PANI/DAO. The surface coverage
( = Q/nFA) of DAO at GCE/CeO2 at 524 K–PANI/DAO was estimated
as 2.783 × 10−6 mol cm−2 which was three fold higher than that
at GCE/CeO2 at 624 K–PANI/DAO (1.346 × 10−9 mol cm−2 ). As the
surface concentration of DAO at GCE/CeO2 at 524 K–PANI/DAO bio-
electrode was higher than that at GCE/CeO2 at 624 K–PANI/DAO,
histamine biomolecules got converted into H2 O2 to a greater extent
and thereby resulted in high IW .
In surface controlled electrode reaction, the current increases
linearly with scan rate whereas in diffusion controlled process,
the current increases gradually but with a square-root depen-
dence on scan rate. The cyclic voltammograms of GCE/CeO2 at
524 K–PANI/DAO at various scan rates were carried out in 0.1 M PBS
(pH 7.4) (Supplementary Fig. S2). Nernstian behaviour of surface-
confined biomolecules always depends on scan rate. And also,
the peak current should be directly proportional to the potential
scan rate. In case of GCE/CeO2 at 524 K–PANI/DAO bioelectrode,
as the scan rate increases from 0.02 to 0.06 V s−1 , cathodic sig-
moidal peak current increased linearly (IW = 38.62 ␮A/V s−1 + 3.047,
R2 = 0.986) (Supplementary Fig. S3) in accordance with the equa-
tion: Ip = nFQ/4RT, where Ip is the peak current, n is number of
electrons transferred, Q is the charge,  is the scan rate, R is the
constant and T is the absolute temperature, suggesting that the
electrochemical process was surface controlled. This also showed
that the reaction occurred only due to surface confined molecules.
The FWHM of GCE/CeO2 at 524 K–PANI/DAO (FWHM = 538 mV)
(Supplementary Fig. S4) was nearly 0.88 times lower than that
of GCE/CeO2 at 624 K–PANI/DAO (FWHM = 637 mV). This implies
that more number of electrons got transferred between DAO and
GCE/CeO2 at 524 K–PANI than that of GCE/CeO2 at 624 K–PANI
in presence of histamine [30]. From Paired t-test (two-tailed), it
was evident that GCE/CeO2 at 524 K–PANI/DAO and GCE/CeO2 at
624 K–PANI/DAO showed a significant difference in net current
before and after addition of histamine (p < 0.05). This was further
confirmed by calculating FWHM for GCE/CeO2 at 524 K–PANI/DAO
and GCE/CeO2 at 624 K–PANI/DAO electrode both in presence and
in absence of 0.45 mM of histamine. In both cases, the FWHM
was found to decrease by nearly 11 times in presence of his-
tamine indicating that the number of electrons transferred between
DAO and working electrode was increased in the presence of
histamine (Supplementary Fig. S5). The electron transfer rate con-
stant (Ks ) was calculated according to the formula, Ks = Ip /Q, where
Ip is the cathodic peak current and Q is the charge consumed.
The Ks of GCE/CeO2 at 524 K–PANI/DAO (Ks = 0.029 s−1 ) was cal-
culated to be 1.8 times higher than GCE/CeO2 at 624 K–PANI/DAO
(Ks = 0.016 s−1 ). It showed that the electron transfer between DAO
and CeO2 –PANI was very fast in GCE/CeO2 at 524 K–PANI/DAO
when compared to that of electron transfer in GCE/CeO2 at
624 K–PANI/DAO (Table 1).
The surface coverage (I* ) of histamine over GCE/CeO2 at
524 K–PANI/DAO and GCE/CeO2 at 624 K–PANI/DAO was cal-
culated (using supplementary Eq. (4)).It was observed that
GCE/CeO2 at 624 K–PANI/DAO (5.899 picomol cm−2 ) showed nearly
1.4 times higher surface coverage of histamine than GCE/CeO2 at
Epc (mV) Iw (␮A)  (␮ mol cm−2 ) Ks (s−1 )
−6
−985 8.95 2.783 × 10 0.029
−1037 8.54 1.346 × 10−9 0.016
524 K–PANI/DAO (4.195 picomol cm−2 ). Employing the Langmuir
