Anal. Methods Environ. Chem. J. 6 (1) (2023) 5-16
Research Article, Issue 1
Analytical Methods in Environmental Chemis try Journal
Journal home page: www.amecj.com/ir
AMECJ
Adsorption behavior of Crys tal Violet dye in aqueous
solution using Co+2 hectorite composite
as adsorbent surface
Ahmed Jaber Ibrahima,*
a Scientic Research Center, Al-Ayen University, ThiQar 64011, Iraq
ABSTRACT
This s tudy focused on the adsorption behavior of the cationic Crys tal
Violet (CV) dye from aqueous solutions using a Co+2
composite as an adsorbent surface. The initial and equilibrium CV dye
concentrations were determined using a UV-Vis spectrophotometer. The
results were discussed and presented for the impacts of pH, primary CV
dye concentration, composite dosage, and temperature. The optimum
conditions were found for eliminating Crys tal Violet dye from the
aqueous solution at a pH 4, ideal temperature 293 K, and 0.5 g L-1
of composite dose. The pseudo-second-order kinetic, intraparticle
in good agreement and in charge of regulating the adsorption reaction.
The adsorption operation was also thermodynamically examined to
o),
oo). The negative
oo) indicated that
the adsorption process was a spontaneous and exothermic reaction.
While the activation energy (Ea) data which fell within the normal
range for physisorption, was discovered to be 22.434 kJ mol-1. This
result proved that physical adsorption occurs between the CV dye and
the adsorbent surface (Cohectorite composite).
Keywords:
Adsorption,
UV-Vis spectrophotometer,
Crys tal violet,
Isotherm,
Thermodynamics,
Kinetics
ARTICLE INFO:
Received 15 Nov 2022
Revised form 1 Feb 2023
Accepted 25 Feb 2023
Available online 30 Mar 2023
*Corresponding Author: Ahmed Jaber Ibrahim
Email: ahmed.jibrahim@alayen.edu.iq
https://doi.org/10.24200/amecj.v6.i01.219
------------------------
1. Introduction
Although synthetic dyes are widely utilized in the
textile sector, 20 to 40 % of these pigments s till
[1-3]. The majority of pigments
contain hazardous and cancer-causing subs tances.
of people and the environment since they are
resis tive and so s table in a recovering ecosys tem [4].
Therefore, before the dye-containing was tewater
is released into the environment, the dyes mus t
be removed to safeguard persons and ecosys tems
from pollution. The elimination of contaminants
textiles has been documented using several physical,
chemical, and biological decolorization processes.
However, these sectors had only welcomed a small
number of them [5–18]. Adsorption is the bes t
among the several dye removal procedures since it
contaminants from aqueous solutions. Adsorption
is preferable to compete for sys tems for utilizing
recycled water regarding low cos t, formability and
s tyling simplicity, ease of use, and sensitivity to
harmful contaminants. According to several s tudies
6Anal. Methods Environ. Chem. J. 6 (1) (2023) 5-16
[19,20], activated charcoal and polymer resins are
the bes t adsorbents for eliminating pigments from
suitably saturated sewage. The adsorption capability
of some reactive dyes by activated carbon is known
to be relatively poor. The sewage treatment process
utilizing clay-basic [21], AC-ZnO nanos tructure [22],
cotton [23], Ultraviolet-activated sodium perborate
[24], halloysite nanotubes [25]
mat [26], chitosan [27], and natural zeolite-basic
[18] has thus been the subject of Previous s tudies.
Because of their large surface area and molecular
sieve composition, Clay-based materials are
[3]. The
generality widely utilized layered silicate is hectorite.
Tetrahedral subs tituted and octahedral subs tituted
are the two s tructural kinds. Hectorite is an excellent
adsorbent for eliminating dye from comparatively
saturated was tewater. This is attributed to the fact that
hectorite has a unique s tructure with internal channels
that permits the passage of solutes and bonded organic
and inorganic ions into the s tructure of hectorite.
This article sugges ted using a Co+2-hectorite
composite as an Adsorbent surface to absorb crys tal
violet (CV) dye from aqueous solutions. The results
were discussed and presented for the impacts of pH,
primary CV dye concentration, composite dosage,
and temperature. The data from the tes ts were
analyzed by the pseudo-second-order kinetic, intra-
adsorption operation was also thermodynamically
examined to determine thermodynamic variables
o o),
o).
2. Materials and Methods
2.1. Ins truments
Thermos tatic Controlled shaker (SHKE4000, Thermo
(UV-3600i Plus, Shimadzu, Japan), pH meter
Germany), Mechanical s tirrer (Euros tar 60 digital,
2.2. Chemicals
All chemicals with high purity were purchased
from the original Company. The chemicals such as,
Crys tal Violet (CV) dye (C25H30ClN3, Mwt 407.986
dalton, CAS N.: 548-62-9, Tokyo Chemical
Fig. 1), hectorite (Na0.3(Mg,
Li)3Si4O10(OH)2, Mwt 360.58 dalton, CAS N.:
12173-47-6, Spectrum Chemical Co., USA),
hydrochloric acid 37% (HCL, CAS N.: 7647-
01-0, Sigma-Aldrich Chemie GmbH Co., USA),
and Cobalt chloride (CoCl2, CAS N.: 7646-79-9,
American Elements Co., USA) were prepared for
this research.
Fig. 1. The s tructural formula of Crys tal Violet dye
2.3. Preparation of Co+2-hectorite composite
The ion-exchanged technique was used to prepare
the sorbent in a single s tep. Two grams of hectorite
were mixed with 0.2 liters of dis tilled water and
swirled for 2 hours. Using hydrochloric acid (1M,
reduced to 6. Cobalt chloride (CoCl2) solution
was added in the calculated amount while s tirring
dis tilled water. At 80oC, the product was dried after
centrifugation.
2.4. Adsorbate
Crys tal Violet (CV) dye was used as the model gues t
to examine the adsorption capability. Using a UV-Vis
spectrophotometer with a range of 200 - 800 nm, the
7
Adsorption of CV Dye by Cobalt-Hectorite Composite Ahmed Jaber Ibrahim
maximum wavelength of 585 nm was determined,
which corresponds to the highes t absorption of the
dye solution, as shown in Figure 2. To create the
s tock solution, dis tilled water was used to dissolve a
carefully weighed quantity of CV dye. The solutions
for adsorption tes ting were made at the necessary
concentrations by applying serial dilutions to the
s tock solution. Firs t, a calibration curve for CV dye
was drawn. In kinetic and thermodynamic s tudies,
this curve was used to translate data on concentration
from absorbance measurements.
2.5. Adsorption process
At various temperatures, adsorption s tudies were
conducted in a controlled thermos tatic shaker.
Up until the point of equilibrium, the shaking
persis ted. The initial and equilibrium CV dye
concentrations were determined using a UV-Vis
spectrophotometer. The adsorption capability of the
adsorbent was determined using these data. It was
possible to determine the quantity of CV adsorbed
(qe) at equilibrium. The mass balance is shown in
Equation 1.
qe=v(C0- Ce )/W (Eq.1)
Where v is the volume of dye solution used (L), is
the primary dye concentration in the liquid phase
(g L-1), is the liquid phase dye concentration at
equilibrium (g L-1), and W is the mass of sorbent
utilized (g). By adding 0.03 g sorbent at various
temperatures to 0.060 L of crys tal violet solution
(0.150 g L-1), kinetic inves tigations were conducted.
The liquid phase crys tal violet concentration was
monitored at predetermined intervals.
3. Results and Discussion
3.1. The inuence of solution pH
of CV dye by Co+2-hectorite complex was s tudied
in the range of 2–12 under the conditions: 0.150
g L-1 CV dye concentration, 0.5 g L-1 Composite
dosages, 293 temperature, and 1 hour Contact
time). The implies of the repeated experimental
outcomes are plotted in Figure 3. The experimental
results showed that the degree of adsorption of CV
dye on the Co+2-hectorite composite reached 95%
when the pH of the solution was 4. Therefore, the
optimal pH was considered to be 4, which achieves
the maximum adsorption of CV dye. On this basis,
the remainder of the subsequent tes ts were carried
out at this optimum pH value. Other inves tigators
Fig. 2. The UV-Vis spectrum of Crys tal violet dye
8
have shown a tendency similar to the adsorption
process of the Congo red azo dye as a function of
pH [28].
3.2. Eect of sorbent composite dose
Between 0.5 and 1.5 g L-1 of Co+2-hectorite
composite, the dose was tes ted under the
conditions: (primary CV dye concentration 0.5 g
L-1, 0.6 g L-1, 0.7 g L-1 , ph=4, 293 K temperature
dye adsorption. Figure 4
a decrease in with an increase in Co+2-hectorite
dosage. Because a greater adsorbent dose decreases
less adsorption at larger adsorbent doses. CV dye
can quickly arrive at the adsorption locations, and
the qe rises when the amount of adsorbent is modes t.
is being utilized with an increase in adsorbent
amount, the correlating increase in adsorption
reaction per unit clus ter is decreased.
Higher adsorbent dosages caused particle
aggregation, reducing the overall surface area
and the multitude of active adsorption locations.
The highes t CV dye adsorption in this research
was accomplished at a Co+2-hectorite dose of
0.5g remaining trials were carried out at this
concentration.
0.0 0.5 1.0 1.5 2.0
0.2
0.3
0.4
0.5
0.6
sorbent compsite dose (g.L
-1
)
qe (g.g
-1
)
0.5 g.L
-1
0.6 g.L
-1
0.7 g.L
-1
0246810 12 14
0.86
0.88
0.90
0.92
0.94
0.96
0.98
Initial pH level
Rate of adsorpation
Fig. 3. +2-hectorite composite
Fig. 4.
Anal. Methods Environ. Chem. J. 6 (1) (2023) 5-16
9
3.3. Eect of primary crys tal violet (CV) dye
concentration and temperature
It is unclear how varied CV dye concentrations
+2-hectorite composite removes
may include varying amounts of dye. This s tudy
examined the adsorption of concentrations of 0.100,
0.125, 0.150, 0.175, and 0.200 g L-1 CV dye for
the Co+2
means are shown in Figure 5, which shows that the
on the adsorption capability of the Co+2-hectorite
composite. Based on the data shown in Figure 5,
the qe of Co+2-hectorite rose at various temperatures
when the primary CV dye concentration was raised.
This is explained by the reality that free adsorption
locations are accessible at the s tart of the tes t and
the CV dye concentration was at its highes t.
The impact of temperature on the CV dye adsorption
equilibrium on the Co+2-hectorite surface is also
depicted in Figure 5. For a primary concentration of
0.100-0.200 g L-1, it can be seen that the qe declined
as the temperature rose, indicating an exothermic
process. Though the impact of temperature on the
adsorption equilibrium was negligible at the low
s tarting concentration of CV dye (0.100 g L-1), it
was s till present.
3.4. The inves tigation of intra-particle diusion
and lm diusion
the Weber-Morris kinetic model was used s as
Equation 2[29].
qt=Kidt0.5 + C (Eq.2)
where represents the removal capacity (mg g-1) at
time(t), Kid
rate cons tant (mg per g min0.5), and C represents a
cons tant whose value is proportionate to the limit
layer (mg g-1).
When the adsorption sys tem corresponds with the
versus t0.5 should have a s traight line with a slope of
Kid and an intercept of C, according to Equation 2.
Figure 6 shows a plot of the means of the replicated
experimental outcomes. There are two dis tinct
zones in Figure 6
linear sections are attributed to macro- and micro-
0.10 0.15 0.20
0.30
0.35
0.40
0.45
0.50
CV primary concentration (g.L-1)
qe (g.g-1)
293 K
303 K
313 K
Fig. 5.
Adsorption of CV Dye by Cobalt-Hectorite Composite Ahmed Jaber Ibrahim
10
phenomenon is linked to an extremely slow CV
Additionally, this promotes the sluggish quiet rate
of adsorbate movement from the liquid s tage to
the surface of the adsorbent. The mass transfer rate
end phases explains why the s traight line deviates
departure from the point of origin sugges ts that pore
[30].
Equation 3.
(Eq.3)
where r0 (m) represents the mean radius of the
adsorbent particles and t0.5 (min) the time needed to
The rate-limiting phase will be intra-particle
[31], if the
level is in the scope 10-15-10-18 m2 per S. According
to Table 1, which was used in this inves tigation,
the computed DP level varied from 1.65x10-14 to
2.47x10-14 m2 s-1 at various temperatures, implying
primary process limiting CV dye adsorption onto
Co+2-hectorite surface.
Fig. 6. Scatter plot of qt versus t0.5 for adsorption of CV dye on Co+2-hectorite composite
at s tudied temperatures
246810
0.20
0.25
0.30
0.35
0.40
t0.5
qt (g.g
-1
)
293 K
303 K
313 K
Table 1.
at the temperatures s tudied
Temperature (k) DP (m2 S-1) DF (m2 S-1) ro(m)
293 77.44 1.65 x 10-14 3.44 x 10-13
6.54 x 10-4
303 63.36 2.02 x 10-14 4.21 x 10-13
313 51.87 2.47 x 10-14 5.63 x 10-13
Anal. Methods Environ. Chem. J. 6 (1) (2023) 5-16
11
Equation 4
adsorption kinetics reactions.
