Sensors and Actuators B 177 (2013) 871878

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Sensors and Actuators B: Chemical

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

Computer-aided sensor design and analysis of thiocarbohydrazide in biological

matrices using electropolymerized-molecularly imprinted polypyrrole modied
pencil graphite electrode
Azizollah Nezhadali a,b, , Raham Shadmehri b
a
b

Department of Chemistry, Payame Noor University (PNU), Mashhad, Iran

Department of Chemistry, Payame Noor University, P.B. 19395-4697, Tehran 19569, Iran

a r t i c l e

i n f o

Article history:
Received 9 September 2012
Received in revised form
26 November 2012
Accepted 27 November 2012
Available online 6 December 2012
Keywords:
Thiocarbohydrazide
Pencil graphite
Polypyrrole
Molecularly imprinted polymer
Computational study

a b s t r a c t
In this work, molecularly imprinted polymer (MIP) of thiocarbohydrazide (TCH) was prepared through
electropolymerization and electrodeposition of pyrrole on a pencil graphite electrode. The ability of the
fabricated MIP to act as a sensor of determining TCH in a few biological matrices was investigated. Preparation of MIP and quantitative measurements were performed by cyclic voltammetry (CV) and differential
pulse voltammetry (DPV), respectively. Several important parameters controlling the performance of
the polypyrrole and the method efciency were investigated and optimized. The molecularly imprinted
layer exhibited selectivity and sensitivity toward TCH. The selection of monomer was performed computationally using ab initio calculations. A computational study based on restricted MP2 was performed
to evaluate the templatemonomer geometry and interaction energy in the prepolymerization mixture.
The calibration curve demonstrated linearity over a concentration range of 0.56 mM with a correlation
coefcient (r2 ) of 0.9805. The detection limit of TCH was obtained 5.0 105 M.
2012 Elsevier B.V. All rights reserved.

1. Introduction
TCH (1,3-diaminothiourea), one of the simplest hydrazine
derivatives of thiocarbonic acid, is an important chemical intermediate in organic synthesis [1] and it is an important analytical
reagent of tremendous utility. TCH posses anticarcinogenic and
antibacterial properties. It exhibits toxicity toward house-y comparable to that of DDT. The fungicidal properties of TCH have also
been tested against different species. TCH is also used as an additive to prevent the excessive loss of cellulose during the alkaline
work-up of wood pulp [2].
A new trend is to develop novel sensors based on molecularly
imprinted polymers (MIPs), which represent a class of materials possessing tailor-made afnity and selectivity for the target
molecule [3]. On the other hand, molecular imprinting is a technique for the creation of highly efcient synthetic molecular
receptors [4]. Wulff et al. rst demonstrated the method by designing a polymeric receptor using reversible covalent bonds between
monomer-print molecules [5] and the method much expanded by
the work of the group of Arshady and Mosbach in 1980s [6]. In
recent years MIP technology matured to a valuable complementary

Corresponding author at: Department of Chemistry, Payame Noor University

(PNU), Mashhad, Iran. Tel.: +98 5118683887; fax: +98 5118683001.
E-mail address: aziz nezhadali@pnu.ac.ir (A. Nezhadali).
0925-4005/$ see front matter 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.snb.2012.11.103

concept for biological recognition with increased applicability in

analytical chemistry, offering a cheap and versatile platform for
creating a polymer matrix with a molecule-specic recognition
properties with applications ranging from purication of racemic
mixtures [4,7] and separation media [811] to chemical sensing
[4,1214], catalytic control of complex chemical reactions [4], catalytic synthesis [15,16] and antibody and enzyme mimics [17].
Their stability, ease of preparation and low cost [1820] for most of
the target analytes make them attractive for numerous applications
[18].
MIPs can be prepared according to a number of approaches
that are different in the way the template is linked to the
functional monomer and subsequently to the polymeric binding sites. Thus, the template can be linked and subsequently
recognized by reversible covalent bonds, metal ion coordination [18] or non-covalent bonds [18,2124]. Some preparation
methods of molecular imprinting polymers (MIPs) have been
reported including chemical grafting, soft lithography technique,
molecular self-assembled approach, electropolymerization [25]
and thermally by the addition of heat and photochemically by
exposure to UV [26] and [27] radiation. Electropolymerization
has been successfully utilized for the preparation of electroactive
and electro-inactive polymers on a variety of conductive surfaces
including platinum, indium tin oxide, carbon, oxidizable metals,
stainless steel, and zinc as examples [28]. Molecular imprinting
has been approached using the electrosynthesis of conducting

