Chemical Engineering Journal Advances 7 (2021) 100118

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Chemical Engineering Journal Advances

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

Molecularly imprinted polymer synthesis and selective solid phase

extraction applications for the detection of ziram, a dithiocarbamate
fungicide
Çiğdem Öter∗, Özlem Selçuk Zorer
Faculty of Science, Department of Chemistry, Van Yuzuncu Yil University, 65080 Van, Turkey

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

Keywords: It is becoming more and more critical to identify and eliminate pesticide residues that are intended to be used
Molecularly imprinted polymers in combating agriculture but pose a threat to all living things due to their unconscious use. In this study, a high-
Pollutants performance molecular imprinted material (MIP) was synthesized to remove ziram, a dithiocarbamate fungicide,
Solid-phase extraction
from its liquid phase. After characterization studies of the printed polymer synthesized by the suspension poly-
Suspension polymerization
merization process, the re-bonding ability and the suitability of the adsorption process to kinetic and isothermal
Ziram
models were compared with an non-imprinted reference polymer (NIP). Selectivity of MIP adsorbent prepared by
imprinting method against template molecule ziram; their adsorption ability against different dithiocarbamate
fungicides (maneb, ferbam, and propineb) was determined by testing. Some extraction parameters were opti-
mized to evaluate the extraction performance of MIPs used as adsorbents in the solid-phase extraction column.
It has been found that under optimized conditions, the developed MIP-SPE method provides high recovery for
ziram. This method was developed; it was determined that MIP-SPE showed higher extraction efficiency against
ziram than commercial C18-SPE and non-print polymer solid-phase extraction (NIP-SPE) processes. Thus, the
MIP-SPE method has proven to be an environmentally friendly, sensitive, easy-to-work, time-saving, efficient
sample pretreatment method that uses less solvent. These findings will provide a fundamentally technical for the
removal of the trace hazardous pollutants.

1. Introduction
Pesticides as agricultural chemicals; It finds widespread use to in-
crease agricultural production by controlling or killing various fungal
diseases, pests and insects. Due to the worldwide increase in pesticide
use in recent years, natural waters are severely polluted by various pes-
ticides. These chemicals applied directly to the groundwork are quickly
carried to groundwater [1,2]. Also, uncontrolled application of pesti-
cides can pose a danger to public health. Ziram, a black spot, is a group
of dithiocarbamate fungicides widely used to control molds, blight, and
rotting and improve the quality of fruits and vegetables during trans-
portation and storage. Also, it is used in the rubber processing industry
as an accelerator or booster that facilitates the increased vulcanization
rate [3]. As a result of ziram residues, serious health problems such
as nausea, headache, and even cancer can occur. Also, it can lead to
asthma, risk of Parkinson’s disease, skin allergies, respiratory and eye
inflammation [4,5]. Due to these adverse effects, USA Environmental
Protection Agency (EPA) has determined ziram residues between 2 and
7 ppm as the tolerable limit for bananas, peaches, strawberries, pears,
and apples [6,7].
Thus, there is a need to develop selective and precision analytical
methods for environmental pollutant determination, monitoring, and
control of pesticide residue levels. The most commonly used analytical
methods for detecting pesticides such as ziram; high performance liq-
uid chromatography (HPLC) [8], HPLC followed by ultraviolet (UV) or
mass spectroscopy [9], liquid chromatography-mass spectrometry (LC-
MS/MS) [10], electrochemical detection [11], biosensors [12], stir bar
sorptive extraction (SBSE) [13], gas chromatography-mass spectrom-
etry (GC–MS) [14], dispersant liquid-liquid microextraction (DLLME)
[15], solid-phase extraction (SPE) [16] and solid-phase microextraction
(SPME) [17].
More specific and innovative methods should be applied with tra-
ditional approaches to remove contaminants from water selectively.
Molecularly printing technology; is based on the synthesis of materials
capable of molecular recognition in the template molecule’s existence.
With memory regions created by the following removal of the template
molecule, fabrics were produced that selectively recognize and rebind

∗
Corresponding author.
E-mail address: cigdemoter@yyu.edu.tr (Ç. Öter).

