Food Chemistry 407 (2023) 135067

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Preparation of an efficient magnetic nano-sorbent based on modified

cellulose and carboxylated carbon nano-tubes for extraction of pesticides
from food and agricultural water samples before GC-FID analysis
Majid Behpour , Mehrdad Shadi , Saeed Nojavan *
Department of Analytical Chemistry and Pollutants, Shahid Beheshti University, Evin, Tehran, Iran

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

Keywords: This paper reports the direct synthesis approach of carboxamide functionalized magnetic nano-composite named
Carbon nano-tube Fe3O4@SiO2-NH2@dialdehyde cellulose (DAC)@CNT-COOH as an effectual sorbent for the co-extraction of
Cellulose seven agricultural insecticides and herbicides from vegetable, fruit, and water samples using the magnetic
Food samples
dispersive micro-solid-phase extraction procedure. Under the optimized extraction conditions (sorbent amount:
Herbicides
Insecticides
18.1 mg; desorption time: 6.5 min; desorption solvent volume: 185 μL; desorption solvent: acetonitrile;
Magnetic dispersive micro-solid-phase extraction time: 9.5 min; pH of sample solution: 7.0, and salt content: 5.0 % w/v sodium chloride), good linearity
extraction within the range 0.5–1200 ng/mL (R2 ≥ 0.998) was achieved. Extraction efficiencies were in the range
63.4–84.1 %, the limits of detection were 0.08–1.0 ng/mL, and acceptable relative recoveries (87.6–103.8 %),
and precisions were also achieved (RSDs < 6.8 %, n = 3). Ultimately, the obtained results showed that the
developed method could be applied to determine trace amounts of desired analytes in various agricultural water
and food samples.

1. Introduction matrices, sample preparation is required to perform accurate analysis

Nowadays, agriculture has become inextricably linked to pesticides
(mainly insecticides and herbicides). Using pesticides has the primary
purpose of increasing yield, improving crop quality, decreasing crop
losses, and extending postharvest life by controlling pests, insects,
weeds, and other unfavorable organisms (Yang et al., 2021; Tian et al.,
2016; Amini, Ebrahimzadeh, Seidi & Jalilian, 2021). However, pesti­
cides are intrinsically toxic to the well-being and health of humans and
wildlife. Due to their widespread applications, bioaccumulation, and
stability, pesticides are considered as the most hazardous and pervasive
chemical substances introduced to the environment. It has been estab­
lished that pesticide exposure leads to diverse human issues such as
cancer, respiratory problems, neurodevelopmental delays in children,
and neurological conditions in adults like Parkinson’s disease (Al-Nasir
et al., 2020; Guo et al., 2021; Salisaeng, Arnnok, Patdhanagul & Bur­
akham, 2016). To decline their earlier-mentioned effects and increase
food safety, the demand for sensitive, high-throughput, and reliable
analytical methods to measure pesticide residues in water and food
samples has grown significantly (Tian et al., 2016). Since pesticides
occur in agricultural products at trace levels and in complicated
(Al-Nasir et al., 2020; Guo et al., 2021; Behpour, Maghsoudi & Nojavan,
2022). The most common sample preparation methods used for various
pesticides are solid-phase extraction (SPE) and liquid–liquid extraction
(Hejabri Kandeh, Amini & Ebrahimzadeh, 2021). Although, owing to
their drawbacks, including large consumption of hazardous organic
solvents, labor-intensive processes, solvent losses, and being time-
consuming, the application of these methods has been restricted (Beh­
pour, Nojavan & Ghaniyari, 2020). To ameliorate extraction methods,
dispersive micro-SPE (D-μSPE) based on the solid sorbent dispersion in
the sample solution has been offered as a QuEChERS (quick, easy, cheap,
effective, rugged, and safe) clean-up technique for a wide range of
sample types (Yang et al., 2021). In this method, centrifugation or
applying an external magnetic field can perform the separation process.
Magnetic D-µSPE (MD-µSPE) is an efficient methodology in which the
separation of adsorbent with magnetic features is carried out in a facile
manner by an external magnet without more centrifugation (Majd &
Nojavan, 2021a). MD-µSPE has excellent potential to make a consider­
able contact interface between the dispersive sorbents and intended
analytes. In this method, selecting a proper sorbent is pivotal to
achieving high enrichment efficiency and better selectivity of target

* Corresponding author.
E-mail address: s_nojavan@sbu.ac.ir (S. Nojavan).

