Microchemical Journal 162 (2021) 105850

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Microchemical Journal
journal homepage: www.elsevier.com/locate/microc

A simple and effective determination of methyl red in wastewater samples

by UV–Vis spectrophotometer with matrix matching calibration strategy
after vortex assisted deep eutectic solvent based liquid phase extraction and
evaluation of green profile
Nihan Atsever , Tülay Borahan , Ayça Girgin , Dotse Selali Chormey , Sezgin Bakırdere *
Yıldız Technical University, Chemistry Department, 34210 İstanbul, Turkey

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

Keywords: In this study, a green, simple and sensitive vortex assisted deep eutectic solvent based liquid phase extraction
Methyl red (VA-DES-LPE) method was developed to preconcentrate methyl red from wastewater samples before its deter­
Deep eutectic solvent mination by UV–Vis spectrophotometry. Deep eutectic solvent (DES) was employed as the extractant for the
Liquid phase extraction
isolation and preconcentration of methyl red from aqueous solution. Important parameters having effect on the
Wastewater
UV–Vis spectrophotometer
extraction efficiency were investigated and optimized carefully using the “one-factor-at-a-time” approach. Under
the determined optimum conditions, limit of detection (LOD) and limit of quantification (LOQ) values were
determined as 2.3 and 7.5 µg/L, respectively. The preconcentration factor was calculated as 35 folds with respect
to the developed method. Linearity of the presented method was determined in the range of 8.0–1000 µg/L. The
developed method was successfully applied for the determination of methyl red in wastewater samples, and
satisfactory percent recoveries (≈100%) were obtained for different spike concentrations.

1. Introduction
Azo dyes are synthetic organic compounds that have a large range of
color spectrum and their structures contain azo groups (-N = N-) by
which they are distinguished [1,2]. They are the most utilized group of
dyes thanks to their good stabilities, synthetic features and broad color
spectra [3]. Due to good stability of the azo groups, azo dyes are
extensively employed in printing, cosmetics and weaving [4]. They have
been widely utilized in several applications, namely, textiles, electro-
optical devices, nonlinear optics (NLO), colorants for foods and dye-
sensitized solar cells [5]. The discharge of wastewater polluted with
dyes into environmental water bodies may pose a threat to aquatic or­
ganisms, and biomagnify as it moves up the food chain and cause
harmful effects to humans and other high trophic animals [6]. Methyl
red is a non-polar mono azo dye that is chemically known as 2-(4-
dimethylaminophenylazo)benzoic acid, and it is used as an indicator in
the pH range of 4.4–6.0 [7]. It is also employed in ink-jet printing, textile
industries and different trading products [8]. It is known to be a toxic
azo dye that causes cytotoxic and mutagenic effects on organisms [9].
Methyl red undergoes reductive cleavage to produce aromatic amines
that have the potential to cause carcinogenic and mutagenic effects [10].
In case of dermal contact, ingestion or inhalation of methyl red, health
effects such as sensitization to the skin and eyes and digestive tract
irritation may occur [11]. The concentration of dyes become diluted
when they mix with water streams, and their concentrations in waste­
water could be in μg/L levels [12]. When wastewater is released into the
environment, some dyes may cause harmful effects to aquatic life even
at very low concentrations. Textile dyes including methyl red when
released into water bodies could hinder photosynthetic and other bio­
logical activities by blocking sunlight penetration and limiting respira­
tion [13]. Thus, wastewater treatment strategies based on the use of
microbial culture such as bacteria have been used to decolorize and
biodegrade methyl red prior to its release into the environment [14,15].
It is therefore very important to develop sensitive and simple analytical
strategies for the determination of methyl red at trace concentration
levels to augment wastewater treatment processes.
Different analytical techniques such as liquid chromatography mass
spectrometry (LC-MS/MS) [16], gas chromatography-mass spectrom­
etry (GC–MS) [17], high performance liquid chromatography (HPLC)
[18], and thin-layer chromatography [19] have been used to determine

* Corresponding Author.
E-mail address: bsezgin@yildiz.edu.tr (S. Bakırdere).