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polymers through galvanostatic, potentiostatic and cyclic voltammetric methods. These methods provide a simple and rapid
technique of controlling the thickness of the conductive polymer
lm grown adherent to a transducer of any size and shape [29].
Polymers such as polyphenols, poly(aminophenylboronate),
poly(phenylenediamine-co-aniline),
poly(phenylenediamine)
[30], polypyrrole [25,2933], polyaniline [30], poly(3-thiophene
acetic acid) [34], poly(o-aminophenol) [35,36] and others have
been used to prepare MIPs, recently. The carbon electrodes
employed frequently for voltammetric measurements. These electrodes have the following advantages [37]: (a) a polarized potential
domain similar to or, at the negative limit, wider than the domains
at platinum or gold electrodes in aqueous solutions; (b) a variety of
easy methods of surface modication by electrochemical, chemical
and mechanical treatment; (c) high stability at slightly degraded
surface activity; and usable as disposable electrochemical tool.
In sensor application, polypyrrole is one of the most explored
conducting polymers. It has been employed in the reagent phase of
a number of potentiometric, amperometric, conductometric and
optical sensors for chemical species [29]. Electrosynthesis of MIP
can be carried out in aqueous or in organic media, but aqueous
media are usually chosen to synthesize imprinted polymers [38].
The development of a MIP still relies very much on experiments,
including the preparation of the media and the use of adsorption
to evaluate the media. This makes the design and development
of MIP time-consuming [39]. So, we used computational quantum
chemistry as well as reported before [3942] to evaluate the best
functional monomer for preparation of TCH MIP sensor.
An MIP was prepared by electropolymerization of pyrrole onto
a stainless steel frit, using ochratoxin A as the template, in order to
make a micro-solid phase preconcentration device [25]. Shustina
et al. [43] reported the chromatographic determination of TCH in
aqueous solutions. The use of MIP modied pencil graphite electrode prepared by electropolymerization of pyrrole to determine
the TCH using differential pulse voltammetry was reported for
the rst time in this work. TCH was chosen as template molecule
because of its popularity and electroactivity [2].
2. Experimental
2.1. Chemicals and reagents
Sodium perchlorate (99102%), boric acid (99.9999 Suprapur), acetic acid (99.5%), pyrrole (97%), were purchased from
Merck (Darmstadt, Germany). Thiourea (>97%, Fluka, Steinheim,
Germany), TCH (98%, Aldrich, St. Louis, MO, United States), ticlopidine hydrochloride (99%, Fluka, Steinheim, Germany), camphor
(95%, SigmaAldrich, St. Louis, MO, United States), thioacetamide
(>98%, Scharlau, Sentmenat, Barcelona, Spain), sodium hydroxide
(98%, Lobachemie, Mumbai, India), and other reagents were commercially available as analytical grade and used without further
purication. Stock solutions of TCH and buffer solution, were prepared by using locally made pure deionized water. Freshly solutions
of TCH were prepared each day owing to its low stability.

microscopy (SEM) in a LEO 1450VP (Cambridge, England) microscope.

