Materials Science and Engineering C 59 (2016) 136–144

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Materials Science and Engineering C

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

Enhanced amperometric detection of metronidazole in drug

formulations and urine samples based on chitosan protected
tetrasulfonated copper phthalocyanine thin-film modified glassy
carbon electrode
S. Meenakshi a, K. Pandian a,⁎, L.S. Jayakumari b, S. Inbasekaran c
a
Department of Inorganic Chemistry, University of Madras, Guindy Campus, Chennai 600025, India
b
Department of Rubber and Plastics Technology, MIT Campus, Anna University, Chennai 600044, India
c
Sensor Division, Central Leather Research Institute (CLRI), CSIR, Adyar, Chennai 600020, India

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

Article history: An enhanced electrocatalytic reduction of metronidazole antibiotic drug molecule using chitosan protected
Received 16 May 2015 tetrasulfonated copper phthalocyanine (Chit/CuTsPc) thin-film modified glassy carbon electrode (GCE) has
Received in revised form 25 July 2015 been developed. An irreversible reduction occurs at −0.47 V (vs. Ag/AgCl) using Chit/CuTsPc modified GCE. A
Accepted 27 August 2015
maximum peak current value is obtained at pH 1 and the electrochemical reduction reaction is a diffusion con-
Available online 3 September 2015
trolled one. The detection limit is found to be 0.41 nM from differential pulse voltammetry (DPV) method. This
Keywords:
present investigation method is adopted for electrochemical detection of metronidazole in drug formulation
Differential pulse voltammetry and urine samples by using DPV method.
Metronidazole © 2015 Elsevier B.V. All rights reserved.
Tetrasulfonated copper phthalocyanine
Chitosan
Urine samples

1. Introduction spectrophotometry [12], polarography [13], liquid chromatography

Metronidazole, 1-(2-hydroxyethyl)-2-methyl-5-nitroimidazole
(MTZ), is one of the nitroimidazole derivatives well-known for its anti-
microbial properties [1,2]. It is effective against Trichomonas [3],
Vincent's organisms [4], and anaerobic bacteria [5,6]. Veterinarians
also use MTZ to treat bacterial infections specifically giardia in dogs
and cats [7]. The chemical structure of the drug is shown in Fig. 1.
Though MTZ is typically well tolerated at b 2 g daily dosage and beyond,
a level of toxicity has been reported [8]. Longer-term use for severe in-
fection is often limited by the development of peripheral neuropathies
[9], as well as optic neuropathy [10], neutropenia and central nervous
toxicities including cerebellar ataxia, encephalopathy and seizures.
Therefore, it is necessary to monitor MTZ concentration in patients
under high antibiotic therapy.
The toxicity of MTZ drug is dose-dependent on biological fluids like
serum, urine and saliva. It is well absorbed in serum as well as tissue
concentrations usually at high levels [11]. Several analytical methods
have been reported for determination of MTZ, which include
⁎ Corresponding author at: Department of Inorganic Chemistry, University of Madras,
Chennai 600025, India.
E-mail addresses: jeevapandian@yahoo.co.uk (K. Pandian), jaya@mitindia.edu
(L.S. Jayakumari).
[14], gas chromatography [15], high performance liquid chromatogra-
phy [16] and gas chromatography–mass spectrometry (GC–MS) [17].
However, the above reported methods could not mention the impor-
tance of selectivity of MTZ determination. Hence, it is primary impor-
tance to develop an alternative method for MTZ determination with a
high degree of selectivity and sensitivity.
Chemically modified electrodes (CMEs) have continued to be a
major concern during the past decade. CME has been widely considered
for selective and sensitive electrochemical determination such as envi-
ronmentally [18–20] and biologically important compounds [21–23].
CMEs can catalyze the electrode reaction by significantly decreasing
the required over-potential, pH independent, fast reacting and stable
in redox behavior [24,25]. Modification of CME with suitable materials
to facilitate electrochemical reactions of redox compounds to proceed
without any hindrance [26,27]. Notably, certain transition metal com-
plexes of porphyrins and metal phthalocyanines are used to catalyze
electrooxidation or electroreduction of chemical and biological com-
pounds. Mostly, metal phthalocyanine modified electrode is very attrac-
tive because of less soluble compounds and its simple modifications of
molecular structure [28,29]. It can be used extensively in catalysts, semi-
conductors and anticancer medicine.
Chitosan is one of the naturally available biopolymers and it is wide-
ly used in various applications, due to its poor solubility and high

