Journal of Electroanalytical Chemistry 878 (2020) 114574

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Journal of Electroanalytical Chemistry

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

Electrochemical sensing of ciprofloxacin and paracetamol in environmental

water using titanium sol based sensor
Aleksandra Pollap, Karolina Baran, Natalia Kuszewska, Jolanta Kochana
⁎
Jagiellonian University, Faculty of Chemistry, Department of Analytical Chemistry, Gronostajowa 2, 30-387 Kraków, Poland

A R T I C L E I N F O A B S T R A C T

Article history: In this work we present a new sensitive electrochemical method for simultaneous determination of ciprofloxacin
Received 25 May 2020 (antibiotic) and paracetamol (painkiller). For this purpose graphite electrode was modified with titanium diox-
Received in revised form 10 August 2020 ide sol as binding agent. TiO2 sol was enriched with gold nanoparticles and CMK-3 type mesoporous carbon,
Accepted 10 August 2020
both increasing the composite conductivity, and Nafion, responsible for signal stabilization. Developed sensor
Available online 14 August 2020
has been characterized via cyclic voltammetry technique presenting LOD equal to 0.108 μM and 0.210 μM and
Keywords:
sensitivity of 15.93 μA μM−1 and 11.56 μA μM−1 for ciprofloxacin and paracetamol, respectively. For both analytes
Ciprofloxacin two linear ranges from 1 to 10 μM and from 10 to 52 μM were found. Developed method enables to determine with
Paracetamol high accuracy and precision ciprofloxacin and paracetamol traces in environmental water matrices as well as in certified
Environment reference standard of waste water matrix.
Sensor
Voltammetry

1. Introduction One of synthetic antibiotics is ciprofloxacin, 1-cyclopropyl-6-fluoro-

Since the penicillin discovery in 1929 by Alexander Fleming, there has
been an intensive medicine development related to the production and ap-
plication of antibiotics [1]. According to recent reports, from 100 to 200
thousand tons of antibiotics are produced around the world throughout
yearly [2]. Despite the fact that these pharmaceuticals are powerful agents
for bacterial infections treatment, increase of its production and consump-
tion results in environmental pollution. Antibiotics residues are detected
in surface water as well as groundwater, sewage and in soil [1,3]. The main
sources of environmental pollution are not only the excretion of antibiotics
and metabolites in the faeces by humans and animals, but also industry
and wastes from hospitals and farms [4–8]. Furthermore, the presence of
antibiotic residues is also dangerous for people consuming food of animal
origin leading to allergic or cancerogenic reactions [9,10]. Nevertheless,
among all negative consequences of antibiotic residues in environment
the most harmful is an increasing drug resistance of microorganisms.
This phenomenon is described as a bacteria ability to develop despite
the antibiotic presence [1]. The pharmaceutical residues issue in the
environment refers not only to antibiotics. Due to the fact, that many
painkillers including substituted acetanilides, like paracetamol, are often
available without a prescription, their residues could be also found in
the environment causing limitation in cell proliferation or reproductive
damages [11].
1,4-dihydro-4-oxo-7-(piperazinyl)quinolone-3-carboxylic acid (CPX). It is
a chemotherapeutic which belongs to fluoroquinolone group [12]. CPX is
characterized by bactericidal activity against both Gram-positive and
Gram-negative bacteria. Thereby, this antibiotic is used for treatment sev-
eral diseases such as digestive and respiratory system infections as well as
urinary tract infections [13]. Bactericidal activity of CPX results from the
decrease in the activity of enzymes taking part in DNA replication in bacte-
ria [12,13]. Considering the environmental aspect, synthetic and semi-
synthetic antibiotics, including fluoroquinolones, are less susceptible to
bacterial decomposition processes and as a consequence are removed
from the environment after much longer period of time in comparison
with other groups of antibiotics [8].
Paracetamol, N-(4-hydroxyphenyl) acetamide) (PA), reported as one of
the most frequently consumed medicines in the world, is a phenacetin me-
tabolite exhibiting a strong analgesic and antipyretic effect [14,15]. It is
often used as an aspirin substitute due to detrimental influence of salicy-
lates on gastric mucosa [16]. The paracetamol effect is mainly based on lim-
itation of prostaglandin synthesis [11].
Considering the consequences of pharmaceutical residues in the envi-
ronment reaching concentrations even mg L−1 [17,18], the necessity to de-
velop new methods for their determination seems to be really important.
The most employed approaches described in literature for simultaneous de-
termination of ciprofloxacin and paracetamol are ultra-high performance