(Eq.4)
Where CS is the concentration of adsorbate in the
solid phases, Cl is the concentration of adsorbate in
the liquid phase, and r0 and t0.5 share the identical
-5m)
[31]
value will fall between 10-10 and 10-12 m2 per second
DF were discovered to be in the arrange of 10-13
m2s-1 (Table 1)
reaction was not the lone phase in the adsorption
process that was rate-limiting. Intra-particle and
kinetic process. The kinetic reaction was governed
high at the beginning of the adsorption process. CV
+2-hectorite
when they were adsorbed on the surface of the
composite, and the adsorption reaction was what
controlled this.
3.5. Thermodynamics s tudy
The pseudo-second-order [32] model has been
inves tigated about kinetic modeling to determine
the adsorption mechanism (Equation 5).
(Eq.5)
Where qe is the equilibrium adsorption capability
(g.g-1), k2 is the pseudo-second-order adsorption
rate cons tant (g min g-1), and qt is the amount
of CV adsorbed at time t (g g-1). To determine
rate parameters, the s traight line plots of t/qt vs.
t for the pseudo-second-order models have also
been inves tigated (Fig. 7). Table 2 contains the
2, k, and qe at numerous
temperatures. The Arrhenius equation (Eq. 6) can
express the pseudo-second-order rate cons tants as
a temperature performance.
ln k = ln A - Ea/RT (Eq.6)
Figure 8 shows a visualization of the means of the
replicated experimental outcomes.
050 100
0
100
200
300
400
t (min)
t/qt
293 K
303 K
313 K
Fig. 7. The pseudo-second-order kinetics model for the adsorption of CV dye onto Co+2-hectorite composite
at s tudied temperatures
Adsorption of CV Dye by Cobalt-Hectorite Composite Ahmed Jaber Ibrahim
12
The Ea is calculated using Equation 6 (Table 2).
The size of the activation energy gives a clue as
to the primary kind of adsorption, either chemical
or physical. Physisorption processes typically
have activation energies between 5 and 40 kJ mol-
1, whereas greater activation energies (between
40 and 800 kJ.mol-1) point to chemisorption. The
dispersive interaction between the crys tal violet
and the Co+2-hectorite surface implies it. The Gibbs
o o), and
o), which are thermodynamic
characteris tics, have been calculated to assess
the viability and exothermic characteris tic of the
adsorption reaction. Equation 7
change in Gibbs free energy to the equilibrium
cons tant (k).
(Eq. 7)
The below formula shows how the s tandard free
energy change at cons tant temperature is also
correlated with enthalpy and entropy changes
(Eq. 8).
(Eq. 8)
Table 2. The Arrhenius activation energy (Ea) and pseudo-second-order kinetics parameter values
for the adsorption process at temperatures s tudied
Temperature (k) (kJ mol-1) r2qe (g g-1)k2
293
22.434
0.9996 0.335 2.087
303 0.9954 0.317 2.794
313 0.9965 0.302 3.587
0.0028 0.0029 0.0030 0.0031 0.0032 0.0033
-7.4
-7.2
-7.0
-6.8
-6.6
-6.4
-6.2
1/T
lnk2
Fig. 8. Arrhenius Scatter plots for the adsorption of CV dye onto Co+2-hectorite at s tudied temperatures
Anal. Methods Environ. Chem. J. 6 (1) (2023) 5-16
13
The slope and intercept of Scatter plots of lnK vs
o and
o (Fig. 9). Table 3 contains the obtained values.
Indicating the viability and spontaneity of the CV
adsorption reaction on the Co+2-hectorite surface,
o) levels were observed
to be decreasingly negative with temperature. It is
o) values
are negative, indicating the exothermic character
o)
value is less than 40 kJ.mol-1 shows that the crys tal
violet adsorption by the composite of Co+2-hectorite
is physisorption. The results from the current s tudy
[28] on
the adsorption reaction of congo red azo dye from
aqueous solution by ODA-hectorite and CTAB-
hectorite as adsorbent surfaces.
4. Conclusion
The results of this inves tigation demons trate
+2-hectorite composite as an
adsorbent surface for eliminating Crys tal Violet
(CV) dye from aqueous solutions. The elimination
of CV worked bes t at a pH of 4. The ideal
temperature and composite dose were 293oK and
0.5 g L-1, respectively. The experimental results
and the pseudo-second-order kinetic model were in
good agreement, as indicated by the s traight lines
were in charge of regulating the adsorption reaction.
The exothermic and spontaneous response of CV
adsorption on Co+2-hectorite composite is revealed
by evaluating the thermodynamic parameters. The
activation energy for adsorption, which fell within
the normal range for physisorption, was discovered
Table 3. Thermodynamic variables for the adsorption process
Temperature (k)Dis tribution coecient (k) ΔGO
(kJ mol-1)
ΔHO
(kJ mol-1)
ΔSO
(kJ mol-1)
293 4.654 -3.745
-31.546 -94.883
303 3.324 -2.926
313 2.067 -1.768
0.0028 0.0029 0.0030 0.0031 0.0032 0.0033
0.0
0.5
1.0
1.5
2.0
1/T
lnk
Fig. 9. Scatter plot of lnK vs. 1/T for CV dye adsorption onto Co+2-hectorite composite.
Adsorption of CV Dye by Cobalt-Hectorite Composite Ahmed Jaber Ibrahim
14
to be 22.434 kJ mol-1
the design of was tewater treatment facilities that
remove the dye.
5. Acknowledgements
The s tructural formula of the Physical Chemis try
Lab., Chemis try Department, College of Education
for Pure Science (Ibn-al Haitham), University of
Baghdad, supports this research.
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Anal. Methods Environ. Chem. J. 6 (1) (2023) 5-16
Anal. Methods Environ. Chem. J. 6 (1) (2023) 17-28
Research Article, Issue 1
Analytical Methods in Environmental Chemis try Journal
Journal home page: www.amecj.com/ir
AMECJ
Preparation of recycled poly s tyrene derivatives to remove
heavy metal ions from contaminated water
Hadi Salman Al-Lamia,*, Hussein Ali Al-Mosawia, and Nadhum Abdulnabi Awada
aDepartment of Chemi s try, College of Science, University of Basrah, Basrah, Iraq
ABSTRACT
In recent years, numerous researchers have concentrated on the
process of turning wa s te into usable materials. Poly s tyrene and its
due to their out s tanding ion exchange behavior toward various
toxic heavy metals in aqueous solutions. Therefore, this s tudy is
resins for the removal of Pb2+, Cd2+, and Fe3+ heavy metal ions from
their contaminated water samples based on the sulfonated single-
used poly s tyrene teacup wa s te (SPS), which was used to prepare
sulfonated poly s tyrene-g-acrylamide monomer (SPS-g-Acryl) and
sulfonated poly s tyrene-g-chitosan (SPS-g-Chit) using commercial
chitosan (DD=85%) originally extracted from shrimp cortex. The
concentrations of the selected heavy metal ions were measured
spectrometer (F-AAS). The analytical s tudies s tarted by exploring
Pb2+, Cd2+, and Fe3+ from their aqueous solutions. The obtained results
revealed that as the pH of the analyzed ion solution is increased, the
the inve s tigated ions, with SPS-g-Chit resin being the be s t in both
batching and column loading methods, and they could be compared
in the following order: SPS-g-Chit > SPS-g-Acryl > SPS, and it could
be reused after regeneration.
Keywords:
Sulfonated poly s tyrene,
Chitosan,
Grafting,
Acrylamide,
Heavy metals,
Flame atomic absorption spectrometer
ARTICLE INFO:
Received 7 Dec 2022
Revised form 17 Feb 2023
Accepted 4 Mar 2023
Available online 30 Mar 2023
*Corresponding Author: Hadi Salman Al-Lami
Email: hadi.abbas@uobasrah.edu.iq
https://doi.org/10.24200/amecj.v6.i01.223
------------------------
1. Introduction
Water is one of the mo s t crucial natural resources
for the survival of all living forms, food security,
economic development, and welfare. De-pollution
is a gift from Mother Nature to the globe that cannot
be replicated for many purposes and co s ts a lot to ship
[1]
water is in rivers and s treams. Despite the small
water is critical. The exponential development of
the population and increased indu s trialization have
led to an enormous rise in freshwater consumption
in recent decades. Heavy metal pollution of the
water environment threatens human health in mo s t
countries. It has increased recently along with
economic development and population growth,
primarily from mining, electroplating, and the
production and shipment of batteries. They damage
the body, including the skin, lungs, head, liver, and
kidneys, and cause tumors, birth defects, and other
conditions. Consequently, the quality of drinking
18
water is declining, and there is a need for better
water management and a method for preventing
water contamination and providing pure water [2].
This explains why, for the government and experts,
water contamination has become a s tudy subject
[3]. Both the water and its geographic and seasonal
supply, as well as their surface and groundwater
content, are very important for climate, economic
growth, and development. Because of increased
population growth, urbanization, expansion of
continue to be long-la s ting. If water is contaminated
in a city, all livelihoods and citizens are confronted
with drinking poisoned water because they have no
alternative. The damage to the human body (skin,
lungs, head, liver, and kidneys) causes tumors, birth
defects, and other conditions. We concluded that
the quality of drinking water is declining because
of increasing population, agricultural practices,
and indu s trialization, and there is a need for better
water management and a method for preventing
water contamination and providing pure water [2].
Due to the issues above, a sub s tantial focus has
less expensive, and long-la s ting wa s tewater
treatment sy s tems that do not add to environmental
s tress or pose a health risk to humans. In recent
years, extensive te s ting has been done to develop
technologies. Coagulation, membrane processes,
photocatalytic degradation are some methods
used to remove hazardous sub s tances from water.
However, as seen in Figure 1, various sources of
pollutants are in the water [4].
Many methods have been s tudied to treat heavy
physical, or biological. The mo s t promising
technologies to overcome these con s traints are
adsorption and ion exchange [5,6]. Given the
various indu s trial applications of large quantities
of ion exchange resins consumed after being used
conditions, they should be replaced by new
ones [7,8]. This prompted us to look for a less
expensive alternative method of providing it. In
Anal. Methods Environ. Chem. J. 6 (1) (2023) 17-28
Fig. 1. Some sources of pollutants in water [4]
19
previous work, sulfonation and grafting acrylamide
monomer and chitosan polymerization synthesized
two ion exchange resins from recycled poly s tyrene
single-used teacup wa s te [9]. They were used as
ion exchangers to determine the total hardness of
tap water. Both prepared resins were characterized
sulfonation and grafting processes on sulfonated
poly s tyrene, respectively. The grafting process
poly s tyrene resin in removing the hardness of tap
water. Poly s tyrene sulfonating reduces the volume
of solid wa s te and contributes to environmental
cleanliness.
The results of our inve s tigation into the technical
viability are presented in this research using
sulfonated poly s tyrene-g-acrylamide (SPS-Acryl).
Sulfonated poly s tyrene-g-chitosan (SPS-Chit)
derived from poly s tyrene single-use teacups after
sulfonation (SPS) for heavy metal removal in batch
experiments for checking their feasibility if applied
in practice are presented in this research. In addition
to reducing environmental degradation, recycling
this material as an ion exchange material also limits
the exploitation of natural resources.
2. Experimental
2.1. Materials and in s trument
Sulfuric acid (96%) was used as a sulfonating
acrylamide monomer were used as grafting
solution were purchased from Sigma-Aldrich.
Cadmium (II) nitrate, iron (III) chloride, and lead
(II) nitrate were purchased from Global Chemical
for heavy metal ions in pollutant samples. The
concentrations of the Pb2+, Cd2+, and Fe3+ heavy
absorption spectrometer (F-AAS, Varian AA240
FS, USA). The IKA magnetic s tirrer was purchased
Z671835, Germany). Digital pH meter purchased
2.2. Sulfonated poly s tyrene preparation (SPS)
The single-use poly s tyrene teacups were collected
and washed several times with tap and di s tilled
water before drying at room temperature. As
mentioned in the literature, sulfonated poly s tyrene
and SPS-g-acrylamide monomers were prepared
[9,10]
chopped wa s te teacups made of poly s tyrene were
added. 50 mL of 96% sulfuric acid was added,
and the mixture was continuously s tirred at room
approximately 4 hours at 60–65°C. Sulfonated
poly s tyrene was obtained through separation and
the reaction. Following a pH-neutralizing wash
with di s tilled water, it was dried at 60°C.
2.3. Synthesis of SPS-g-acrylamide resin (SPS-
g-Acryl)
SPS-g-acrylamide was prepared by grafting
acrylamide monomers onto the prepared SPS.
This was done by weighing 5.0 g of SPS resin and
a magnetic s tirrer. After that, 5 g of acrylamide
monomer was gradually added. The mixture was
s tirred at 40 °C for 3 hours to complete the grafting
process. The resin was then dried at 60 °C after
being rinsed with di s tilled water to remove any
non-grafted monomer [10,11]. Scheme 1 shows the
grafting reaction of the acrylamide monomer on the
sulfonated poly s tyrene.