2.3. Hardware and software
All computations were carried out on computers with 2 GB
memory and 150 GB hard disk. Molecular structures were drawn by
ChemBioDraw Ultra and ChemBio3D Ultra 2008 (Cambridgesoft).
Quantum calculations have been carried out using Gaussian 09 [44]
software. The optimized 3-dimensional molecular structures were
produced by Gauss View 5.0 (Gaussian, Inc.).
2.4. Preparation of MIP and NIP electrodes
Pencil graphite electrode (Owner, HB, 0.7 mm diameter, Korea)
was washed by water and dried at room temperature before
experiments. Then, pencil graphite electrode was immersed in the
polymerization solution. The MIP was obtained by electrodeposition on the surface of the pencil graphite electrode using cyclic
voltammetry in the potential range between 1.0 and 1.9 V during 25 cycles (scan rate 100 mV/s) in aqueous solution of 0.1 M
NaClO4 , 0.02 M pyrrole and 0.010 M TCH. After the electropolymerization process, the embedded TCH was extracted to give a surface
complimentary in shape and functionality to the original template
TCH. The imprinted polymer was conditioned in BrittonRobinson
buffer solution using differential pulse potential scanning between
0.0 and 1.0 V. This process provided to remove the template from
inside the polymer. A control electrode (non-imprinted polymer
modied electrode, NIP) was prepared under the same experimental conditions but without adding TCH, to check the reliability of
the measurements. All of the MIP and NIP measurements in this
work were done triplicates.
2.5. Electroanalytical measurements
Differential pulse voltammetric measurements were carried out
in a three-electrode cell, in BrittonRobinson buffer at pH 5.0. The
BrittonRobinson buffer was prepared by adjusting the pH of a solution of 0.04 M H3 BO3 , 0.04 M H3 PO4 , and 0.04 M CH3 COOH to 5.0
by 0.2 M NaOH solution monitoring by pH-meter. Current measurements were performed using differential pulse voltammetry (DPV)
in the potential range between 0.0 and 1.0 V. To record differential pulse voltammograms, the following instrumental parameters
were used: step potential 8 mV, modulation amplitude 50 mV and
scan rate 16 mV/s. All electroanalytical measurements were made
at room temperature.
2.6. Sample preparation in biological matrices
Twenty milligrams of separate samples of beef, camel meat,
chicken, lamb meat and sh meat hydrolyzed by 50 ml of an
ethanol/HCl 1 M mixture (1:1 v/v), based on the work of Curcio
et al. [45]. The samples were then neutralized and the nal volumes were 100 ml. The neutralized sample matrices were spiked
with TCH solution. The human blood serum sample was analyzed
by standard addition technique applied without any pretreatment.

2.2. Instrumentation
3. Results and discussion
Electrochemical studies were performed using Autolab PGSTAT
12 potentiostatgalvanostat controlled by GPES 4.9 software (Ecochemie, The Netherlands). Three-electrode system was used for all
measurements; a pencil graphite electrode as the working electrode and a platinum auxiliary electrode. All measurements carried
out with a silver/silver chloride reference electrode. Metrohm 744
pH-meter (Metrohm, Switzerland) was employed for pH measurements. Surface evaluations were performed by scanning electron

3.1. Selection of the functional monomer

Formation of a complex between the template molecule and
functional monomers by self-assembly process is the rst step in
the preparation of MIP. The monomer that can interact with the
template most intensively will give the complex the highest stability. This will facilitate the subsequent preparations including