⁎ Corresponding author.
E-mail address: kochana@chemia.uj.edu.pl. (J. Kochana).

http://dx.doi.org/10.1016/j.jelechem.2020.114574
1572-6657/© 2020 Elsevier B.V. All rights reserved.
A. Pollap et al.
liquid chromatography coupled to tandem mass spectrometry [19,20], UV–
Vis detector [21] or photodiode array detector [22]. Also the capillary elec-
trophoresis with UV detection [23] and spectrophotometry technique [24]
are applied in this field. Due to a number of advantages characterizing elec-
trochemical methods such as low cost, simplicity, no sample preparation
necessity and short analysis time, they are considered as interesting alterna-
tive to previously mentioned methods. Electrochemical sensors for determi-
nation of antibiotics of different classes were developed for tetracyclines
[25,26], sulfonamides [27,28], β-lactams [29,30], phenicols [31,32], quin-
olones [33] and fluoroquinolones [34]. Immunoassays designated for anti-
biotics electrochemical quantification was reported as well [35].
Electrochemical sensing was also performed for painkillers, such as paracet-
amol [36–38], diclofenac [39] or morphine [40].
The best to our knowledge, so far only two electrochemical sensors were
developed for simultaneous ciprofloxacin and paracetamol determination.
Santos et al. proposed electrochemical sensor based on glassy carbon elec-
trode modified with graphene oxide and nickel oxide [14]. Authors applied
square wave voltammetry for analytes determination in enriched biological
samples, including urine and serum. Another approach was investigated by
Kingsley et co-workers [41]. For sensor construction authors modified a
carbon paste electrode with copper zinc ferrite nanoparticles. Developed
method was examined in determination of CPX and PA in urine and
serum samples, as well as in pharmaceutical formulations using adsorptive
stripping voltammetry.
In the present work, for the first time we propose an electrochemical
sensor based on TiO2 sol for simultaneous determination of ciprofloxacin
and paracetamol in environmental water samples. One of the most mean-
ingful steps during development of a new electrochemical sensor is the se-
lection of composite modifying the electrode surface. Its role is to improve
electrochemical sensor behaviour by increasing its conductivity and ensur-
ing the optimal rate of substrate penetration to the sensor surface and, as a
consequence, to better its analytical parameters. Among materials applied
as components of modifying layers carbon materials and metal nanoparti-
cles could be listen. The construction of proposed sensor was based on mod-
ification of graphite electrode with titanium dioxide sol enriched with
mesoporous carbon CMK-3, gold nanoparticles and Nafion. CMK-3 type
mesoporous carbon was applied due to its large surface area and high con-
ductivity [42,43]. The similar properties of increasing the conductivity are
characteristic to applied gold nanoparticles (AuNPs) [30,44]. Nafion, as a
conductive polymer, was chosen because of its conductivity increasing ca-
pability, as well as enhancing sensor long-term stability [27,45]. Electro-
chemical characteristics of developed sensor via cyclic voltammetry was
discussed for both analytes and the analytical parameters such as linear
range, sensitivity, limit of detection, selectivity and reproducibility were
designated. The functioning of proposed method was confirmed during de-
termination of ciprofloxacin and paracetamol in two types of environmen-
tal water matrices, as well as in certified reference material (CRM) of waste
water matrix.
2. Experimental
2.1. Chemicals and materials
Amoxicillin (>90% purity), ciprofloxacin (>98% purity), paracetamol
(>98% purity) sulfamethoxazole, HAuCl4·3H2O solution (1%), Nafion
((5% (w/v) solution in mixture of low aliphatic alcohol and water), K3[Fe
(CN)6] and titanium(IV) isopropoxide were purchased from Sigma-Aldrich
(USA); L-(+)-ascorbic acid, KH2PO4, Na2HPO4 ·2 H2O, potassium hydrogen
phthalate (C8H5KO4), ethanol (96%), HCl (35%), KHCO3, 2-propanol, and
trisodium citrate were obtained from Avantor Performance Materials
Poland S.A. (Poland); acetone and HNO3 (65%) were obtained from
LACHNER (Czech Republic); 0.3 mm alumina powder (for polishing graphite