https://doi.org/10.1016/j.microc.2020.105850
Received 2 November 2020; Received in revised form 1 December 2020; Accepted 8 December 2020
Available online 15 December 2020
0026-265X/© 2020 Elsevier B.V. All rights reserved.
N. Atsever et al.
dyes in various samples. Nonetheless, these techniques are relatively
expensive and require experts to operate the instruments [20]. Beside
these methods, UV–Vis spectrophotometry is commonly used because it
is simple, rapid, cheap, accurate and precise [21]. However, it is chal­
lenging to perform direct determinations at trace levels by UV–Vis
spectrophotometry. The main reason arises from the instrument’s low
sensitivity and selectivity, alongside interfering species in complex
matrices that have effects on the analyte’s signal [22]. These drawbacks
can be altered by applying an appropriate sample treatment process
before analysis to mitigate the interferants and preconcentrate the target
analyte(s) [20]. Extraction methods such as liquid–liquid extraction
(LLE) [23], solid phase extraction (SPE) [24], cloud point extraction
(CPE) [25], solid phase microextraction (SPME) [26], dispersive liq­
uid–liquid microextraction [6], and silica coated magnetic nanoparticles
[27] have been used in the determination of dyes in different matrices.
Different studies have reported the toxicity of organic solvents and their
harmful effects to the environment and humans [28,29]. In recent years,
analytical chemists seek to find eco-friendly, cheap, and adaptable sol­
vents that can be used in extraction processes [30]. Deep eutectic sol­
vents (DESs) are one of the most trending green solvents due to their
advantages over other toxic organic solvents such as ease of preparation,
high extraction efficiency and low toxicity. Hydrogen bond donors
(HBDs) and hydrogen bond acceptors (HBAs) come together at a
particular molar ratio to form DESs [31,32]. Additionally, different
combinations of hydrogen bond acceptors and hydrogen bond donors
can be adjusted depending on the extraction needs [33]. The suitability
of DESs for separation/preconcentration of both inorganic and organic
analytes has been confirmed by different reported studies [30,34,35].
Some recent studies have reported the use of biphasic extraction
systems that utilize hydrophilic DES phase and hydrophobic DES phase
to improve the overall extraction efficiency for high and low polarity
compounds, respectively [36,37].
This study was aimed at developing an extraction/preconcentration
method for the determination of methyl red in wastewater samples at
very low concentration levels by UV–Vis spectrophotometry. Applica­
bility and accuracy of the developed method was tested on wastewater
samples.
2. Materials and methods
2.1. Instrumentation
Absorbance measurements were performed with a Shimadzu UV-
120–02 model UV–Vis spectrophotometer, using 1.0 cm quartz cells.
Hettich EBA 20 model centrifuge was used for centrifugation. Mixing
processes were carried out using a vortex (Wise Mix) and a HAPA model
ultrasonic bath. A Hanna HI 2211 pH/ORP meter was used for pH
measurements throughout the experiments.
2.2. Chemicals and reagents
All reagents/chemicals used throughout this study were of analytical
grade. Stock solution of methyl red (1000 mg/L) was prepared in
ethanol. Calibration standards for direct UV–Vis measurements were
prepared by diluting the stock methyl red solution with ethanol, and
working/calibration standard solutions for the extraction procedure
were prepared in 1.0% ethanol solution (Isolab, Germany) by diluting
with ultrapure water obtained from a Milli-Q® Reference Ultrapure
Water Purification System. DES was produced by mixing phenol (HBD)
with choline chloride (HBA) (ChCl, Acros Organics) at an optimum
molar ratio. pH 4.0 buffer solution was prepared with 0.10 M potassium
hydrogen phthalate solution and 0.10 M hydrochloric acid solution. The
wastewater sample used in the recovery experiment was obtained from
an Advanced Biological Wastewater Treatment Plant in İstanbul.
Tetrahydrofuran, potassium hydrogen phthalate, hydrochloric acid,
phenol and methyl red were all obtained from Merck (Darmstadt,
2
Microchemical Journal 162 (2021) 105850
Germany).
2.3. Synthesis of DES
Deep eutectic solvent was synthesized by mixing phenol with choline
chloride (a quaternary ammonium salt). 1:2, 1:3, 1:4 and 1:5, (ChCl:
PhOH) molar ratios of DESs were prepared and tested to determine the
optimal ratio. The synthesis procedure includes mixing of the two
chemicals in a 50 mL falcon tube by vortex at room conditions. The
mixing process was continued until a homogenous eutectic liquid was
obtained.
2.4. Extraction procedure
In the extraction procedure, 30.0 mL of standard/sample solution
and 1.0 mL of pH 4.0 buffer solution were added to a 50.0 mL falcon
tube. 1.5 mL of hydrophilic DES was injected into this solution with a
syringe and the obtained solution was mixed with vortex for 30 s for
homogeneity of DES in the sample solution. Afterwards, aggregation of
DES molecules was achieved by the addition of THF (4.5 mL) and the
turbid solution formed was mixed by vortexing for 30 s. For the last step
of the extraction procedure, centrifugation was applied to the final so­
lution for phase separation. Then, 1.8 mL of the upper turbid DES-rich
phase was transferred into a clean centrifuge tube, and 50 µL ethanol
was added to obtain a clear and transparent DES-rich phase for UV–Vis
measurements.
3. Results
All experimental parameters including the type and volume of buffer
solution, deep eutectic solvent molar ratio and volume, mixing period,
amount of THF and ethanol volume that influence the performance of
the method were optimized to acquire the highest extraction efficiency
for the analyte. Optimizations were carried out with 3 replicates using
the “one-variable-at-a-time” approach. Optimum values were selected
based on the highest absorbance signal obtained.
3.1. Absorption of spectra
In spectrophotometric measurements, working with the wavelength
of maximum absorbance (λmax) value is of great importance for accurate
and reliable results. For this reason, the wavelength range of 230–500
nm was scanned using a standard solution of methyl red against a blank
reagent. Maximum absorbance value was obtained at 430 nm. There­
fore, 430 nm was used as the maximum absorption wavelength for direct
determinations by the UV–Vis spectrophotometer. Furthermore, ab­
sorption profile was investigated in the range of 380–620 nm against a
blank reagent to determine the maximum absorption wavelength of
methyl red when it is extracted to the DES-rich phase. Results showed
that the maximum absorption wavelength was shifted to 505 nm in the
DES medium. As reported by Kumar et al. and Skulcova et al., the ab­
sorption maximum of the analyte varied from the initial maximum
absorbance probably due to the DES extract’s pH value [38,39]. Thus,
505 nm was used for the measurements after the extraction procedure
was applied.
3.2. Effect of pH and volume of buffer
Generally, extraction efficiency is affected by the pH of the aqueous
solution as it determines the form in which the analyte remains in so­
lution [40]. In this case, interactions between the target analytes and
DES molecules can be influenced according to the solution’s pH [41]. To
examine this effect, buffer solutions in the pH range of 2.0–13 were
added to standard solutions and extracted under equivalent conditions.
According to the results obtained (Fig. 1), interactions between the
analytes and DES were stronger in the acidic medium, while the
N. Atsever et al. Microchemical Journal 162 (2021) 105850