https://doi.org/10.1016/j.foodchem.2022.135067
Received 3 August 2022; Received in revised form 4 November 2022; Accepted 23 November 2022
Available online 28 November 2022
0308-8146/© 2022 Elsevier Ltd. All rights reserved.
M. Behpour et al.
analytes in environmental and food analyses (Salehpour, Bayatloo &
Nojavan, 2021). Investigations demonstrate that the use of nano-
structured materials with a large surface-to-volume ratio has resulted
in significant advancements in assaying trace level pollutants (Pang
et al., 2020).
Carbon nano-tubes (CNTs) consist of graphene layers with numerous
oxygen-containing functional groups (epoxy, hydroxyl, carbonyl, and
carboxylic) on the edges and planes of graphene layers (Jabbari, Veleta,
Zarei-Chaleshtori, Gardea-Torresdey & Villagrán, 2016). Due to their
advantages like non-toxicity, low cost, ease of functionalization with
different materials, CNTs are considered a promising candidate for the
fabrication of magnetic sorbents. However, nano-sized dimensions and
high surface tension of CNTs culminate in the aggregation of CNTs in
aqueous solutions and decrease their capability to extract analytes (Yang
et al., 2009). Concerning the faults mentioned for CNTs, polymeric
structures like cellulose can be combined with CNTs to stabilize the
CNTs suspension by forming infrastructure with high mechanical sta­
bility and adsorption capacity (Yazdanpanah & Nojavan, 2019).
Cellulose, an abundant and widely available biodegradable resource,
can be extracted from various natural sources, including wood pulp and
cotton (Klemm, Heublein, Fink & Bohn, 2005). Due to the non-toxicity,
eco-friendly, renewability, and cost-effectively Properties, Cellulose
scaffolds have multiple applications in many fields, such as tissue en­
gineering, bioimaging, wound dressing, cosmetic, wearable epidermal
sensors, and pharmaceutical industries (Siró & Plackett, 2010). The
hydroxy-functional group on the cellulose surface demonstrates the
massive potential for the adsorption of many molecules, including metal
ions and pesticide molecules (Wang, Lu & Zhang, 2016). The shortage of
several functionalities on the cellulose and its particular structure has
created the conditions for cellulose surface modification to take
advantage of the maximum potential structural properties (Bahri, Shadi,
Mohammadian, Javanbakht & Shaabani, 2021).
Materials functionalization and combination by multicomponent
reactions (MCRs) have received significant attention due to reducing
waste production, minimizing the number of operations, decreasing
energy consumption, and increasing molecular complexity in a one-step
reaction (Shaabani et al., 2019). Material functionalization via MCRs is
an ingenious technique that includes high productivity synthetic stra­
tegies with the minimum number of chemical steps to create high value-
added scaffolds from accessible materials (Jadhav et al., 2019). Recently
notable development has been made in extending the territory of this
technique in several fields, such as drug delivery, biosensors, enzyme
immobilization, and polymer chemistry; however, there are limited re­
ports in this area, and this strategy is still in its infancy (Zhu, Wang &
Wang, 2014).
Considering all these in mind, here the new synthesis of magnetic
composite (Fe3O4@SiO2-NH2@dialdehyde cellulose (DAC)@CNT-
COOH) was presented, and its application as an effective and novel
sorbent in magnetic D-µSPE was investigated by simultaneous extraction
of seven agricultural pesticides, including atrazine (ATR, Log P = 2.61),
ametryn (AMT, Log P = 2.98), metribuzin (MTR, Log P = 1.70), etho­
fumesate (ETF, Log P = 2.70), diazinon (DZN, Log P = 3.81), chlor­
pyrifos (CPS, Log P = 4.96) and profenofos (PFS, Log P = 4.68) from
various agricultural water and food samples before GC-FID analysis.
Vital factors that affect the extraction efficiencies of the MD-µSPE
method based on Fe3O4@SiO2-NH2@DAC@CNT-COOH were thor­
oughly evaluated and optimized. The Fe3O4@SiO2-NH2@DAC@CNT-
COOH nano-composite merges the superiorities of three components all
in one: (i) magnetic properties of the composite dedicate strong
magnetism to the sorbent, which simplifies the extraction process; (ii)
incorporation of CNT and DAC (the structure of cellulose should be
modified by utilizing a potent oxidizing agent for use in the adsorbent
synthesis procedure) results in the preparation of sorbent with large
specific surface area and remarkable adsorption capacity. In this nano-
composite, the unique morphology and porous structure, combined
with aromatic rings and many different functional groups, allow a wide
2
Food Chemistry 407 (2023) 135067
variety of pesticides to be adsorbed with satisfactory adsorption effi­
ciency. To our knowledge, there is no study on using the composite of
Fe3O4@SiO2-NH2@DAC@CNT-COOH as the adsorbent for the extrac­
tion of analytes.
2. Experimental
2.1. Chemicals and reagents
ATR (98 %), AMT (99 %), MTR (97 %), ETF (96 %), DZN (95 %), CPS
(95 %), PFS (96 %) and, phosalone (PZL) (97 %) were acquired from
Ariashimi Company (Tehran, Iran). Iron (III) chloride hexahydrate
(FeCl3⋅6H2O), Iron (II) chloride tetrahydrate (FeCl2⋅4H=O), cellulose
nano-crystals (CNCs), potassium periodate (KIO4), nitric acid (HNO3),
sulfuric acid (H2SO4), ammonia solution 25 % (NH4OH), hydrochloric
acid (HCl), sodium hydroxide (NaOH), sodium chloride (NaCl) were
purchased from Merck (Darmstadt, Germany). Also, cyclohexyl iso­
cyanide (C7H11N), tetraethyl orthosilicate (TEOS) (SiC8H20O4), (3-
Aminopropyl) triethoxysilane (APTES) (C9H23NO3Si) were obtained
from Sigma-Aldrich (St. Louis, MO, USA). HPLC grade methanol
(MeOH) and acetonitrile (ACN) were purchased from Dr. Mojallali
(Tehran, Iran). Toluene, acetone, ethanol, and 2-propanol (2-PrOH)
were achieved from Caledon (George-town, Ont., Canada). Multi-walled
CNT with carboxylic acid functionalization (MWCNT-COOH) had an
inside diameter of 3–5 nm, an outside diameter of 5–15 nm, and 50 µm
lengths were obtained from Iranian Nano-materials Pioneers Company
(Mashhad, Iran). A Milli-Q water purification system was used to pre­
pare all solutions and buffers for HPLC (Millipore, Milford, MA, USA).
2.2. Solutions
The standard stock solutions (1000.0 mg/L) of seven pesticides were
prepared separately in HPLC-grade methanol and kept in a refrigerator
at 4 ◦ C to prevent the decomposition of analytes. To optimize parameters
and other experiments, mixed working solutions at different concen­
trations containing the seven pesticides were prepared daily by diluting
the standard solutions with HPLC-grade water. Also, the pH value of the
solutions was adjusted using NaOH or HCl solutions (0.1 M). Further, the
internal standard PZL (300.0 mg/L) was prepared in methanol and kept
at 4 ◦ C.
2.3. Real samples collection and preparation
Two different agricultural water samples (Shahriar, Iran) were
collected and stored in brown glass bottles in a fridge at 4 ◦ C. Before use,
the water samples were filtered via a millipore 0.22 µm cellulose acetate
membrane filter. Afterward, the pHs were adjusted, and subsequent
extraction was done with the MD-µSPE method without dilution. The
fresh fruits (apple and orange) and vegetables (tomato and carrot) were
purchased from local markets (Tehran, Iran). In order to be used, each
food sample was homogenized individually with a high-speed homog­
enizer after being cut up into bits. Then, 10.0 g of fruits or vegetables
was accurately weighed and extracted by 10.0 mL of the acetonitrile
through ultrasonication for 30 min. Next, all spiked samples were
centrifuged for 3 min at 6000 rpm to eliminate their pulps. Finally, the
achieved supernatant was vaporized to dryness by nitrogen purging and
redissolved with 25 mL HPLC grade water in a centrifuge tube for the
subsequent analysis (Majd & Nojavan, 2021a).
2.4. Instrumentation and GC-FID conditions
An Agilent Technology 7890B GC-FID system with a capillary col­
umn (HP-5, 30 m × 0.32 mm × 0.25 μm) was used to analyze seven
pesticides in this study. The column temperature was set at 100 ◦ C
initially and then increased by 10 ◦ C/min to 250 ◦ C, which was held for
11 min. Injection port and FID temperatures were set at 200 and 300 ◦ C,
M. Behpour et al.
Fig. 1. Preparation process of the Fe3O4@SiO2-NH2@DAC@CNT-COOH nano-composite.
respectively. The split injection mode was fixed at 5:1, and nitrogen gas
(99.999 %) was used as the carrier gas at 1.2 mL/min at a constant flow.
Measurement of pH values of all sample solutions was done with an 827
Metrohm digital pH meter and a glass calomel electrode (Herisau,
Switzerland). GHB Fixette II centrifuge (Wehingen, Germany) was
applied to treat real samples. An MS3 digital agitator from IKA Company
(Staufen, Germany) was used to mix the sample solutions. The synthesis
process was conducted utilizing a heating-magnetic stirrer (Heidolph
MR 3001 K, Schwabach, Germany). Fourier transform infrared (FT-IR)
characterization was acquired with a Thermo Nicolet FT-IR spectro­
photometer model NEXUS 470 (Waltham, MA, USA) within a scan range
of 400–4000 cm− 1 employing KBr pellets. To examine the X-ray
diffraction (XRD) patterns, an X’Pert Pro diffractometer instrument
(PANalytical, Almelo, The Netherlands) containing a Cu Kα radiation
source was used. To examine the morphology of the synthesized nano-
composite and to perform energy-dispersive X-ray spectroscopy (EDX),
a field emission scanning electron microscope (FE-SEM) model
MIRA3TESCAN-XMU (Czech Republic) was used. To monitor magneti­
zation produced nano-composite, a vibrating sample magnetometer
(VSM) model LBKFB (Meghnatis Daghigh Kavir Co.; Kashan Kavir; Iran)
was used. The thermogravimetric analysis was conducted using a BÄHR-
Thermoanalyse GmbH model BAHR STA 503 thermogravimetric
analyzer. A Belsorp mini II instrument (Microtrac Bel Corp, Japan) was
used to perform the Barrett-Joyner-Halenda (BJH) and Brunauer-
Emmett-Teller (BET) analyses.
2.5. Preparation process of the Fe3O4@SiO2-NH2@DAC@CNT-COOH
nano-composite
Our ongoing interest in developing magnetic structures as an
adsorbent (Majd & Nojavan, 2021a; Salehpour, Bayatloo & Nojavan,
2021; Majd & Nojavan, 2021b; Yazdanpanah & Nojavan, 2019) and the
attractive property of MCRs as a material functionalization strategy to
improve materials feature prompted us to synthesize a novel adsorbent
using the MCR strategy to improve adsorption capacity and properties.
In this regard, an efficient magnetic adsorbent (Fe3O4@SiO2-
NH2@DAC@CNT-COOH) was introduced based on CNT and DAC via the
one-pot four-component Ugi reaction. This reaction offered great po­
tential for the preparation of carboxamide-functionalized scaffolds on
the adsorbent, which is used for MD-μSPE. As shown in Fig. 1, the
strategy for preparing Fe3O4@SiO2-NH2@DAC@CNT-COOH includes
3
Food Chemistry 407 (2023) 135067
three steps: (i) Preparation of Fe3O4@SiO2-NH2 core/shell nano-
particles (NPs); (ii) Cellulose surface modification using a potent
oxidizing agent to DAC (cellulose structure must be modified for use in
Ugi-MCR); (iii) The coating of CNT-COOH with peptidomimetic skeleton
from available starting materials, including DAC, Fe3O4@SiO2-NH2,
cyclohexyl isocyanide via the Ugi-MCR.
2.5.1. Synthesis of Fe3O4@SiO2-NH2
For the preparation of the Fe3O4@SiO2-NH2, Fe3O4 nano-particles
were synthesized according to the previously reported procedure
(Zhang et al., 2014). Firstly, FeCl3⋅.6H2O (4.72 g) and FeCl2⋅.4H2O
(1.72 g) were dissolved in pure water (100.0 mL). Then, NH4OH (10.0
mL) was added dropwise to the reaction mixture and stirred for 1 h at
500 rpm under a gentle nitrogen stream at 80 ◦ C. Finally, the resulting
precipitate was magnetically separated, and washed with pure water to
pH 7.0, then dried at 60 ◦ C. In the next step, Fe3O4 nano-particles (1.00
g) were dispersed in a mixture of deionized water (10.0 mL) and ethanol
(40.0 mL) in an ultrasonic manner for 30 min. Then, the produced
suspension was added to the mixture of the tetraethyl ortho silicate
(TEOS, 0.5 mL) and NH4OH (2.0 mL); afterwards, the mixture was
stirred at 50 ◦ C for 6 h. In the following, 1.0 g of the resulted Fe3O4@­
SiO2 nano-particles were ultrasonically dispersed in the mixture of the
25.0 mL toluene/methanol (1:1, v/v) for 15 min. Eventually, 2.0 mL of
3-triethoxysilylpropylamine (APTES) was added to the mixture and then
stirred for 4 h. The product was completely washed with ethanol and
then dried at 60 ◦ C under vacuum for 12 h (Liu, Li, Li & He, 2013).
2.5.2. Synthesis of dialdehyde cellulose (DAC)
DAC was synthesized as described in the literature (Sirvio, Hyvakko,
Liimatainen, Niinimaki & Hormi, 2011). KIO4 (0.41 g) and cellulose
(0.50 g) were added to a 50 mL flask involving deionized water (5.0 mL).
The reaction vessel was covered with aluminum foil and was stirred for
12 h at 85 ◦ C. On completion, the product was filtered and completely
washed with deionized water and dried under vacuum at 70 ◦ C.
2.5.3. Synthesis of Fe3O4@SiO2-NH2@DAC@CNT-COOH
For the preparation of the Fe3O4@SiO2-NH2@DAC@CNT-COOH,
DAC (1.0 g) and Fe3O4@SiO2-NH2 NPs (1.0 g) were dissolved in EtOH
(15.0 mL) and stirred for 30 min at 50 ◦ C. Then, cyclohexyl isocyanide
(1.00 mmol) and CNT-COOH (1.00 g) were added to the reaction
mixture and stirred for 24 h at 50 ◦ C in a closed vessel. The product was
M. Behpour et al. Food Chemistry 407 (2023) 135067