electrode surfaces) was purchased from Buehler Micropolish (USA);
CH3COOH (100%), MgSO4 ·7H2O, NaNO3, and NH3(aq) (25%) were ob-
tained from Merck (Germany); CaCl2 was obtained from CHEMPUR
(Germany). CMK-3 type mesoporous carbon was synthesized by Chemical
2
Journal of Electroanalytical Chemistry 878 (2020) 114574
Technology Department at Faculty of Chemistry at Jagiellonian University
in Cracow, Poland. Certified reference material of waste water (EnviroMAT
Waste Water) was purchased from SCP Science (Canada). Abrasive papers
of various roughness (Norton, Poland) were applied for polishing the elec-
trode surface. 0.1 M phosphate buffer solutions (PB) of pH 5.0, 6.0 and 7.0
were prepared by mixing appropriate volumes of 0.1 M KH2PO4 and 0.1 M
Na2HPO4 solutions, while 0.1 M potassium hydrogen phthalate-HCl buffers
of pH 3.0 and 4.0 were prepared using the appropriate volumes of
C8H5KO4 and HCl solutions. Ciprofloxacin and paracetamol solutions as stan-
dard solutions were prepared in 0.1 M HCl. All chemicals were of analytical
grade. Aqueous solutions were prepared with Milli-Q water.
2.2. Apparatus and measurements
M161 electrochemical analyzer (Mtm-Anko, Poland) was used for cyclic
voltammetry measurements in potential range from −500 to 1200 mV at
scan rate of 62.5 mV s−1. The traditional approach involving the three-
electrode cup with developed sensor as working electrode, reference elec-
trode (Ag/AgCl/saturated KCl) and platinum wire as counter electrode
(both purchased from Mtm-Anko, Poland) was applied. The 0.1 M phosphate
buffer of pH 5 acted as supporting electrolyte. Sonic 3 ultrasonic bath
(Polsonic, Poland) and vortex (IKA, Germany) were used to prepare nano-
composite modified graphite electrode. For electrode construction pencil-
type graphite (Carbon, Poland) with a diameter of 6 mm and 0.28 cm2 geo-
metric surface area, cooper wires and Teflon bodies were used.
ERH-11 combined glass electrode (HYDROMED, Poland) and CP-501
pH-meter (Elmetron, Poland) were applied for buffers preparation.
The river water samples were characterized using the CX-701 portable
multifunction meter (Elmetron, Poland), an IJ44A pH electrode (Elmetron,
Poland), an ECF-1t conductivity cell of K = 0.45 cm−1 with metal electrode
and temperature sensor (Elmetron, Poland) and a COG-1t oxygen sensor
(Elmetron, Poland).
2.3. Measurement procedure
The cyclic voltammetry measurements were carried out in three-
electrode glass cup of volume 10 mL. The appropriate portions of samples
and standard solutions of ciprofloxacin and paracetamol were added into
5 mL of supporting electrolyte recording the three voltammograms (till sig-
nal stabilization) for given concentrations of analytes. The analytical char-
acteristics towards analytes was described based on maximum of
recorded two oxidation waves at potentials ca. 400 mV and ca. 700 mV
for ciprofloxacin and paracetamol, respectively. As calibration method the
standard addition method was applied.
2.4. Sensor preparation
Before modification of graphite electrode with appropriate nanocom-
posite, it was necessary to clean, polish and activate the electrode surface.
Thus, the abrasive papers of different roughness and alumina slurry
(0.3 mm) were applied for levelling and polishing its surface. For surface ac-
tivation purposes, the graphite electrodes were immersed and sonicated
successively in following solutions: ethanol, HNO3, NH3(aq), L-(+)-ascorbic
acid, acetone and finally in water [30].
For TiO2 sol preparation the previous described procedure based on
acid hydrolysis and titanium(IV) ispopropoxide polycondensation was ap-
plied [46]. Suspension of gold nanoparticles (AuNPs) was synthesized by
tetrachloroauric acid reduction applying sodium citrate solution [47]. Syn-
thesis of CMK-3 mesoporous carbon was based on hard templating method
with application of SBA-15 type mesoporous silica [48].
Nanocomposite preparation included mesoporous carbon CMK-3
weighing and addition of proper volumes of titanium dioxide sol mixed
with AuNPs (9:4, v/v), Nafion and 0.1 M phosphate buffer solution of
pH 6. After addition of each component the nanocomposite was shacked
using vortex and sonicated to obtain homogeneous. To modify electrode sur-
face composite was put on it using pipette.
A. Pollap et al.