Fig. 1. Absorbance results after extraction with different pH values of buffer solution, n = for error bar (λmax: 430 nm, 30 mL of 250 μg/L standard solution, 1.0 mL
buffer solution, 1.5 mL DES (1:3, ChCl:PhOH), 30 s mixing (vortex), 4.5 mL THF, 30 s mixing (vortex) and 100 µL ethanol).

extraction efficiency decreased as the pH of the solution increased. The
highest extraction efficiency was obtained at pH 4.0, and it was selected
as the optimum to be used in the subsequent experiments. The obtained
results confirmed that the separation of methyl red from the aqueous
solution depends on the pH of the solution.
After determining the optimum pH of the buffer solution, its volume
was optimized by testing 0.50, 1.0, 1.5 and 2.0 mL. A decrease in the
absorbance values was observed with increasing buffer solution volume,
and this can be attributed to the dilution effect. High absorbance values
were obtained with 0.50 and 1.0 mL buffer volumes. As a result, 1.0 mL
of buffer solution was selected as the optimum in order to achieve a
higher buffer capacity for the 30 mL sample volume.
3.3. Effect of deep eutectic solvent (DES) molar ratio and volume
In this optimization step, the effect of molar ratio of DES on the re­
covery of the methyl red was investigated and optimized. In the prep­
aration of eutectic mixtures, peculiar molar ratios of HBA and HBD were
mixed with the aim of forming strong hydrogen bonds. Hence, ratios of
1:2, 1:3, 1:4 and 1:5, (ChCl:PhOH) were prepared and tested under
equivalent conditions. A decrease in the viscosity of the eutectic mix­
tures was observed when the phenol content increased, and the highest
absorbance values were obtained at the ratios of 1:3 and 1:5. In order to
avoid surplus chemical consumption, the optimum ratio was chosen as
1:3.
After examining the effect of DES molar ratio on absorbance values,
the effect of its volume on extraction output was investigated. It is
important to optimize the volume of DES to find a sufficient amount for
an efficient recovery of the analyte. Thus, 1.5, 2.0, 2.5 and 3.0 mL DES
volumes were tested. There was a significant decrease in the absorbance
values with increasing volume due to dilution of the analyte in the DES-
rich phase, as can be seen in Fig. 2. Thus, 1.5 mL was chosen as the
optimum value. Lower volumes of DES were not tested due to the vol­
ume restriction of the instrument. A mixing process (vortex) was applied
after DES addition. Optimum mixing period by vortex was examined by
testing 0.0, 15, 30 and 60 s. 30 s vortexing period gave the highest
absorbance value and was chosen as the optimum value.

Fig. 2. Absorbance results obtained after extraction with different volumes of deep eutectic solvent, n = 3 for error bars (λmax: 430 nm, 30 mL of 250 μg/L standard
solution, 1.0 mL buffer solution, 1:3, (ChCl:PhOH) DES, 15 s mixing (vortex), 4.0 mL THF, 15 s mixing (vortex) and 300 µL ethanol).