https://doi.org/10.1016/j.ceja.2021.100118
Received 26 February 2021; Received in revised form 23 April 2021; Accepted 24 April 2021
2666-8211/© 2021 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/)
Ç. Öter and Ö.S. Zorer
the original template molecule from a mixture. A few synthesis processes
have been improved for MIPs, such as precipitation polymerization, bulk
polymerization, suspension polymerization, in-situ polymerization, and
surface polymerization. [18]. Molecularly imprinted polymers (MIPs)
have numerous properties such as easy synthesis, low cost, excellent
reusability in harsh physical and chemical conditions, and high stabil-
ity.
MIPS; recently used as selective solid-phase extraction (SPE) sorbents
to pre-concentrate and selectively remove contaminants in environmen-
tal applications. Also, selective solid-phase extraction, where molecu-
larly imprinted polymers were used as sorbents, is used for the produc-
tive separation of trace compounds in complicated matrices resistant
to diverse solvents and temperature. There has been much research on
the use of MIPs in wastewater treatment. Some of the studies in which
MIPs are applied successfully; pesticides [19], serum [20], urine [21],
food [22], pharmaceuticals [23], personal care products [24], biolog-
ical samples [25]. Solid-phase extraction combined with UV-–vis spec-
trophotometry is recommended as it provides advantages in terms of low
cost, simplicity, and miniaturization for small volume samples [26].
In this study, using suspension polymerization technique (0.5: 4: 20;
template: monomer ratio: crosslinker molar ratio), ziram printed poly-
mer (MIP), and non-imprinted polymer (NIP) were synthesized. The sus-
pension polymerization method has been used to get ahead of irregular
particles’ formation and prevent unnecessary labor and time loss. Syn-
thesized polymers; It was characterized by scanning electron microscopy
(SEM) and Fourier conversion infrared spectrophotometer (FT-IR). Bind-
ing experiments examined the adsorption properties of MIPs. Selective
analysis of ziram in contaminated waters was performed by using MIP
as adsorbents in the MISPE-UV process.
2. Experimental
2.1. Materials and apparatus
Zinc dimethyl dithiocarbamate (Ziram), Iron (III) dimethyl
dithiocarbamate (Ferbam), Manganese ethylene dithiocarba-
mate (Maneb), Propylenebis(dithiocarbamate) acid (Propineb),
methacrylic acid (MAA) 99%, Polyvinyl alcohol (PVA), 2,2′- azo-
bis (2-methylpropionitrile) (AIBN), ethylene glycol dimethacrylate
(EGDMA) 98%, chloroform, acetic acid (CH3 COOH), methanol (MeOH)
were obtained from Sigma-Aldrich (USA). All their agents used were of
analytical grade.
FT-IR spectrum of the samples was saved on a Bio-Rad-Win-IR Model
FTIR spectrometer. The polymers’ morphology was monitored by scan-
ning electron microscopy (SEM, Zeiss Sigma 300 FESEM). Optizen-POP
model UV spectrophotometer was used for absorbance measurements.
An 620 Lab pH Meter was used for the pH settings. VisiprepTM SPE
Vacuum Manifold-Standard, 12-port model with was purchased from
Sigma-Aldrich. Empty SPE cartridge (3 mL cartridge) and SPE-C18 car-
tridge (3 mL cartridge) were supplied from Teknokroma.
2.2. Synthesis of MIP and NIP
Polymers were synthesized by the suspension polymerization
method [27]. MIP synthesis was prepared with a stationary tem-
plate/monomer /crosslinker ratio (0.5: 4: 20). The template molecule
Ziram (0.5 mmol) and functional monomer MAA (4 mmol) were stirred
at room temperature in 20 mL of chloroform for about 60 min to form
the pre-complex in a round bottom flask. In a different beaker, 200 mg
of polyvinyl alcohol (PVA) was dissolved in 25 mL of ultrapure wa-
ter by heating to ensure homogeneity. Then, the crosslinker EGDMA
(20 mmol), 20 mg initiator AIBN, and polyvinyl alcohol solution were
added to the polymerization flask and purged with a nitrogen stream
for 10 min. It was put in an oil bath at 60 °C for 24 h to realize the
polymerization process. Besides being a fungicide, ziram is used as an