adsorption isotherm [31] in combination to Nernst behaviour (Sup-
plementary Eqs. (5) and (6)), amount of histamine (H2 O2 ) adsorbed
onto GCE/CeO2 at 524 K–PANI/DAO bioelectrode for an each his-
tamine concentration at different time intervals were calculated
(Supplementary Fig. S6). Supplementary Fig. S7 shows the plot of
amount of adsorbed histamine (H2 O2 ) onto the surface of GCE/CeO2
at 524 K–PANI/DAO bioelectrode as a function of histamine concen-
tration. Langmuir linearized plot (Supplementary Fig. S8) shows
R2 value nearly equal to 0.96 also confirmed Langmuir adsorption
model. The calculated equilibrium constant (K) from the Langmuir
linearized plot was found to be 2.6 × 1012 M−1 .
Even though GCE/CeO2 at 624 K–PANI/DAO showed high sur-
face coverage of histamine, the electron transfer between DAO and
CeO2 –PANI was very slow. Owing to this further studies were car-
ried using GCE/CeO2 at 524 K–PANI/DAO.
4.5. Voltammetric detection of histamine based on modified
GCE/CeO2 at 524 K–PANI/DAO
Fig. 3(a) represents the cyclic voltammograms and (b) calibra-
tion plot of GCE/CeO2 at 524 K–PANI/DAO in 0.1 M PBS (pH 7.4)
with increasing concentration of histamine ranging from 0.45 to
1.05 mM. Owing to production of hydrogen peroxide (H2 O2 ) [32],
an increase in cathodic peak current (Ipc ) was observed with an
increase in histamine concentration. The cyclic voltammograms
showed well resolved peaks at concentration ranging from 0.45 to
1.05 mM of histamine with a significant peak-to-peak separation
and no shift in cathodic peak potential (Epc ) was observed. This
showed that DAO activity was not affected due to immobilization
at the GCE electrode surface.
A good sensitivity of 724.94 ␮A cm−2 mM−1 was observed
throughout the linear range (0.45–1.05 mM) with R2 of 0.96 (Sup-
plementary Fig. S9). The lower fractional saturation of DAO active
sites by histamine was attained in <57%. As a result of more active
sites for histamine to bind on to DAO and fast electron trans-
fer property of CeO2 at 524 K–PANI, the sensitivity got increased
(Supplementary Fig. S10). Levenberg–Marquardt fitting between
current density (J) and histamine concentration helps in determina-
tion of Jmax which was observed to be 1858.67 ␮A cm−2 indicating
high DAO loading and activity. From the Hill plot, the degree of
co-operativity between histamine and DAO was observed to be
positive as its value of 6.56 is greater than 1 and it showed that
there exists a good binding of histamine to active site of DAO that
results in the enhancement of the affinity for further incoming
histamine molecules (Supplementary Fig. S11). The other char-
acteristic parameter evaluated was KM whose value of 0.798 mM
indicates that there exist a good affinity between histamine and
DAO. Limit of detection (LOD) and limit of quantification (LOQ) of
GCE/CeO2 at 524 K–PANI/DAO were estimated to be 48.7 ␮M and
132.4 ␮M.
The biosensor efficiency of histamine detection was calculated
using Supplementary Eq. (7) and it was observed to be 321.3%
which indicated that the developed biosensor had a better abil-
ity to convert histamine to H2 O2 and thereby showed an increase
in sensitivity [34].
As histamine poisoning is considered to be a dose-dependent
poisoning, it is important to estimate the dose response. From the
dose response analysis (Supplementary Fig. S12), the histamine
 M.B. Gumpu et al. / Sensors and Actuators B 199 (2014) 330–338 335