3
N. Atsever et al.
3.4. Effect of emulsifier agent
Aprotic solvents (emulsifier agents) are not able to form intra­
molecular hydrogen bonds since they do not have N–H or O–H bonds,
however they can accept H-bonds. These types of solvents are required
for the self-aggregation of DES molecules in aqueous solution to form a
separate water insoluble phase [42]. In this regard, THF was used as an
emulsifier agent to initiate phase separation and mass transfer of target
analytes into the DES phase. Various volumes of THF were tested
including 3.0, 3.5, 4.0 and 4.5 mL. Although higher absorbance values
were obtained when 3.0 and 3.5 mL of THF volumes were used, their
standard deviation values were quite high, therefore, 4.5 mL of THF was
chosen as the optimum volume to obtain more precise results. Addi­
tionally, after THF addition, mixing time period was optimized and 30 s
vortex period was chosen as the optimum value.
3.5. Effect of ethanol volume
Addition of ethanol to the turbid DES-rich phase was very critical as
it helped to obtain a clear/transparent phase for absorbance measure­
ments. Amount of ethanol added to the DES-rich phase was optimized in
order to determine a sufficient volume without causing excess dilution.
The volumes of ethanol tested under similar conditions were 50, 100,
200 and 300 µL. A decreasing trend in absorbance signals was observed
as the ethanol volume increased. The absorbance values recorded for the
50 and 100 µL volumes were close to each other, but 50 µL was chosen as
the optimum volume because excess chemical usage is undesirable.
3.6. Analytical figures of merit
The analytical performance of the analytical method developed was
evaluated under the optimized conditions given in Table 1, and the
values obtained are compared to some literature studies in Table 2.
Analytical performance of the presented method was evaluated ac­
cording to detection limit (LOD), quantification limit (LOQ), recovery,
repeatability and linearity. LOQ and LOD values for the UV–Vis and VA-
DES-LPE-UV–Vis systems were calculated using the equations given
below, where S represents the standard deviation of the lowest con­
centration in the calibration plot with six replicate measurements.
LOD = 3xS/slope(1)
LOQ = 10xS/slope(2)
Under the optimum conditions, calibration graphs were constructed
and compared for both measurements using 430 nm and 505 nm
wavelengths. LOD and LOQ values were calculated for the developed
method as 2.3 and 7.5 µg/L, respectively. The developed method
exhibited a broad calibration range of 8.0–1000 µg/L, with a coefficient
of determination value of (R2) 0.9998. Combination of the developed
method with the UV–Vis spectrophotometer showed a 35-fold
improvement in detection power (LOD comparison) when compared
to the direct UV–Vis measurements. Overlay absorbance signals of 100,
250 and 500 µg/L standard solution extracts under the optimum con­
ditions is depicted in Fig. 3.
Table 1
Optimized parameters of the VA-DES-LPE-UV–Vis method.
Parameters Value
Wavelength 505 nm
pH of buffer solution, volume 4.0, 1.0 mL
DES molar rate, volume ChCl:PhOH; 1:3, 1.5 mL
Vortex period after DES addition 30 s
Emulsifier agent, volume THF, 4.5 mL
Vortex period after THF addition 30 s
Ethanol volume 50 µL
4
Microchemical Journal 162 (2021) 105850
Table 2
Analytical performance of the method and comparison to other literature
studies.
Method LOD, LOQ, µg/ Linear Range, RSD Ref.
µg/L L µg/L %
UV–Vis 80 250 250–25000 12.1 This
work
VA-DES-LPE- 2.3 7.5 8.0–1000 11.6 This
UV–Vis work
*DLLME-UV–Vis 5000 15,000 15 mg/L − 10 g/ – [6]
L
**DSPE-UV–Vis 1400 4600 100–25000 1.0 [43]
*DLLME: Dispersive liquid–liquid microextraction
** DSPE: Dispersive solid-phase extraction
3.7. Recovery
Investigation of the applicability and accuracy of the VA-DES-LPE
method was carried out in wastewater samples by spiked recovery ex­
periments. Firstly, wastewater samples were filtered, and blank sample
analyses were performed to check whether methyl red was present in the
wastewater samples. No analytical signal was observed in the samples,
suggesting methyl red was not present in the sample or below the
method’s LOD. Afterwards, wastewater samples were spiked at con­
centrations of 25, 50, 250, 500 and 1000 µg/L. Under the optimum
conditions, the spiked wastewater samples were extracted and
compared with the calibration standards solutions. The calculated
percent recoveries were lower than 89% when the external calibration
strategy was employed. The low percent recovery results proved inter­
ference effects emanating from the complex nature of the wastewater
matrix. Thus, matrix matching calibration strategy was applied to
eliminate the matrix effects, and to improve the accuracy of the results.
Matrix matched calibration standards were prepared in a separate
wastewater matrix cleared not to contain methyl red, and a four-point
calibration graph was constructed. Percent recoveries calculated for
the concentrations of 250 and 500 µg/L were 101 ± 1.0% and 99 ±
1.9%, respectively. Matrix matching method improved the accuracy of
the results in wastewater samples, and the low %RSD values established
high precision.
3.8. Evaluation of green profile
“Eco-friendly Analytical Chemistry” has caught great attention since
it was first introduced in 1999 [44]. In this regard, green chemistry was
first introduced by Anastasin [45] and Green Analytical Chemistry
(GAC) was born as a consequence [46]. The goals of GAC include the
reduction of energy consumption, eliminating/reducing the use of toxic
solvents and lowering waste generation [47]. In 2012, the GAC
approach proposed by Galuszka et al. outlined penalty points by which
analytical methods are graded according to greenness [48]. In this study,
penalty points of the VA-DES-LPE method were calculated for the
evaluation of its green profile (Table 3). In this respect, penalty points
taken against the volumes of sample solution, buffer solution, DES, THF
and ethanol were summed and subtracted from the base value of 100.
Analytical eco-scale of the method was calculated as 92, which repre­
sents ‘excellent green analysis’ according to the volumes of sample and
reagents, and generated waste per analysis [48].
4. Conclusion
In this study, a simple liquid phase extraction method was developed
using deep eutectic solvent to extract methyl red from wastewater
samples prior to determination by UV–Vis spectrophotometry. The
developed analytical method facilitated the determination of methyl red
at very low concentrations in wastewater samples with high accuracy
and precision. Phenol and choline chloride were used to obtain the eco-
N. Atsever et al. Microchemical Journal 162 (2021) 105850