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873

polymerization and the formation of the cavity of a complementary

structure to that of the template molecule [39]. The selection of suitable functional monomers for imprinting process may be aided by
molecular modeling and computational methods [41]. Monomers
with the highest binding scores are subsequently selected to produce full scale MIPs with hopefully superior recognition properties.
In this work, ve commercially available functional monomers
which were reported to be electrochemically polymerized, i.e. thiophene [4648], 3-methylthiophene [46], pyrrole [25,2931,33],
1-methylpyrrole [49,50] and furan [51] were selected. The electronic stabilization energy, E, was calculated by the equation
[42]:
E = E(templatemonomer complex) E(template)
E(monomer)
A complex with higher E value predicts the higher binding
energy following most stable lowest energy conformation [41]. Correlation can be added as a perturbation from the HartreeFock (HF)
wave function. This is called MllerPlesset (MP) perturbation theory. In mapping the HF wave function onto a perturbation theory
formulation, HF becomes a rst-order perturbation. Thus, a minimal amount of correlation is added by using the second order MP2
method [52]. So, we chose the restricted MP2 method to compute
electronic energies. The applied basis set was 631 G. Table 1 shows
the chosen monomers and their computed energies in the absence
and in the presence of TCH in gas phase. The results indicate that
the TCHpyrrole complex stabilization energy is greater than the
other evaluated functional monomers. So, we selected pyrrole to
prepare the MIP sensor.
3.2. Electropolymerization of molecularly imprinted polypyrrole
The electrochemical behavior of pyrrole was investigated in
aqueous solution of 0.1 M NaClO4 using potential cycling between
1.0 and +1.9 V (versus Ag/AgCl). Electrooxidation of pyrrole
monomer occurs at the anode, and the resulting polymer deposits
onto the surface of pencil graphite electrode. Fig. 1 demonstrates
four cycles obtained by electropolymerization of pyrrole on the
surface of pencil graphite electrode. For imprinted electropolymerizations, TCH was added to the electrochemical cell at a
concentration of 10 mM.
During the electrodeposition of the conductive polymer, TCH
template molecules are trapped in the polymer matrix as a result of
the ability of these molecules to interact with the pyrrole units. The
Schematic representation of imprinting and removal of TCH from
TCH imprinted polypyrrole modied pencil electrode is shown in
Fig. 2. The sulfur atom in the C S group of the TCH molecule could
form a hydrogen bond [53] with the hydrogen atom in the N H

Fig. 1. Four cycles obtained by electropolymerization of pyrrole on the surface of

pencil graphite electrode in an aqueous solution of 0.1 M NaClO4 and 0.02 M pyrrole. The rst peak (A) is obtained by the rst CV scan and is attributed to the
electropolymerization of pyrrole. Peak (B) is optained by the second CV scan.

group of the pyrrole units (Fig. 2a). Hydrogen bonding could occur
between the hydrogen in the N H and/or NH2 group of TCH structure and the nitrogen of the N H group of pyrrole units (Fig. 2a).
There may be a hydrogen bond between the nitrogen of N H
and/or NH2 groups of TCH and hydrogen of N H group of pyrrole
units (Fig. 2a). To remove the template from inside the polypyrrole
lm, the imprinted polymer was conditioned in BrittonRobinson
buffer solution (pH = 5) using differential pulse scanning potential
between 0.0 and 1.0 V in this work.
3.3. Effect of pH
The effect of pH on the peak potential and peak current was studied using 2 mM TCH in BrittonRobinson buffer from pH 3.0 to 8.0
(Fig. 3). The optimal pH (5) was obtained with the polypyrrole modied pencil graphite electrode according to the highest response of
the current for the constant TCH concentration (2 mM).
3.4. Electrochemical oxidation of TCH
The electrochemical oxidation of TCH has been evaluated by
Mamatha et al. [2]. By the suggested mechanism of electrochemical oxidation of TCH based on the work of Mamatha et al. [2],
an oxidation reaction happens on the graphite electrode surface
and releases two electrons. The mentioned electrochemical reaction produces an unstable intermediate which could be oxidized
by a chemical oxidation reaction which releases an electron. The
latter electron releasing would lead to another releasing reaction
according to the unstable positive charged intermediate produced
on the last oxidation process [2].

Table 1
The chosen monomers and their computed energies in the absence and in the presence of TCH in gas phase.
Molecules
b

TCH
Furan
Pyrrole
1-Methylpyrrole
Thiophene
3-Methylthiophene
TCHfuran
TCHpyrrole
TCH1-methylpyrrole
TCHthiophene
TCH3-methylthiophene
a
b

1 Hartree = 2625.5 kJ mol1 .

TCH: thiocarbohydrazide.