accelerator or booster that facilitates the increased vulcanization rate in
2
Chemical Engineering Journal Advances 7 (2021) 100118
the rubber processing industry. The fact that the ziram fungicide used
as a template also has an accelerating effect increased the overall re-
action rate in the radical polymerization reaction. The formed polymer
was extracted using 100 mL of 90:10 (v:v) methanol: acetic acid with a
Soxhlet apparatus for 72 h until no ziram was detected. Subsequently,
the polymer was washed using methanol and dried at 50 °C for 24 h.
Non-imprinted polymers (NIP) were synthesized following the same
procedure but without the template molecule ziram.
2.3. Rebinding experiments
For research on the rebinding ability of the synthesized polymers,
the adsorption of ziram in an aqueous solution was evaluated using the
same amount of MIP, and NIP. 10 mg MIP and NIP adsorbents were
mixed with 50 mg/L Ziram solutions at room temperature. The amount
of ziram remaining in the solution at the end of the adsorption process
was observed at 295 nm using a UV/vis spectrophotometer. pH change is
an essential parameter in terms of adsorption efficiency as it affects both
the polymer surface and the ziram molecule. To determine the effect of
pH change on the adsorption features of polymer molecules; The pH
values of 50 mg/L ziram solutions were changed from 4 to 9, and their
adsorption on MIP and NIP was studied. Experimental data, adsorption
capacity (g) of polymers and adsorption capacity (Qt ) per unit mass, and
adsorption capacity of the polymers at equilibrium (Qe ) were calculated
by the following equations Eq (1)-(2):
( )
𝐶0 − 𝐶𝑡
𝑄𝑡 = 𝑥𝑉 (1)
𝑊
( )
𝐶0 − 𝐶𝑒
𝑄𝑒 = 𝑥𝑉 (2)
𝑊
Here; Co , Ce , and Ct are the initial concentration, equilibrium con-
centration, and concentration at time t (mg/L), respectively. V (mL) is
the volume of the solution, and w (mg) is the mass of the polymer par-
ticles.
In studies conducted for MIP and NIP, a rise in adsorption efficiency
was determined between pH 3–6, and maximum adsorption efficiency
was obtained at pH 6 (86.44%, 50.38% for MIP and NIP, respectively).
Hydrophobic interactions and hydrogen bonds have been a driving force
for the ziram molecule to be absorbed into the particular binding sites
of the MIP [28]. A decrease in adsorption efficiency was observed when
pH> 7. As the pH increased, there was an interaction between the ziram
molecule and the polymer adsorbents, while competition between elec-
trostatic repulsion and hydrophobic interactions occurred. Because of
the repulsive interactions arising from this competition, the adsorption
efficiency has decreased [29]. 6 pH values at which maximum adsorp-
tion efficiency was observed were chosen for further studies.
2.4. Adsorption kinetics and isotherms
Experiments with different contact times (between 5 and 150 min)
were performed at room temperature for kinetic studies. 10 mL of ziram
solutions with a concentration of 50 mg/L (pH 6) were mixed with 10 mg
MIP or NIP. At the end of the period, the absorbance was eliminated by
filtration, and the amount of ziram remaining in the solution was exam-
ined using a UV/Vis spectrophotometer. Kinetic parameters were deter-
mined using adsorption kinetics models, and the adsorption process was
estimated. In the study, the kinetics of ziram adsorption on MIP and NIP
particles were analyzed using four diverse kinetic models: the Pseudo-
first-order, Pseudo-second-order, Weber-Morris intra-particle diffusion
models, and Elovich kinetic model [30,31,32,33].
Experimental data tests were applied on the adsorption isotherm
models of Langmuir, Freundlich, Temkin, and Dubinin-Radushkevich to
examine the affinity and surface properties f polymer particles as well as
the analysis of adsorption mechanisms [34,35,36,37]. Adsorption stud-
ies were performed at various ziram concentrations (between 10 and
150 mg/L) using 10 mg of MIP or NIP.
Ç. Öter and Ö.S. Zorer Chemical Engineering Journal Advances 7 (2021) 100118