Fig. 3. (a) Cyclic voltammogram and (b) calibration plot for varying concentration studies of histamine on modified GCE/CeO2 at 524 K–PANI/DAO in 0.1 M PBS (pH 7.4),
(c) amperometric response for varying concentration studies of histamine on modified GCE/CeO2 at 524 K–PANI/DAO in 0.1 M PBS (pH 7.4) at −0.1 V and (d) second order
derivative estimation.

concentration (EC 50) at which histamine biosensor showed 50%
of response was found to be 0.76 mM. This value was in good
match with KM (0.8 mM) indicating that there exists a good affinity
between histamine and DAO.
4.6. Amperometric studies
Fig. 3(c) shows the amperometric current–time response of
GCE/CeO2 at 524 K–PANI/DAO in 0.1 M PBS (pH 7.4) with increas-
ing concentration of histamine ranging from 0.45 to 0.9 mM. The
applied potential was set at −0.1 V for prolonged DAO stabil-
ity. From the second order derivative of amperometric response
(Fig. 3(d)), it was found that from 0.45 to 0.6 mM of histamine,
an increase in amperometric response with a fractional satura-
tion <55% was observed. When the histamine concentration was
increased further to 0.75 and 0.9 mM, a decrease in amperomet-
ric response with a fractional saturation lesser than 65% was
observed. These results showed that only up to 0.6 mM, the amper-
ometric response depends on histamine concentration, while the
amperometric response to histamine at concentrations greater
than 0.6 mM depends on DAO activity. The response time of the
designed histamine biosensor was found to be <1 s which was con-
sidered to be very fast when compared to other existing biosensors
for histamine. The comparison of characteristics of the developed
histamine biosensor with existing biosensors is shown in Table 2.
4.7. Repeatability and reproducibility (precision)
Reproducibility and repeatability of GCE/CeO2 at 524 K–PANI/
DAO with 0.45 mM histamine in 0.1 M PBS (pH 7.4) for
inter-assay (n = 7) and intra-assay (n = 7) were observed for every
1 h and the results were analysed using RSD and ANOVA. The RSD of
intra-assay and inter-assay were found to be 0.10 and 0.05 respec-
tively. One-way ANOVA results showed that the mean Ipc (n = 7)
for GCE/CeO2 at 524 K–PANI/DAO was not significantly different at
0.05 level (p > 0.05), indicating good embedding between DAO and
CeO2 –PANI. From Two-way ANOVA, it was found that the mean
difference between the reproduced measured mean Ipc (n = 7) was
not significantly different at 0.05 level (p > 0.05) signifying that the
fabrication technique was good.
4.8. Accuracy
Accuracy of GCE/CeO2 at 524 K–PANI/DAO towards histamine
was calculated by taking 0.6, 0.75 and 0.9 mM of histamine in 0.1 M
PBS (pH 7.4). For 0.6, 0.75 and 0.9 mM of histamine, the predicted
histamine concentration ± relative error (n = 3) were found to be
0.64 ± 0.07, 0.77 ± 0.1 and 0.92 ± 0.03 mM respectively. These low
relative errors implied good accuracy of measurement of histamine.
4.9. Interference studies
Interference studies were conducted to test the percentage
inhibition of DAO immobilized on CeO2 –PANI. Inhibition studies
were carried out using 0.01 mM of ascorbic acid (1.7 ␮g mL−1 ),
0.01 mM of urea (0.06 mg mL−1 ), 1 mM of sucrose (0.34 mg mL−1 ).
These are the common interfering agents that were used at phys-
iological concentrations, because these bio-chemicals can interact
with the developed bio-electrode to a greater extent while sens-
ing histamine [12,33]. Percentage inhibition was calculated using

Table 2
Comparison of characteristics of the developed histamine biosensor with existing biosensors.