cooled to 25 ◦ C after the completion of the reaction and completely agricultural water samples:
washed with ethanol/water (20:80, v/v), and separated by centrifuga­ ( ) ( )
Vfinal Cfinal
tion (20000 rpm, 6 times, and 20 min). ER% = × × 100 (1)
Vinitial Cinitial
2.6. Magnetic dispersive micro-solid-phase extraction (MD-μSPE) ( )
Cfound − Creal
workflow RR% = × 100 (2)
Cadded

As illustrated in Figure S1, the MD-μSPE procedure was performed as
follows: To 25.0 mL of the sample solution containing 1.25 g of sodium
chloride (pH = 7.0), which had been placed into a centrifuge tube, the
appropriate amount of the synthesized Fe3O4@SiO2-NH2@DAC@CNT-
COOH nano-composite (≈ 18 mg) was dispersed to the mentioned tube
as the adsorbent. Afterward, the mixture was shaken at a constant rate
utilizing a vortex for 9.5 min to aid the adsorption of the target pesti­
cides onto the nano-particles. After the extraction, the sorbent was iso­
lated quickly from the suspension using an external magnet at the
bottom of the centrifuge tube, and the supernatant was decanted. Then,
the analytes were desorbed from the isolated magnetic nano-composite
by 185 μL of acetonitrile by shaking the sample tube for 6.5 min. Next,
the resulting desorption solvent was separated and filtered using
disposable syringe filters (0.22 μm). Finally, after adding the internal
standard, 1.0 µL of the eluent was injected into the GC-FID device for
further analysis.
2.7. Optimization of extraction parameters
The MD-μSPE experimental factors, including sorbent type
(Fe3O4@SiO2-NH2, CNT-COOH, DAC, and Fe3O4@SiO2-
NH2@DAC@CNT-COOH nano-composites), eluent type (methanol, 2-
propanol, acetonitrile, ethanol, and acetone), sample solution pH
(5.0–10.0), salt content (0–10 % w/v), amount of sorbent (8–27 mg),
extraction time (5 to 15 min), desorption time (3 to 9 min), and elution
volume (150–450 µL) were examined to attain the highest efficiency for
extraction of some insecticides and herbicides from agricultural waters
and food samples. Also, the GC-FID instrument was applied for the
detection and quantification of analytes. Optimizing the sorbent type,
eluent type, pH of the sample solution, and salt content was achieved
using the one variable at a time method. In order to optimize the other
four factors, the response surface methodology (RSM) was applied.
2.8. Quality control and quality assurance
To assess the quality features of the MD-μSPE method and Fe3O4@­
SiO2-NH2@DAC@CNT-COOH sorbent together in the extraction of
target insecticides and herbicides, quantitative parameters, including
the limits of detection (LODs) and quantification (LOQs), coefficients of
determination (R2), linear dynamic ranges (LDRs), intra-day and inter-
day relative standard deviations (RSDs%), preconcentration factors
(PFs), and extraction recoveries (ERs%) were obtained under optimal
extraction conditions. The linearity was investigated in the range of
0.5–1200.0 ng/mL using eleven different concentration levels (0.5, 1.0,
2.5, 5.0, 10.0, 50.0, 250.0, 500.0, 750.0, 1000.0 and 1200.0 ng/mL) of
the pesticides, and calibration plots were attained. The Intra-day of the
proposed method was obtained by carrying out five replicated extrac­
tions of the desired analytes at four concentration levels (25.0, 100.0,
250.0, and 750.0 ng/mL) during a day, and also the inter-day based on
three replicated extractions at similar concentration levels in three
subsequent days. The LOQ and LOD were obtained by determining the
minimum concentration at which analytes could be identified and
quantified. The preconcentration factor (PF) was defined by the slope of
the extraction calibration curve of each analyte after the MD-µSPE
method to the slope of the direct calibration curve of each analyte. Ac­
cording to Eqs. (1) & (2), ER calculations were conducted using a con­
centration level of 25.0 ng/mL with five replications, and RR
calculations were performed using concentration levels of 75.0, 250.0,
and 750.0 µg/kg for food samples and 25.0, 100.0, and 500.0 ng/mL for
In Eq. (1), Vfinal is the volume of the desorption solvent, and Vinitial is
the volume of the sample solution. Also, Cfinal and Cinitial represent the
concentration of pesticides in the desorption solvent and in the sample
solution, respectively. In Eq. (2), Cfound represents the concentration of
pesticides after adding the known amount, Creal represents the concen­
tration of target analytes in the food and water samples in advance, and
Cadded represents the concentration of pesticides added into the samples.
Additionally, the matrix effect value (ME%) was studied under the
optimized conditions by analyzing water samples and real samples by
the Eq. (3):
Rm
ME% = (( ) − 1) × 100 (3)
Rw
Rm is the recovery of the desired analytes in the real matrix, and Rw is
the recovery in ultra-pure water.
Moreover, the obtained results of the added concentration (certified
value) were compared with the found concentration through statistical
analysis utilizing a Student’s t-test (at a 95 % level of confidence) using
Eq. (4):
(x − μ) √̅̅̅
tcalculated = n (4)
s
In Eq. (4), x and n are the average value and the number of records in
a sample set, respectively, while µ is the known value and s is the
standard deviation of the sample set.
The adsorption capacity of the nano-composite (mg/g) was assessed,
conforming to the Eq. (5):
C0 − Ce
Qe (mg g− 1 ) = ×V (5)
M
Where M (g) is the Fe3O4@SiO2-NH2@DAC@CNT-COOH amount; C0
(mg/L) is the initial concentration of analytes in aqueous solution; Ce
(mg/L) is the equilibrium concentration of intended analytes, and V (L)
is the volume of the aqueous solution.
In order to fit the isotherm data, the Freundlich isotherm was
selected according to the Eq. (6):
Log Qe = m Log Ce + log α (6)
Where Qe (mg/g) is the adsorption amount of analytes at the equilib­
rium; m is the heterogeneity index; Ce (mg/L) is the equilibrium con­
centration of analytes and α is the Freundlich constant.
2.9. Experimental design
Experimental designs and data analysis were carried out using
Design-Expert 13.0.1.0 version. In this study, the central composite
design (CCD) was combined with a desirability function (DF) to deter­
mine the optimal values for factors influencing adsorption and desorp­
tion (sorbent amount, extraction time, solvent volume, and desorption
time). As a result, the 30 experiments with six central points were
conducted in random order. The regression equation and the levels of
factors are summarized in Table S1. The experimental response was
based on the recovery of analytes in each run. The ANOVA analysis was
performed on the collected data; the results can be found in Table S2. As
can be seen, all terms (linear, quadratic, and cross-product) were sig­
nificant (P < 0.05), and there is no evidence of Lack of Fit (LOF) at the
95 % confidence level, meaning that the model is significant and illus­
trates the observed differences in the response variable. A response