The final nanocomposite used for preparation of developed sensor
consisted of (per electrode): 8.6 10−5 g of CMK-3, 4.6 μL of titanium diox-
ide sol, 1.9 μL of AuNPs, 6.0 μL of Nafion and 7.5 μL of PB (pH 6).
Before use, the constructed sensor was stored above saturated disodium
hydrogen phosphate diluted with water (1:1, v/v) for 20 h in 4 °C and later
for 20 h in 0.1 M phosphate buffer solution of pH 6 in 4 °C in order to TiO2
sol polymerization on the electrode surface [30].
2.5. Sample preparation
To verify the possibility of sensor application, ciprofloxacin and paraceta-
mol were simultaneously determinated in matrices of environmental water
from Wisła and Racławka rivers (Poland), as well as CRM of waste water. At
the sampling site, the following parameters of river waters were measured:
temperature (20.2 °C, 18.3 °C), pH (7.26 ± 0.12, 7.14 ± 0.09), conductivity
(1.27 ± 0.5 mS cm−1, 1.14 ± 0.5 mS cm−1) and oxygen percentage
(49.7 ± 3.8%, 52.4 ± 4.2%) for Wisła and Racławka, respectively (each pa-
rameter was measured three times). For electrochemical analysis no special
sample preparation was needed. CPX and PA were dissolved in 0.1 M HCl
and added to given type of environmental water and certified reference mate-
rial of waste water (1:24, v/v) to reach the proper analytes concentration.
3. Results and discussion
3.1. Preliminary voltammetric studies. Choice of nanocomposite composition
A very important issue during selection of electrode modifying layer is
both the composition of nanocomposite and the ratio of particular compo-
nents. Thus, a variety of nanocomponents were examined for graphite elec-
trode modification. The CV measurements were carried out for Fe(II)/Fe
(III) redox couple verifying the sensitivity as well as mechanical stability
of proposed sensors. The electrodes were modified with following nano-
composites: TiO2/Nafion/CMK-3, TiO2/AuNPs/Nafion/CMK-3 and TiO2/
AuNPs/PB/Nafion/CMK-3 with different ingredients ratio. The amount of
CMK-3 mesoporous carbon was fixed on constant level of 8.6 10−5 g for
each composite, chosen during previous experiments [30]. As could be
seen in Fig. 1 the matrix composite significantly affected the shape of regis-
tered CV curves. Enriching the TiO2/Nafion/CMK-3 composite (curve
A) with AuNPs (curve B) caused the increase of capacitive current which
was demonstrated in one of the previous reports [49]. This phenomenon
was probably due to increased active area of electrode [50].
The influence of PB of pH 6.0, being diluting agent, on recorded CVs
was noticeable. The addition of PB to the electrode composite changed
the signal for tested redox probe and allowed the formation of observable
oxidation and reduction signals (see curves B and C in Fig. 1). A higher de-
gree of composite dilution using PB could have contributed to greater distri-
bution of the modifying layer components and to exposure of more active
centers. Changing the component ratio in TiO2/AuNPs/PB/Nafion/CMK-
3 composite from (1: 1: 1: 1 (v/v/v/v)) to (2.40: 1: 3.96: 3.17 (v/v/v/v))
(curves C and D in Fig. 1, respectively) resulted in the highest sensitivity to-
wards tested redox probe (see insert of Fig. 1). Taking into account shapes
of recorded signals, sensitivity towards model depolarizer and mechanical
durability of modification mixture, the nanocomposite consisting of TiO2/
AuNPs/PB/Nafion/CMK-3 with ratio of 2.40: 1: 3.96: 3.17 (v/v/v/v) was
chosen as optimal layer modifying graphite electrode. In our previous re-
port it has been demonstrated that modification of electrode with devel-
oped composite enabled for increase of electroactive area of ca. 20% in
comparison to bare graphite electrode [30].
3.2. Electrochemistry of developed sensor towards ciprofloxacin and paracetamol
Both pharmaceuticals, ciprofloxacin and paracetamol are recognized as
electroactive compounds [14]. Firstly, to verify the possibility of simulta-
neous determination of both analytes, electrochemical measurements
were conducted for each of them separately. Recording the voltammetric
response of sensor in ciprofloxacin solution, the signals in potential range
3
Journal of Electroanalytical Chemistry 878 (2020) 114574
Fig. 1. Cyclic voltammograms recorded in PB of pH 6 containing 4.13 mM K3[Fe(CN)6]