2.4. Synthesis of SPS-g-chitosan resin (SPS-g-Chit)
was dissolved in 75 mL of 2% acetic acid, and
the solution was gradually added to 5 g of SPS.
The mixture was then heated for 3 hours at 60–65
times with deionized water and acetone after
allowing the liquid to cool to room temperature.
The white product was dried using a vacuum
desiccator [12,13]. The grafting reaction of the
chitosan on the sulfonated poly s tyrene is depicted
in Scheme 2.
Removal heavy metal by Cationic Polymeric Resins Hadi Salman Al-Lami et al
20
2.5. Preparation of heavy metal ion s tandard
solutions
of the element ions (Pb2+, Cd2+, and Fe3+) were
made by diluting 1000 mg L-1 s tokes solutions of
their salts to 50 mg L-1 with double-di s tilled water.
The salts of these elements used were FeCl3.6H2O,
Cd(NO3)2.4H2O, and Pb(NO3)2 [14]. All the
concentrations of heavy metal ion solutions were
measured before and after each experiment with a
Flame atomic absorption spectrometer (Sens, Japan).
2.6. Heavy metal ions removal in batch method
Batch-mode ion exchange experiments were
performed in beakers under con s tant time and
temperature conditions [15,16]
SPS-g-Acryl, and SPS-g-Chit resins was s tudied
changeability and bonding of heavy metal ions
(Pb2+, Cd2+, and Fe3+) to the resins.
Anal. Methods Environ. Chem. J. 6 (1) (2023) 17-28
Scheme 1. The chemical equation of grafting acrylamide onto SPS resin
Scheme 2. The grafting reaction of SPS resin with Chitosan
21
2.7. Study the eect of the pH on the resins by
batch method
One gram of each resin (SPS, SPS-g-Acryl, and SPS-
g-Chit) was treated with 25 ml of prepared aqueous
solutions of Pb2+, Cd2+, and Fe3+ in pH ranges of 2,
4, 6, and 8 [17,18]. Each bottle containing resins
was left at room temperature for an hour of shaking
(175 rpm min-1
collected in 50 mL of a pla s tic bottle.
2.8. Heavy metal ions removal by column
method
This experiment was carried out to inve s tigate the
practicability of using SPS, SPS-g-Acryl, and SPS-g-
Chit resins for heavy metal removal in a continuous
[19,20]. The water solution containing
a mixture of the three heavy metal ions (Pb2+, Cd2+,
and Fe3+) was continuously passed through a vertical
glass column with a height of 20 cm and a diameter of
rate of 1 mL min-1, an inlet heavy metal concentration
of 50 mg L-1, and 10 g of each prepared resin, as
shown in Scheme 3. All te s ts were run continuously
Finally, the concentrations of the Pb2+,
Cd2+, and Fe3+
atomic absorption spectrometer (F-AAS)
2.9. Regeneration of the loaded resins
For regeneration experiments, the be s t-loaded resin
with metal ion removal obtained at pH 8 from a
column method was chosen. One gram of the
loaded resin was treated for a half-hour in a column
mL of 0.1 N HNO3 before being washed directly
with 30 mL of deionized water and collected in
pla s tic bottle samples [21].
2.10. Examination of single and mixture heavy
metal ions by column method
A column with a 15 cm length and a 2.5-mm inner
diameter was chosen for the analytical s tudies. It
was then loaded with 1.0 g of SPS, SPS-g-Acryl, and
SPS-g-Chit resins, and 25 mL of metal ion solutions
(Pb2+, Cd2+, and Fe3+-1
with a 50 mg L-1 concentration at pH 8. A mixture of
the three ions (Pb2+, Cd2+, and Fe3+) was prepared in
25 ml with a 16.6 mg L-1, and the pH was adju s ted
to 8. The column was charged with 1 g of each
resin. The mixture of ion solutions was allowed to
min-1. The descending solution was collected in
portions for each component over an approximately
30-minute interval. The concentrations of the heavy
metals were measured with F-AAS.
Removal heavy metal by Cationic Polymeric Resins Hadi Salman Al-Lami et al
Scheme 3. The Schematic diagram for heavy metal ions removal by column method
22
3. Results and Discussion
The sulfonation reaction by sulfuric acid provides
sulfonic groups SO3H+ attached to the poly s tyrene
backbone chains, giving ion exchange capacity to
the poly s tyrene, which will be a center for the grafted
acrylic monomer and chitosan polymer material
to make up copolymers attached to the sulfonated
poly s tyrene [10-12]. One advantage of this approach
is the ability to precisely modify characteri s tics by
adju s ting the grafting or sulfonating conditions. The
activation of the backbone polymer, the grafting
of a monomer onto the produced polymer and the
subsequent functionalization of the grafted polymer
are all s teps in the graft copolymerization technique
of ion-exchange synthesis, yielding a product with
high chemical selectivity [22,23]. Because heavy
metals are typically produced by known sources,
removing them rather than releasing them into
the environment is preferable. The ion exchange
capacity of the made resins is determined by the
nature of the active groups, whose selectivity
varies depending on the heavy metal ions and
pH, the temperature and concentration employed,
the nature of those ions, and the number of active
dependent changeability. The pH was controlled
by dilute solutions of 0.01 N nitric acid and 0.01
N ammonia solution [10].
the changeability of the three heavy metal ions
(Pb2+, Cd2+, and Fe3+) with the active groups of the
Chit, was inve s tigated by changing the pH from 2
to 8 with an initial concentration of ion solutions of
50 mg L-1. Since metal cations attached to -SO3H in
functional groups were released and H+ ions were
added after the ion exchange reaction, the pH after
equilibrium was slightly reduced. Therefore, the
metal precipitation did not take place [24,25].
3.1. Eciency for heavy metal removal in the
batch method
3.1.1.Sulfonated poly s tyrene resin (SPS)
by measuring the concentration of the unabsorbed
the metal ions separately. The results obtained are
shown in Table 1 and Figure 2. It was found that
Pb2+
L-1
L-1)
at pH ) mg L-1
at pH 8 (11.7 mg L-1), and Fe3+ has the highe s t
) at all pH values.
prepared resins to work as ion exchange resins for
with a low initial concentration. Using sulfonated
poly s tyrene resin to separate Zn2+, Cu2+, and Cd2+
heavy metal ions from their solutions, Tran and his
coworkers came to the same conclusion [6].
Anal. Methods Environ. Chem. J. 6 (1) (2023) 17-28
Table 1.
-1)
pH Pb %Eciency Pb Fe %Eciency Fe Cd %Eciency Cd
250.00 0.00 0.41 99.18 12.77 74.46
440.76 18.48 0.64 98.72 13.51 73.00
646.34 7.32 0.72 98.56 13.11 73.80
816.73 66.54 0.70 98.60 11.70 77.00
23
Removal heavy metal by Cationic Polymeric Resins Hadi Salman Al-Lami et al
Fig. 3.
Fig. 2.
Table 2.-1)
pH Pb Eciency Pb% Fe Eciency Fe% Cd Eciency Cd%
247.03 5.94 45.96 8.10 17.18 65.64
440.23 19.54 34.25 31.50 14.87 70.30
645.80 8.40 2.60 95.00 14.16 71.70
812.53 75.00 0.52 99.00 9.47 81.70
24
3.1.2.Sulfonate poly s tyrene-g-acrylamide (SPS-
g-Acryl)
It is widely under s tood that mo s t polymers used
in water and wa s tewater treatment are acrylamide-
based. The basic acrylamide monomer may be
combined to provide polymers of varying iconicity.
properties based on their monomer characteri s tics
and how the copolymerization reaction takes place.
SPS-g-acrylamide for water treatment due to its high
cationic charge content. This polymer is primarily
nonionic at pH <4, but deprotonation to the cationic
form (SPS-g-Acryl) occurs at increasing pH (8)
[26]
4, 6, and 8, respectively, was s tudied for removing
three heavy metal ions by SPS-g-Acryl resin, and
the results are shown in Table 2 and Figure 3. It is
noted that the Pb2+
at pH 2 (47.03 mg L-1
pH 8 (12.53 mg L-1). On the other hand, for Cd2+,
-1)
and high (81.70%) at pH 8 (9.47 mg L-1). While for
Fe3+
(45.96 mg L-1
(0.52 mg L-1).
3.1.3.Sulfonated poly s tyrene-grafted-chitosan
(SPS-g-Chit)
Sulfonated poly s tyrene is a polymerized s tyrene
monomer by the sulfonation process. This process
produces a grafted polymer with improved properties
of this material and increases the crosslinking
process, which leads to the expansion of the
polymeric network [27.28]. When chitosan is grafted
onto this network, it will lead to better properties due
to the high functionality of the chitosan polymer, in
addition to relying on the active groups whose work
increases with increasing pH [29,30]
inve s tigated, and the results are shown in Table 3 and
Figure 4. Pb2+
pH 2 (27.31 mg L-1
at pH 8 (7.5 mg L-1). Whereas, Cd2+ and Fe3+ ions
slightly higher than 99% (Table 3).
3.2. Treatment of single heavy metal ions by
column method
Chromatography is a physical method of analysis
and separation that uses a s tationary phase with a
through and generally contains the sample. The
ion exchange column belongs to the (solid-liquid)
chromatography category. When a mixture of two
or more ions runs through the column in quantities
exchange capacity. It was completely absorbed by
the resin and then separated from its con s tituents
to elute out the weakly
on. Elution is the process of separating these ions
Anal. Methods Environ. Chem. J. 6 (1) (2023) 17-28
Table 3.
-1)
pH Pb %Eciency Pb Fe %Eciency Fe Cd %Eciency Cd
2 27.31 45.38 0.02 99.96 2.53 95.00
415.96 68.08 0.003 99.99 0.37 99.26
615.38 69.24 0.01 99.98 0.35 99.30
87.50 85.00 0.01 99.98 0.35 99.30
25
until they are separated quantitatively. The Flame
atomic absorption spectrometer method is. The
optimum pH of 8 is favorable due to the partial
hydrolysis of metal ions. As previously s tated, the
method revealed that the SPS-g-Chit resin was the
metal ions s tudied. Therefore, it was used for the
removal of single metal ions by the column method.
The pH 8 for the SPS-g-Chit resin is an exchange
2+ and 99.76% for
Cd2+3+ is
reduced to 78.36%.
3.3. Treatment of heavy metal ions mixture
Table 4
Pb3+ and Cd2+. The grafting process of SPS with
chitosan added another active group to increase the
due to its high content of amine groups in a rational
manner, where the exchange mechanism depends
on each of the protons of these amine groups or
metal ions, as well as cross-linking, which tends to
enlarge the polymeric network, resulting in better
[31]
when the active group concentration increased
[8,10,11]. The resin presented no exchange
3+. This may be due to the ferric
exchange.
Removal heavy metal by Cationic Polymeric Resins Hadi Salman Al-Lami et al
Fig. 4.
Table 4.
at other times (Unit: mg L-1)
Time (min) Pb %Eciency Pb Fe %Eciency Fe Cd %Eciency Cd
30 0.417 98.00 16.6 0 0.733 95.00
60 0 100 16.6 0 0.05 99.60
90 0 100 16.6 0 0.037 99.70
120 0 100 16.6 0 0.022 99.80
150 0 100 16.6 0 0.01 99.90
26
3.4. Regeneration of Loaded Resins
The SPS-g-Chit resin was selected for the ion
exchange regeneration resin experiment because
it has the greate s t changeability toward the three
metal ions used in this s tudy (Pb2+, Cd2+, and Fe3+).
Table 5 shows the outcomes that were attained. It
demon s trates that after around an hour of treatment,
the lead and cadmium ions showed the maximum
capacity and the ability to use the SPS-g-Chit resin
as an ion exchanger in indu s trial processes.
4. Conclusion
Recycling single-use teacups and shrimp cortex
chitosan as ion exchange materials limits the
extraction of natural resources and reduces
environmental pollution. Ion exchange resins
can be produced and used with less expense
poly s tyrene wa s te that has been sulfonated, and its
grafted acrylamide monomer and chitosan polymer
derivatives have the potential to have a higher ion
exchange capacity. H+s competing with metal ions
at lower pH levels were discovered to be responsible
for better results in treating metal ions at higher pH
levels of 8. In terms of their active groups, chemical
s tructures, and s tereotypical s tructures, the
produced resins varied in performance depending
on the metal ion and resin type used. In conclusion,
as the raw material used to create ion exchange
resin is made from wa s te materials, sulfonated
poly s tyrene and using chitosan originally extracted
from shrimp cortex are thought to be technically
and environmentally possible to remove heavy
metals at a reasonable co s t.
5. Acknowledgements
The authors thank the Department of Chemi s try,
College of Science, University of Basrah, Basrah,
Iraq for supporting this work.