4
M. Behpour et al.
Fig. 2. FE-SEM image of (a) CNT-COOH, (b) DAC, (c) Fe3O4@SiO2-NH2, and (d) Fe3O4@SiO2-NH2@DAC@CNT-COOH.
surface methodology (RSM) was also used to analyze the effects of in­
dependent variables on the response. The RSM method graphically de­
scribes the relationship between factors and responses and identifies the
optimum conditions. Figure S2 shows the 3D-response surfaces and
contour lines of the CCD, which investigate the relationship between the
average extraction recoveries and two variables.
3. Results and discussion
3.1. Characterization of sorbent
3.1.1. FT‑IR analysis
The samples were analyzed by FT-IR spectroscopy to affirm the
successful synthesis of sorbent. The FT-IR spectra of the cellulose, DAC,
Fe3O4 and Fe3O4@SiO2-NH2 were in excellent agreement with the pre­
vious report (Figure S3) (Javanbakht et al., 2020). The FT-IR spectrum
of the Fe3O4@SiO2-NH2@DAC@CNT-COOH displayed the maximum
absorption peak at 3400 cm-1 that is related to –NH and –OH groups.
Other main bonds for the Fe3O4@SiO2-NH2@DAC@CNT-COOH nano-
composite are 2918 cm− 1 and 2850 cm− 1 (C–H stretching, for
5
Food Chemistry 407 (2023) 135067
vibrations the methylene and methyl groups, medium), 1650 cm− 1
(C–– O stretching, medium), 1630 cm− 1 (N–H bending, for amine
groups, medium), 1595 cm− 1 (C– – C stretching, weak), 1435 cm− 1 and
1377 cm− 1 (–CH2- and –CH3 bending, weak), 1330 cm− 1 (C–N
stretching, weak), 1160 cm− 1 (C–O bending, strong), 1110 cm− 1
(C–O–C stretching, strong), 1063 cm− 1 and 752 cm− 1 (Si-O-Si un-
symmetric and symmetric linear stretching, strong), 451 cm− 1 and
790 cm− 1 (vibrations peaks of Si-O-Si and Si-OH, weak) and 573 cm− 1
(Fe-O vibration, strong).
3.1.2. X-ray diffraction analysis
The X-ray diffraction (XRD) pattern of the pure Fe3O4@SiO2-NH2,
DAC, and Fe3O4@SiO2-NH2@DAC@CNT-COOH are shown to investi­
gate the structure of the synthetic adsorbent (Figure S4). The XRD pat­
terns of the pure Fe3O4 and Fe3O4@SiO2-NH2, DAC, and CNT-COOH
were in excellent agreement with the previous report (Wang, Zhang,
Liu & Li, 2014). For the XRD pattern of the synthetic Fe3O4@SiO2-
NH2@DAC@CNT-COOH composite, the diffraction peaks at 2θ = 22.3◦ ,
30.2◦ , 35.5◦ ,43.0◦ , 57.2◦ and 62.7 are respectively well corresponding
to (0 2 0), (2 2 0), (3 1 1), (4 0 0), (5 1 1) and (4 4 0). The XRD pattern of
M. Behpour et al.
the Fe3O4@SiO2-NH2@DAC@CNT-COOH specifies the presence of both
Fe3O4@SiO2-NH2 and DAC in the adsorbent without change in the
structure of each component during the reaction.
3.1.3. Morphological and elemental analysis
The morphology of the Fe3O4@SiO2-NH2, DAC, and Fe3O4@SiO2-
NH2@DAC@CNT-COOH NPs was studied via FE-SEM analysis. The FE-
SEM image of CNT-COOH displays the nano-tube structure of this par­
ticle with low dispersity (Fig. 2(a)). According to the literature, in
contrast to the FE-SEM image of cellulose fibers that emerge to have
relatively flat and smooth structures, excrescences are on the surface of
DAC fiber (Fig. 2(b)) (Bahri, Shadi, Mohammadian, Javanbakht &
Shaabani, 2021). The FE-SEM image of the Fe3O4@SiO2-NH2 shows
spherical core–shell structures (Fig. 2(c)). After surface modification via
a one-pot four-component Ugi reaction, morphology emerged with a
well-separated and spherical surface without aggregation (Fig. 2(d)).
Also, the primary morphology of the starting materials has been
maintained.
The elemental analysis (chemical composition) was also examined
by energy-dispersive X-ray spectroscopy (EDX) analysis (Figure S5). As
expected, C, O, Si, Fe, and N atoms were detected in the sorbent struc­
ture, which further proves the potential of the Ugi MCR for producing
worthy functionalization on the sidewalls CNTs without any obvious
changes in the morphology and structure.
3.1.4. Magnetization studies
The synthesized nano-composite must have sufficient magnetic
properties that allow it to be easily and rapidly separated from the
sample solution in the MD-μSPE process. Hence, magnetic character­
ization via a magnetometer 300 K was studied and displayed magneti­
zation saturation (Ms) values of 59.46, 46.25, 19.63 emu/g for the
Fe3O4@SiO2-NH2, and Fe3O4@SiO2-NH2@DAC@CNT-COOH, respec­
tively. Fe3O4@SiO2-NH2@DAC@CNT-COOH compared to the Fe3O4@­
SiO2-NH2 and Fe3O4 displayed a decrease in the magnetic power that
can be related to the coating of a nonmagnetic DAC and CNT-COOH
structure on the magnetic section (Figure S6).
3.1.5. Thermogravimetric analysis
TGA analysis was carried out by heating Fe3O4@SiO2-
NH2@DAC@CNT-COOH up to 600 ◦ C at a heating rate of 10 ◦ C min − 1
under the air atmosphere for studying the weight percentage of car­
boxamide functional moieties grafted onto the CNTs Surface (Figure S7).
In the first step, initial weight loss up to 100 ◦ C was attributed to
removing the physically adsorbed solvent molecules and moisture,
which led to a release of CO2 and H2O. The obvious weight loss was
observed at 200–500 ◦ C, arising from the decomposition of the cova­
lently grafted carboxamide skeleton bonded on the surface.
of the adsorbent structure and the deformation of functional groups
of the starting materials. The residual mass at 600 ◦ C indicates Fe3O4
NPs that the amount in Fe3O4@SiO2-NH2@DAC@CNT-COOH is about
53 %.
3.1.6. Specific surface area properties
The nitrogen adsorption–desorption isotherms of Fe3O4@SiO2-
NH2@DAC@CNT-COOH composite are shown in Figure S8. The BET
surface area of the nano-adsorbent was calculated at 72.017 m2/g. The
pore size distributions were calculated via the Barrett–Joyner–Halenda
method for Fe3O4@SiO2-NH2@DAC@CNT-COOH studied to confirm the
presence of well-developed mesoporosity with the excellent pore-size
distribution. The result showed that the BJH pore size of the Fe3O4@­
SiO2-NH2@DAC@CNT-COOH was 29.48 nm which ensures the com­
posite is in the category of mesoporous materials. Besides, the pore
volume was obtained 0.5395 cm3/g.
6
Food Chemistry 407 (2023) 135067
3.2. Optimization of extraction conditions
3.2.1. Sorbent type
The type of sorbent used is an important parameter that affects the
efficiency of the extraction. In this work, the extraction capabilities of
Fe3O4@SiO2-NH2, CNT-COOH, DAC, and Fe3O4@SiO2-
NH2@DAC@CNT-COOH nano-composites were evaluated. Compared to
the other three sorbents, Fe3O4@SiO2-NH2@DAC@CNT-COOH showed
the highest ability to adsorb to the desired analytes (Figure S9). A
notable reason is the excellent dispersibility and solubility of Fe3O4@­
SiO2-NH2@DAC@CNT-COOH in aqueous solutions, owing to the pres­
ence of DAC in the sorbent structure. Moreover, higher specific surface
area and the presence of many functional groups on the Fe3O4@SiO2-
NH2@DAC@CNT-COOH can improve the extraction recoveries via
hydrogen bonding and π-π accumulation interactions. Additionally, the
Fe3O4 NPs present in the sorbent structure increased its stability, and its
collection with an external magnet could be rapidly completed. Thereby,
Fe3O4@SiO2-NH2@DAC@CNT-COOH was utilized in the following
experiments.
3.2.2. Type of desorption solvent
According to previous studies, desorption solvent has been shown to
play a crucial role in obtaining good extraction efficiency and should be
optimized. It is essential that the solvent chosen has a high affinity for
the desired analytes and must be powerful enough to desorb most of the
analytes from the sorbent (Majd & Nojavan, 2021a). Based on earlier
research, several organic solvents were used for the elution study,
including methanol, 2-propanol, acetonitrile, ethanol, and acetone. As
illustrated in Figure S10, the desorption ability of acetonitrile seemed to
be higher than the other eluents for the elected pesticides. The cause
might be that these insecticides and herbicides have proper solubility in
acetonitrile. Hence, acetonitrile was accepted as the desorption solvent
in the subsequent studies.
3.2.3. pH effect
An important parameter that influences the adsorption of the target
analytes onto the sorbents is the pH value of the sample solution since
both the surface charge of the sorbent and the ionic state of pesticides
are pH-dependent. It has been determined that the highest efficiency in
the sorption process occurs when the polarity of the functional groups on
the sorbent and analyte is the same and the pH of the sample solution is
controlled (Ramezani, Yamini & Nazraz, 2021). A study of the effect of
pH on pesticides extraction performance was carried out in order to
obtain an optimal interaction between the analyte and sorbent, and
extractions were done by varying the pH value from 5.0 to 10.0 in the
sample solutions using dilute HCl/NaOH solutions with the aid of a pH
meter. The results show that the highest efficiency of the extraction for
analytes is achieved at a pH of 7.0. Most pesticides are not ionizable in
aqueous solutions and cannot be stable in highly acidic and basic solu­
tions (Shakourian, Yamini & Safari, 2020). Hence, it seems that at pH
7.0, the target analytes can be adsorbed onto the Fe3O4@SiO2-
NH2@DAC@CNT-COOH via π-π accumulation interactions. Moreover,
the COOH groups in the CNTs structure can interact with the desired
analytes to fabricate hydrogen bonds. Therefore, as illustrated in
Figure S11, the highest extraction recovery of pesticides was at pH 7.0,
and this point was selected for further investigations.
3.2.4. Effect of ionic strength
The salt concentration of the sample solution has a considerable in­
fluence on the sorption of the desired analytes. On the basis of kinetic
and thermodynamic studies, sodium chloride (NaCl) can improve or
decrease the extraction efficiency (Fan et al., 2020). At high salt con­
centrations, analytes solubility in the sample solution reduces due to the
“salting out” effect (Siró & Plackett, 2010). This phenomenon could
enhance extraction performance. Although, adding salt can increase the
sample solution viscosity and consequently reduce the mass transfer of
M. Behpour et al. Food Chemistry 407 (2023) 135067