Fig. 3. Overlay absorbance signals of 100, 250 and 500 µg/L standard solution extracts under the optimum conditions.

Declaration of Competing Interest

Table 3
The penalty points (PPs) for methyl red determination in wastewater samples by
The authors declare that they have no known competing financial
VA-DES-LPE-UV–Vis.
interests or personal relationships that could have appeared to influence
Reagents the work reported in this paper.
Penalty Points

Methyl red standard (30 mL) 2

Buffer solution (1.0 mL) 1
DES (1.5 mL) 2
THF (4.5 mL) 1 Supplementary data to this article can be found online at https://doi.
Ethanol (50 µL) 1 org/10.1016/j.microc.2020.105850.
Waste (1.8 mL) 1
∑8
Instrument
UV–Vis 0
Total Penalty Points: 8
Analytical Eco-Scale Total Score: 92
friendly eutectic mixture, distinguished by its cheapness and low
toxicity. About 35 folds increment in the detection power of the UV–Vis
system was obtained with the combination of the developed VA-DES-
LPE method, corresponding to a detection limit of 2.3 µg/L. Spike re­
covery experiments were used to validate the accuracy and applicability
of the developed method to real sample matrix. Matrix matching cali­
bration strategy was used to mitigate interference from the wastewater
matrix to obtain satisfactory results (≈100%). This method recorded an
analytical eco-scale point of 92, validating it as an excellent green
analysis method. The proposed method is rapid, simple, economic,
sensitive and can be applied for routine laboratory analysis.
CRediT authorship contribution statement
Nihan Atsever: Conceptualization, Methodology, Validation,
Writing - original draft. Tülay Borahan: Conceptualization, Methodol­
ogy, Validation, Writing - original draft. Ayça Girgin: Conceptualiza­
tion, Methodology, Validation, Writing - original draft. Dotse Selali
Chormey: Conceptualization, Methodology, Validation, Writing - orig­
inal draft. Sezgin Bakırdere: Supervision, Project administration,
Writing - review & editing.
Appendix A. Supplementary data
References
[1] M.R. Maliyappa, J. Keshavayya, N.M. Mallikarjuna, I. Pushpavathi, Novel
substituted aniline based heterocyclic dispersed azo dyes coupling with 5-methyl-
2-(6-methyl-1, 3-benzothiazol-2-yl)-2, 4-dihydro-3H-pyrazol-3-one: Synthesis,
structural, computational and biological studies, J. Mol. Struct. 1205 (2020)
127576, https://doi.org/10.1016/j.molstruc.2019.127576.
[2] J. Jiao, J. Wang, M. Li, J. Li, Q. Li, Q. Quan, J. Chen, Simultaneous determination
of three azo dyes in food product by ion mobility spectrometry, J. Chromatogr. B.
1025 (2016) 105–109, https://doi.org/10.1016/j.jchromb.2016.05.002.
[3] B. Babür, N. Seferoğlu, E. Aktan, T. Hökelek, E. Şahin, Z. Seferoğlu,
Phenylazoindole dyes 3: Determination of azo-hydrazone tautomers of new
phenylazoindole dyes in solution and solid state, J. Mol. Struct. 1081 (2015)
175–181, https://doi.org/10.1016/j.molstruc.2014.09.082.
[4] G. Palanisamy, T. Pazhanivel, K. Bhuvaneswari, G. Bharathi, G. Marimuthu,
T. Maiyalagan, Spinel oxide ZnCr2O4 incorporated with ZnS quantum dots for
application on visible light driven photocatalyst Azo dye degradation, Colloids
Surf., A 590 (2020) 124505, https://doi.org/10.1016/j.colsurfa.2020.124505.
[5] M.R. Maliyappa, J. Keshavayya, M. Mahanthappa, Y. Shivaraj, K.V. Basavarajappa,
6-Substituted benzothiazole based dispersed azo dyes having pyrazole moiety:
Synthesis, characterization, electrochemical and DFT studies, J. Mol. Struct. 1199
(2020) 126959, https://doi.org/10.1016/j.molstruc.2019.126959.
[6] S. Khodadoust, M. Ghaedi, Application of response surface methodology for
determination of methyl red in water samples by spectrophotometry method,
Spectrochim, Acta Part A Mol. Biomol. Spectrosc. 133 (2014) 87–92, https://doi.
org/10.1016/j.saa.2014.04.119.
[7] M.R. Plutino, E. Guido, C. Colleoni, G. Rosace, Effect of GPTMS functionalization
on the improvement of the pH-sensitive methyl red photostability, Sensors
Actuators B Chem. 238 (2017) 281–291, https://doi.org/10.1016/j.
snb.2016.07.050.
[8] Z. Zaheer, A. AL-Asfar, E.S. Aazam, Adsorption of methyl red on biogenic Ag@Fe
nanocomposite adsorbent: Isotherms, kinetics and mechanisms, J. Mol. Liq. 283
(2019) 287–298, https://doi.org/10.1016/j.molliq.2019.03.030.
[9] S.C.D. Sharma, Q. Sun, J. Li, Y. Wang, F. Suanon, J. Yang, C.-P. Yu, Decolorization
of azo dye methyl red by suspended and co-immobilized bacterial cells with