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Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
Research Article, Issue 1
Analytical Methods in Environmental Chemis try Journal
Journal home page: www.amecj.com/ir
AMECJ
arsenic: arsenate and arsenite
Madhawa Pradeepa Nawarathne a, Ruvini Lakmali Weerensinghe b, and Chathuranga Dharmarathne c,*
a Pograduate Initute of Science, University of Peradeniya, Sri Lanka
b Faculty of Graduate Studies, University of Sri Jayewardenepura, Sri Lanka
c Department of Biological Sciences, Macquarie University, Sydney, NSW, Auralia
ABSTRACT
s tates in nature, including arsenate and arsenite, as common inorganic
health and the environment. Therefore, the detection of arsenic is
critical. Exploring new approaches with low detection ranges and
high sensitivity is crucial. This review paper consi s ts of optical
-1). Initially proposed colorimetric
approaches such as the Gutzeit and molybdenum blue method can
easily to use. However, the production of toxic sub s tances limits their
nanoparticle-
Fluorometric methods also have noticeable attention to arsenic
detection. Fluorescent approaches reported in this paper are based
on semiconductor nanomaterials, other nanomaterials, and their
activity on enzyme activity can be used to detect arsenic through
This review highlighted the
advantages, disadvantages, comparisons, and uses of colorimetric
Keywords:
Arsenate,
Arsenite,
Colorimetric,
Fluorometric,
Nanoparticles
ARTICLE INFO:
Received 6 Dec 2022
Revised form 24 Jan 2023
Accepted 27 Feb 2023
Available online 28 Mar 2023
*Corresponding Author: Chathuranga Dharmarathne
Email: chathurangadharma@gmail.com
https://doi.org/10.24200/amecj.v6.i01.224
------------------------
1. Introduction
Arsenic is the 20th mo s t abundant and widely
as a metal and as a metalloid [1]. Arsenic occurs in
several chemical oxidation s tates in nature, such as
As (V), As (III), As (0), and As (-III) [2]. Also, it can
exi s t in both organic and inorganic forms. Inorganic
arsenic exi s ts as arsenate [As (V)] and arsenite [As
(III)], while organic forms are monomethyl arsenic
acid (MMA), dimethyl arsenic acid (DMA), dithiol
arsenate (DTA), etc. [3]. Even the trace concentration
of arsenic shows higher toxicity than mo s t other heavy
metals. Both inorganic forms of arsenate and arsenite
are hazardous and toxic. Among them, arsenite is
considered the mo s t toxic form [4]. Contaminated
groundwater, wa s tewater, and drinking water are the
main sources of arsenic that enter the environment
through indu s trial operations, agricultural activities,
etc. [4-6]. Exposure to arsenic in the long term can
cause
cancers, skin lesions, neurotoxicity, cardiovascular
disease, diabetes, etc. [5]. Therefore, the detection
and removal of arsenic are critical to human health.
The World Health Organization (WHO) and US
Environmental Protection Agency (USEPA) have
s tipulated some guidelines with a 0.01 mg L-1
30 Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
L-1 permissible limit of arsenic for drinking water
[6]. Therefore, a series of techniques are developed
to monitor the arsenic concentration. Numerous
laboratory techniques, including Atomic Absorption
Spectroscopy (AAS) [7], Atomic Fluorescence
Spectroscopy (AFS) [8], Inductively Coupled Plasma
Atomic Emission Spectroscopy (ICP-AES) [9],
Inductively Coupled Plasma Mass Spectroscopy (ICP-
MS) [9,10], Graphite Furnace Atomic Absorption
(GFAA), Hydride Generation Atomic Adsorption
(HGAA) and Neutron Activation analysis [11] are
some of the methods which currently being used.
They provide accurate detection of arsenic even at
trace concentrations. High expensive in s trumentation,
the requirement for sophi s ticated laboratory setup,
producing highly toxic chemicals during the process
are the drawbacks of these methods. Also, the inability
limitations [12]. Therefore, various chromatographic
methods, electrochemical methods [13, 14], and
optical methods are developed to overcome these
limitations [15]. Among them, optical methods are
essential due to their high sensitivity, selectivity,
simplicity in operation, fa s t response, co s t-
optical methods are considered as promising
techniques for arsenic detection [14,16,17,18]. This
review presents current optical methods including
arsenate and arsenite.
2. Experimental
analyse arsenic (arsenite and arsenate) in various
matrixes.
2.1. Colorimetric detection of arsenite and
arsenate
2.1.1.Advancement of colorimetric methods
and detection based on s tructurally modied
molecules.
Colorimetric methods are based on the Gutzeit
reaction, which detects arsenic quantitatively. This
method relies on the reaction of arsenic gas with
hydrogen ions to form a yellow s tain on mercuric
chloride paper in the presence of reducing
agents. However, certain limitations are followed
when analyzing the groundwater samples, and it
generates highly toxic arsine gas and byproducts
of mercuric compounds. Therefore, protective
equipment is required for the analysis procedure
[19,20]. In addition, traditional methods such
as molybdenum blue [21-25], ethyl violet, and
gallocyanin can be used for sensing arsenic, yet,
the interferences intervened with the results, and
low sensitivity hinders its applicant the resulting
[26]. Therefore, scienti s ts have continuously
tried to remove the generation of toxic chemicals
and interferences in the detection. Some arsenic
detection methods are involved with s tructurally
based rhodamine monomers (Nor-Rh) and their
polymer form (PNor-Rh) are used for very
selective detection of arsenite colorimetrically
in the presence of potassium
iodate and hydrochloric acid. They used an open
form of rhodamine which is pink and highly
luminescent. When arsenite was present in the
sample, the reaction led to the oxidation of As (III)
to As (V) by iodate reduction reaction to iodine.
As a result, the solution changed its pink color to
brown colorimetrically and observed green color
concentration, the intensity of the brown color
increased. As a result, they developed a simple
and easy practical device based on a polymer-
synthesized a thiol-based norbornene monomer
(Nor-Th) and its polymer form (PNor-Th) for
arsenite removal from water [27]. Aptamers are
single- s trand RNA or DNA oligonucleotides that
can bind with target ions or molecules. Recently
aptamers are used as a recognition element for
developing arsenic detection methods [28-33].
For example, Zhou and colleagues [33] proposed
a method for colorimetric detection of arsenite
based on regulating hemin peroxidase catalytic
activity using arsenic binding aptamers. They used
31
Review: Colorimetric and Fluorometric Detection of Arsenic Madhawa Pradeepa Nawarathne et al
an arsenic-binding DNA aptamer called Ars-3.
Hemin acts as a cataly s t that can catalyze many
oxidation reactions. But the catalytic activity
of hemin is very slow in the aqueous medium.
Catalytic activity can be improved by hemin
binding onto the surface of nanosheets or covered
with guanine-rich oligonucleotide by forming an
active form of the G-quadruplex s tructure. Arsenic
binding aptamers can inhibit hemin catalytic
activity temporarily. In the presence of arsenite,
As (III) binds to Ars-3 and forms an aptamer-As
(III) complex. Therefore, an oxidation reaction
occurs in TMB molecules to generate yellow
diamine products. While the absence of arsenite,
Ars-3 aptamers complex with pyrrole rings of
hemin. As a result, the catalytic activity of hemin
is decreased and generates blue products of cation
radicals. Therefore, this method is susceptible
and selective for arsenite. Figure 1 shows the
schematic description of the As (III) detection by
arsenic binding aptamers.
Traditional methods are easy to perform and
inexpensive, yet re s trained by the generation of
due to complex redox reactions and complex
separations. Therefore, other anions and cations
can interfere with the analysis. The same assay
analyte. Therefore, the involvement of other ions
can be negligible. These assays/polymer-coated
[33,34].
2.1.2. Usage of metal nanoparticles
Researchers enhanced the involvement of novel
methods with non-toxic product generation.
Recently, they have used metal nanoparticles
including arsenic, to obtain a visual color change in
the analysis [35]. It is based on the excellent optical
properties of nanoparticles, such as high sensitivity,
regions. Gold nanoparticles [36-40] and silver
nanoparticles [41-49] are mainly used for analysis
procedures. Color change occurs due to the
aggregation of nanoparticles with the target analyte
and size-dependent Surface Plasmon resonance
(SPR) properties of nanoparticles [48,49].
Fig. 1. Schematic description of the colorimetric detection of As (III) based on the inhibition
of hemin peroxidase activity by arsenic-binding aptamers [33]
32
2.1.2.1. Detection based on the modied silver
nanoparticles.
Recently, nanotechnology has been commonly
used in optical detection due to the uniqueness of
its chemical, biological, and physical properties.
[50].
Thiol-based ligands are commonly used because
they have an excellent capacity to bind with
arsenic [51,52]. As an example, a multiligands
assay was synthesized for the detection of arsenite
based on silver nanoparticles (AgNPs) which were
(GSH), dithiothreitol (DTT) and asparagine
(Asn). Thiol-based ligands bind to the surface of
AgNPs, which can va s tly enhance the selectivity
toward the arsenite. Aggregation occurs due to
the formation of As-O and As-S linkages between
GSH/DTT/Asn-AgNPs and As (III). Color change
occurs from yellow to light pink and purple with
increased arsenite concentration. Figure 2 shows
the GSH/DTT/Asn-AgNPs complex formation
and aggregation with arsenite. This method is
promising for the colorimetric detection of arsenite
with high sensitivity, low co s t, and facility for rapid
onsite detection [52].
In addition, arsenic adsorbents such as iron (III)
oxide are widely used for detection procedures.
Siangproh and his group [53,54] developed
silver nanoplates (AgNPls) to detect arsenite and
with the detection. Therefore, they synthesized
and applied ferrihydrite-coated silica gel (SiO2-
arsenic (arsenate and arsenite). SiO2-Fh has a high
When arsenic adsorbs onto the SiO2-
Fh, the dark blue color of AgNPls was changed to
purple, pink, orange, and yellow respective to the
concentrations of arsenic present in the sample.
Both arsenate and arsenite show similar behavior
on the color changes of AgNPls. Therefore, this
method is suitable for the determination of total
inorganic arsenic.
2.1.2.2. Detection based on the modied gold
nanoparticles
Gold nanoparticles (AuNPs) have numerous
applications in the optical detection of heavy
metals, including arsenate, arsenite, and aromatic
compounds [54,55]. Mo s t of the AuNPs based
ligands. Priyadarshani et al. [55] synthesized gold
nanorod (GNR) based sensor GNR-PEG-DMSA. It
rapidly detecting arsenite and arsenate. GNR-PEG-
DMSA sensor is synthesized through conjugation
of GNR with poly (ethylene glycol) methyl ether
thiol (mPEG-SH) followed by the addition of
meso-2,3-dimercaptosuccinic acid. CTAB is
Fig. 2. Illus tration of synthesis of GSH/DTT/Asn-AgNPs used as a colorimetric probe
for As (III) detection [52]
Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
33
used as a surfactant which is capped with GNR,
and the s tability of the sensor is maintained by
PEG. DMSA is a ligand that covalently binds to
arsenite and arsenate bind with free –SH groups of
DMSA, which are present on the surface of GNR.
Aggregation initiates with the formation of As-
thiolate complexes between nanorods. The Colour
of the solution changes from dark bluish-purple to
almo s t colorless. This sensor has a regenerating
ability using s trong chelating agents like EDTA.
Also, it can be used to quantitatively determine the
total arsenic in a sample using a small amount of
sensor materials. Figure 3 shows the synthesis of
the GNR-PEG-DMSA sensor and its interactions
with As (III) and As (V).
The same group proposed Europium functionalized
single gold nanoparticle-based new sensor
GNP-MMT@Eu for colorimetric detection
of trace concentration of both As (III) and As
(V). Nanosensor is synthesized by chemical
conjugation of GNPs with 2-mercapto-4-methyl-5-
europium chloride [Eu (III)]. Aggregation occurs in
the presence of arsenite or arsenate by binding to
the surface of GNP-MMT@Eu. It occurs through
coordinated Eu-OH groups consi s ting on the surface
of GNP-MMT@Eu via electro s tatic attraction and
covalent-type interactions. Afterward, it forms
the GNP-MMT@Eu-As (III)/As (V) complex.
Arsenate shows rapid and more sensitive color
changes for the nanosensor than the arsenite. The
initial color of the sensor gradually changes from
red to blue. Sensor-based paper s trips can detect
importantly, the sensors can be regenerated. Here,
Figure 4 shows the synthesis of gold nanosensors
and its interaction with As (III) and As (V) [56].
Zhang and co-workers [57] proposed an arsenate
arsenate on acid phosphatase (ACP) bioactivity
using citrate-capped AuNPs as the optical reporter
[57-59]
(AMP) as the sub s trate and prevented AuNPs from
aggregation. The activity of ACP hydrolyses the
charged nucleotide into the uncharged nucleoside.
The presence of ACP dephosphorylation of AMP
to adenosine and resulting adenosine leads to the
aggregation by nucleoside binds to AuNPs through
metal-ligand interaction by replacing weakly bound
citrate. As a result, the color change occurred from
red to purple to blue. But in the presence of As (V),
the color of the solution reversed from blue through
purple to the initial red color due to the inhibitory
Fig. 3.
with As (III) and As (V) [55]
Review: Colorimetric and Fluorometric Detection of Arsenic Madhawa Pradeepa Nawarathne et al
34
AMP in the enzymatic reaction. The color change
occurred respectively with the As (V) concentration
increment. This sensor has a remarkable sensitivity
towards As (V). However, there are some limitations
in this assay. Their visible detection limit is a bit
higher than the s tandard limit of WHO, and certain
concentrations of Cu2+, F-, and H2PO4
- can interfere
with arsenate detection. Figure 5 shows the catalytic
activity of ACP on AMP with As (V) and without
As (V) and color changes in AuNPs.
developed to rapidly detect arsenite in low
concentrations with a low detection range. The
nanoparticles (Au-TA-TG), that can rapidly interact
with arsenite to produce a visible dark bluish-black
precipitate at the interfacial zone. Figure 6 depicts
gold nanoparticle solution and arsenic sample
separately. Due to the capillary action of the “Y”-
When Au-TA-TG solution meets arsenite ions, a
bluish-black precipitate can be observed quickly.