E (Hartree)

E (Hartree)

E (kj/mol)a

657.0398947
229.0001464
209.1871613
248.2907469
551.6025606
590.7186053
886.0521442
866.2392034
905.3404299
1208.6489635
1247.7693611

0.0121031
0.0121474
0.0097883
0.0065082
0.0108611

31.7766890
31.8929987
25.6991816
17.0872791
28.5158180

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Fig. 2. Schematic representstion of (a) imprinting and (b) removal of thiocarbohydrazide from thiocarbohydrazide imprinted polypyrrole modied pencil graphite electrode.

TCH, the lms were grown in solutions of constant concentration

of TCH and varying pyrrole concentrations in the range of 550 mM
by cycling potential between 1.0 and 1.9. Fig. 4 shows the result
of the mentioned comparison. The monomer concentration should
be proportional to the thickness of the deposit and amount of the

Fig. 3. The effect of pH on the MIP modied pencil graphite electrode response for
2 mM thiocarbohydrazide in Britton-Robinson buffer solutions.

3.5. Effect of monomer concentration

Although the synthesis of non-covalent MIPs is relatively easy
and a large library of functional monomers is available, optimization of MIP formulation implies the selection of the best functional
monomer and the most adequate polymerization for each selected
template, among other reagents [54].
The effect of monomer concentration on the analytical behavior
of the sensor during polymerization was determined. To determine
the effect of monomer concentration on the response of MIP to

Fig. 4. The effect of the monomer concentration on the MIP modied pencil graphite electrode for 2 mM thiocarbohydrazide. The response was measured
through DPV in 2 mM thiocarbohydrazide in Britton-Robinson buffer medium at pH
5.0.

Fig. 5. The SEM micrographs of: (a) the molecularly imprinted and (b) nonimprinted polypyrrole - pencil graphite electrode prepared using 25 CV cycles by
immersing the unmodied electrode in 0.01 M and 0 M solutions of TCH, respectively; Both containing 0.02 M pyrrole and 0.1 M NaClO4 .

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875

Fig. 6. The effect of number of cycles to MIP- modied pencil graphite electrode
response in Britton-Robinson buffer medium at pH 5.0.

imprint molecule in the polymeric matrix. The response of the

MIP electrode to TCH was found to increase with increasing pyrrole concentration up to 20 mM. According the results (Fig. 4) the
optimal monomer concentration was chosen 20 mM for the next
experiments.
Fig. 5a and b shows scanning electron micrographs of polypyrrole formed on pencil graphite electrodes at optimal conditions that
investigated in this work. The optimum conditions are: 0.01 M template, 25 CV cycles, 0.02 M pyrrole and 0.1 M supporting electrolyte
(NaClO4 ). As Shown in Fig. 5 the coatings are porous and grainy. It is
believed that the porosity and grain size of the electropolymerized
polypyrrole are dependent on the supporting electrolyte and other
experimental conditions, used in the method [55].
3.6. Effect of the electropolymerization cycles
The optimum number of CV cycles used to form the polypyrrole
layer on the pencil graphite electrode was evaluated by comparison of MIP electrode responses to the constant TCH concentration
prepared by different number of CV cycles. The medium applied to
fabricate the MIP electrodes and optimize the number of CV cycles
was an aqueous solution containing 0.1 M NaClO4 , 0.01 M TCH and
0.02 M pyrrole. The applied cycling potential was between 1.0
and 1.9 V. The response of the MIP electrodes to TCH was found
to increase with increasing the number of cycles up to 25 (Fig. 6).
The thickness of the polypyrrole was obtained 197 nm by using the
charge passed through the electropolymerization process [56].
3.7. Effect of the template concentration
The quantity and quality of the MIP recognition sites is a direct
function of the mechanisms and extent the monomertemplate

Fig. 8. The most stable geometry of template-monomer complex in the prepolymerization mixture, obtained by the restricted MP2 method calculation in gas phase:
TCH(pyrrole)3 .