Besides, the applicability of every model for adsorption systems is Table 1

also examined by analyzed the sum of error squares (SSE) as given in Kinetic parameters of ziram adsorption on polymers.
(Eq. (3)) below. Models Parameters MIP NIP
√
∑( )2 Pseudo-first order Qe (mg/g) 7.51 4.85
𝑄𝑒,𝑒𝑥𝑝 − 𝑄𝑒,𝑐𝑎𝑙
SSE = (3) k1 (1/min) 0.112 0.062
𝑁 R2 0.9864 0.9330
SSE 3.5364 2.3238
where; Qe,exp (mg/g) and Qe, cal (mg/g) respectively are the adsorption
Pseudo-second order Qe (mg/g) 45.25 25.84
capacity of the ziram at equilibrium obtained from the experiment and k2 (g/mg.min) 0.037 0.064
calculated from the model. N is the number of data points [38]. R2 0.9994 0.9988
SSE 0.4186 0.3905
Elovich 𝛼 (mg/g.min) 4.94 × 1015 4.37 × 105
2.5. MIP-SPE procedure 𝛽 (g/mg) 0.92 0.62
R2 0.9952 0.8866
The 40 mg MIP was packaged dry in 3 mL SPE cartridges, and 10 mL SSE 4.039 3.2666
of methanol and 10 mL of deionized water were used for condition- Weber-Morris k (mg/g.min1/2 ) 1.59 1.005
intra-particle diffusion I (mg/g) 37.4 20.63
ing, respectively. Then 10 mL of ziram solution at a concentration of
R2 0.9771 0.9574
50 mg/L was loaded and washed with 3 mL of wash solution. Finally, SSE 1.5948 1.2062
the cartridge was air-dried for 5 min. It was separated from the solid
phase with the elution solvent (6 mL). Ziram in the eluent was detected
spectrophotometrically in solution at 295 nm (using an Optizen POP
and 2953 cm−1 are the stretching vibrations of =C–H and C–H, respec-
UV–vis spectrophotometer). Using the same procedure for comparison,
tively. 1726 cm−1 and 1641 cm−1 were, respectively, C = O and C = C
commercial C18-SPE and NIP SPE analyzes.
stretching vibrations. The peaks at 1242 cm−1 and 1147 cm−1 were the
stretching vibrations of C–O in carboxyl and ester, respectively, showing
3. Results and discussion the presence of MAA and EGDMA The peaks from 3600cm−1 to 3400
cm−1 were the tensile vibrations of O–H and N–H, respectively [43].
3.1. Synthesis of MIP for ziram FTIR absorbance changes are because ziram forms hydrogen bonds with
the functional monomer MAA in the reaction process in the imprinted
In this study, by using the suspension polymerization process, im- material [44]. It can be seen that the polymerization procedure per-
printed polymer synthesis has been made to remove ziram, which is a formed successfully as the chemical spectra of both NIP (Fig. 4b) and
type of DTC fungicide. The scheme of the synthesis is shown in Fig. 2. As MIP (Fig. 4c) after extraction were similar.
a functional monomer, MAA, capable of forming hydrogen bonds with
the template molecule ziram, capable of acting both as a hydrogen bond 3.3. Adsorption kinetics
donor and as a hydrogen bond acceptor, was chosen. When selecting
the porogen; Solvents capable of promoting the formation of template- The adsorption capacity of ziram (50 mg/L) for MIP and NIP was ex-
functional monomer complexes are preferred. Solvents such as acetoni- amined, and the data are shown in Fig. 5. Accordingly, while the adsorp-
trile, chloroform, and toluene are widely used to prepare printed poly- tion capacity of polymer particles rises in the first 15 min, it decreases
mers. When a porogen solvent with a high dielectric constant is applied; after reaching equilibrium. The rapid uptake of dissolved ziram during
The bonding features of imprinted polymers are significantly developed the initial times may be due to many usable areas on the adsorbent and
compared to the unimprinted polymer [39]. Chloroform was used as a the high concentration gradient between ziram in solution. At the end
porogen in this study. EDGMA was chosen as a crosslinker because it of a certain period, the retention of ziram in the remaining empty ar-
has sufficient mechanical stability and provides high crosslinking rates. eas became difficult, and the adsorption capacity decreased due to the
Another critical parameter in imprinted polymer design is the type of thrust between the ziram and the surface of the adsorbent. Adsorption
initiator. First, AIBN was used, which produces fewer side reactions and equilibrium was achieved at 20 min for MIP and 30 min for NIP. In
a free radical with stable properties [40]. In the synthesis reaction, the Fig. 5, it was observed that the adsorption capacity of the printed poly-
temperature and reaction time were 60 °C and 24 h, respectively; fur- mer particle is higher than the unimprinted polymer particle. The higher
ther, the molar ratio of template/ monomer/ crosslinker was fixed at binding capacity is due to particular imprinting sites in the MIP. By Sri-
0.5: 4: 20. vastava et al., in the study conducted to evaluate ziram’s adsorption
The formed polymer was extracted using 100 mL of 90:10 (v:v) properties on different soil types in the Allahabad region, the optimum
methanol: acetic acid with a Soxhlet apparatus for 72 h until no ziram contact time was found to be 2 h [45]. Another study for the adsorption
was detected. As a result of the reaction, a high affinity and selectivity of ziram to the hanging mercury drop; the maximum contact time was
ziram imprinted polymer synthesis was performed using the suspension 1500 s (25 min) by Giannakopoulos and Deligiannakis [46].
polymerization method (Fig. 2). Experimental kinetic adsorption data to determine the ziram uptake
rate of adsorbents during the adsorption process; The pseudo-first-order,
3.2. Characterization of polymers pseudo-second-order, were fitted to the Weber-Morris intra-particle dif-
fusion model and Elovich model. Kinetic parameters got from the models
The morphology of MIP and NIP particles was studied using SEM. As are shown in Table 1. When the R2 in the table is examined, it is seen that
seen in Fig. 3, MIP and NIP particles exhibit irregular spherical shapes the R2 value of the pseudo-second-order kinetic model for polymer par-
with heterogeneous sizes. MIP particles have a diameter of about 30 ticles is higher than the other models and this model fits the experimen-
𝜇m (Fig. 3b) and spherical morphology. The MIP surface is rougher and tal data better. Accordingly, ziram adsorption was mainly controlled by
denser, offering more bonding sites for templates than the NIP surface. chemical adsorption [47]. The Weber-Morris intraparticle diffusion plot
When the morphological structures of polymer particles are examined, defines in Fig. 6 was used to identify whether intraparticle diffusion
due to their dence and porous nature, MIP particles are thought to pro- or outer diffusion is the rate-limiting step. Since the intraparticle dif-
vide better field access for both ziram uptake and diffusion [41,42]. fusion graph does not pass through the starting point, this model was
FTIR analysis was performed to determine functional groups and examined, and it was seen that intra-particle diffusion was not the dom-
bond types in printed and unprinted polymers. The FT-IR spectra of (a) inant step, and the adsorption mechanism was governed by more than
MIP, (b) NIP, and (c) MIP after extraction are shown in Fig. 4. 2995 cm−1 one mechanism [48]. Based on the 𝛼 values calculated from the Elovich