Thioctic acid (TA) and thioguanine (TG) are used to
to bind and interact with arsenite. The color change
occurs due to the aggregation of gold nanoparticles
suitable for environmental analysis [60].
Tetradecyl (tri hexyl) phosphonium chloride has a
s trong interaction with arsenite. This ionic liquid is
Fig. 4. Schematic of the synthesis of the gold nanosensor, GNP-MMT@Eu,
and its aggregation with arsenate and arsenite [56]
Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
35
sensitive visual observations for arsenite detection,
with the probe color changing from red to blue in the
presence of arsenite. The total amount of inorganic
arsenic can be determined using this probe. The
low co s t and high tolerance to common ions make
Figure 7 shows
the behavior of the probe in the presence of arsenite
and arsenate [61].
Fig. 5. Illus tration of bioactivity of ACP on AMP with arsenate and without arsenate
and color changes in AuNPs with an increment of arsenate concentration [57]
Fig. 6.
[60]
Review: Colorimetric and Fluorometric Detection of Arsenic Madhawa Pradeepa Nawarathne et al
36
many advantages including simple, low co s t,
less time-consuming, nontoxic, and ease of data
increase selectivity and sensitivity. Moreover, some
sensors can regenerate and are reusable. However,
there are some disadvantages such as complicated
preparation processes, and long reaction times.
maintenance is essential [55-61].
2.1.2.3. Detection based on unmodied gold
nanoparticles
Gold nanoparticle-based colorimetric assays
synthesized through expensive processes.
Therefore, scienti s ts were encouraged to develop
[62,63]. For example, a simpler
adsorption properties on AuNPs between random
coil G-/T-rich ssDNA and folded DNA bound
to arsenite was synthesized. While arsenite can
easily attach to the G-/T-rich ssDNA via hydrogen
AuNPs that prevent the salt-induced aggregation
by enhancing the electro s tatic repulsion between
ssDNA-adsorbed AuNPs and maintain the s tability
of AuNPs. Therefore, adding enough salt leads to
changes from red to blue resulting in ssDNA
becoming compact and folded DNA. Figure 8
represents the colorimetric s trategy of arsenite
detection. Visual inspection can be used for semi-
quantitative detection of arsenite, while UV/Vis
absorbance spectroscopy technique can be used for
quantitative detection [63].
Peptide ligands are promising materials for desired
target analytes, including metals, biomolecules,
and drugs [64,65]. Yang et al. [65]
As (III)-binding heptapeptide sequences of T-Q-S-
Y-K-H-G through phage display peptide library
techniques using a biopanning process. The
sensing sy s tem contains a unique peptide sequence
the sensing probe. Due to slow aggregation and the
prerequisite of high concentration of heptapeptide,
cy s teine residue (C terminal) is conjugated to the
end of the heptapeptide sequence resulting in an
octapeptide sequence of T-Q-S-Y-K-H-G-C which
In the absence of As (III), the octapeptide can
AuNPs changes from wine red to blue. Nitrogen-
containing functional groups (-NH, -N=), –OH
groups, and sulfur-containing groups (-SH), which
are present in the peptide sequence, can bind
Fig. 7. Illu s tration of gold nanoparticle probe and its behavior with the presence of arsenic [61]
Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
37
with As (III) via s trong hydrogen bonds. With As
(III) presence, octapeptide binds to As (III) and
remains red. UV/Vis spectroscopic technique can
determine As (III) concentration. As (V) and other
(III) detection. Therefore, this method is unique
for the determination of arsenite. Operation is
easier and more convenient than other complicated
can be used for naked-eye detection without
complicated in s trumentation and much knowledge.
Therefore, they are more economical, convenient,
simple, sensitive, selective, reliable, and co s t-
slightly interfere with the detection [62-65]
2.1.2.4. Detection based on arsenate adsorption
on nanozymes
Some nanomaterials, such as metal oxide
nanoparticles, noble metal nanoparticles, carbon-
based nanomaterials, and two-dimensional
nanomaterials, have a natural enzyme-mimicking
activity called nanozymes. Among them, two-
dimensional nanomaterials show excellent
enzyme-mimicking activity. Therefore, researchers
are intere s ted in using nanozymes due to their ease
of mass production with low co s t, robu s tness, high
s tability, etc. For example, cobalt oxyhydroxide
used in catalysis material for dual-mode assay
of arsenate detection, rely on its peroxidase-like
colorimetric detection of arsenate. This excellent
(ABTS) into its green color oxidized product
(ABTSOX). The presence of arsenate, As (V),
adsorbs onto the CoOOH surface, and interaction
through electro s tatic and covalent interaction
(As-O) to attenuate the peroxidase-like activity of
CoOOH. As a result, catalytic activity decreased,
and the green color solution changed to very pale
green or close to colorless. Sensitivity can increase
The assay is highly selective for arsenate and can
detect total inorganic arsenic. The advantages are
that it can be used for handy, onsite, sensitive,
Fig. 8. AuNPs-based colorimetric s trategy for arsenite detection [63]
Review of Colorimetric and Fluorometric Detection of Arsenic Madhawa Pradeepa Nawarathne et al
38
to detect trace As (V) with obviously improved
sensitivity and detection limit. Figure 9 shows the
dual-mode assay for arsenate detection based on
[66].
2.1.2.5. Detection of dual heavy metal ions
together, including arsenite
Researchers developed simple chemical sensors
to detect toxic heavy metals, including arsenic,
on humans and the environment [67-69]. A simple
designed for the naked eye detection of Hg2+ and
As3+ using a simple s tep reaction. They used Isatin
with 3,3-dihydroxybenzidine to obtain binding
sites in the chemosensors that bind mercuric ions
and arsenite. Three chemosensors, CS1, CS2, and
The color change of CS1 and CS3 changes from
orange to colorless in the presence of Hg2+ and
from orange to aqua-blue for As3+. CS1 shows
2+ and As3+. However, it
other cations. CS2 changes its color from yellow
to pink with the addition of Hg2+. Figure 10 shows
the binding mechanism of CS1, CS2, and CS3 with
Hg2+ and AsO2
-. These sensors can monitor the
mercury and arsenic level in the environment [68].
Neety Yadav and colleagues [69] developed a single
chemical sensor capable of detecting both arsenite
and cyanide ions, with detection limits in the micro-
and nano-range. The probe was synthesized by the
reaction of thiosemicarbazide dissolved in ethanol
with 2-hydroxy-1-naphthaldehyde (Fig. 11). This
probe has two acidic protons that are deprotonated
through the interaction of arsenite and cyanide
ions, which are indicated by the color change that
occurs in this sensor. In the presence of arsenite and
cyanide ions, the color of the probe changes from
light yellow to dark yellow due to deprotonation
and s trong hydrogen bonding between the probe
Fig. 9.
for arsenate detection (A) and illu s tration of electrochemical assay of CoOOH
[66]
Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
39
using the same probe, utilizing the increase in
hydrogen bonding. The probe has the advantage of
detecting the high toxicity of two anions, simply
When considering the above methods, mo s t are
shown low detection limits. Also, sensitivity and
selectivity towards arsenate, arsenite or both forms
are very high. It is especially high in nanomaterials
compared to traditional methods. Table 1 shows the
overview of above mentioned colorimetric methods,
limit of WHO. Many ways discussed in the text are
used for the detection of As (III) due to the excellent
oxidation property of As (III) which oxidizes into As
(V). Therefore, total inorganic arsenic concentration
can be detected using these methods.
Fig. 10. Schematic representation of the binding mechanism of CS1, CS2 and CS3 with Hg2+ and AsO2
- [68]
Review of Colorimetric and Fluorometric Detection of Arsenic Madhawa Pradeepa Nawarathne et al
40
2.2. Fluorometric determination of arsenite
Fluorometric methods have tremendous attention
for detecting arsenic due to their simplicity,
less expensiveness, ease of operation, non-
de s tructive, and fa s t response with low detection
limits. Therefore, it is a promising technique for
the detection of arsenic and it is very useful for
environmental analysis [70-73].
2.2.1.Detection based on arsenite interacts with
thiolated nano s tructures.
Recently, quantum dots (QDs) have been used as one
of the promising materials to detect arsenic due to
their low toxicity and unique optical properties. QDs
properties include high quantum yield, size-tunable
narrow symmetrical emission peaks. Also, the
preparation of QDs is very simple, s traightforward,
and does not require toxic precursors and organic
solvents [73-77]. For example, environmentally
friendly dithiothreitol (DTT) functionalized
water-soluble carbon quantum dots (CQDs) were
synthesized by microwave pyrolysis of citric
Fig. 11. Diagram of the synthesis of the probe for the detect arsenite and cyanide ions [69]
Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
Table 1.
of molecules and usage of metal nanoparticles.
Materials Arsenic LR
(μg L-1)
LOD
(μg L-1)References
Aptamer/ Hemin-H2O2 sy s tem As (III) - 6.00 [33]
GSH/DTT/Asn- AgNPs As (III) 0.4-20 0.36 [52]
AgNPls/SiO2-Fh As (III) & As (V) 500-30000 0.50 [54]
GNR-PEG-DMSA As (III) & As (V) 0.001-10 1.00 [55]
GNP-MMT@Eu As (III) & As (V) 1-1000 1.00 [56]
AuNPs/ACP/AMP As (V) 7.5-7520 7.50 [57]
AuNPs/G-T- rich DNA As (III) 5-2000 2.00 [63]
Heptapeptide/ AuNPs As (III) - 4.00 [65]
As (V) 4-500 3.72 [66]
LOD: Detection limit
LR: Linear range
41
turn-on detection of arsenite. DTT exhibits on the
surface of CQDs using S-S bonds to impart –SH
functionalities on their surface formed DTT-CQDs
complex. In the presence of As (III), arsenite
binds with the sulfur group of CQDs through the
–SH group of DDT ligand to form a s table As
(III)-DDT-CQDs complex, and as a result, CQDs
Figure 12 shows the
synthesis of functionalized CQDs and arsenite
binding processes on CQDs. Compared with metal-
based semiconductor QDs such as CdS-QDs, ZnS-
QDs, and CdTe-QDs, the mentioned approach
is useful for real-world applications, including
environmental analysis. Because metal-based
human health due to their elemental composition
and toxicity [77]
brightness with limited quantitative capability.
In s tead of the brightness changes, the detection
of color variation is the mo s t important factor for
These analyses are low-co s t, easy to operate,
and portable. For example, a color multiplexing-
dosage-sensitive detection of As (III) with clear
probes, which can achieve a wide color variation
from red to cyan. They have synthesized cyan CDs
and red CdTe QDs-based cyan and red probes by
hydrothermal and classical methods, respectively.
ligands, enhancing the aqueous solubility and
surface of QDs. As-S bonds are formed byAs (III)
addition and trigger the aggregation of GSH/DTT-
from red to cyan. A wide range of color variations
Fig. 12. Schematic representation of the synthesized DTT functionalized CQDs
for detection of arsenite and binding mechanism of arsenite [77]
Review of Colorimetric and Fluorometric Detection of Arsenic Madhawa Pradeepa Nawarathne et al
42
can be observed with As (III) concentration.
They realized dosage-sensitive visualization of
arsenite detection by the ink of sensory solution
printed on the te s t paper. Figure 13 represents the
various color variations from red to cyan for
arsenite detection [78].
Nanoclu s ters containing fewer atoms show a high
quantum yield and can be used to sense toxic metal
ions. Gold nanoclu s ters and silver nanoclu s ters are
approaches. Quantum yield can be increased by
adding capping ligands such as thiol-based and
dipeptide ligands. Compared with other capping
materials such as glutathione, dipeptide ligand-
(L-cy s teinyl-Lc s teine) capped water-soluble
synthesized using a core etching pathway through
a “Top-down” mechanism for the detection
of arsenite (Fig. 14). Synthesized dicy s teine
enhances gradually. The reason may be due to
the positive charge of As (III) interacting with the
negative charge thiolated gold clu s ter and due to
electron poor As (III) ions. Meanwhile, in the
presence of arsenite, the radiative decay rate of
can detect As (III) at low concentrations with high
reused by adding succinic acid to chelates As (III)
through complexation [79].
Selectively sensitive silver-doped hollow CdS/
ZnS bi-layer nanoparticles (Ag-h-CdS/ZnS) are
detect arsenite. AgBr nanoparticles were used as the
core to synthesize Ag-h-CdS/ZnS nanoparticles.
Also, L-cy s teine is used to functionalize the
nanoparticles. In the presence of arsenite, cy s teine
Fig. 13. Fluorescent colorimetry te s t paper for obtaining various color variations
from red to cyan using CDs and QDs dual probe [78].
Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
43
quenching of the nanoparticles due to changes
in the electronic s tructure and accelerating the
non-radioactive nature of excitons. This sensor
has many advantages, including ease of use,
[80]. A cy s teine-functionalized tetraphenyl
was developed for the highly selective detection
of arsenite with a low detection limit. Free-SH
groups in cy s teine can bind with arsenite through
As-S bonds, forming As (Cy s tPE)3 s tructure. TPE
present in this complex promotes the formation
induced emission feature (AIE), thus creating a
complex. Figure 15 depicts the formation of As
(Cy s tPE)3
[81].
There are many biological and chemical sensors
developed for arsenite detection due to their redox
properties and s trong thiophilicity. The usage
of thiol ligands can obtain many advantages,
including high sensitivity and selectivity due to the
presence of a va s t number of sulphur that utilizes
As-S bonds with arsenic in aqueous solutions.
Also, CQDs show many advantages, including
superior chemical s tability, high aqueous solubility,
tuneable surface functionalities, and resi s tance to
photobleaching. However, complicated thiolated
applications [75-81].
Fig. 14. Illu s tration of synthetic route for the formation of gold clu s ters
through a top-down mechanism [79]
Fig. 15. Illu s tration of Asv(Cy s tPE)3[81]
Review of Colorimetric and Fluorometric Detection of Arsenic Madhawa Pradeepa Nawarathne et al
44
2.2.2.Detection based on arsenite interacts with
biologically functionalized nanomaterials.
Nano s tructure-based sensors provide many
advantages including rapid and sensitive responses
to detecting arsenic in cell living. For example,
Mesoporous Silica Nanoparticles (MSNs) are
considered a promising material for arsenic
detection due to their high inner surface area and
[82-86].
Therefore, MSNs can be functionalized by capping
materials which enhance the detection capability.
Aptamers are DNA sequences and act as capping
for determining arsenite [83]. Oroval et al. [86]
fabricated an arsenite sensing sy s tem using aptamer-
capped MSNs. They used MCM-41 mesoporous
silica nanoparticles and pores of MCM-41 inorganic
support were loaded with rhodamine B. Rhodomine
B was capped by Arsenite aptamer (Ars-3) in
MSNs pores. As (III) has a high potential to bind
with aptamer and then displace it from the MSN
be used for quantitative detection of As (III). This
analysis due to its simplicity. Figure 16 shows the
functionalized Ars-3 aptamer in the sensory sy s tem
for arsenite detection.
The dye-labeled G/T rich single- s trand DNA-
wrapped single-wall carbon nanotubes (SWCNT)-
arsenite quantitatively at the femtogram level. Here,
was used to label the ssDNA, and that s tructure
is the wrapping material of SWCNT. Arsenite can
bind with the G/T bases of ssDNA in living cells,
SWCNTs. As a result, ssDNA can be dissociated
Fig. 16.
of arsenite detection [86]
Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
45
from the surface of SWCNTs and condensed in
the live cells. The condensed s tructure of ssDNA
facilitates the HEX to interact with G/T bases bound
quenching of HEX dye. Figure 17 illu s trates the
nanoprobe interaction with arsenite in the lysosome
of a living cell [87].
2.2.3.Detection based on arsenite interacts with
chemosensor.
was fabricated for “turn on” detection of arsenite
through the intermolecular hydrogen bonding
process. Arsenite selective HL probe is synthesized
by condensation of 2,6-diformyl-p-cresol with
4-aminoantipyrine. The absence of arsenite ions
higher than in the absence of arsenite. Fluorescence
of arsenite. When arsenite ions are present in the
sample, intermolecular hydrogen bonds are formed
between arsenite and the probe to form HL-As
intensity increases with the increment of As (III)
concentration. Arsenite ions interact with phenolic
O-H to form s trong hydrogen bonds, which can
(CHEF) process. This probe can be used to imagine
Fig. 17. Illu s tration of nanoprobe interaction with arsenite in living cells [87]
Review of Colorimetric and Fluorometric Detection of Arsenic Madhawa Pradeepa Nawarathne et al
46
arsenite contributions in living cells, such as
cancer cells, and is applicable for detecting trace-
enhancement. Figure 18
formation of HL in the presence of arsenite [88].
2.3. Fluorometric determination of arsenate
2.3.1.Detection based on the interaction of
arsenate with iron-modied materials.
Nowadays, various types of solid nanomaterials are
used as an adsorbent for arsenic removal processes.
limitations [89-93]. Liu and co-workers [93]
proposed an arsenate detection method that relies
on the s trong interaction between As (V) and
the surface of metal oxides. They synthesized
labeled DNA with nanoparticles. DNA adsorbs
on ferric oxide nanoparticles through phosphate
presence of arsenate competes with adsorbed DNA
for binding sites and displaces adsorbed DNA
by removing adsorbed DNA from the surface of
The sensitivity can be improved by increasing
to adsorbing properties with high density and ease
of desorption. In addition, they enhanced their
scope for increasing adsorption capacity by using
2, CePO4, and
Fe3O4, which contain hard Lewis acids and a
bonding preference for phosphate in DNA. Among
them, CeO2 nanoparticles perform better than the
other two, achieving a ten times lower detection
limit compared to Fe3O4 nanoparticles. The
advantages of this method included the requirement
for small sample volumes for the analysis and
being highly sensitive to shorter DNA. They can
Figure 19
the induction of arsenate into the sensory sy s tem.
The same group proposed a s tudy based on
oxide nanoparticles for arsenate detection in
environmental analysis. Polyphosphate present
surface of iron oxide. As the nature of DNA and
Fe3O4 nanoparticles, salt induces the sy s tem for
Figure 20 shows the schematic of sensing arsenate
by DNA-functionalized iron oxide nanoparticles.
Fig. 18. [88]
Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
47
yield. In contra s t, in the presence of arsenate,
arsenate competes for the binding sites of DNA
signal recovered in the sy s tem. The intensity of
dependent [94].
In addition, Metal-Organic Framework (MOF)
materials are considered sensing materials,
facilitating many advantages in analyzing target
materials. Because it has attractive characteri s tics
such as the presence of organic binding ligands
or metal centers and a large surface area that
ions [95,96]. For example, amino-functionalized
iron-containing MOFs were synthesized with
detecting and removing arsenate. They fabricated
NH2-MIL-88(Fe) nano octahedra by solvothermal
treatment of FeCl3.6H2O and NH2-BDC in DMF.
Unsaturated iron sites in the sensory sy s tem have a
As-O-Fe bonds. Synthesized MOFs show weak
NH2-BDC organic linker to Fe3-3-oxo clu s ters.
introduction of arsenate into the sy s tem. Moreover,
of arsenate concentration. In the presence of
then aggregation occurs between iron-oxo-clu s ters
and arsenate. This method has great potential for
sensing arsenate compared to other methods, such
2,
of fabrication, rapid response, high sensitivity
Fig. 19.
with the presence of arsenate [93]
Fig. 20. Illu s tration of sensing arsenate by DNA functionalized iron oxide nanoparticles [94]
Review of Colorimetric and Fluorometric Detection of Arsenic Madhawa Pradeepa Nawarathne et al
48
nature, and anti-interference ability. And applicable
for polluted environmental analysis due to the
[96].
2.3.2. Detection based on the interaction of
arsenate with enzyme activity
Enzymes are biological molecules that speed
up the reaction rate by binding to the reactant/
sub s trate. Some arsenic detection processes
[97-99]. For example, a highly selective
enzymatic catalysis sy s tem was synthesized
using inexpensive glyceraldehyde 3-phosphate
dehydrogenase (GAPDH) to detect a low arsenate
concentration with a low detection limit. GAPDH
catalytic sy s tem contains an enzyme (GAPDH),
a coenzyme (NAD+), and a sub s trate (G3P). In
the reaction, cy s teine in the holoenzyme interacts
with the sub s trate to form an acyl-enzyme
intermediate through hydride transfer from
intermediate hemithioacetal to NAD+. As a result,
NADH is released, and another NAD+ has an
NAD+-acyl-enzyme. Arsenate can act as an acyl
acceptor, and present arsenate interacts with
NAD+-acyl-enzyme to form s table 1-arseno-3-
phosphoglycerate (APG). The better performance
of arsenate as a nucleophile leads to hydrolysis
of arsenoglycerate, and the resulting holoenzyme
re-enters the catalytic cycle. The result is that
sy s tem. In addition, APG hydrolyzed rapidly to
regenerate arsenate, which leads the catalytic
cycle by continuously generating NADH with
excess NAD+. Fluorescence yield increases with
the amount of arsenate are increased in the sample.
Furthermore, the catalytic sy s tem is inexpensive,
rapid response, highly sensitive, and useful for
arsenate in environmental samples and safety
applications. Figure 21 illu s trates the enzyme
catalytic sy s tem for sensing arsenate and APG
hydrolysis reaction [97].
Fig. 21. Schematic illu s tration of catalytic enzyme mechanism for sensing
of arsenate and APG hydrolysis reaction [97]
Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
49
The involvement of arsenate inhibitory activity on
phosphatase enzymes shows remarkable pathways
to arsenate detection. As an example, Jian-Ding
Qie and co-authors [98]
nanoprobe containing CdSe/ZnS quantum dots
(QDs) coated with the terbium (III) complex of
guanosine monophosphate (Tb-GMP) using one-
pot adaptive self-assembly process. Fabricated
QD/TB-GMP composite which exhibited dual
excitation properties are used for ratiometric
determination of arsenate. The presence of acid
phosphatase enzyme (ACP) can catalyze the
hydrolysis of GMP and as resulting phosphate ions
in the presence of arsenate, As (V) inhibits the
catalytic activity of ACP. Therefore, hydrolysis
and its intensity increased with the increment
of arsenate concentration. This visual analysis
method can be used to quantitatively determine
-1 detection limit and high
selectivity toward As (V). High solubility, facile
preparation process, excellent single excitation,
analysis are advantages of nanoprobe. And it is
a feasible method for environmental analysis.
Figure 22 shows the synthesis of QD/Tb-GMP and
As (V).
2.3.3. Detection of arsenate that present in the
living cell using chemosensors
Arsenate detection is essential in living cells due to
on cells. As an example, de s troy the conversion
of ATP into ADP permanently. Arsenate selective
by condensing salicylaldehyde with 4-amino-
antipyrine. In the presence of arsenate, s trong
hydrogen bonds are created between APSAL
and As (V) to form APSAL-As (V) complex.
The molecular level interaction between As
(V) and APSAL can be described using density
functional theory (DFT). The presence of arsenate
ions. APSAL is highly selective for arsenate and
obtains micromolar range detection limits. Optimal
sensor. This sy s tem applies to detect intracellular
arsenate in living cells [99].
Fig. 22. Schematic representation of the formation of QD/Tb-GMP
[98].
Review of Colorimetric and Fluorometric Detection of Arsenic Madhawa Pradeepa Nawarathne et al
50
Abu et al.
probe to detect arsenate and arsenite in living cell-
imaging applications [100]. Detection relies on
formatting nano/micro s tructures by H-bonding
interactions in the presence or absence of arsenate
and arsenite. Arsenic (arsenate and arsenite)
interacts with 2,6 diformyl-p-cresol-dioxime
(DFC-DO) ligand, which has a remarkable sensing
capacity for arsenite and arsenate detection and
forms DFC-DO-H2AsO4
- and DFC-DO-AsO2
-
respectively through intermolecular hydrogen
bonding. As a result, the quantum yield of the free
ligands increases. Above proposed methods exhibit
low detection limits below the WHO permissible
-1 with high selectivity towards either
arsenate or arsenite. Table 2 shows an overview of
determining arsenate and arsenite. Among them,
MOF materials showed the lowe s t detection limit.
3. Conclusion
This review has covered colorimetric and
and arsenite. Discussed colorimetric methods
highlighted the evolution of colorimetric methods,
molecules, and usage of nanoparticles with/without
showed better performance compared to traditional
methods. Discussed all methods are reliable, easy
selectivity and further indicated lower detection
limits (below the WHO recommendations). Thiol-
functionalized AgNPs showed the lowe s t detection
-1
mo s tly based on nano s tructural probes and
facilitated high sensitivity and selectivity. Those
are reliable, simple, non-de s tructive, and co s t-
low detection limits. Among them, MOF materials
-1).
sensitive to arsenic.
4. Acknowledgement
We thank Dr. Arjuna Wijekoon for his kind
guidance throughout the process. We acknowledge
the Wallumattagal clan of the Dharug nation as the
traditional cu s todians of the Macquarie University
land. We s trongly support equity, diversity, and
inclusion in science [101]. The authors come from
Anal. Methods Environ. Chem. J. 6 (1) (2023) 29-57
Table 2. Comparing Fluorometric methods for the detection of As (III) and As (V)
Materials Arsenic DR
(μg L-1)
LOD
(μg L-1) References
DTT/CQDs As (III) 5-100 0.086 [77]
GSH/DTT-QDs/CDs As (III) 5-100 1.7 [78]
Dipeptide-AuCs As (III) - ~4.04 [79]
Aptamer/ MSNs As (III) - 0.9 [86]
HL probe As (III) - 4.1 [88]
DNA/CeO2 NPs As (V) 0-150 2.2 [96]
NH2-Fe-MOF As (V) 0.1-50 0.056 [95]
Enzyme catalytic activity As (V) 0-200 10 [97]
QD/Tb-GMP As (V) 0.5-200 0.39 [98]
APSAL probe As (V) - 5 [99]
LOD: Limit of Detection
DR: Detection Range
51
Ph.D. s tudent, and ECR). The authors have no
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Anal. Methods Environ. Chem. J. 6 (1) (2023) 58-68
Research Article, Issue 1
Analytical Methods in Environmental Chemis try Journal
Journal home page: www.amecj.com/ir
AMECJ
Study of the behavior and determination of phenol Based on
with nickel oxide-nitrogen
carbon quantum dots using cyclic voltammetry
Khalil Ibrahim Alabid a,* and Hajar Naser Nasser a
aAnalytical Chemi s try - Department of Chemi s try - Faculty of Science - Tishreen University – Syria.