http://dx.doi.org/10.1016/j.msec.2015.08.063
0928-4931/© 2015 Elsevier B.V. All rights reserved.
 S. Meenakshi et al. / Materials Science and Engineering C 59 (2016) 136–144
Fig. 1. Structure of MTZ.
thermal stability [30,31]. Nowadays, chitosan is considered as one of the
most important polymer to stabilize modified electrode substrates.
Chemically modified chitosan is an efficient sorbent material for heavy
metals, carbon electrode coated with carbon nanotube/chitosan com-
posites for detection of polyphenols [32], paracetamol and uric acid
[33]. Furthermore, the electrochemical reduction of MTZ at deoxyribo-
nucleic acid (DNA) modified GCE [34], metalloporphyrin modified car-
bon paste electrode [35], activated GCE [36], nanomaterial thin film
coated GCE [37], carbon fiber microdisk electrode [38], Ni/Fe-LDH mod-
ified GCE [39], MIP/MWCNT/GCE [40] and Cu-Poly(Cys) [41] has been
reported.
With this above background, in the present study we report a novel
voltammetric sensor based Chit/CuTsPc/GCE which has been developed
for selective determination of MTZ. The prepared MTZ sensor was char-
acterized in terms of selectivity, sensitivity and reproducibility. A rela-
tively low applied potential and improved selectivity for MTZ
detection has been realized. In addition, the designed sensor was suc-
cessfully applied for determination of MTZ in commercially available
tablets and urine samples.
2. Experimental
2.1. Chemicals
All reagents and chemicals used were of analytical grade and with-
out further purification. Metronidazole was obtained from Vanavile
Chemicals, Pvt. Ltd., Cuddalore, Tamil Nadu, India. Flagyl tablets at
200 mg and 400 mg were purchased from local medical stores. Copper
phthalocyanine tetrasulfonic acid tetrasodium salt (CuTsPc) and chito-
san were purchased from Sigma Aldrich, USA. Sodium hydroxide was
purchased from Fisher Scientific, India. All other chemicals used in this
study were purchased from SRL Chemicals, Pvt. Ltd., India.
2.2. Instrumental methods
UV–Visible spectral studies were observed by using a Shimadzu UV–
Visible Spectrophotometer, Japan (Model UV-1800). Morphological and
structural investigations were carried out with field emission microsco-
py (JEOL-JSM-6360 Instrument, USA) and atomic force microscopy im-
ages were obtained using AFM Park Systems, XE-70, South Korea. The
cyclic voltammetric experiment was carried out using a CHI 660A elec-
trochemical instrument, Gamry model 330, USA. A conventional three
electrode system comprising of a glassy carbon electrode (GCE) 3 mm
in diameter was purchased from BAS. Pvt. Ltd., USA. Ag/AgCl and plati-
num wire were used as a reference electrode and counter electrode, re-
spectively. A Bioanalytical System (BAS, USA) polishing kit was used to
polish the GCE surface.
Electrochemical impedance spectroscopy (EIS) measurements were
performed with the use of the CHI-660A electrochemical instrument.
Electrolyte was prepared by using 0.1 M KCl containing a 5 mM
[Fe(CN)6]3−/4– redox probe. All solutions were purged with high purity
nitrogen gas for about 10 min before performing all electrochemical ex-
periments. All experiments were carried out at room temperature.
137
2.3. Preparation of the Chit/CuTsPc thin-film
The Chit/CuTsPc composite was prepared by two different methods
viz. infiltration and direct mixing method. The 1% (w/v) chitosan solu-
tion was poured onto a glass plate and left to dry through evaporation
for 24 h and then heated at 60 °C for 6 h. Later, the thin film was neutral-
ized with 1 M NaOH, washed thoroughly and dried. The dry film was
immersed in 0.01 M CuTsPc solution for 24 h followed by washing
with deionized (DI) water and stored in deionized water prior to differ-
ent measurements. Alternatively, CuTsPc solution (2 mg mL− 1) was
prepared by dissolving 10 mg CuTsPc in 10 mL water. Chitosan (1% w/
v) was prepared by dissolving 1 g of chitosan in 100 mL of DI water
and then 1 mL of glacial acetic acid was added. The mixture was allowed
to stir for few minutes until the complete dissolution of chitosan. The
whole reaction mixture was concentrated by using a rota-evaporator
to reach a final volume of 10 mL. The viscous matrix was drop-casted
over GCE and used for the electrochemical studies.
2.4. Preparation of the Chit/CuTsPc modified GCE
The GCE surface was mechanically cleaned by polishing with 500 μm
alumina powder, washed with double distilled (DD) water and then
sonicated for 5 min in ethanol, followed by DD water. GCE substrate
was modified with Chit/CuTsPc solution (5 μL) by drop-casting method
using a micro-syringe and dried under room temperature before use.
3. Results and discussion
Recently, polymer stabilized metal phthalocyanine was demonstrat-
ed for construction of many types of chemically modified electrodes
which are widely used in various applications. For example, chitosan
[42], poly-L-lysine [43], polyallylamine [44] and natural gum [45] are
used to stabilize metal phthalocyanine and these hybrid nanocomposite
modified electrodes can be used for electrochemical determination of
biologically important molecules. In the present study, chitosan is cho-
sen as a dispersing agent to stabilize the CuTsPc film because of the
strong electrostatic interaction between chitosan with CuTsPc [46]. Sta-
bility and film forming performance of CuTsPc dispersed chitosan were
investigated through UV–Visible spectral studies. CuTsPc peak appeared
at 614 nm and 695 nm corresponding to dimeric and monomeric spe-
cies due to the π–π* transition on macrocycle [47]. In Fig. S1, the short
band position is slightly red shift when compared to the CuTsPc solution
because of molecular aggregation. The molecules were uniformly dis-
tributed and formed CuTsPc nanocrystals within the chitosan polymer
matrix. So the drop-casting method [48] is highly preferred for loading
copper phthalocyanine type of electron transfer mediators within the
Fig. 2. SEM image of Chit/CuTsPc.
138 S. Meenakshi et al. / Materials Science and Engineering C 59 (2016) 136–144