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Ç. Öter and Ö.S. Zorer Chemical Engineering Journal Advances 7 (2021) 100118

Fig. 1. Structure of used dithiocarbamate

fungicides.

Table 2 surface. The maximum monolayer adsorption coefficient (Qmax) value

Isotherm parameters of ziram adsorption on polymers. was found as 56.98 mg/g for MIP and 17.75 mg/g for NIP. RL values (di-
Models Parameters MIP NIP mensionless separation factor) were seen as 0.024 and 0.66 for MIP and
NIP, respectively. These values in the range 0 <RL <1 show that the ad-
Langmuir Qmax (mg/g) 56.18 17.75
sorption process is suitable, and the binding affinity of the ziram to the
b (L/mg) 0.157 0.013
RL 0.24 0.66 selective regions of the MIP is higher. Since the n values calculated from
R2 0.9929 0.9912 the Freundlich isotherm equation are in the range of 1<n<10 for both
SSE 0.8052 5.1876 polymer particles, it can be concluded that both adsorptions are applica-
Freundlich Kf [(mg/g)(L/mg)1/n )] 16.39 0.65
ble. Temkin bT constant is the change constant of the adsorption energy,
n 3.54 1.91
R2 0.9826 0.9156
which demonstrates whether the adsorption reaction is endothermic (bT
SSE 6.2223 7.2355 <1) or exothermic (bT >1). bT >1 indicates that the adsorption reaction
Temkin bT 9.44 13.8 of ziram on polymer particles is exothermic [49]. Small E values in the
KT (L/g) 3.69 0.196 Dubinin-Radushkevich model; show that physical forces are influential
R2 0.9689 0.9223
in adsorption phenomena.
SSE 6.9593 5.6925
Dubinin-Radushkevich Qmax (mg/g) 45.17 6.4 These interactions result from van der Waals forces, hydrogen bonds,
𝛽 (1/mol2 J2 ) 0.978 4.55 hydrophobic interaction, and electrostatic forces. The results showed
E (J/mol) 0.105 0.104 that MIP had more pronounced binding sites and higher binding con-
R2 0.8409 0.7826
stants to non-print polymer than NIP. This state demonstrated that the
SSE 2.4232 6.5833
printed polymer’s binding affinity was due to the printing process’s spe-
cific positions.