Sensitivity Stability (%, days) KM (mM) Linear range LOD Response time References

– 20–50, 60 at 278 K – Up to 6000 ␮M 25 ␮M 1.5 min [35]

– – 0.27 Up to 6000 ␮M 50 ␮M – [6]
3.2 mA M−1 cm−2 – – 1–50 ␮M 0.5 ␮M – [36]
359.8 mA M−1 cm−2 50, 14 at 310 K 0.22 Up to 20 ␮M – – [37]
– – 1.3 25 to 4 mM – <3 s [38]
0.19 mA M−1 cm−2 60, 30 at 277 K – 10–100 ␮M 5 ␮M – [39]
6.1 & 3.5 mA M−1 cm−2 – – – 200–1000 mg L−1 & 20–100 mg L−1 – [40]
1.5 mA M−1 cm−2 87, 21 – 6–100 ␮M 6 ␮M – [41]
5.56 nA ppm−1 30 days at 278 K – 0–60 ppm 0.65 ppm <1 min [42]
– – – 0.2–1.6 ␮M 0.18 ± 0.01 ␮M – [43]
724.94 ␮A cm−2 mM−1 86% (18 days) at 298 K 0.798 450–1050 ␮M 48.7 ␮M <1 s Present work
336 M.B. Gumpu et al. / Sensors and Actuators B 199 (2014) 330–338

Table 3
Recovery characteristics of histamine biosensor.

Sample (60 mg of fish) Histamine added (mM) Mean histamine detected (mM) Recovery (%) RSD (%) (n = 3)

1 0.460 0.467 101.5 1.0

2 0.480 0.486 101.25 0.8
3 0.520 0.551 105.9 0.04
4 0.540 0.557 103.1 0.14
5 0.560 0.579 101.7 0.23
6 0.580 0.582 100.3 0.1
7 0.720 0.725 100.6 0.4
8 0.980 0.987 100.7 0.5

Supplementary Eq. (8). Only ascorbic acid showed percentage inhi-

bition of 5.5% while urea and sucrose showed percentage inhibition
<5% (Supplementary Fig. S13). Usually interference contribution
higher than 5% is not accepted. Further, it may change Imax and
KM , so a prediction band at 0.5 confidence level with an upper and
lower limit for measured net current was introduced to improve
the specificity of a histamine biosensor (Supplementary Fig. S14).
Selectivity for an interferent (SI %) should be always zero for any
biosensor application and it was calculated using Supplementary
Eq. (9). Selectivity studies were carried out with 0.45 mM of urea
(0.027 mg mL−1 ), 0.45 mM of sucrose (0.15 mg mL−1 ) and 0.45 mM
of ascorbic acid (0.07 mg mL−1 ) in 0.1 M PBS (pH 7.4). The relative
selectivity of histamine detecting biosensor for histamine, urea,
ascorbic acid and sucrose were found to be 100%, 9.6%, 9.2% and
8.2% respectively (Supplementary Fig. S15). These results showed
that GCE/CeO2 at 524 K–PANI/DAO electrode was selectively spe-
cific towards histamine.
Generally permeability of interferent (P [I]) [34] should be low Fig. 4. Comparative study of specificity studies of histamine, cadaverine and com-
and permeability of substrate (P [S]) should be high for any biosen- bination of histamine, cadaverine.
sor application. The permeability (using supplementary Eqs. (10)
and (11)) of 0.45 mM of histamine through biofilm (66 mg mL−1 )
was estimated to be 586.79% which was four times higher than
0.45 mM of urea, 0.45 mM of sucrose and 0.45 mM of ascorbic selective based on the potential alone. As this sensor was calibrated
acid (Supplementary Fig. S16), indicating that histamine permeates for the potential of −985 mV, the developed sensor is selective to
through biofilm faster than all the other interfering agents. histamine, inspite of the presence of cadaverine.
The dry stability of the developed biosensor was checked for 18 Fig. 4
days at room temperature. The stability was evaluated using Sup-
plementary Eq. (12) and was observed to be 86% (Supplementary
Fig. S17). 5. Conclusion