Fig. 3. AFM images of chitosan protected CuTsPc.

chitosan polymer matrix. Another advantage of using chitosan as a pro-
tective layer is its poor solubility in water. Finally, the water soluble
CuTsPc is uniformly dispersed within the chitosan matrix.
The surface morphology of CuTsPc dispersed chitosan film was in-
vestigated by AFM and SEM studies. Fig. 2 shows that the morphology
of the CuTsPc film was uniformly distributed within the polymer matrix.
The average size of the Chit/CuTsPc film was 5.47 μm measured under a
resolution of 100 μm. Elemental composition was confirmed by energy
dispersive X-ray analysis (EDAX) as shown in Fig. S2. In addition, Chit/
CuTsPc is confirmed with AFM studies and the average roughness of
the polymer film was found to be 30 nm as shown in Fig. 3.
FT-IR spectra of chitosan and Chit/CuTsPc are shown in Fig.4. From
chitosan, the strong band at 3351 cm− 1 was observed which can be
owed to presence of the\\OH hydroxyl group. The absorption peaks ac-
quired at 1651 cm−1 and 1074 cm−1 correspond to stretching vibration
of C_O and skeletal vibration of the C\\O group [49]. The bands at
1374 cm−1 and 2867 cm−1 are characteristic of CH3 and C\\H function-
al groups. As compared to Chit/CuTsPc, the absorption band at
3429 cm−1 is attributed to stretching vibration of NH2 (amine) group
[50]. In FT-IR spectra of Chit/CuTsPc, the C\\O stretching vibration of
chitosan intensity is found to be faded due to the interaction between
chitosan and pthalocyanine [51]. Thus, these results confirmed the cop-
per phthalocyanine presence on chitosan polymer matrix.
Fig. 5 shows the cyclic voltammogram of a) bare, b) Chit/CuTsPc/GCE
in the presence of 0.1 M KCl containing PBS (pH 7) at a scan rate of
100 mV/s within potential ranges from −1.0 V to +0.8 V. There is no