5
N. Atsever et al.
mediators anthraquinone-2,6-disulfonate and Fe3O4 nanoparticles, Int.
Biodeterior. Biodegradation. 112 (2016) 88–97, https://doi.org/10.1016/j.
ibiod.2016.04.035.
[10] M.A. Ahmad, N.B. Ahmed, K.A. Adegoke, O.S. Bello, Sorption studies of methyl red
dye removal using lemon grass (Cymbopogon citratus), Chem. Data Collect. 22
(2019) 100249, https://doi.org/10.1016/j.cdc.2019.100249.
[11] U. Guyo, N. Matewere, K. Matina, T. Nharingo, M. Moyo, Fabrication of a
sustainable maize stover-graft-methyl methacrylate biopolymer for remediation of
methyl red contaminated wasters, Sustain. Mater. Technol. 13 (2017) 9–17,
https://doi.org/10.1016/j.susmat.2017.05.001.
[12] J. Riu, I. Schönsee, D. Barceló, C. Ràfols, Determination of sulphonated azo dyes in
water and wastewater, TrAC Trends in Anal. Chem. 16 (7) (1997) 405–419,
https://doi.org/10.1016/S0165-9936(97)00034-4.
[13] D.A. Yaseen, M. Scholz, Treatment of synthetic textile wastewater containing dye
mixtures with microcosms, Environ Sci Pollut Res 25 (2) (2018) 1980–1997,
https://doi.org/10.1007/s11356-017-0633-7.
[14] P.K. Wong, P.Y. Yuen, Decolorization and biodegradation of methyl red by
Klebsiella pneumoniae RS-13, Water Res. 30 (7) (1996) 1736–1744, https://doi.
org/10.1016/0043-1354(96)00067-X.
[15] P.P. Vijaya, S. Sandhya, Decolorization and Complete Degradation of Methyl Red
by a Mixed Culture, Environmentalist. 23 (2003) 145–149, https://doi.org/
10.1023/A:1024839805387.
[16] E. Guerra, M. Celeiro, J.P. Lamas, M. Llompart, C. Garcia-Jares, Determination of
dyes in cosmetic products by micro-matrix solid phase dispersion and liquid
chromatography coupled to tandem mass spectrometry, J. Chromatogr. A. 1415
(2015) 27–37, https://doi.org/10.1016/j.chroma.2015.08.054.
[17] U. Balçık, D.S. Chormey, M.F. Ayyıldız, S. Bakırdere, Liquid phase microextraction
based sensitive analytical strategy for the determination of 22 hazardous aromatic
amine products of azo dyes in wastewater and tap water samples by GC-MS system,
Microchem. J. 155 (2020) 104712, https://doi.org/10.1016/j.
microc.2020.104712.
[18] Z. Lian, J. Wang, Molecularly imprinted polymer for selective extraction of
malachite green from seawater and seafood coupled with high-performance liquid
chromatographic determination, Mar. Pollut. Bull. 64 (12) (2012) 2656–2662,
https://doi.org/10.1016/j.marpolbul.2012.10.011.
[19] ]. Oka, Y. Ikaia, T. Ohno, N. Kawamura, J. Hayakawa, K.-I. Harada, M. Suzuki,
Identification of unlawful food dyes by thin-layer chromatography-fast atom
bombardment mass spectrometry, J. Chromatogr. A 674 (1-2) (1994) 301–307,
https://doi.org/10.1016/0021-9673(94)85235-9.
[20] S.H. Hashemi, M. Kaykhaii, A. Jamali Keikha, E. Mirmoradzehi, Box-Behnken
design optimization of pipette tip solid phase extraction for methyl orange and acid
red determination by spectrophotometry in seawater samples using graphite based
magnetic NiFe2O4 decorated exfoliated as sorbent, Spectrochim. Acta Part A Mol.
Biomol. Spectrosc. 213 (2019) 218–227, https://doi.org/10.1016/j.
saa.2019.01.049.
[21] M. Dehghani Mohammad Abadi, N. Ashraf, M. Chamsaz, F. Shemirani, An
overview of liquid phase microextraction approaches combined with UV–Vis
spectrophotometry, Talanta 99 (2012) 1–12, https://doi.org/10.1016/j.
talanta.2012.05.027.
[22] S.H. Hashemi, M. Kaykhaii, A.J. Keikha, Z. Sajjadi, Application of Box-Behnken
design in response surface methodology for the molecularly imprinted polymer