ABSTRACT
carbon pa s te electrode (MCPE) with nickel oxide nanoparticles doped
by nitrogen carbon quantum dots as nanoadsorbent (NiO-NCQD) and
cyclic voltammetry (CV). The MCP electrode was manufactured
Cyclic voltammetry can provide behavior
trans
trans) by increasing the
phenol concentration in the solution, and increasing of con s tant K0
when the concentration of phenol increased in the solution. Also,
the highe s t occupied molecular orbital (HOMO), lowe s t unoccupied
calculated. In this s tudy, EHOMO=4.92eV, ELUMO
were considered. The drinking water samples from Latakia city were
analyzed based on NiO-NCQD adsorbent using the MCPE method
(NiO-NCQD/MCPE). The phenol concentration in the drinking water
sample in Latakia was achieved less than the quantitative detection
limit (LOQ), and the proposed procedure was validated by spiking
samples.
Keywords:
Phenol,
Cyclic voltammetry,
Nickel oxide nanoparticles,
Nitrogen carbon quantum dots,
Kinetic
ARTICLE INFO:
Received 2 Dec 2022
Revised form 7 Feb 2023
Accepted 29 Feb 2023
Available online 29 Mar 2023
*Corresponding Author: Khalil Ibrahim Alabid
Email: khalilibrahimalabid@gmail.com
https://doi.org/10.24200/amecj.v6.i01.227
------------------------
1. Introduction
Phenol is described as an aromatic organic
compound C6H5OH. Phenol and its derivatives
are the main pollutants in water sources [1]. It
is highly toxic [2-4] and enters the human body
through inge s tion, inhalation, or contact with the
severe damage. Among these damages: Damage to
the lungs, liver, kidneys, urinary and reproductive
tracts, cardiovascular disease, shortness of breath,
neurological problems, as well as severe abdominal
pain, ga s trointe s tinal irritation, nausea, vomiting,
diarrhea, sweating, coma, and death. Inge s tion of
1g of phenol is a lethal dose [5-9]
oxidative s tress in biological materials, disrupting
endocrine metabolism and promoting cancer
[10]
according to the world health organization (WHO)
that its concentration does not exceed one µg L-1
in drinking water [11]. Phenol and total phenol
can be e s timated spectrophotometrically in the
visible (VIS) [12-19], in the ultraviolet (UV) [20],
and High-Performance Liquid Chromatography
(HPLC) [21-22]. Schema 1 showed the phenol
oxidation (one-electron oxidation) and reaction
process [23,24].
59
Cyclic voltammetry is one of the mo s t important
electrochemical techniques that help provide
information about the kinetics, mechanics, and
behavior of the s tudied material [25]
possible from cyclic voltammetry to know if the
reaction is subject to oxidation, reduction, or both.
It has three cases: reversible, quasi-reversible, or
irreversible [26]. Cyclic voltammetry can provide
[27-30], mass transport
(mtrans) [31-33]),
[34], and con s tant K0 [35], the highe s t occupied
molecular orbital (HOMO), lowe s t unoccupied
molecular orbital (LUMO) [36, 37], Gibbs free
[38] and interface trap density (Dit)
[39].
Randles-Sevcik irreversible Equation 1 [27-30].
The mass transport is given by Equation 2 [31-33].
by Equation 3 [34]. Con s tant (k0
s tandard rate con s tant (k0) ratio to mass transfer. It
is given by Equation 4 [35]. The HOMO-LUMO
values are given by Equations 5 and 6 [36-37].
Equation 7 [38].
The interface trap density (Dit) can be obtained by
Equation 8[39].
(Eq.1)
Where, ip: Peak current (A), n: Number of electrons,
–1), A: electrode area
(cm2
C: concentration (mol. cm–3), R: gas con s tant
(J. mol–1 K–1
2s–1), v: Scan rate (V s–1).
(Eq.2)
(Eq.3)
represents a measure of the symmetry barrier in a
of electrons involved in the rate-determining s tep.
Schema 1. The oxidation and reaction of phenol
Study and Determination of Phenol by NiO-NCQD and CV Khalil Ibrahim Alabid et al
60 Anal. Methods Environ. Chem. J. 6 (1) (2023) 58-68
(Eq.4)
Where: io: exchange current density (A m–2), in the
case where the oxidation is irreversible it mu s t be:
k0 ≪ mtrans, as for according to Nicholson, mu s t be
k0 < 3.5 ×10-4 × v 1/2.
EHOMO eV= [ Eox - E1/2 + 4.8] (Eq.5)
ELUMO = (EHOMO – Eg) (Eq.6)
Where: Eox: oxidation potential (From CV), E1/2:
half-oxidation potential for peak, Eg: Optical
Bandgap (from absorption s tudies).
ox - Ered - Eg + C (Eq.7)
Where: Eox: Oxidation potential, Ered: Redaction
potential, Eg: the excited singlet s tate energies, C: is
the electro s tatic interaction energy for the initially
formed ion pair, generally considered negligible in
polar solvents.
(Eq.8)
A, q, Cox, V, and Eg are the gate area, electron
voltage shift, and bandgap.
Carbon/graphite pa s te electrodes (CPE) are
important for being chemically inert, easy to
fabricate, electrode surface renewability, low ohmic
resi s tance, low co s t, and environmentally friendly.
However, its kinetics, s tability, and selectivity are
[40]
relied upon, as they are more selective and sensitive
to organic compounds. This research is one of the
critical research s tudies on the behavior of phenol
in the electrochemical cell and determines the
concentration of phenol in a drinking water sample
using a selective electrode for carbon pa s te with
nanoparticles by cyclic voltammetry.
2. Experimental
2.1. In s truments
Voltammetry sy s tem for trace analysis and education.
Complete accessories with VA Computrace software
and all electrodes for a complete measurement
sy s tem: Multi-Mode Electrode pro (MME pro),
Ag/AgCl reference electrode, and Pt auxiliary
electrode. In this s tudy, a modern voltammetric was
connected to a PC based on a USB port (Metrohm
cell). Sartorius pH meter type PB-11 was used from
Data Weighing Sy s tem Company (pH meter and
2.2. Reagents and Materials
All chemicals with high purity were purchased
from Sigma or Merck Company (Germany). Phenol
C6H6O purchased from Acros Organics Company
(AC221755000, molecular weight 94.11g mol-1,
-3, high purity 99%).
The monopotassium dihydrogen phosphate
(KH2PO4) was prepared from Sigma, Germany
(CAS No.: 7778-77-0). The boiled and cooled
Sigma).
2.3. Synthesis of NiO-NCQD nanocomposite
Take 0.6 g of NiO Nanoparticles (20nm) are added
with 30 mL of nitrogen quantum carbon dot after
then washed three times with di s tilled water and
dried in an oven at 60 for 12h to get NiO-NCQD
nanocomposite.
2.4. General procedure
2.4.1.Fabrication of selective electrode
The selective electrode is made (in the laboratory). It
consi s ts of a glass tube that is open at both ends and
conducting electric current is connected between the
carbon pa s te using NiO-NCQD nanocomposite (12 %),
61
Study and Determination of Phenol by NiO-NCQD and CV Khalil Ibrahim Alabid et al
the electrode body is made of glass. Symbolizes the
factory electrode (NiO-NCQD/MCPE) shown in
Figure 1. Then the electrode is connected to the volt-
amperometric cell (VA), which consi s ts of a working
electrode (WE) and a comparison electrode, and it is
usually an Ag/AgCl electrode where its potential is
2.4.2.Preparation of s tock solution and
monopotassium phosphate buer
To prepare a 0.1036 M phenol solution, take 0.974
g of phenol, then dissolve it into 100 ml di s tilled
prepared from KH2PO4 at a concentration of 0.1 M
and a solution of KOH potassium hydroxide at 0.1
obtain a pH of 4 and 7.
3. Results and discussion
3.1. Eect of pH
on the current intensity I(µA) of a s tandard phenol
solution shown in Figure 2.
From the previous drawing curve, Through the
values of and U(V), it is noted that it two peaks
Fig. 1. Schematic of factory electrode components (NiO-NCQD/MCPE)
Fig. 2.
62
and achieves the highe s t value of peak current =
49.5µA, =72µA at pH =7 respectively, so these
two values are adopted. In the case of phenol,
when used CV method, it undergoes an oxidation
process only without reduction, so the sy s tem is
irreversible, phenol concentration is s tudied with
ranges of phenol (10 - 250 - 500 – 750 – 1000) µM
(4,7), scan rate = 100mv.s-1 = 0.1v. s-1 both of pH
(4,7), s tep voltage is 0.04166V and 0.05991V for
both (pH =4,7), respectively, using the electrode
(NiO-NCQD/MCPE).
Cyclic voltammetry can provide behavior
(D ), the mass
transport (mtrans), and the values of each are
calculated (Table 1).
3.2. Eect of phenol concentration
o, and mass transport
and interface trap density (Dit) are s tudied for the
phenol concentrations, as in Figures 3(A-D).
From previous curves, the mass transport and
concentration, probably due to increasing
phenol concentration, cause the blockage of the
electrode surface. In the case where the oxidation
is irreversible, it mu s t be: Ko<m trans, according
to Nicholson, mu s t be k0 <3.5 ×10-4× v(1/2), from
previous curves, In this research, K0<m trans and
K0 <3.5 × 10-4×0.1= 3.5×10 -5, the HOMO-LUMO
values are s tudied from cyclic voltammetry
nanocomposite, where Eox= 0.43 V, and E1/2= 0.31
V, the gap from absorption s tudies at 270 nm=4.6
from (UV) so, EHOMO = 4.92 ev, optical band, so
Table 1.
of phenol at pH=4 and pH=7 using (NiO-NCQD/MCPE)
pH CµM I (µA) D×109
(m2·s-1)nα α. mtrans Ko×107(Dit)
eV−1 cm−2
4
1000 90 0.036278 1.615581 2.20006E-05 1.485325000 2.95483E+13
750 72 0.033772 1.974600 2.1227E-05 1.18826000 2.85093E+13
500 64 0.060039 1.974600 2.83026E-05 1.056231000 3.80124E+13
250 56 0.194084 1.870673 5.08868E-05 0.924202000 6.83445E+13
0 45 88.63469 1.653153 0.001087458 0.742663000 1.46053E+15
7
1000 104 0.048443 1.615581 2.54229E-05 1.716375904 3.41447E+13
750 99 0.067397 1.870673 2.99869E-05 1.633857832 4.02745E+13
500 92 0.151635 1.615581 4.4979E-05 1.518332531 6.04099E+13
250 83 0.594651 1.341237 8.90721E-05 1.369800001 1.1963E+14
0 69 220.0211 1.565762 0.001713337 1.138749398 2.30113E+15
Anal. Methods Environ. Chem. J. 6 (1) (2023) 58-68
63
Fig. 3.
D) mass transport,
F) and interface trap density on the surface of the proposed electrode NiO-NCQD/MCPE
Study and Determination of Phenol by NiO-NCQD and CV Khalil Ibrahim Alabid et al
64
ELUMO= 0.32 ev, As for the value of Gibbs free
is spontaneous in the direction with electric current.
The interface trap density (Dit) of the electrode has
a value within (2.95483×10+13- 1.46053×10+15) eV
cm at pH=4 and (3.41447×10+13- 2.30113×10+15)
eV cm at pH=7. The large values indicate
good corresponding and, as noted, interface trap
density (Dit) decrease in value with increasing
initially in the pre-measurement s tage at a rate
of 2000 rpm, where the motion of a chemical
compound in solution inside the electrochemical
cell are, principally three :(convection, migration,
homogenizing the solution in addition, s tirring
of unwanted motion (migration and convection),
that expresses the behavior of phenol within the
nitrogen gas gurgles inside the electrochemical cell
behavior where the temperature of the solution was
oC.
3.3. Application on drinking water samples by
the proposed electrode (NiO-NCQD/MCPE)
A drinking water sample from Latakia city was
analyzed using the proposed method, and it was
found that the sample was less than the detection
limit (<LOD) of the method. The s tandard addition
method found that the sample does not contain
phenol, according to Table 2 and Figures 4-5. Due
to previous curves, the results can be placed in
Table 2.
It is noted from the above that the phenol concentration
in the drinking water sample in Latakia is less than
the quantitative detection limit (LOQ) of the method,
less than 10µM (0.9411 mg L-1).