in 0.1 M KCl at graphite electrode modified with different nanocomposites: A: TiO2/
Nafion/CMK-3 (2: 1 (v/v)), B: TiO2/AuNPs/Nafion/CMK-3 (1: 1: 1 (v/v/v)), C: TiO2/
AuNPs/PB/Nafion/CMK-3 (1: 1: 1: 1 (v/v/v/v)), D: TiO2/AuNPs/PB/Nafion/CMK-3
(2.40: 1: 3.96: 3.17 (v/v/v/v)) and their sensitivities taking into account oxidation
signal (for K3[Fe(CN)6] in 0.1 M KCl up to 4.13 mM); scan rate 62.5 mV s−1.
between 250 and 450 mV and between −100 and 100 mV were observed,
anodic and cathodic, respectively (see Fig. 2A). For paracetamol solution
two oxidation signals at 300–500 mV (poorly developed) and at
600–800 mV as well as wide reduction wave between 300 and 500 mV
were recorded in the same measuring conditions (Fig. 2B). Comparing the
recorded CV curves it could be observed that CPX oxidation signal, assumed
as analytical one, partially overlapped with the first, flat anodic wave of PA
at 300–500 mV. However, mentioned signal for paracetamol was recorded
in solutions of higher concentration (starting from 100 μM), much higher
than the content of paracetamol in environmental water [17]. In the next
step, the possibility of simultaneous CPX and PA determination was exam-
ined. Exemplary voltammogram recorded for solution of 25 μM CPX and
40 μM of PA is shown in Fig. 2C. The developed TiO2/PB/AuNPs/CMK-
3/Nafion sensor exhibited noticeable electrochemical response towards
both analytes. On voltammetric curve two anodic signals at 300–400 mV
and 600–800 mV were observed, the first being assumed to be the analyti-
cal signal for CPX and the second the for PA. Afterwards, the influence of
one analyte on the signal registered for the other was investigated. For
this the signals for three CPX solutions of increasing concentrations were re-
corded, then the portion of both analites (CPX and PA) was added and fur-
ther CPX additions were continued. Such measurements were conducted
for three PA concentrations: 2.5 μM, 5.0 μM and 10.0 μM. CPX calibration
curves thus obtained are presented in Fig. 3. Analyzing the graphs shown,
it could be seen that the addition of PA in concentration of 10 μM brought
on the increase of CPX signals higher than that resulting from the calibra-
tion relationship (see Fig. 3C). Application of adequate calibration method
as standard addition method could overcome this kind of interference effect
[51]. The measurements carried out to examine the effect of CPX on cali-
bration curves for PA did not confirm this influence (data not shown).
The verification of usefulness of standard addition method for analysis of
synthetic two-components samples was described in Paragraph 3.5.
The effect of scan rate on electrochemical response towards both analytes
is presented in Fig. 4A. It was noticed that as the scan rate increases the anodic
waves shifted to more positive potentials indicating an irreversible process.
Poorly formed anodic wave observed at higher scan rates for PA suggests oc-
currence of slow electrode reactions. The linear dependence of the oxidation
current on square root of the scan rate for both analytes shown on Fig. 4B sug-
gests that that oxidation process was diffusively controlled. However, the ob-
tained slopes values of plots log I vs. log v (see Fig. S1 in Supplementary
A. Pollap et al.
Fig. 2. Exemplary voltammograms recorded for 0.059 mM CPX solution (A), 0.1 mM
PA solution (B) and 0.025 mM CPX and 0.040 mM PA solutions (C) in 0.1 PB of
pH 5.0 at scan rate 62.5 mV s−1.
Information), 0.59 for PA and 0.68 for CPX, may indicate the diffusion-
controlled processes with a minor contribution of adsorption [52].
Based on the conducted research, considerations on the mechanisms of
electrode reactions were undertaken. PA molecule contains a benzene ring
substituted with hydroxyl and acetylated amino group in positions 1 and 4.
Because two oxidation signals were registered (see Fig. 2B) it can be as-
sumed that oxidation process comprises two separated steps. The first one
is associated with formation of radical which finally, as a result of delocal-
ization, forms N-acetyl-p-quinone imine as final product [41]. It is
suspected that the first step of PA oxidation is slow, that is why radical
form not immediately creates the final product, the imine. Thus, the first