Fig. 5. Cyclic voltammogram of (a) bare and (b) Chit/CuTsPc modified GCE in the presence
Fig. 4. FT-IR spectra of chitosan and Chit/CuTsPc. of 0.1 M KCl containing phosphate buffer solution (pH 7) at a scan rate of 100 mV/s.
 S. Meenakshi et al. / Materials Science and Engineering C 59 (2016) 136–144 139

Fig. 6. Electrochemical impedance studies (EIS) of a) bare GCE and b) Chit/CuTsPc/GCE in

the presence of 0.1 M KCl and 5 mM K3[Fe(CN)6]3−/4−.
oxidation and reduction peaks which appeared in bare GCE and chito-
san. As seen in Fig. 5b, a pair of two redox peaks were obtained. The ox-
idation peaks appeared at − 0.28 V and − 0.10 V while the reduction
peaks appeared at −0.23 V and −0.69 V. This pair of redox peaks is as-
cribed to Cu(I)/Cu(0) and Cu(II)/Cu(I) coupling of CuTsPc which sug-
gests that CuTsPc strongly bonds with chitosan and its enhanced a
better electrochemical behavior of Chit/CuTsPc/GCE [52].
3.1. Electrochemical impedance studies (EIS)
Fig. 7. Cyclic voltammogram of MTZ on (a) bare GCE, (b) bare GCE with 0.1 M of 50 μL MTZ
and (c) Chit/CuTsPc/GCE in 0.1 M of 50 μL MTZ in the presence of 0.1 M KCl containing
phosphate buffer solution at a scan rate of 20 mV/s.
values. The response of the high current is due to the electrocatalytic re-
duction of MTZ and the peak current value was almost three times
higher than that of bare GCE.
The electrochemical behavior of MTZ was investigated in an acidic
medium using chitosan protected CuTsPc modified GCE. The modified
electrode was prepared in two different ways; either by direct mixing
of CuTsPc in chitosan solution to form a stable suspension or by

Electrochemical impedance spectroscopic (EIS) measurement was

carried out in order to understand electron transfer of modified elec-
trode surface. EIS can be applied to preparing modified electrodes
which could provide useful information [53]. The curve of EIS includes
a semicircular part and a linear part. The semicircular part at higher fre-
quencies corresponds to a electron-transfer-limited process and its di-
ameter is equal to that of electron transfer resistance (Rct) which
controls the electron transfer kinetics of the redox probe at the elec-
trode interface. Meanwhile, the linear part at lower frequencies corre-
sponds to the diffusion process. Fig. 6 shows EIS curves of different
modified electrodes during the stepwise fabrication procedure. Bare
GCE exhibited an almost straight line which is characteristic of a diffu-
sion limited electrochemical process (Fig. 6a). The semicircle diameter
of well conducting substrates is equal to the electron transfer resistance
(Rct). The value of Rct was measured after modification of the electrode
surface with Chit/CuTsPc in the presence of K3[Fe(CN)6]3 −/4- as the
redox probe (Fig. 6b). At GCE, Rct value is relatively high which indicates
that electron transfer process is slow whereas in case of Chit/CuTsPc/
GCE, the Rct value is 100 Ω which implied that the electrochemical
redox behavior of the probe to the substrate electrodes was accelerated.
These results show that the Chit/CuTsPc film can act as an effective elec-
tron conduction pathway between electrode and electrolyte.

3.2. Electrochemical reduction of MTZ

MTZ is a nitro group functionalised imidazole derivative [33] and the

nitro group undergoes a single step reduction reaction on the cathodic
side [35]. According to previous reports, a cathodic peak is obtained
due to four electron reduction of the nitro group in MTZ which corre-
sponds to hydroxylamine [54]. The electron transfer behavior is sluggish
on bare GCE electrode than the Chit/CuTsPc film modified GCE under an
identical experimental condition. As can be seen in Fig. 7, the reduction
peak was observed at −0.44 V vs. Ag/AgCl for bare GCE whereas, a re-
duction peak appeared at − 0.47 V vs. Ag/AgCl for the Chit/CuTsPc/ Fig. 8. Influence of a) pH of the medium on electrochemical reduction of MTZ in Chit/
GCE. A well-defined sharp peak was obtained for reduction of MTZ in CuTsPc/GCE in the presence of 0.1 M KCl containing phosphate buffer solution at a scan
Chit/CuTsPc modified GCE with enhanced reduction peak current rate of 20 mV/s (pH 1, 3, 5, 7 and 9) and b) peak potential of MTZ.
140 S. Meenakshi et al. / Materials Science and Engineering C 59 (2016) 136–144