pipette-tip solid phase extraction of methyl red from seawater samples and its
determination by spectrophotometery, Mar. Pollut. Bull. 137 (2018) 306–314,
https://doi.org/10.1016/j.marpolbul.2018.10.037.
[23] S. Elumalai, G. Muthuraman, Removal and Recovery of Methyl Violet Dye from
Industrial Wastewater by Liquid-Liquid Extraction BT - Water Quality
Management, in: V.P. Singh, S. Yadav, R.N. Yadava (Eds.), Springer Singapore,
Singapore, 2018, pp. 231–240.
[24] X.-R. Shi, X.-L. Chen, Y.-L. Hao, L. Li, H.-J. Xu, M.-M. Wang, Magnetic metal-
organic frameworks for fast and efficient solid-phase extraction of six Sudan dyes
in tomato sauce, J. Chromatogr. B. 1086 (2018) 146–152, https://doi.org/
10.1016/j.jchromb.2018.04.022.
[25] E. Heidarizadi, R. Tabaraki, Simultaneous spectrophotometric determination of
synthetic dyes in food samples after cloud point extraction using multiple response
optimizations, Talanta 148 (2016) 237–246, https://doi.org/10.1016/j.
talanta.2015.10.075.
[26] A. Asfaram, M. Ghaedi, A. Goudarzi, Optimization of ultrasound-assisted dispersive
solid-phase microextraction based on nanoparticles followed by spectrophotometry
for the simultaneous determination of dyes using experimental design, Ultrason.
Sonochem. 32 (2016) 407–417, https://doi.org/10.1016/j.ultsonch.2016.04.009.
[27] M. Shariati-Rad, M. Irandoust, S. Amri, M. Feyzi, F. Ja’fari, Removal,
preconcentration and determination of methyl red in water samples using silica
coated magnetic nanoparticles, J. Appl. Res. Water Wastewater. 1 (2014) 6–12.
https://arww.razi.ac.ir/article_44.html.
Microchemical Journal 162 (2021) 105850
[28] A. Levet, C. Bordes, Y. Clément, P. Mignon, C. Morell, H. Chermette, P. Marote,
P. Lantéri, Acute aquatic toxicity of organic solvents modeled by QSARs, J Mol
Model 22 (12) (2016), https://doi.org/10.1007/s00894-016-3156-0.
[29] C.-H. Lin, C.-H. Lai, Y.-P. Peng, P.-C. Wu, K.-Y. Chuang, T.-Y. Yen, Y.-K. Xiang,
Comparative health risk of inhaled exposure to organic solvents, toxic metals, and
hexavalent chromium from the use of spray paints in Taiwan, Environ Sci Pollut
Res 26 (2019) 33906–33916, https://doi.org/10.1007/s11356-018-2669-8.
[30] B. Doğan, A. Elik, N. Altunay, Determination of paracetamol in synthetic urea and
pharmaceutical samples by shaker-assisted deep eutectic solvent microextraction
and spectrophotometry, Microchem. J. 154 (2020) 104645, https://doi.org/
10.1016/j.microc.2020.104645.
[31] N.D. Oktaviyanti, Kartini, A. Mun’im, Application and optimization of ultrasound-
assisted deep eutectic solvent for the extraction of new skin-lightening cosmetic
materials from Ixora javanica flower, Heliyon 5 (11) (2019) e02950, https://doi.
org/10.1016/j.heliyon.2019.e02950.
[32] W. Tang, Y. An, K.H. Row, Recoverable deep eutectic solvent-based aniline organic
pollutant separation technology using choline salt as adsorbent, J. Mol. Liq. 306
(2020) 112910, https://doi.org/10.1016/j.molliq.2020.112910.
[33] F. Pena-Pereira, M. Tobiszewski, The application of green solvents in separation
processes, Elsevier, 2017.
[34] M. Pirsaheb, N. Fattahi, Development of a liquid-phase microextraction based on
the freezing of a deep eutectic solvent followed by HPLC-UV for sensitive
determination of common pesticides in environmental water samples, RSC Adv. 8
(21) (2018) 11412–11418, https://doi.org/10.1039/C8RA00912K.
[35] R. Akramipour, M.R. Golpayegani, M. Ghasemi, N. Noori, N. Fattahi, Development