The influence of ceria nano-interface in improving the efficiency

4.10. Verification of recovery of histamine sensor
of GCE working electrode while sensing histamine was successfully
investigated. The figure of merits of the developed biosensor with
To test the recovery of developed biosensor, 100 ␮L of histamine
nano-interface namely sensitivity, recovery, linearity, response
extract (2 mg mL−1 ) along with histamine concentrations of 0.46,
time and stability were found to be 724.94 ␮A cm−2 mM−1 ,
0.48, 0.52, 0.54, 0.56, 0.58, 0.72 and 0.98 mM was added and detec-
100.3–105.9%, 0.45–1.05 mM, <1 s and 86% (up to 18 days) respec-
tion was done in triplicates. Recoveries of histamine from giant fish
tively. The faster response time of <1 s indicated the rapid sensing
prawn ranged from 100.3 to 105.9% with an RPES of 0.125% and
process and found to be superior to the existing histamine
RMSCEV of 0.004% with an RSD of <0.23% which was within the
biosensors. Significantly, the developed biosensor showed surface
acceptable limit (Table 3). Recovery within the range from 100.3 to
controlled reaction and hence helped to estimate exactly the con-
105.9% indicated that the developed bio-electrode had the ability
centration of histamine present in the tiger prawn samples.
to overcome potential interferents.
In order to prove the selectivity of histamine in the recov-
ery studies, selectivity was evaluated using, cadaverine (biogenic
amine). To perform the selectivity studies, histamine, cadaverine Acknowledgements
and a combination of histamine, cadaverine was taken. Among
these combinations, histamine, cadaverine showed a reduction The authors are grateful to the Nano Mission Council, India
potential at −985 mV, −650 mV whereas combination of histamine (No. SR/NM/PG-16/2007).The authors also wish to express their
and cadaverine showed two reduction peaks (−985 mV for his- sincere thanks to Department of Science & Technology, New
tamine and −650 mV for cadaverine) as shown in Fig (4). In this, Delhi, India for their financial support (DST/TSG/PT/2008/28), and
even though cadaverine is present along with histamine, the devel- (SR/FST/ETI-284/2011(C)) for CH electrochemical analyzer. We also
oped sensor showed significantly different peaks for both with acknowledge SASTRA University, Thanjavur for extending infras-
same magnitude of current. This says that the developed sensor is tructural support to carry out the study.
 M.B. Gumpu et al. / Sensors and Actuators B 199 (2014) 330–338
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.snb.2014.04.009.
References
[1] L. Maintz, N. Novak, Histamine and histamine intolerance, Am. J. Clin. Nutr. 85
(2003) 1185–1196.
[2] M.L. Latorre-Moratalla, T. Veciana-Nogues, S. Bover-Cid, M. Garriga, T.
Aymerich, E. Zanardi, A. Ianieri, M.J. Fraqueza, L. Patarata, E.H. Drosinos, A.
Laukova, R. Talon, M.C. Vidal-Carou, Biogenic amines in traditional fermented
sausages produced in selected European countries, Food Chem. 107 (2008)
912–921.
[3] V. Venugopal, Biosensors in fish production and quality control, Biosens. Bio-
electron. 17 (2002) 147–157.
[4] Dietary Management of Histamine Intolerance, Vickerstaff Health Services Inc.
Fact sheet, 2014, pp. 1–9.
[5] U.D. Kantaria, R.H. Gokani, Quality and safety of biogenic amines, Int. J. Res.
Pharm. Biomed. Sci. 2 (2011) 1461–1468.
[6] P. Bouvrette, K.B. Male, J.H.T. Luong, B.F. Gibbs, Amperometric biosensors for