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Journal of Electroanalytical Chemistry 878 (2020) 114574
oxidation signal for PA is visibly smaller than the second one as it was
shown in Fig. 2B. Proposed mechanism of CPX oxidation refers to two elec-
trons of nitrogen atom from piperazine ring (see Fig. 5). As described in the
literature, the –NH–CH2– group can be oxidized to form –N=CH– group
[41,53]. The proposed mechanisms of electrode reactions of both analytes
are presented in Fig. S2.
3.3. Optimization study
Type of supporting electrolyte has a significant influence on electro-
chemical determinations. Thus, optimization tests have begun with selec-
tion of supporting electrolyte for CPX determination. The range of pH
tested was from 3.0 to 7.0 applying 0.1 M potassium hydrogen phthalate-
HCl buffers (pH 3.0 and 4.0) and 0.1 M phosphate buffer solutions
(pH 5.0, 6.0 and 7.0). The measurements were carried out in antibiotic so-
lutions of concentration of 0.48–1.67 mM. Definitely the highest sensitivity
towards CPX was noticed for buffer of pH 5.0, while the lowest sensitivity
was determined for buffer of pH 3.0 (Fig. S3A). White precipitate was ob-
served during the measurements carried out in PB at pH 7.0, which was
probably associated with limited CPX solubility [54]. Therefore, as the op-
timal supporting electrolyte for CPX determination the PB of pH 5.0 was
chosen. For paracetamol determination in concentration range between
0.48 and 1.4 mM PB of pH 5.0, 6.0 and 7.0 were examined. For this analyte
the highest sensitivity was recorded for PB of pH 7.0, while the lowest for
pH 5.0 (see Fig. S3B). Taking into account the aim of the work, i.e. the de-
velopment of analytical method for two-component analysis, the presented
above results as well as the limited solubility of CPX at pH of 7.0, the PB of
pH 5.0 was choosen as supporting electrolyte for simultaneous CPX and PA
determination.
Another important issue for examination of sensor functionality is its re-
sponse time. This parameter was tested in CPX concentration range be-
tween 0.105 and 0.503 mM recording the signal after 180, 300 and 600 s
from addition of given portion of analyte. For 180-s measurements the no-
ticed sensitivity was the highest, while for the longest measurements the
sensitivity was the lowest. For paracetamol, no effect of response time on
measurement sensitivity was observed. Thus, as optimal response time for
developed sensor 180 s was chosen.
3.4. Evaluation of analytical parameters
The developed method for simultaneous ciprofloxacin and paracetamol
determination was characterized for each analyte by analytical parameters
such as linear range, sensitivity, limit of detection (LOD), reproducibility
and selectivity.
The linear range was determined based on the signals registered at ca.
400 mV (for CPX) and ca.700 mV (for PA) in solutions containing both
analites (Fig. 5). The developed sensor exhibited linear response (correlation
between oxidation current and analyte concentration) in ranges between 1
and 10 μM and from 10 to 52 μM for both analytes (Fig. 6). The higher sensi-
tivities were observed in lower CPX and PA concentration range.
The sensitivities (mean values of 5 sensors) were found to be
15.93 μA μM −1 and 11.56 μA μM −1 (56.89 μA μM −1 cm −2 and
41.29 μA μM −1 cm −2 ), in lower concentration range, for CPX and
PA, respectively. For higher concentrations, up to 52 μM, the sensitiv-
ities were about two times lower for both analytes, probably due to
blocking of sensor surface by reaction products. LOD values were cal-
culated taking into account measurements precision estimated for
0.1 μM analytes solutions [30]. Based on 3SD/a formula, where SD
is standard deviation of five current signals and a is average sensitiv-
ity, the LOD values were found to be 0.108 and 0.210 μM for CPX and
PA, respectively. To examine reproducibility of proposed method, the
measurements for solution of 5.10 μM CPX and 2.60 μM PA were car-
ried out using five independently prepared sensors. The lower RSD
values were noticed for CPX determination, RSD value was found to
be 2.1% while for PA 4.5% value was calculated. Therefore, it could
be concluded that developed method exhibited good reproducibility
A. Pollap et al. Journal of Electroanalytical Chemistry 878 (2020) 114574