Table 1
Analytical data of the seven pesticides obtained by the proposed method.
a
Analytes LOD LDR Regression equation R2 PF ERb (%) Intra-day RSD (%)c Inter-day RSD (%)c

25.0 100.0 250.0 750.0 25.0 100.0 250.0 750.0

ATR 0.30 1.0–1000.0 y = 3.0206x + 0.286 0.9995 101.9 75.4 4.2 4.0 3.7 3.1 7.2 6.4 6.6 5.3
AMT 0.15 1.0–1000.0 y = 3.4303x + 0.344 0.9990 97.7 72.3 5.1 4.5 4.2 4.4 8.6 7.5 7.3 6.6
MTR 0.30 1.0–1000.0 y = 2.8533x + 0.275 0.9993 98.5 72.9 4.5 3.9 4.0 3.4 5.9 6.4 6.1 6.3
ETF 0.08 0.5–1000.0 y = 4.7232x + 0.4032 0.9992 113.6 84.1 4.6 5.3 3.7 3.9 8.1 7.7 7.2 6.9
DZN 0.75 3.0–1200.0 y = 1.911x + 0.269 0.9989 93.8 69.4 5.4 4.1 3.2 3.3 8.7 7.4 7.0 5.9
CPS 0.75 3.0–1200.0 y = 1.8266x + 0.246 0.9995 85.7 63.4 4.8 3.9 4.1 3.6 6.6 7.1 6.3 5.2
PFS 1.0 5.0–1200.0 y = 1.6943x + 0.2847 0.9994 91.1 67.4 5.5 4.2 3.8 4.2 8.3 6.5 5.4 5.0