diamine using diamine oxidase purified from porcine kidney, Enzyme Microb.
Technol. 20 (1997) 32–38.
[7] H.C. Chen, Y.R. Huang, H.H. Hsu, C.S. Lin, W.C. Chen, C.M. Lin, Y.H. Tsai,
Determination of histamine and histamine forming bacteria in tuna dumpling
implicated in a food-borne poisoning, Food Control. 21 (2010) 13–18.
[8] M. Nanda, Tamim, W. Lloyd, Bennett, A. Timothy, Shellem, A. John, Doerr,
High performance liquid chromatographic determination of biogenic amines
in poultry Carcasses, J. Agric. Food Chem. 50 (8) (2002) 5012–5015.
[9] C.-F. Chow, M.H.W. Lam, W.-Y. Wong, Design and synthesis of heterobimetallic
Ru(II)–Ln(III) complexes as chemodosimetric ensembles for the detection of
biogenic amine odorants, Anal. Chem. 85 (17) (2013) 8246–8253.
[10] A.M. Ayesha, M.N. Ibraheim, A.E. El-Hakim, E.A.H. Mostafa, Exploring the con-
tamination level by biogenic amines in fish samples collected from markets in
Thuel-Saudi Arabia, Afr. J. Microbiol. Res. 6 (2011) 1158–1164.
[11] M. Peeters, P. Csipai, B. Geerets, A. Weustenraed, B. Van Grinsven, R. Thoelen,
J. Gruber, W. De Ceuninck, T.J. Cleij, F.J. Troost, P. Wagner, Heat-transfer-
based detection of l-nicotine, histamine and serotonin using molecularly
imprinted polymers as biomimetic receptors, Anal. Bioanal. Chem. 405 (2013)
6453–6460.
[12] Y. Itoh, R. Oishi, M. Nishibori, K. Saeki, Characterization of histamine release
from the rat hypothalamus as measured by in vivo microdialysis, J. Neurochem.
56 (1991) 769–774.
[13] C.M. Kneow, F.A. Bakar, A.B. Salleh, L.Y. Heng, R. Wagiram, L.S. Bean, An amper-
ometric biosensor for the rapid assessment of histamine level in tiger prawn
(Penaeus monodon) spoilage, Food Chem. 105 (2007) 1636–1641.
[14] D. Telsing, A. Terzic, T. Krenn, V. Kassarnig, K. Kalcher, A. Ortner, Development
of a voltametric amine oxidase-modified biosensor for the determination of
biogenic amines in food, Int. J. Electrochem. Sci. 7 (2012) 6893–6903.
[15] M.A.A. Lomillo, O.D. Renedo, P. Matos, M.A.A. Martinez, Disposable biosen-
sors for determination of biogenic amines, Anal. Chim. Acta 665 (2010)
26–31.
[16] C.P. McMahon, G. Rocchitta, S.M. Kirwan, S.J. Killoran, P.A. Serra, J.P. Lowry, R.D.
O’Neill, Oxygen tolerance of an implantable polymer/enzyme composite gluta-
mate biosensor displaying polycation-enhanced substrate sensitivity, Biosens.
Bioelectron. 22 (2007) 1466–1473.
[17] E. Bitziou, D. O’Hare, B.A. Patel, Simultaneous detection of pH changes and his-
tamine release from Oxyntic glands in isolated stomach, Anal. Chem. 80 (2008)
8733–8740.
[18] M.M. Rahman, A.J.S. Ahammad, J.H. Jin, S.J. Ahn, J.J. Lee, A comprehensive review
of glucose biosensor based on nanostructured metal-oxides, Sensors 10 (2010)
4855–4886.
[19] P.R. Solanki, A. Kaushik, V.V. Agrawal, B.D. Malhotra, Nanostructured metal
oxide-based biosensors, NPG Asia Mater. 3 (2011) 17–24.
[20] U.N. Devi, J.B.B. Rayappan, B.G. Jeyaprakash, Confirmation of ceria–polyaniline
nanostructure: an optical approach, J. Appl. Sci. 12 (2012) 1750–1753.
[21] J. Stejskal, R.G. Gilbert, Polyaniline preparation of a conducting polymer (IUPAC
technical report), Pure Appl. Chem. 74 (5) (2002) 857–867.
[22] S. Benramache, F. Chabane, B. Benhaoua, F.Z. Lemmadi, Study on the correla-
tion between crystallite size and optical gap energy of doped ZnO thin film, J.
Semicond. 34 (2013) 023001-1–023001-4.
[23] I. Šeděnková, M. Trchová, N.V. Blinova, J. Stejskal, In-situ polymerized polyani-