Fig. 9. Electrochemical mechanism of MTZ.

infiltration approach in an aqueous solution of CuTsPc exposed into chi- rates. As shown in the inset Fig. 10b, peak current has a linear relation-
tosan film. Fig. S3 displays the electrochemical reduction peak for MTZ ship with the square root of the scan rate and the regression equation of
using two different modified electrodes and reduction peak potentials Ipa (μA) = −3.5956x + 0.5640 (R2 = 0.9950). This shows that the re-
were close to − 0.47 V vs. Ag/AgCl in both cases and observed higher duction of MTZ on Chit/CuTsPc/GCE was a predominantly diffusion con-
peak current values in the case of CuTsPc dispersed chitosan film trolled one. Moreover, the cathodic peak potential of MTZ increases
which attributed to higher loading of catalyst than the infiltration meth- linearly with the natural logarithm of the scan rate (log υ) and the linear
od. Thus, electrochemical studies showed that CuTsPc dispersed chito- regression equation of Epa (V) = − 0.0115x + 0.4461 (R2 = 0.9710)
san film modified electrode to be efficient in the electrocatalytic (inset Fig. 10c). Since the reduction reaction of MTZ is irreversible in na-
reduction of MTZ. ture, a linear relationship between Epa and the natural logarithm of the
scan rate (log υ) obeyed Eqs. (1) and (2);
3.3. Influence of pH on reduction of MTZ " !   #
0
RT D1=2 αnFυ 1=2
The pH of solution is an essential parameter which influences the re- Ep ¼ Eθ − 0:780 þ ln þ ln ð1Þ
αnF k
0 RT
duction rate of MTZ. The influence of pH on reduction peak current
value was measured at different pH ranges (Fig. 8a). The maximum
RT
peak current was obtained within acidic pH medium with a maximal ¼kþ logυ ð2Þ
2αnF
current response obtained at pH = 1. The slope value is 59 mV/pH,
which confirmed that the mechanism is pH dependent on acidic and
where Ep is the formal redox potential, α is the electron transfer co-
neutral media. If pH values are beyond 8, the reduction reaction is pH in-
efficient, D is the diffusion coefficient, k0 is the standard heterogeneous
dependent, and it is difficult to analyze the electrochemical reduction of
MTZ [35,36]. Nitro group derivatives correspond to reduction of the
nitro group (R-NO) to form hydroxylamine (RNHOH), involving 4 elec-
trons and 4 protons followed by a two-electron reduction of hydroxyl-
amine to amine. However, MTZ shows weakly basic (or strong acid)
properties at pH values higher than 8, this depicts a decrease in proton-
ation of hydroxylamine with the increase in pH [55]. The redox electro-
chemical mechanism of MTZ is shown in Fig. 9.

3.4. Effect of scan rate and concentration on the peak current of MTZ

The effect of scan rate on peak current and peak potential of MTZ on
Chit/CuTsPc/GCE, investigating the reaction kinetics was studied by cy-
clic voltammetry (CV). The CV for 50 mM MTZ in acidic pH solution
(pH 1) with scan rates that vary from 5 to 100 mV/s is shown in Fig.
10a. The reduction peak current was increased with increasing scan

Fig. 10. (a) Cyclic voltammogram of Chit/CuTsPc/GCE in the presence of 0.1 M KCl contain- Fig. 11. (a) Cyclic voltammogram of Chit/CuTsPc/GCE in the presence of 0.1 M KCl contain-
ing PBS (pH 1) at different scan rates (5–100 mV/s). Inset: (b) Plot of the peak current vs. ing PBS (pH 1) at a scan rate of 20 mV/s with increasing concentrations of MTZ. (b) Cali-
square root of the scan rate. Inset: (c) Plot of Epa vs. logυ. bration plot of the peak current vs. concentration.
 S. Meenakshi et al. / Materials Science and Engineering C 59 (2016) 136–144 141

scan rate of 20 mV/s. The cathodic peak current of MTZ was increased
by increasing to various concentrations. A linear response was observed
in the peak current against various concentrations of MTZ and linear re-
gression equation of y = −3.5956 (μM) + 0.5640 with a correlation co-
efficient of (R2) = 0.9916 as shown in Fig. 11b. This implies that the
catalytic peak current has a linear relationship with the concentration
of MTZ.