Fig. 3. Influence of the PA addition (marked with an arrow) on CPX calibration curves; PA concentrations: 2.5 μM (A), 5.0 μM (B) and 10 μM (C).

Fig. 4. The effect of scan rate (from 10 to 70 mV s−1) on CV curves recorded in solution containing 0.6 mM CPX and 0.4 mM PA (A) and the anodic currents as a function of
square root of applied scan rate (B).

towards both analytes. Sensors were used for analysis within 2 weeks
of preparation and no deterioration in their analytical parameters
was observed during this time.
The selectivity of each analytical method is a very important issue, as it en-
ables for accurate determination of analyte in complex matrices. Considering
the presence of different compounds in environmental waters, the influence
− − −
of inorganic ions (Ca2+, K+, Mg2+, Na+, SO2− 4 , Cl , HCO3 and NO3 ) on sig-
nals recorded for CPX and PA was examined. Furthermore, amoxicillin and
sulfametoxazole, two different antibiotics were also taken into account as po-
tential interfering agents. The sensor selectivity was investigated by monitor-
ing electrochemical response of developed sensor in solution containing
20 μM CPX and 20 μM PA in presence of potential interferent at different

5
A. Pollap et al. Journal of Electroanalytical Chemistry 878 (2020) 114574

Comparing the analytical characteristic of sensors reported for si-

Fig. 5. Voltammograms recorded in ciprofloxacin and paracetamol solutions of
various concentration (up to 52 μM for both analytes) in PB of 5.0 at scan rate
62.5 mV s−1.
concentrations. It was found that for inorganic ions 15-fold excess did not sig-
nificantly affect the signal for CPX, however caused the 14% signal increase
for paracetamol suggesting additive interference effect. For 50-fold excess
of inorganic ions the signals changes were significant causing the 31% and
35% increase of signals for CPX and PA, respectively. Considering the influ-
ence of 2-fold lower concentration of amoxicillin, in comparison with
analytes concentrations no significant differences for CPX and PA were ob-
served. Nevertheless at higher amoxicillin concentration (20 μM) 18% signal
decrease for PA was noticed, simultaneously no signal changes for CPX were
observed. 2-fold lower concentration as well as the same concentration of
sulfametoxazole caused ca. 30% signal increase for ciprofloxacin and no sig-
nificant signal change for PA. In order to obtain accurate results of analysis
various calibration approaches could be employed, e.g. integrated calibration
[55], or generalized calibration strategy [56].
multaneous determination of CPX and PA and developed TiO2 /
AuNPs/PB/Nafion/CMK-3 electrode (see Table 1) it can be seen that
the lowest LOD towards CPX exhibited sensor based on copper zinc fer-
rite nanoparticles [41], while for PA determination the lowest LOD was
calculated for NiONP-GO-CTS:EPH/GCE sensor [14]. Although the de-
tection limit of proposed sensor was higher in comparison with another
sensors described in literature, proposed method of simultaneous CPX
and PA determination is characterized by very good sensitivity, signifi-
cantly higher than for sensors based on nickel oxide nanoparticles or
copper zinc ferrite nanoparticles. Additionally, proposed sensor does
not require application of harmful compounds during its construction
like epichlorohydrin, being recognized as carcinogen [57]. Thus, pro-
posed sensor based on titanium dioxide sol is in good agreement with
requirements set out by Green Chemistry Principles [58].
3.5. Determination of CPX and PA in synthetic and natural samples
To verify functioning of proposed method CPX and PA were simulta-
neously determined in synthetic samples with different analytes concentra-
tion. For calibration purposes standard addition method was applied using
five additions of standard solutions. The synthetic samples were analysed
three times at three different concentrations of CPX and PA (Table 2).
Obtained.
results showed that developed sensor allowed to simultaneous determine
both analytes at different level concentrations with high accuracy. The rela-
tive error values were within the range 0.6–2.4% and 2.6–3.6% for ciproflox-
acin and paracetamol, respectively. It could be seen that higher accuracy was
noticed towards ciprofloxacin. The proposed sensor demonstrated also good
measurement precision as evidenced by low RSD values. The relative stan-
dard deviation did not exceed 5% for CPX and PA as well. It is worth to men-
tioning that applied standard addition method enabled for limitation of
multiplicative effect discussed earlier (Paragraph 3.2).

Fig. 6. Calibration curves showing linear relationship between current and ciprofloxacin concentration (A) and paracetamol concentration (B).

Table 1
Comparison of analytical characteristics of electrochemical sensors designated for simultaneous determination of ciprofloxacin and paracetamol.
Sensor Technique Analyte Linear range [μM] Sensitivity LOD Sample Reference
[μA μM−1] [μM]

CZF/CPE ASV CPX 0.909 - 4.70 103 0.657 2.58 10−3 urine, serum, pharmaceutical preparations [41]
PA 0.185 - 0.0248 8.85 10−2
4.76 102
NiONP-GO-CTS: SWV CPX 0.040–0.97 9.87 6.0 10−3 urine, serum [14]
EPH/GCE PA 0.10–2.9 8.88 6.7 10−3
TiO2/PB/ CV CPX 1–10 and 15.93 1.08 10−1 environmental water this work
AuNPs/ PA 10–52 11.56 2.10 10−1
CMK-3/
Nafion/GE

CPE – carbon paste electrode, CST – chitosan, CZF - copper zinc ferrite nanoparticles, EPH – epichlorohydrin, GCE - glassy carbon electrode, GE – graphite electrode,
GO – graphene oxide, NiONP - nickel oxide nanoparticles, SWV - square wave voltammetry.