model, it was found that the 𝛼 value of the MIP (4.94 × 1015 mg/g.min)
has a higher initial sorption rate compared to the 𝛼 value of the NIP
(2.2 × 105 mg/g.min). Obtained results prove that the molecularly im-
printed polymer shows faster binding kinetics than the non-imprinted
polymer [38].
3.4. Adsorption isotherms
By investigating the suitability of the data obtained by the adsorp-
tion of the ziram solutions in different concentrations on MIP and NIP
(Fig. 7), the isotherm models of Langmuir, Freundlich, Temkin, and
Dubinin Radushkevich; adsorption isotherm parameters are shown in
Table 2. When the R2 values are examined, it is seen that the most suit-
able isotherm model for MIP (R2 = 0.9929) and NIP (R2 = 0.9912) is the
Langmuir isotherm model. These data show the adsorption of ziram to
the binding sites homogeneously distributed on the polymer particles’
3.5. The selectivity of MIP
Ferbam, Maneb, and Propineb are a kind of dithiocarbamate (DTC)
fungicides like Ziram, and their chemical structures are analogous to
Ziram (the chemical structures of these molecules are shown in Fig. 1).
The maximum adsorption wavelengths for the other three fungicides se-
lected were determined as 362 nm for Maneb, 594 nm for Ferbam, and
272 nm for Propineb. Since the fungicide species’ maximum absorp-
tion wavelengths are far from each other, we can use the UV–vis spec-
trophotometer to evaluate the adsorption capacity over a broad spec-
trum range. For selective binding studies, each fungicide in the concen-
tration of 50 mg/L; in pH 6, at 20 min, at room temperature, mixed
with 10 mg MIP or NIP. At the end of the study, MIP’s target molecule
can be seen in Fig. 8, where the adsorption efficiency is higher than
the other fungicide types. In the experiments conducted under the same

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Ç. Öter and Ö.S. Zorer Chemical Engineering Journal Advances 7 (2021) 100118

Fig. 2. Schematic representation of ziram im-

printed polymer.

Fig. 3. Scanning electron microscopy of polymers: (a, b) MIP

and (c, d) NIP. Scale bar: 200 nm for (a, c); 100 nm for (b, d).

5
Ç. Öter and Ö.S. Zorer
Fig. 5. Equilibrium profiles of MIP and NIP for ziram (pH= 6, C0 = 50 mg/g,
T = 25 °C, w = 10 mg).
conditions, 89.65% of Ziram, 39.26% of Maneb, 45.32% of Ferbam, and
37.68% of Propineb were adsorbed by MIP (Fig. 8). These results proved
that the MIP has higher molecular recognition selectivity for template
ziram. Successful printing gaps are formed in the polymerization process
based on the shape, functionality, and size interaction with the template
molecules.
As expected, low adsorption yields were obtained in experiments
with NIP. The NIP adsorbent’s adsorption capacity, which does not have
specific binding sites, is lower than that of the MIP, and the Ziram is not
selectively adsorbed. No significant difference was observed between
fungicide molecules in adsorption processes with NIP. Adsorption ef-
ficiency of non-selective NIP for Ziram; ranges between 44.02% and
23.11% (Fig. 8). It has been found that the adsorption rates of each
compound are close to each other due to possible hydrogen bonds be-
tween the amine groups of the fungicides and the carboxylic groups of
the MAA.
The MIP can only selectively recognize Ziram because of imprinted
sites that match the template molecule. The imprinting factor (𝛼), de-
fined as the ratio of adsorption quantities of imprinted polymer (QMIP )
to non-imprinted polymer (QNIP ); and selectivity factor (𝛽), defined as
6
Chemical Engineering Journal Advances 7 (2021) 100118
Fig. 4. FT-IR spectra of (a) MIP (b) NIP and (c) MIP after extraction.
Fig. 6. Intra-particle diffusion kinetic model plots of MIP and NIP.
the ratio of adsorption amount of template (QZ ) to that of the com-
petitor reference fungicide molecules (QRF ), were defined by following
equations, respectively Eqs. (4)-(5) [50]:
𝑄𝑀 𝐼 𝑃
𝛼= (4)
𝑄𝑁 𝐼 𝑃
𝑄𝑍
𝛽= (5)
𝑄𝑅𝐹
Factors 𝛼 and b were calculated from adsorption experiments and
shown in Table 3. According to Table 3; The adsorption amount of the
imprinted polymer (MIP) for the template molecule ziram is higher than
the other three molecules (ferbam, maneb, and propineb) as reference
fungicides. These results show that the MIP has significant adsorption
selectivity for the template ziram.
In another study conducted by Yılmaz and Soylak on magnetic solid-
phase extraction of Ziram in food and water samples using magnetic
carboxylated nanodiamonds, a very high rate of ziram recovery was ob-
tained from synthetic mixtures [51]. In the study by Malik et al.; (1,2′
Ç. Öter and Ö.S. Zorer Chemical Engineering Journal Advances 7 (2021) 100118