3.5. Differential pulse voltammetry studies on MTZ reduction

The DPV method was used for studying sensitive electrochemical re-
duction of MTZ using the Chit/CuTsPc film modified electrode as shown
in Fig. 12a. A better sensitivity was achieved when the modified elec-
trode system was used instead of bare GCE. A sharp peak was observed
within a reduction peak potential of −0.45 V vs. Ag/AgCl. This value is
very close to the cyclic voltammetry peak potential. A calibration
graph was plotted against the concentration of MTZ with respect to re-
duction peak current values (Fig. 12b). A linear relationship was obtain-
ed with a regression equation of y = −0.7340 (nM) −0.7611 and R2 =
0.9942. The detection limit value (3σ/s, σ is a standard deviation) was
found to be 0.41 nM. The detection limit of the present method is
comparable with previously reported experimental values as shown in
Table 1.

3.6. Amperometric detection of MTZ

Amperometry experiment was carried out on the electrochem-

Fig. 12. (a) Differential pulse voltammetry behavior for MTZ at Chit/CuTsPc/GCE in the
presence of 0.1 M KCl containing PBS (pH 1) at various concentrations ranging from
6.0 × 10−9 M to 8.0 × 10−8 M. (b) Calibration plot of peak current vs. concentration.
rate constant, R is the universal gas constant, T is the absolute tempera-
ture, and F is Faraday's constant. The slope line is equal to RT/2αnF and
α is found to be 0.7 in the electrode reaction of MTZ due to the irrevers-
ible electrode process. In the present study, the number of protons and
electrons involved during the reduction process of MTZ is almost equal.
Fig. 11a shows a cyclic voltammogram of the electrocatalytic reduc-
tion of MTZ at the Chit/CuTsPc modified GCE in 0.1 M PBS (pH 1) at a
ical detection of MTZ under hydrodynamic conditions to obtain
the maximum sensitivity. Here applied potential was fixed at
− 0.45 V vs. Ag/AgCl (Fig. 13a). In each addition of MTZ, the peak
current values were measured by a constant electrode potential
value. The steep rise in peak current value was calculated and a cal-
ibration plot was made by plotting measured peak current values
with respect to the addition of MTZ. The calibration was followed
by a linear relationship of y = − 0.2106 (nM) − 0.0386 and corre-
lation coefficient of (R2) = 0.9976 (Fig. 13b). These results strongly
suggested the selective and sensitive determination of MTZ by
Chit/CuTsPc/GCE.
Several compounds are selected to analyze interference studies by
the amperometric method. If tolerance limit was taken as the maximum
concentration of substances, which caused an approximately 5% relative
error, for 10 mol L−1 DA, AA, and UA, no interference was observed for
subsequent compounds like Na+, K+, Ca2+, Mg2+, Fe3+, Zn2+ and NO-3.

Table 1
Determination of MTZ in different electrodes.

Electrode Technique Linear range (mol/L) R2 LOD (mol/L) Ref.

Activated GCE LSV 2 × 10−6–6 × 10−4 0.9970 1.1 × 10−6 [36]

MWNT/GCE DPV 2.5 × 10−8–1 × 10−5 0.9970 6 × 10−9 [37]
Carbon fiber microdisk electrode SWASV 5 × 10−6–1.6 × 10−5 0.9950 5 × 10−7 [38]
DNA/GCE DPV 5 × 10−8–60 × 10−6 0.9955 2 × 10−8 [56]
SWCNT/GCE CV 1 × 10−7–2 × 10−4 0.9971 6.3 × 10−8 [57]
Au electrode CV 5 × 10−7–1 × 10−5 0.9975 1.5 × 10−7 [58]
HMDE DPV 2.3 × 10−7–1.8 × 10−6 0.9998 3.6 × 10−8 [59]
MIP-CPE DPV 3.3 × 10−10–1.5 × 10−8 0.9970 1.5 × 10−10 [60]
Gr-IL/GCE DPV 1 × 10−7–2.5 × 10−5 0.9990 4.7 × 10−8 [61]
3D GNT/CPE SWASV 1 × 10−9–2 × 10−6 0.9969 0.1 × 10−9 [62]
Cysteic acid/PDDA-GN/GCE LSV 1 × 10−8–800 × 10−6 0.9940 2.3 × 10−9 [63]
MMIP/MGCE DPSV 5 × 10−8–1 × 10−6 0.9946 0.16 × 10−7 [64]
Chit/CuTsPc/GCE DPV 0.8 × 10−9–72 × 10−7 0.9976 4.1 × 10−10 This work