line films. Preparation in solutions of HCl, sulfuric or phosphoric acid, Thin solid
films 515 (2006) 1640–1646.
[24] E. Kumar, P. Selvarajan, D. Muthuraj, Preparation and characterization of
polyaniline/cerium oxide (CeO2 ) nanocomposite via in situ polymerization, J.
Mater. Sci. 47 (2012) 7148–7156.
[25] D. Valechha, S. Lokhande, M. Klementova, J. Subrt, S. Rayalu, N. Labhsetwar,
Study of nanostructured ceria for catalytic CO oxidation, J. Mater. Chem. 21
(2011) 3718–3725.
[26] V. Kumar, S. Singh, S. Aggarwal, Synthesis of 1-dimensional polyani-
line nanofibers by reverse microemulsion, Colloid. Polym. Sci. 287 (2009)
1107–1110.
337
[27] J. Coates, Interpretation of infrared spectra, A practical approach, in: R.A Meyers
(Ed.), Encyclopedia of Analytical Chemistry, John Wiley & Sons Ltd., Chichester,
2002, pp. 10815–10837.
[28] A. Burger, Mechanisms of release of biogenic amines, J. Med. Chem. 10 (4) (1967)
744.
[29] N. Nesakumar, K. Thandavan, S. Sethuraman, U.M. Krishnan, J.B.B. Rayappan,
An electrochemical biosensor with nanointerface for lactate detection based on
lactate dehydrogenase immobilized on zinc oxide nanorods, J. Colloid Interface
Sci. 414 (2014) 90–96.
[30] A.L. Eckermann, D.J. Feld, J.A. Shaw, T.J. Meade, Electrochemistry of redox active
self-assembled monolayers, Coord. Chem. Rev. 254 (2010) 1769–1802.
[31] M. Jacob, J. Goran, Stevenson, Electrochemical behavior of flavin adenine dinu-
cleotide adsorbed onto carbon nanotube and nitrogen-doped carbon nanotube
electrodes, Langmuir (2013), http://dx.doi.org/10.1021/la403020y.
[32] K.B. Male, J.H.T. Luong, P. Bouvrette, B.F. Gibbs, Amperometric biosensors for
diamine using diamine oxidase purified from porcine kidney, J. Food Sci. 61
(1996) 1012–1016.
[33] S. Shanmugam, K. Thandavam, S. Gandhi, S. Sethuraman, J.B.B. Rayappan, U.M.
Krishanan, Development and evaluation of a highly sensitive rapid response
enzymatic nanointerfaced biosensor for the detection of putrescine, R. Soc.
Chem. 136 (2011) 5234–5240.
[34] S.A. Rothwell, S.J. Killoran, R.D. O’Neill, Enzyme immobilization strategies and
electropolymerization conditions to control sensitivity and selectivity param-
eters of a polymer-enzyme composite glucose biosensor, Sensors 10 (2010)
6439–6462.
[35] K.B. Male, P. Bouvrette, J.H.T. Luong, Amperometric biosensors for total his-
tamine, putrescine and cadaverine using diamine oxidase, J. Food Sci. 61 (1996)
1012–1016.
[36] R. Draisci, G. Volpe, L. Lucentini, A. Cecilia, R. Fedrico, G. Palleschi, Determination
of biogenic amines with an electrochemical biosensor and its application to
salted anchovies, Food Chem. 62 (1998) 225–232.
[37] M. Wimmerova, L. Macholan, Sensitive amperometric biosensor for the deter-
mination of biogenic and synthetic amine using pea seedlings amine oxidase:
a novel approach for enzyme immobilization, Biosens. Bioelectron. 14 (1999)
695–702.
[38] K. Zeng, H. Tachukawa, Z. Zhu, V.L. Davidson, Amperometric detection of his-
tamine with a methylamine dehydrogenase polypyrrole-based sensor, Anal.
Chem. 72 (10) (2000) 2211–2215.
[39] D. Compagnone, G. Isoldi, D. Moscone, G. Palleschi, Amperometric detection of
biogenic amines in cheese using immobilized diamine oxidase, Anal. Lett. 34
(2006) 841–854.
[40] O. Norazlina, A.B. Fatimah, B.S. Abu, Y.H. Lee, A. Rahman, A preliminary
investigation on a histamine biosensor constructed from diamine oxidase
immobilised onto an oxygen probe, Malaysia J. Anal. Sci. 10 (1) (2006)