of an efficient sample preparation method for the speciation of Se (IV)/Se (VI) and
total inorganic selenium in blood of children with acute leukemia, New J. Chem.
43 (2019) 6951–6958.
[36] W. Tang, K. Ho Row, Evaluation of CO2-induced azole-based switchable ionic
liquid with hydrophobic/hydrophilic reversible transition as single solvent system
for coupling lipid extraction and separation from wet microalgae, Bioresour.
Technol. 296 (2020) 122309, https://doi.org/10.1016/j.biortech.2019.122309.
[37] W. Tang, K.H. Row, Design and evaluation of polarity controlled and recyclable
deep eutectic solvent based biphasic system for the polarity driven extraction and
separation of compounds, J. Cleaner Prod. 268 (2020) 122306, https://doi.org/
10.1016/j.jclepro.2020.122306.
[38] A. Skulcova, V. Majova, M. Kohutova, M. Grosik, J. Sima, M. Jablonsky, UV/Vis
Spectrometry as a quantification tool for lignin solubilized in deep eutectic
solvents, BioResources. 12 (2017) 6713–6722.
[39] A.K. Kumar, B.S. Parikh, E. Shah, L.Z. Liu, M.A. Cotta, Cellulosic ethanol
production from green solvent-pretreated rice straw, Biocatal. Agric. Biotechnol. 7
(2016) 14–23, https://doi.org/10.1016/j.bcab.2016.04.008.
[40] L. Jia, X. Huang, W. Zhao, H. Wang, X. Jing, An effervescence tablet-assisted
microextraction based on the solidification of deep eutectic solvents for the
determination of strobilurin fungicides in water, juice, wine, and vinegar samples
by HPLC, Food Chem. 317 (2020) 126424, https://doi.org/10.1016/j.
foodchem.2020.126424.
[41] M. Faraji, M. Mahmoodi-Maymand, F. Dastmalchi, Green, fast and simple
dispersive liquid-liquid microextraction method by using hydrophobic deep
eutectic solvent for analysis of folic acid in fortified flour samples before liquid
chromatography determination, Food Chem. 320 (2020) 126486, https://doi.org/
10.1016/j.foodchem.2020.126486.
[42] N. Altunay, A. Elik, R. Gürkan, A novel, green and safe ultrasound-assisted
emulsification liquid phase microextraction based on alcohol-based deep eutectic
solvent for determination of patulin in fruit juices by spectrophotometry, J. Food
Compos. Anal. 82 (2019) 103256, https://doi.org/10.1016/j.jfca.2019.103256.
[43] G.R. Bardajee, S. Azimi, M.B.A.S. Sharifi, Application of central composite design
for methyl red dispersive solid phase extraction based on silver nanocomposite
hydrogel: Microwave assisted synthesis, Microchem. J. 133 (2017) 358–369,
https://doi.org/10.1016/j.microc.2017.03.037.
[44] D.S. Hage, Reference module in chemistry, molecular sciences and chemical
engineering, in: Elsevier Inc., 2013.
[45] Paul T. Anastas, Green Chemistry and the Role of Analytical Methodology
Development, Crit. Rev. Anal. Chem. 29 (3) (1999) 167–175, https://doi.org/
10.1080/10408349891199356.
[46] A. Gałuszka, Z. Migaszewski, J. Namieśnik, The 12 principles of green analytical
chemistry and the SIGNIFICANCE mnemonic of green analytical practices, TrAC,
Trends Anal. Chem. 50 (2013) 78–84, https://doi.org/10.1016/j.trac.2013.04.010.
[47] I. Pacheco-Fernández, V. Pino, Green solvents in analytical chemistry, Curr. Opin.
Green Sustainable Chem. 18 (2019) 42–50, https://doi.org/10.1016/j.
cogsc.2018.12.010.
[48] A. Gałuszka, Z.M. Migaszewski, P. Konieczka, J. Namieśnik, Analytical Eco-Scale
for assessing the greenness of analytical procedures, TrAC, Trends Anal. Chem. 37
(2012) 61–72, https://doi.org/10.1016/j.trac.2012.03.013.