All concentrations are based on ng/mL.

a
Preconcentration factor.
b
Extraction recovery percentage.
c
Relative standard deviation (n = 5 samples for intra-day and n = 3 days for inter-day).

the analytes (Majd & Nojavan, 2021a). To reach the optimum ionic
strength of the aqueous media, various extractions were done by
different NaCl concentrations, ranging from 0 to 10.0 % w/v. As the
results can be observed in Figure S12, the extraction recovery of the
target pesticides was increased to a maximum level by adding NaCl
concentration to 5.0 % w/v; nevertheless, more concentrations led to an
increase in the viscosity and density of the solution, and as a result
reduce the mobility of desired analytes which, in turn, provides to a
notable decline in the mass transfer of the analytes (Majd & Nojavan,
2021b). Therefore, other extractions were performed utilizing the
sample solution’s 5.0 % w/v of sodium chloride.
3.2.5. Multivariate optimization
A critical factor in the adsorption process and extraction efficiency is
magnetic sorbent dosage. As displayed in Figure S2(a&b), the higher
extraction efficiency of the intended analytes was reached by enhancing
the nano-sorbent amounts since it increases the sorption capacity. It
should be mentioned that excessive sorbent dosage decreases pre­
concentration because of the higher eluent volume needed to desorb the
analyte (Majd, Nojavan & Maghsoudi, 2022). Hence, the low sorbent
amount in this work illustrates that the Fe3O4@SiO2-NH2@DAC@CNT-
COOH nano-sorbent has a short diffusion path and a large productive
surface area that leads to increased extraction kinetics and efficiency.
The effect of a period of time on extraction performance was altered
from 5.0 to 15.0 min. As MD-μSPE is an equilibrium-based technique,
enough time is obliged to reach high desirability. Thus, conforming to
the final results, which are represented in Figure S2(b&c), and since long
extraction times lead to back-extraction, apparently 9.5 min would be
enough to extract the desired analytes. To achieve the high desorption of
the target analytes, the desorption time, and eluent volume as essential
factors for obtaining a good recovery were optimized. Considering π-π
interaction between pesticides containing aromatic rings and Fe3O4@­
SiO2-NH2@DAC@CNT-COOH, enough time is required to attain com­
plete desorption. Also, a sufficient volume of eluent should be available
for soaking the magnetic sorbent and desorbing the analytes. However, a
large amount of eluent might reduce preconcentration factors. Consid­
ering these issues, and as shown in Figure S2(d), 185.0 µL of eluent and
6.5 min of desorption time were sufficient to achieve the best desorption
performance.
COOH improved, but saturated after a particular concentration.
Adsorption isotherm studies for the Langmuir, Freundlich, and Temkin
isotherms were fitted and the related parameters were summarized in
Table S3. By comparing the correlation coefficients (R2), Freundlich
isotherm could better fit experimental data, which show that the
adsorption of target pesticides onto the magnetic Fe3O4@SiO2-
NH2@DAC@CNT-COOH is multilayer adsorption. Hence, according to
Eq (6), Freundlich isotherm was chosen to fit the isotherm data. In the
following, the binding kinetics of target pesticides were examined at
various times, varying from 5 to 40 min after adding 20.0 mg of
Fe3O4@SiO2-NH2@DAC@CNT-COOH sorbent to 25.0 mL aqueous
sample solution. After that, the precipitates were separated by a magnet,
and the retained concentration of the analytes in the supernatant solu­
tion was detected using GC-FID devices. The adsorption kinetics was
studied using pseudo-first-order and pseudo-second-order kinetic
models. Table S4 details these models and corresponding parameters.
The results indicated that the Pseudo-second order kinetic model has a
higher R2 and is more appropriate for fitting the kinetic data. It can be
inferred that the sorption of the target analytes onto the nano-composite
may be owing to sharing or exchanging of the electrons between mag­
netic Fe3O4@SiO2-NH2@DAC@CNT-COOH and analytes.
3.4. Reusability of the magnetic adsorbent
The recyclability of the synthesized sorbent is a vital feature in
practical applications. In this work, to assess the reusability of the
magnetic Fe3O4@SiO2-NH2@DAC@CNT-COOH, the extraction re­
coveries of the nano-composite were calculated after several times used
in the spiked sample solutions. To ensure pesticides were not left on the
sorbent after each extraction, the sorbent was washed three times with
acetonitrile and deionized water. The results of extraction revealed that
Fe3O4@SiO2-NH2@DAC@CNT-COOH could be used for 15 extraction
cycles without a noticeable loss (<10 %) of the extraction recoveries
(Figure S13). Furthermore, the reusability of the sorbent was assessed in
the real samples. According to the acquired results, the Fe3O4@SiO2-
NH2@DAC@CNT-COOH can be reused up to 6 times for the samples
with a complicated matrix without significant performance loss.
3.5. Assessment of cellulose effect in the nano-composite

3.3. Binding performance of the v@DAC@CNT-COOH
To evaluate the binding capacity of target pesticides on the
Fe3O4@SiO2-NH2@DAC@CNT-COOH nano-composite, the saturation
binding experiment was conducted. For that purpose, the binding
isotherm of Fe3O4@SiO2-NH2@DAC@CNT-COOH to intended analytes
was achieved at different pesticides concentrations of the initial solution
in the range of 25.0 to 500.0 mg/L under the optimized conditions.
According to obtained results by raising the primary concentration of
analytes, the adsorption amount of the Fe3O4@SiO2-NH2@DAC@CNT-
To evaluate the cellulose effect in the nanocomposite, it was
compared with Fe3O4@SiO2-NH2@CNT-COOH, synthesized according
to the reported procedure (Xu et al., 2008). To this end, the two nano-
composites were examined for their mechanical stability, adsorption
capacity, and extraction efficiency. 20.0 mg of each nano-composites
was dispersed in 15 mL of water to compare their stability over a
period of 15 days. Figure S14 illustrates that Fe3O4@SiO2-
NH2@DAC@CNT-COOH displayed a significantly more stable suspen­
sion. The main reason is that CNTs tend to form bundles that disperse
poorly in aqueous solutions. Hence, covalent and non-covalent