137–142.
[41] D. Carelli, D. Centonze, C. Palermo, M. Quinto, T. Rotunno, An interfernce free
amperometric biosensor for the detection of biogenic amines in food products,
Biosens. Bioelectron. 23 (2007) 640–647.
[42] C.M. Kneow, A.B. Fathima, A.B. Salleh, Y.H. Lee, W. Rahman, S.B. Low, An
amperometric biosensor for rapid assessment of histamine level in tiger prawn
(Pneaeus monodon) spoilage, Food Chem. 105 (2007) 1636–1641.
[43] M.A. Alonso-Lomillo, O. Dominguez-Renedo, P. Matos, M.J. Arcos-Martinez, Dis-
posable biosensors for determination of biogenic amines, Anal. Chim. Acta 665
(2010) 26–31.
Biographies
Manju Bhargavi Gumpu received BTech Biotechnology in 2011, from Jawaharlal
Nehru Technological University, Anantapur and MTech in Medical Nanotechnology
from SASTRA University, Thanjavur in 2013. She is currently working as Research
Scholar in the Centre for Nanotechnology and Advanced Biomaterials (CeNTAB)
and School of Electrical and Electronics Engineering, SASTRA Univeristy, Thanjavur,
India. Her current research interests are fabrication of nanostructured electrochem-
ical enzyme based biosensor for water quality analysis. Her research areas include
biosensors, Electrochemistry, Enzyme technology
Noel Nesakumar gained a BE in Bio-medical Instrumentation at Anna University
in 2006 and a MTech degree in Medical Nanotechnology from SASTRA University
in 2010. Currently he is a Ph.D. candidate at the Centre for Nanotechnology and
Advanced Biomaterials (CeNTAB) and School of Electrical & Electronics Engineering,
SASTRA University, Thanjavur, India. His area of focus on a researcher is the devel-
opment of electrochemical biosensors for early detection of pesticide residues in
rice and guar gum.
Swaminathan Sethuraman is the Director, Centre for Nanotechnology and
Advanced Biomaterials (CeNTAB). He obtained his Bachelor’s degree in Chemical
Engineering from Bharathidasan University, Trichy, India in 1998 and PhD, in Chem-
ical and Biological Engineering from Drexel University, Philadelphia, USA, in 2005.
He then worked as a research associate at University of Virginia, Charlottesville, USA
in Orthopaedic Surgery. His research areas of interest include regenerative medicine,
stem cells, drug delivery system.
Uma Maheswari Krishnan is a Professor at the Centre for Nanotechnology &
Advanced Biomaterials (CeNTAB) and School of Chemical and Biotechnology, SAS-
TRA University, India. She received her PhD degree in Applied Chemistry from
338 M.B. Gumpu et al. / Sensors and Actuators B 199 (2014) 330–338

Bharathiar University, India in 2000 and worked as research associate at Univer-
sity of Texas, USA till 2002 and had training on thin film processing, Clean room
processes at University of Arkansas, USA. Since, 2003 she is in SASTRA University.
Her present field of interest includes smart drug delivery systems, nanobiosensors,
electrophysiology and mesoporous materials.
currently working as a Professor in School of Electrical & Electronics Engineering and
Centre for Nanotechnology & Advanced Biomaterials (CeNTAB), SASTRA University.
His current research interests include lattice dynamics, fabrication of thin film based
chemical & biosensors and functional nanomaterials. He is also working in the field
of embedded systems and steganography.

John Bosco Balaguru Rayappan received his MSc and PhD degrees in Physics from
Bharathidasan University, Tiruchirapalli, India in 1996 and 2003, respectively. He is