GCE — Glassy carbon electrode

HMDE — Hanging mercury drop electrode
MIP-CPE — Molecularly imprinted polymer-carbon paste electrode.
SWASV — Square wave adsorptive stripping voltammetry
Gr-IL — Graphene-ionic liquid
DPSV — Differential pulse stripping voltammetry
MMIP — Magnetic molecularly imprinted polymer
LSV — Linear sweep voltammetry
142 S. Meenakshi et al. / Materials Science and Engineering C 59 (2016) 136–144

agreement with the reported values (Table 2). Recovery of MTZ was
found between 94.13 and 103.4%, indicating the applicability of this
method to determine MTZ in pharmaceutical samples. From the above
results, the developed sensor has good practical applicability.

3.8. Determination of MTZ in urine samples

The applicability of the Chit/CuTsPc/GCE electrode for measuring

concentrations of MTZ was carried out in urine samples from patients.
5 mL of urine samples was taken from different 25 mL standard flasks.
Different quantities of MTZ were added to the urine samples and then
quantitatively diluted using supporting electrolyte. DPV were recorded
and unknown concentrations were determined on a calibration graph.
The expected results of Chit/CuTsPc film modified sensor in Table S5.
The MTZ spiked real sample solutions were prepared by adding MTZ
to the final concentrations (87, 96, 110, and 123 μM) in two time diluted
urine samples. Recovery of MTZ from spiked urine samples was calcu-
lated. The relative standard deviations were less than 2.9% (n = 4).
Based on the results, recovery of MTZ can be detected in biological sam-
ples with more precision and hence it's applied for clinical usage.

4. Conclusion

The voltammetric behavior of MTZ was investigated through

Fig. 13. (a) Amperometric response obtained for reduction of MTZ at Chit/CuTsPc/GCE in
the presence of 0.1 M KCl containing PBS (pH 1) upon addition of 0.02 mL respectively.
(b) Calibration plot of peak current vs. concentration.
Chit/CuTsPc modified GCE by using CV and DPV methods. The re-
duction of MTZ was an irreversible reduction process. An enhanced
electrocatalytic peak current value was observed in Chit/CuTsPc/
GCE when compared with bare GCE in acidic medium (pH 1). Influ-
ence of pH was investigated on reduction peak current and peak
potential, the maximum reduction peak current was acquired at
pH 1. The present investigated method is suitable for analytical de-
termination of MTZ, because it is simple, low cost, highly accurate,
selective and sensitive. The developed sensor was successfully ap-
plied for voltammetric determination of MTZ in pharmaceutical
formulations and urine samples.

Acknowledgments
In addition, a 200-fold concentration of cysteine, starch and caffeine and
Dr. K. Pandian is grateful to Prof. R. Jayavel, Director, Centre for
a 100-fold concentration of glucose, sucrose, fructose, dextrin, oxalic
Nanoscience and Technology, Anna University, Chennai 600025, for
acid and paracetamol do not interfere with the reduction signal of
providing AFM facility. The authors would like to thank the National
MTZ (signal change b 5%) as shown in Fig. S4. However, some com-
Centre for Nanoscience and Nanotechnology, University of Madras,
pounds containing the nitro group such as nitrophenols, chloramphen-
Guindy, Chennai-25, for providing the SEM facility.
icol and orinidazole interfere with signals of MTZ because of the same
electroactive species. This further depicts that the Chit/CuTsPc/GCE
electrode can be successfully used for determination of MTZ with excel- Appendix A. Supplementary data
lent sensitivity and selectivity.
Supplementary data to this article can be found online at http://dx.
doi.org/10.1016/j.msec.2015.08.063.
3.7. Determination of MTZ in tablets
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