7
M. Behpour et al. Food Chemistry 407 (2023) 135067

Table 2
Analytical results of the proposed method for the determination of pesticides in real samples.
Sample Analyte Addeda,b Founda ± SDa RSDc (%) RR (%) MEd (%) t-valuef
e
Apple ATR 0.0 n.d. – – – –
75.0 67.1 ± 4.1 6.1 89.4 − 10.6 3.3
250.0 227.0 ± 10.4 4.6 90.8 − 9.2 3.8
750.0 702.0 ± 36.5 5.2 93.6 − 6.4 2.2
AMT 0.0 n.d. – – – –
75.0 70.1 ± 3.7 5.3 93.5 − 6.5 2.2
250.0 225.0 ± 10.8 4.8 90.0 − 10.0 4.0
750.0 708.0 ± 30.4 4.3 94.4 − 5.6 2.3
MTR 0.0 n.d. – – – –
75.0 67.6 ± 4.6 6.8 90.1 − 9.9 2.8
250.0 221.8 ± 109 5.3 88.7 − 11.3 4.1
750.0 704.3 ± 39.4 5.6 93.9 − 6.1 2.0
ETF 0.0 n.d. – – – –
75.0 70.7 ± 4.2 5.9 94.2 − 5.8 1.8
250.0 235.8 ± 12.5 5.3 94.3 − 5.7 1.9
750.0 690.0 ± 33.1 4.8 92.0 − 8.0 3.1
DZN 0.0 n.d. – – – –
75.0 65.9 ± 3.5 5.7 87.8 − 12.2 4.2
250.0 228.8 ± 10.3 4.5 91.5 − 8.5 3.5
750.0 691.5 ± 34.6 5.0 92.2 − 7.8 2.9
CPS 0.0 n.d. – – – –
75.0 65.7 ± 3.5 5.8 87.6 − 12.4 4.2
250.0 228.5 ± 11.0 4.8 91.4 − 8.6 3.3
750.0 681.8 ± 29.3 4.3 90.9 − 9.1 4.0
PFS 0.0 n.d. – – – –
75.0 69.4 ± 4.3 6.2 92.5 − 7.5 2.2
250.0 234.5 ± 13.4 5.7 93.8 − 6.2 2.0
750.0 698.3 ± 32.1 4.6 93.1 − 6.9 2.7
Tomato ATR 0.0 102.3 4.1 – – –
75.0 170.6 ± 8.7 5.1 91.1 − 8.9 1.3
250.0 338.6 ± 14.9 4.4 94.5 − 5.5 1.5
750.0 810.3 ± 38.1 4.7 94.4 − 5.6 1.9
AMT 0.0 n.d. – – – –
75.0 69.7 ± 2.2 3.2 92.9 − 7.1 4.1
250.0 232.8 ± 10.7 4.6 93.1 − 6.9 2.8
750.0 728.3 ± 22.6 3.1 97.1 − 2.9 1.6
MTR 0.0 n.d. – – – –
75.0 67.2 ± 2.9 5.0 89.6 − 10.4 4.0
250.0 231.8 ± 10.7 4.6 92.7 − 7.3 2.9
750.0 684.8 ± 26.7 3.9 91.3 − 8.7 4.2
ETF 0.0 n.d. – – – –
75.0 70.9 ± 4.3 6.0 94.5 − 5.5 1.6
250.0 232.3 ± 11.8 5.1 92.9 − 7.1 2.6
750.0 741.0 ± 28.2 3.8 98.8 − 1.2 0.5
DZN 0.0 n.d. – – – –
75.0 67.6 ± 2.8 4.9 90.1 − 9.9 3.8
250.0 223.3 ± 9.8 5.0 89.3 − 10.7 4.1
750.0 694.5 ± 25.0 3.6 92.6 − 7.4 3.8
CPS 0.0 38.8 6.1 – – –
75.0 108.6 ± 6.0 5.5 93.0 − 7.0 1.5
250.0 279.3 ± 16.2 5.8 96.2 − 3.8 1.0
750.0 749.1 ± 33.7 4.5 94.7 − 5.3 2.0
PFS 0.0 n.d. – – – –
75.0 66.2 ± 3.0 5.4 88.3 − 11.7 4.2
250.0 227.8 ± 9.1 4.2 91.1 − 8.9 4.0
750.0 708.8 ± 26.2 3.7 94.5 − 5.5 2.7
750.0 714.8 ± 25.7 3.6 95.3 − 4.7 2.3
Agricultural water-1 ATR 0.0 n.d. – – – –
25.0 24.3 ± 1.0 4.3 97.2 − 2.8 1.1
100.0 99.4 ± 4.9 4.9 99.4 − 0.6 0.2
500.0 510.5 ± 18.4 3.6 102.1 2.1 0.9
AMT 0.0 n.d. – – – –
25.0 24.6 ± 0.9 3.5 98.3 − 1.7 0.8
100.0 97.2 ± 4.0 4.1 97.2 − 2.8 1.2
500.0 501.5 ± 21.6 4.3 100.3 0.3 0.1
MTR 0.0 n.d. – – – –
25.0 23.7 ± 1.3 5.6 94.8 − 5.2 1.6
100.0 95.5 ± 4.5 4.7 95.5 − 4.5 1.7
500.0 488.0 ± 24.9 5.1 97.6 − 2.4 0.8
ETF 0.0 n.d. – – – –
25.0 24.6 ± 1.5 6.3 98.4 − 1.6 0.4
100.0 100.4 ± 5.3 5.3 100.4 0.4 0.1
500.0 519.0 ± 24.9 4.8 103.8 3.8 1.3
DZN 0.0 n.d. – – – –
(continued on next page)

8
M. Behpour et al. Food Chemistry 407 (2023) 135067

Table 2 (continued )
Sample Analyte Addeda,b Founda ± SDa RSDc (%) RR (%) MEd (%) t-valuef

25.0 23.4 ± 1.3 5.7 93.6 − 6.4 2.0

100.0 94.4 ± 4.2 4.4 94.4 − 5.6 2.3
500.0 480.5 ± 20.2 4.2 96.1 − 3.9 1.6
CPS 0.0 n.d. – – – –
25.0 23.3 ± 1.4 6.1 93.3 − 6.7 2.0
100.0 96.7 ± 3.5 3.6 96.7 − 3.3 1.6
500.0 476.0 ± 20.0 4.2 95.2 − 4.8 2.0
PFS 0.0 28.9 – – – –
25.0 52.3 ± 3.1 5.9 93.7 − 6.3 0.8
100.0 123.4 ± 6.4 5.2 94.5 − 5.5 1.4
500.0 505.9 ± 24.3 4.8 95.4 − 4.6 1.6
a
The food samples and agricultural water concentrations are based on µg/kg and ng/mL, respectively.
b
The concentrations in the fruits and vegetables are reported in initial homogenized samples, so the spiked concentrations with factor 2.5 are reduced in the final
sample solution.
c
RSDs were obtained by three replicate measurements.
d
Matrix effect was calculated based on Eq. (3).
e
n.d not detected.
f
The critical t-value for the degree of freedom of 2 is 4.3.

functionalizations have been used to modify CNTs to improve their
dispersion in water (Lu et al., 2012). The presence of cellulose in the
proposed nano-composite forms an infrastructure with CNTs, leading to
creating a high level of mechanical stability. Besides, the adsorption
capacity of the synthesized nano-composites and extraction recoveries
of desired analytes at the concentration of 250 ng/mL in water samples
were assessed (Table S5). In light of the results, it can be concluded that
Fe3O4@SiO2-NH2@DAC@CNT-COOH has a larger specific surface area
due to the presence of DAC in the sorbent structure, and has a greater
number of functional groups, which has led to an improvement in
extraction recoveries.
3.6. Evaluation of the analytical performance of the proposed method
optimized method. The RRs of target analytes in three spiked levels were
obtained in 87.6–103.8 %. The RSDs values were within the range of
3.1–6.8 %, revealing the accuracy and repeatability of the introduced
method. Fig. 3 and Figure S15 indicate the acquired un-spiked and
spiked real samples. Additionally, the matrix effects were examined in
this study. The calculated ME% values are displayed in Table 2 and
Table S6. This study shows that the method was not influenced
remarkably by the intricate matrix in the agricultural water samples. In
contrast, more complex samples, such as fruit and vegetable, indicated
soft matrix effects. Overall, the results corroborated that this method
based on using Fe3O4@SiO2-NH2@DAC@CNT-COOH as a sorbent could
be applied to the detection and quantification of trace levels of the target
herbicides and insecticides in complicated samples.