6
A. Pollap et al. Journal of Electroanalytical Chemistry 878 (2020) 114574

Table 2 Funding sources

Results of determination ciprofloxacin and paracetamol in synthetic samples.
Sample Analyte Concentration [μM] RE [%] RSD [%] This research did not receive any specific grant from funding agencies in
Expected Found (n = 3)
the public, commercial, or not-for-profit sectors.

1 CPX 2.55 2.61 2.4 1.1

PA 2.60 2.66 2.6 3.0 Credit authorship contribution statement
2 CPX 5.10 5.22 2.4 1.5
PA 2.60 2.57 −3.0 3.5
3 CPX 20.40 20.28 −0.6 4.7 Aleksandra Pollap: Conceptualization, Methodology, Validation,
PA 20.80 21.54 3.6 2.0 Formal analysis, Resources, Data Curation, Writing - Original Draft, Writing
- Review & Editing. Karolina Baran: Validation, Formal analysis, Investi-
gation, Resources. Natalia Kuszewska: Validation, Formal analysis, Inves-
Table 3
tigation. Jolanta Kochana: Conceptualization, Methodology, Data
Results of determination ciprofloxacin and paracetamol in environmental water Curation, Supervision, Writing - Review & Editing.
matrices.
Type of matrix Analyte Concentration [μM] RE [%] RSD [%] Declaration of Competing Interest
Expected Determined⁎

Wisła river CPX 18.20 18.46 1.4 2.1 The authors have declared that do not have any conflict of interest.
PA 13.24 12.98 2.1 3.3
Racławka river CPX 18.20 18.06 0.7 3.6 Appendix A. Supplementary data
PA 13.24 13.98 5.3 2.5
CRM of waste water CPX 20.00 19.99 −0.1 0.5
PA 15.05 14.96 −0.6 2.7
Supplementary data to this article can be found online at https://doi.
*
org/10.1016/j.jelechem.2020.114574.
Average value (n = 3).

Constructed sensor was also tested in analyses of natural samples. They
were carried out in matrices of environmental water from two polish rivers
as well as in CRM of waste water. Firstly, the signals for unmodified samples
were recorded. As no ciprofloxacin and paracetamol were detected,
analysed waters and CRM were spiked with CPX and PA. The samples
were analysed three times at different concentrations of CPX and PA. Ob-
tained results are shown in Table 3. From data presented in Table 3 it
could be concluded that accuracy of proposed sensor could be regarded
as satisfactory. RE values were low, but for PA determination in Racławka
river the RE value was comparatively higher (5.3%). RSD values describing
method precision were low and did not exceed 3.6% and 3.3% for CPX and
PA, respectively. It was confirmed that developed sensor has a potential to
be applied for environmental water analysis as well as waste water with re-
spect to CPX and PA quantity.
4. Conclusions
In presented paper, the new electrochemical method for simultaneous
determination of ciprofloxacin and paracetamol was proposed. So far elec-
trochemical methods for simultaneous CPX and PA determinations were
designated for biological and pharmaceutical samples. For the first time
the method for simultaneous determination of CPX and PA was verified
in environmental water analysis. To construct a sensor the graphite elec-
trode was modified with TiO2 sol enriched with CMK-3 mesoporous car-
bon, gold nanoparticles and Nafion to improve its electrochemical
response. Developed sensor exhibited a good analytical parameters, espe-
cially high sensitivity. Moreover, the analysis of real samples showed that
developed sensor could be applied for simultaneous determination of cipro-
floxacin and paracetamol in environmental waters and waste water with
high accuracy and precision. It was demonstrated that employing standard
addition calibration method no special sample preparation is needed. Tak-
ing into account time and cost of analysis, no need for hazardous reagents
during sensor construction as well as no special samples preparation, devel-
oped method exhibits many advantages over other analytical methods for
simultaneous determination of CPX and PA such as chromatographic and
electrophoretic methods. The further research on the sensor presented in
the work will be focused on the possibility of miniaturization of the system
using screen-printed electrodes. Also the possibility of simultaneous deter-
mination of CPX and PA in environmental samples with complex matrices
e.g. municipal and hospital sewage will be verified.
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