interactions present in the pre-polymerization mixture [57]. The

effect of the template concentration on the lm electrodeposition
is shown in Fig. 7. The response of the MIP electrode to TCH standard
solution was found to increase with increasing TCH concentration
up to 10 mM during the lm electrodeposition (Fig. 7).
3.8. Optimization of geometries and calculation of energies of
templatemonomer complexes in the prepolymerization mixture
The computational method developed was based on the
use of an ab initio method to evaluate the stability of the
templatemonomer complexes in the prepolymerization mixture
of the template and monomer and their electronic stabilization
energy relative to isolated fragments. The following equation was
applied to evaluate the stabilization energy of TCHpyrrole complexes [42]:
E = E(templatemonomer complex) E(template)
nE(monomer)
where n refers to the number of monomers in the
templatemonomer complex.
minimal
energy
of
the
template(monomer),
The
template(monomer)2 and template(monomer)3 complexes for
the selected monomer (pyrrole) obtained by restricted MP2 calculations were 2,274,311.0285, 2,823,577.1692 and 3,372,
848.2678 kJ mol1 , respectively. The E value for the
templatemonomer complexes, i.e., TCH(pyrrole), TCH(pyrrole)2
and TCH(pyrrole)3 in the prepolymerization mixture were
31.8930, 77.1416 and 127.3483 kJ mol1 respectively. The
results indicate that the templatemonomer complex stability
order is: TCH(pyrrole)3 > TCH(pyrrole)2 > TCH(pyrrole). The most
stable geometry of templatemonomer complex in the prepolymerization mixture, obtained by the restricted MP2 method
calculation is illustrated in Fig. 8.
3.9. Calibration curve

Fig. 7. The response of the MIP electrode fabricated by different concentrations of

templete (TCH) was measured through DPV for 2 mM thiocarbohydrazide standard
solution in Britton-Robinson buffer at pH 5.0.

The calibration curve for the DPV peak current observed for
TCH oxidation versus TCH concentration at MIP electrode at the
optimal amounts of the parameters shows linearity over a concentration range of 0.56 mM. The calibration linear equation
of A = 1.798C + 4.336 (r2 = 0.9805) was obtained in the optimal

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Table 2
Effect of the interferents on the differential pulse voltammetric response on the detection of 2 mM thiocarbohydrazide using the MIP electrode.
Change in current responsea for 2 mM TCH (A)

Interferent molecule

Concentration of interferent (mM)

Thioacetamide

2
4
8

0.432
0.656
1.806

8.946
9.170
10.32

Thiourea

2
4
8

0.593
1.236
1.407

9.107
9.750
9.921

Camphor

1
2
4

0.681
1.070
1.864

7.833
7.444
6.650

Ticlopidine
hydrochloride

1
2
4

0.267
1.082
1.491

8.247
7.432
7.023

Methimazole

1
2
4

0.390
0.896
2.708

8.904
9.410
11.222

Current response (A)

The current in the absence of any interferent was 8.514 A (with MIP).

conditions. The experimental detection limit was obtained

5.0 105 M TCH.
3.10. Effect of interferents
The selectivity of the MIP electrode in this work was evaluated
in the presence of different interfering molecules. Voltammetric
responses of TCH imprinted polypyrrole lm were examined in
the presence of some interfering substances like thioacetamide,
thiourea, methimazole, ticlopidine hydrochloride and camphor.
The differential pulse voltammograms were taken for the oxidation
of TCH (2 mM) after addition of varying concentration of each interferent. The obtained currents in the absence of any interferent were
8.514 A with MIP electrode. The results are given in Table 2. The
comparison of the TCH currents (Table 2) before and after the addition of interferent chemicals shows there is no serious interfering
effect for the method.
3.11. Analysis of real samples
The differential pulse voltammograms were recorded after samples preparation under exactly identical conditions that were
employed while recording differential pulse voltammograms for
plotting calibration plot. DPV measurements demonstrated no peak
for the prepared meat samples (in the no spiking of TCH) in the scan
range of potential Fig. 9.
According to the suppression of the analytical signal of the electrode in human serum matrix, the standard addition method was

Fig. 10. The standard addition calibration curve plotted in the optimized conditions
for determination of TCH in human blood plasma.

chosen only for this real sample matrix (Fig. 10) to evaluate the
applicability of the TCHMIP sensor in such a complex media. It
could be applied to the other real samples matrix. Table 3 shows the
results obtained by MIP-DPV after spiking of TCH standard solution
to the prepared biological matrices.
3.12. Repeatability and reproducibility of the MIP electrodes
The repeatability and reproducibility of the molecularly
imprinted modied pencil graphite electrode was investigated for
2 mM TCH solution. The peak current response of TCH was determined by three measurements of using single electrode resulted
Table 3
The results obtained for the analysis of TCH by MIP using differential pulse voltammetry in a few biological matrices.