The results of the investigation of the figures of merit are reported in
Table 1. All seven target analytes demonstrated excellent linearity
(0.50–1200.0 ng/mL) along with satisfactory coefficients of determi­
nation (R2) above 0.9989. As shown in Table 1, based on the minimum
concentration of analyte that can be identified and quantified, the LODs
and LOQs for the intended analytes are 0.08–1.0 ng/mL and 0.50–5.0
ng/mL, respectively. The Intra-day and inter-day of the proposed
method (n = 5 samples for within day and n = 3 days for between days)
were in the range of 3.1–8.7 % at four concentration levels (25.0, 100.0,
250.0, and 750.0 ng/mL) of target analytes. Also, the PFs were achieved
from 85.7 to 113.6 (EFmax = 135.1), contributing to the recoveries
ranging from 63.4 % to 84.1 %.
3.7. Application of the developed method in real samples
To evaluate the practical efficiency of the suggested method based on
the Fe3O4@SiO2-NH2@DAC@CNT-COOH in the analysis of real sam­
ples, several different un-spiked (blank) and spiked samples involving
agricultural waters, fruits (apple and orange), and vegetables (tomato
and carrot), were prepared as discussed in section 2.3 and were exam­
ined by MD-µSPE method prior to GC-FID analysis. The final results are
listed in Table 2 and Table S6. As a result of the widespread use of
pesticides in agricultural fields, the target herbicides and insecticides
were found in real samples. The results displayed that none of the target
analytes were not seen in agricultural water-2, apple, and orange. On the
contrary, 102.3 μg/kg of ATR and 38.8 μg/kg of CPS were observed in
the tomato and 50.7 μg/kg of DZN in the carrot sample. Also, 28.9 ng/
mL of PFS was found in the agricultural water-1. Furthermore, to study
the method’s accuracy, the real samples were spiked with the pesticides
at the concentrations of 75.0, 250.0, and 750.0 μg/kg for fruit and
vegetable samples and also spiked at the concentrations of 25.0, 100.0,
and 500.0 ng/mL for agricultural waters, and they were examined by the
3.8. Comparative study
To assess the ability of the proposed method based on the Fe3O4@­
SiO2-NH2@DAC@CNT-COOH sorbent, the present work was compared
to some previously reported MD-SPE methods used for monitoring
herbicides and insecticides in various food and water samples at trace
levels (Table 3). Conforming to the acceptable statistical results, the
introduced method has a wider linear range than previous MD-μSPE
methods. The LODs and the standard deviations of the method are the
same as most methods. Besides, this work required a limited amount of
organic solvent to synthesize adsorbent, making it an eco-friendly sor­
bent. Overall, as can be easily understood according to this comparison,
the current method as, a sensitive, uncomplicated, and practical pro­
cedure, can determine the different pesticides in food and water
samples.
4. Conclusion
In summary, this research aimed to introduce Fe3O4@SiO2-
NH2@DAC@CNT-COOH as an effective nano-magnetic adsorbent that
was synthesized in quite a facile manner through covalent surface
functionalization of CNTs via a four-component one-pot Ugi reaction.
The synthesized Fe3O4@SiO2-NH2@DAC@CNT-COOH was character­
ized by FT-IR, XRD, FESEM, VSM, TGA, BET, BJH, and EDX mapping
analysis. The excellent properties of Fe3O4, CNTs, and DAC, all gathered
in Fe3O4@SiO2-NH2@DAC@CNT-COOH nano-sorbent, have led to
substantial magnetic effects, an extensive surface area, fast extraction
dynamic, and good porosity. In this regard, the introduced method re­
veals significant figures of merits. Relevant results were attained, as
evidenced by the adequate LDRs, low LODs, favorable RSDs, and satis­
factory extraction recoveries. In the Fe3O4@SiO2-NH2@DAC@CNT-
COOH nano-composite, the unique morphology and porous structure,

9
M. Behpour et al.
Fig. 3. Typical chromatograms achieved after MD-µSPE from un-spiked and spiked apple, and tomato samples (peaks names: (1) ATR, (2) DZN, (3) MTR, (4) AMT,
(5) ETF, (6) CPS, (7) PFS).
combined with aromatic rings and many different functional groups,
allows a wide variety of pesticides in complicated matrices to be
adsorbed via hydrogen bonding and π-π accumulation interactions with
satisfactory adsorption efficiency. Furthermore, it was demonstrated
that this covalent functionalization increases the solubility and dis­
persibility of CNTs in organic solvents and water. Hence, the integration
procedure of various types of materials, such as DAC and Fe3O4@SiO2-
NH2, in the CNTs substrate is a low-cost, simple, and efficient method for
designing effective adsorbents. Future challenges include the prepara­
tion of the other worthy MCRs functionalized nano-materials and
application in diverse fields of MD-μSPE technique. Eventually, results
indicated that the synthesized sorbent could be usefully employed in the
MD-μSPE procedure to determine the pesticides at trace concentration
amounts in agricultural waters and different forms of food samples such
as fruit and vegetables.
10
Food Chemistry 407 (2023) 135067
Funding
This research did not receive any specific grant from funding
agencies in the public, commercial, or not-for-profit sectors.
CRediT authorship contribution statement
Majid Behpour: Conceptualization, Investigation, Methodology,
Software, Validation, Writing – original draft, Writing – review & edit­
ing. Mehrdad Shadi: Investigation, Methodology. Saeed Nojavan:
Conceptualization, Funding acquisition, Investigation, Project adminis­
tration, Supervision, Writing – review & editing.
Declaration of Competing Interest
The authors declare that they have no known competing financial
M. Behpour et al. Food Chemistry 407 (2023) 135067

Table 3
Comparison of proposed method with other MSPE methods for determination of pesticides.
Sorbent Analytes Matrix Detection Linearity (ng/ LOD (ng/ RSD Extraction time Ref
mL) mL) (%) (min)

Fe3O4@SiO2–C18 Insecticide Water samples HPLC–UV 0.5–10.0 0.05 2.7 7 Maddah et al., 2017
IL-MG Herbicide Water samples HPLC–UV 0.55–500.0 0.09–0.12 ≤ 3.5 20 Zhang et al., 2018
rGOQDs@ Fe Insecticide Fruit juice, water GC–MS 0.19–520.0 0.07 ≤ 4.6 8.5 Akbarzade et al., 2018
samples
Fe3O4@MIL-100(Fe) Herbicide Vegetable, water HPLC–UV 0.07–70.0 2.6–5.3 ≤ 7 Nasrollahpour &
samples 10.2 Moradi, 2018
Magnetized carbon sheet Insecticide Vegetable, water GC-IMS 2.0–10.0 0.85 ≤8 27 Kermani et al., 2019
nanocomposites samples
Magnetic Insecticide Vegetable GC–MS 0.43–1300 0.13–1.49 ≤ 15 Dozein et al., 2021
graphene@Fe3O4@SiO2@TiO2 4.12
Fe3O4@TGA@TMU-6 Insecticide Rice, Water samples HPLC–UV 10.0–150.0 0.5–1.0 ≤ 5.1 40 Shakourian et al.,
2020
mGO/β-CD/MD Herbicide Vegetable HPLC–UV 10.0–700.0 0.01–0.05 ≤ 7.5 30 Majd & Nojavan,
2021b
Fe3O4@SiO2-NH2@DAC@CNT- Herbicide Fruit, vegetable, GC-FID 0.5–1000.0 0.08–0.3 ≤ 5.3 20 This work
COOH Insecticide water samples 3.0–1200.0 0.75–1.0 ≤ 5.5

mGO/β-CD/MD maltodextrin‑ and β‑cyclodextrin‑functionalized graphene oxide, HLB-MPNP hydrophilic–lipophilic balanced magnetic nanoparticle, magnetic poly
(divinylbenzene-co-N-vinylpyrrolidone), rGOQDs@ Fe Zero valent Fe-reduced graphene oxide quantum dots, IL-MG ionic liquid-magnetic grapheme, Fe3O4/rGO
Fe3O4/reduced graphene oxide.

interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
Acknowledgements
Financial support from the Research Affairs of Shahid Beheshti
University is gratefully acknowledged.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.foodchem.2022.135067.
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