was determined 90%. It was found that despite the increasing amount
of MIP, there was not much difference in the amount of ziram extracted.
Therefore, the optimum adsorbent amount was chosen as 40 mg for SPE-
MIP methods.
The washing step is an important procedure to maximize the spe-
cific printing interaction between the loaded sample and the binding
sites and remove non-target molecules. In the polymerization of MIP,
the selected porogen shape and character recognition level are very
high, so it is generally used as a washing solvent [53]. In this pro-
cess, 3 mL of washing solvent; Different mixtures of water-chloroform
(100:0, 80:20, 50:50, 20:80, 0:100 v/v) were used. Washing solutions;
it has been evaluated to achieve the minimum and maximum recov-
ery. The higher the percentage of organic solvent used, the greater the
recovery of ziram. Maximum recovery was achieved using an (80:20,
v/v) chloroform-water solution. Methanol was selected as an eluent
solvent for MIP-SPE; it has strong hydrogen bonds, high analyte per-
meability, and elution efficiency-enhancing properties [54,55]. The ex-
traction capacity increases with acetic acid added to methanol in fixed
proportions for elution [29]. Therefore, the recovery percentages were
Fig. 7. Adsorption isotherms of MIP and NIP for ziram (pH=6, T = 25 °C, tested using solvents (Methanol/Acetic acid; 100:0, 90:10, 80:20, 70:30,
w = 10 mg, C0 =10–150 mg/L). 60:40, 50:50, v/v) formed by adding CH3 COOH to methanol in differ-
ent proportions.The results show that the analyte recovery is increased
Table 3 by adding acetic acid (Fig. 9B). The highest percentage of recovery was
The imprinting factor and selectivity coefficient data. observed in Methanol/Acetic acid (90:10 v/v) solvent. Therefore, the
Fungicide Type QMIP QNIP 𝛼 𝛽 Methanol/Acetic acid (90:10 v/v) solvent was chosen as the optimum
elution solvent for MIP-SPE. Different volumes of elution solvents (3, 6,
Ziram 56.9 21.63 2.63 -
Maneb 19.29 12.37 1.56 QZ /QM 2.95 9, 12 and 15 mL) were used to optimize the elution volume. According
Ferbam 22.27 15.82 1.41 QZ /QF 2.55 to the SPE results, the extracted ziram recovery showed the maximum
Propineb 18.51 11.35 1.63 QZ /QP 3.07 value (94.1%) when the solvent volume was 6 mL (Fig. 9C). Besides, as
the elution volume was increased, a decrease in analyte recovery was ob-
served. Therefore, the optimum elution volume for the MIP-SPE method
Pyridylazo) −2-naphthol (PAN)-Naphthalene was used as adsorbate af- was determined as 6 mL.
ter mixing with other dithiocarbamate solutions, the ziram solution was Different loading volumes ranging from 5 to 25 mL were used for
analyzed by the general procedure. It has been reported that ziram was ziram solutions at a concentration of 50 mg/L. Samples loaded in differ-
detected at a high rate even in the presence of other dithiocarbamates ent volumes were washed with 3 mL of the chloroform-water mixture
[52]. (80:20, v/v) and eluted with 6 mL of elution solution. Recoveries for
sample volumes selected as 5, 10, 15, 20 and 25 mL were 71.2%, 97.6%,
65.1%, 58.4% and 49.7%, respectively. Recovery is maximum when the
3.6. Optimization of the MISPE procedure
loading volume is 10 mL (Fig. 9D). Recovery decreased as the loading
volume increased. This result indicates that the analyte is completely
MIP-SPE process applied for ziram determination to determine its
bound to the active sites when the loading volume is selected as 10 mL.
precision and good sensitivity; optimized in terms of adsorbent amount,
At loading volumes >10 mL, recovery was reduced as the analyte could
wash/elution solutions and volumes, sample loading volumes.
not be absorbed despite the increased loading volume, presumably due
Recovery percentages were determined by testing MIP amounts
to the adsorbent’s active sites’ saturation.
ranging from 20 to 100 mg (Fig. 9A). Accordingly, in the experiments us-
ing 40 mg MIP, the recycling amount increased compared to 20 mg and