Fig. 9. The calibration curve plotted in the optimized conditions (n = 3): BrittonRobinson pH 5.0, the electrode fabricated in 0.01 M thiocarbohydrazide and 0.02 M
pyrrole.

Sample

Founda

Added TCH
(mM)

Detectedb
(mM)

Recovery (%)

Beef
Lamb meat
Fish meat
Camel meat
Chicken
Human blood serum

0.00
0.00
0.00
0.00
0.00
0.00

2.00
2.00
2.00
2.00
2.00
2.00

1.81
1.65
1.83
1.79
1.87
2.29

90.50
82.50
91.50
89.50
93.50
114.5

a
Detected amount of thiocarbohydrazide in the real sample matrices by the
present technique (no spiking of TCH).
b
Detected amount of thiocarbohydrazide in the real sample matrices by the
present technique after spiking.

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in the relative standard deviation of 4.2%. The reproducibility was

investigated by measuring the peak current response of three
electrodes for the mentioned standard solution of TCH. The calculated relative standard deviation demonstrating reproducibility
was 14.3%. The sensor can retain its properties for 10 days if
stored in air at room temperature. No apparent decreases in current responses occurred during the rst day, and 9% decrease was
observed after the electrode was stored for 10 days (n = 3, RSD 5%).
The results were acceptable for such a sensor considering the literature [58]. The response of the electrode remained constant during
21 measurements (7 h totally) storing in solution at the optimized
conditions.
4. Conclusion
As a promising technology with inherent high selectivity and
afnity, molecular imprinting could thus afford an efcient complementary approach for quantitative analysis of organic materials.
The MIP modied pencil graphite electrode has been successfully
applied as a sensor for fast and accurate determination of TCH
in standard solutions and a few biological materials spiked with
TCH. In this work, the effects of template concentration, number of
CV cycles during the electropolymerization of pyrrole on the surface of pencil graphite electrode, monomer concentration and pH
of TCH electrooxidation on the DPV response of TCHMIP were
evaluated. The advantages of very simple instrumentation, easy
preparation of the sensor and low cost of the pencil graphite electrode make the system useful in fabrication of simple devices for
determination of TCH without any usage of organic toxic solvents
as applied before [43] which are hazardous in environment.
This provides the opportunity for further widening the scope of
molecularly imprinted recognition materials in applications such
as sensors and assays operating in, or with, aqueous based, and
environmental media.
Acknowledgement
The authors acknowledge Payame Noor University (PNU)
Research Council for nancial support.
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Biographies
Azizollah Nezhadali is an associate professor of Department of Chemistry, Payame
Noor University in Mashhad, Iran. He received his PhD in Department of Chemistry
(2000) from Ferdowsi University of Mashhad, Iran and the University of Sydney,
Australia (guest PhD student for six months). Now he is a faculty member of Department of Chemistry, Payame Noor University, Mashhad, in Iran. His research interests
cover separation (micro and bulk), electroanalytical chemistry and GC/MS. He has
written a chapter of Macrocylic chemistry book, Nova Publisher, NY, 2010.
Raham Shadmehri is a PhD candidate in Department of Chemistry, Payame Noor
University of Mashhad. He received his bachelor degree in chemistry (2006) from
Payame Noor University of Mashhad, Iran. He received his master degree in analytical chemistry (2009) from Islamic Azad University of Mashhad, Iran. His research
interest is mainly in molecularly imprinted polymers and electroanalytical chemistry. Now he works with research Group of Dr. A. Nezhadali.