Fig. 8. Adsorption performance of MIP and NIP for ziram, maneb,

ferbam and propineb compounds (Contact time: 20 min, pH; 6.0 tem-
perature: 25 °C, C0 = 50 mg/L, w = 10 mg).

7
Ç. Öter and Ö.S. Zorer Chemical Engineering Journal Advances 7 (2021) 100118

Fig. 9. The effect of different parameters on the recovery of ziram by the MIP-SPE method: (a) varying adsorbent masses in mg (b) different elution solvents (c)
elution volumes and (d) effect of sample loading volumes.

Table 4 4. Conclusion
Statistical method (t-test) and of recovery (%) comparison and p-value t-test of
(MIP-SPE and NIP-SPE) and (MIP-SPE and C18 -SPE). It is imperative today to examine and eliminate the residues of pesti-
Analyte Recoveries (%) Recoveries (%) cide species that are desired to be used in combating agriculture but pose
p-Value p-Value a threat to all living things due to their unconscious use. In this study;
MIP-SPE NIP-SPE MIP-SPE C18 -SPE
a molecularly imprinted polymer that can selectively adsorb ziram, a
Ziram 96.76 59.63 0.0013 96.76 82.38 0.0081 fungicide, was synthesized by the suspension polymerization process.
Besides being a fungicide, the ziram molecule is used as a crosslinking
accelerator in rubber processing. Thus, the ziram template also has an
accelerating effect that has increased the overall reaction rate in the
radical polymerization reaction. The fact that the template molecule se-
3.7. Statistical comparison of SPE methods lected in radical polymerization reactions has an accelerator effect can
be considered an innovation in the imprinting process. It was clearly
The t-test, a statistical hypothesis test, was used to identify the proba- confirmed that the synthesized polymer exhibited remarkable adsorp-
bility that the MIP-SPE method would differ significantly from commer- tion performance, selectivity, stability, and reusability. The ability of
cial C18-SPE and NIP-SPE methods and compare them ziram solutions synthesized MIP to rebinding was investigated. The adsorption mecha-
with a concentration of 50 mg/L were used under optimum conditions. nism’s suitability to the kinetic models and isotherm models was eval-
The p-value and the recovery are shown in Table 4.Recovery values uated by comparing it with the non-suppressed polymer (NIP). The ad-
were 96.76% for MIP-SPE, 59.63% for NIP-SPE, and 82.38% for C18- sorption mechanism for MIP showed agreement with the pseudo-second-
SPE. The p-value for the comparison of MIP-SPE and NIP-SPE for the order kinetic model and the Langmuir isotherm, and the Qmax value was
independent t-test was 0.0013, while the p-value for comparing MIP- calculated as 56.98 mg/g. The selectivity studies using different DTC
SPE with the commercial C18-SPE was determined to be 0.0081. The fungicides (maneb, propineb, and ferbam) determined that the MIP ad-
null hypothesis was refused because p values were less than a = 0.05 in sorbent showed high selectivity against ziram. Besides, the extraction
both compare situations [56]. Accordingly, it was found that MIP-SPE performance of MIPs used as adsorbents in the solid phase extraction
datasets differ significantly for C18-SPE and NIP-SPE methods. (SPE) column was evaluated, and some extraction parameters were op-

8
Ç. Öter and Ö.S. Zorer
timized. Excellent stability and low-cost MIP; compared to NIP-SPE and
a commercial sorbent (C18-SPE), it was determined to be a useful and
highly selective synthetic material for ziram extraction.
The MIP-SPE method developed in this study; can be widely used
in the easy, effective, and selective monitoring and analysis of pesti-
cide species. It is also essential and valuable in terms of environmental
protection.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have influenced the work
reported in this paper.
Acknowledgments
This work was supported by the Van Yuzuncu Yil University Scien-
tific Research Projects Support Unit (Project Number: 8444).
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