4. Conclusion
This paper deals with fabricating a phenol-selective
Oxide nanoparticles (NiO) doped with Nitrogen
Carbon Quantum Dots (NCQD) using Cyclic
voltammetry. The electrode was manufactured in
a laboratory. Results be s t conditions are obtained
at pH= 7.0 and 4.0 using KH2PO4,
and the behavior of a phenol solution is s tudied in
an electrochemical cell (Cyclic voltammetry) using
NiO-NCQD/MCPE. The phenol concentration in
Fig. 4. Determination of phenol concentration in drinking water using
the proposed electrode (NiO-NCQD/MCPE) at pH = 4
Anal. Methods Environ. Chem. J. 6 (1) (2023) 58-68
65
the drinking water sample in Latakia is less than the
quantitative detection limit (LOQ) of the method,
that is, less than 10µM (0.9411 mg L-1).
5. Acknowledgments
The authors thank the Faculty of Science-Tishreen
University- Syria and the Higher In s titute for
Environmental Research - Tishreen University-
Syria for their help and support during this work.
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Anal. Methods Environ. Chem. J. 6 (1) (2023) 69-78
Research Article, Issue 1
Analytical Methods in Environmental Chemis try Journal
Journal home page: www.amecj.com/ir
AMECJ
Chemical analysis of essential oils of Thymus
Carmanicus Jalas by gas chromatography-mass
spectrometry and toxicity activity agains t the major
Iranian malaria vector, Anopheles Stephensi
Nazanin Sadat Mousavi a, Alireza Sanei-Dehkordi b,, Ismaeil Alizadeh c,d, Ali Faghihi Zarandi e, Mohsen
Mehdipour Rabori a, Nasrollah Saberi f and Mohammad Amin Gorouhi d *
a Department of Environmental Health Engineering, Faculty of Public Health, Kerman University
of Medical Sciences, Kerman, Iran
b Department of Biology and Control of Disease Vectors, Faculty of Health, Hormozgan University
of Medical Sciences, Bandar Abbas, Iran
c Research Center of Tropical and Infectious Diseases, Kerman University of Medical Sciences, Kerman, Iran
d Department of Vector Biology and Control, Faculty of Public Health, Kerman University of Medical Sciences, Kerman, Iran
e Department of Occupational Health Engineering and Safety at Work, Faculty of Public Health, Kerman University
of Medical Sciences, Kerman, Iran
f Bam University of Medical Sciences, Bam, Iran
ABSTRACT
In the la s t few years, using chemical insecticides to control the
malaria vector has caused environmental pollution and resi s tance
to chemical insecticides. This s tudy aimed to inve s tigate the
chemical analysis of essential oils of Thymus carmanicus Jalas
by gas chromatography and mass spectrometry (GC-MS) and
toxicity activity again s t the major Iranian malaria vector, Anopheles
s tephensi. The essential oil of Thymus carmanicus Jalas was
prepared from dried leaves using the hydro-di s tillation method. Gas
chromatography mass spectrometer (GC-MS) was used to analyze
and identify thyme essential oil compounds. Bioassay was performed
using World Health Organization (WHO) s tandard te s t. The T.
Carmanicus Jalas essential oil consi s ted of 15 compounds, with
the major components by volume. The LC50 and LC90 of thyme oil
were 20.37 and 41.38 mg L-1 at 24h after application, respectively.
the toxicity of 5%, 20%, 25%, 40%, 50%, and 80% concentrations
of Thyme essential oil (P<0.05). The 80% concentration of Thyme
essential oil exhibited 100% toxicity again s t A. s tephensi larvae at
24h after application. T. Carmanicus has a rich source of bioactive
compounds for use as a mosquito larvicide.
Keywords:
Chemical analysis,
Gas chromatography-mass
spectrometry,
Thymus Carmanicus Jalas,
Essential oil,
Malaria
ARTICLE INFO:
Received 13 Nov 2022
Revised form 22 Jan 2023
Accepted 10 Feb 2023
Available online 28 Mar 2023
*Corresponding Author: Mohammad Amin Gorouhi
Email: amingruhi@gmail.com
https://doi.org/10.24200/amecj.v6.i01.225
------------------------
1. Introduction
Malaria is among the mo s t important parasitic
diseases transmitted to humans by Anopheles
mosquitoes. This disease has been reported in
91 countries worldwide [1]. Five Plasmodium
species are transmitted by the bite of Anopheles
mosquitoes [2]. More than 90% of malaria cases
were reported in three southea s tern provinces of
Iran, including Si s tan and Baluchi s tan, Kerman,
and Hormozgan [3]. Out of the 476 Anopheles
70 Anal. Methods Environ. Chem. J. 6 (1) (2023) 69-78
species identified in the world, 70 species are
capable of transmitting malaria, and 40 species
are known as the main vectors. In addition to
malaria transmission, Anopheles could transmit
filariasis and some arboviruses. In Iran, 7
Anopheles species are disease carriers, the mo s t
important of which is Anopheles s tephensi, found
in the southern regions [4]. Insecticide resi s tance
has become a threat to the effectiveness of
chemical vector control methods. This issue is
of particular importance considering the malaria
elimination program in Iran [5]. Therefore,
special attention has been paid to plants as
natural reservoirs with fewer side effects to fight
again s t disease vectors. Currently, controlling
mosquito larvae using larvicides is a major part
of controlling mosquito-borne diseases. The mo s t
common larvicides contain organophosphorus
compounds such as temephos, fenthion, and
chlorpyrifos. However, their toxicity for aquatic
organisms and the environment as well as the
phenomenon of insecticide resi s tance and acute
and chronic toxicity for humans are increasingly
reported. Therefore, it seems necessary to find
new larvicides from alternative sources such
as plants [6, 7]. Herbal insecticides have been
recently used to control vectors due to their
relatively high efficiency, degradability, and
lack of adverse effects on the environment.
Several plant products have been reported
as insecticides for mosquito control. Various
vegetable oils have shown a wide range
of insecticidal activity again s t pe s ts. Also,
these oils have anti-nutritional and repellent
properties, reduce oviposition and disrupt the
natural growth process of pe s ts [7]. Thyme is
among the important genera of the Lamiaceae
family and has been widely used in food indu s try,
pharmaceutical and cosmetic due to its various
biological activities [8]. According to recent
s tudies, Lamiaceae family consi s ts of 4,000 plant
species with 220 genera. Thyme, with 300-400
species, is among the mo s t important species of
this family. This genus includes 18 perennial and
aromatic species that grow in different regions
of the country. Thymus caramanicus is among
the species of this genus [9]. Thyme branches
contain essential oil, tannin, basic bitter
sub s tances, saponin and herbal disinfectants.
The plant essential oil contains variable amounts
of phenolic compounds such as thymol and
carvacrol [10]. Various con s tituents of the oil of
T. carmanicus were reported in previous s tudies.
Although carvacrol is the mo s t abundant
con s tituent in the oils obtained from all the
mentioned s tudies, percentage of the compound
varied, based on the origins of the plant.
While it has been reported as 47.55-96.2.7%
from T. carmanicus cultivated in Kerman,
42.1-93.4 % in Isfahan, 42% in Shahroud,
and 19.8-81.1% Semnan [11-15]. Generally,
there are two devices including Soxhlet and
Clevenger to essential oil extraction [16]. To
analysis the composition of an essential oil
sample, use Gas chromatography columns
with different polarities [17]. The previous
s tudies used several methods to essential oil
analysis, such as Gas Chromatography and
Mass Spectrometry (GC/MS) and capillary gas
chromatography (GC) [18]. Netopilova et al. in
2021 used the GC– flame ionization detector
(FID)/MS for analysis the Origanum vulgare
and Thymus vulgaris [19]. In the present s tudy,
we aimed to inve s tigate the chemical analysis
of essential oils of Thymus carmanicus Jalas
by GC-MS and also the toxicity activity of this
essential oil was evaluated again s t the major
Iranian malaria vector, Anopheles s tephensi as
an environmentally friendly method.
2. Materials and methods
2.1. Plant collection
The fresh leaves of the Thymus carmanicus
Jalas plant were collected from the Hezar
in southea s t of Iran (Fig. 1). Collected Thyme
specimens were identified by the Department
of Pharmacognosy in Kerman University of
Medical Sciences.
71
Chemical Analysis of Essential Oils by GC-MS Nazanin Sadat Mousavi et al
2.2. Extraction of eessential oil
To extract essential oils, 100 g of dried leaves of
Thymus caramanicus
[10]. Then, 600 cc of deionized water was added to
in a clevenger-type apparatus for 3.5 to 4 h. The
essential oil was extracted approximately 3.5 to 4
h at 60 °C [20] (Fig. 2). Then, the extracts were
exsiccated by anhydrous sodium sulphate and
s tored in a dark glass vial at 4 °C in a refrigerator
for further experiment [11, 12].
2.3. Gas chromatographic-mass spectral
analysis
Gas chromatography-mass spectrometer (GC-MS)
thyme essential oil compounds (Hewlett-Packard
6890, Agilent Technology, Santa Clara, California,
USA) (Fig. 2). It is equipped with HP–5MS
column (30 m× 0.25 mm× 0.25 µm). The initial
temperature was 40 °C for 1 min and later was
raised to 270 °C for 5 min at a rate of 20 °C per
minute. Other parameters of the GC-MC machine
included carrier gas Helium (99/999%), injector
temperature (260 °C), detector temperature (FID,
270 °C), split-less mode, the ionization potential of
70eV, scan rate of 1 scan per sec, the scan range of
m/z 40–48 was used for all analysis. The essential
retention indices, and mass spectra fragmentation
with those in a s tored Wiley 7n.1 mass computer
library and those of National In s titute of s tandards
and Technology (NI s t) [21-23].
Fig 1.
Fig. 2. The procedure of collecting plants, extracting and GC-MS analysis of essential oils.
72
2.4. Larvae collection and Toxicity assay
Anopheles s tephensi larvae were collected from
the Paykam area, Bam, south of Kerman Province.
Bioassay was performed using World Health
Organization (WHO) s tandard te s t [24]
concentrations (20, 40, 80, 160, and 320 mg L-1)
of the essential oils obtained from the s tudied plant
were prepared using ethanol as the solvent. Thus,
rd or 4rd in s tar larvae of Anopheles
s tephensi were exposed to these concentrations
in each 400 mL beaker. The experiments were
replicated four times for any concentrations of
thyme essential oil and ethanol.
2.5. s tati s tical Analysis
Probit analysis was used to calculate the LC50
and LC90. Toxicity indices were compared using
analysis of (ANOVA) followed by the Dunnett te s t
to di s tinguish between the treatments. All s tati s tical
analyses were performed using the SPSS version.
16. A p-value of less than 0.05 was considered
3. Results and Discussion
Thyme essential oil was found to contain 15
compounds using GC-MS analyses. Carvacrol,
thymol, and beta-caryophyllene had the highe s t
frequency with 61.92%, 6.13%, and 5.55%,
respectively. The mo s t common compounds are
shown in Table 1. In addition, the chemical analysis
of the essential oil of Thymus carmanicus Jalas is
shown in Figure 3.
Table 1. Con s tituents of Thymus carmanicus Jalas essential oil by GC-MS analyses.
Major Con s tituents (%)Retention timeThyme essential oil components
0.5810.029
5.5511.099
1.111.187Copaene
0.9912.274Limonene
2.6314.224Isoledene
1.2614.634
1.9215.251terpinene
0.4116.609Sabinene
1.9517.932
61.9218.541Carvacrol
0.8724.681
1.2641.959P-cymene
0.3245.059
6.1348.209Thymol
3.3454.345Naphthalene
9.77-Other compounds
Anal. Methods Environ. Chem. J. 6 (1) (2023) 69-78
73
The results of the dose-response te s t are shown
with the calculation of toxicity lethal concentration
as ppm (mg L-1) essential oil (LC50 and LC99) in
Table 2. They were 20.37 and 41.38 mg L-1 for LC50
and LC90 at 24h after application, respectively. The
calculated dose-response curve for Thyme essential
oil after 24h is shown in Figure 4.
in toxicity between 5%, 20%, and 25% of Thyme
essential oil (P>0.05). As well as, there was no
and 80% of Thyme essential oil (P>0.05). At 24h after
between the toxicity of 5%, 20%, 25%, and 40%,
Table 2. Lethal doses of thyme essential oil again s t Anopheles s tephensi larvae.
PChi-Square*** (df) **
LC90 (CL*)
mg L-1
LC50 (CL*)
mg L-1
Time
0.001>
1.16
(4)
40.6
(0.005)
41.38
(38.26-45.41)
20.37
(18.02-22.52)
After 24 h
LC50 and LC90: Lethal dose necessary to kill 50% and 90% of larvae, respectively.
** s tandard error.
***Chi-square (degree of freedom).
Fig. 3. A typical GC-MS chromatogram showing the chemical analysis
of essential oil from Thymus carmanicus Jalas.
Chemical Analysis of Essential Oils by GC-MS Nazanin Sadat Mousavi et al
74
50%, and 80% concentrations of Thyme essential oil
(P<0.05). The 80% concentration of Thyme essential
oil exhibited 100% toxicity again s t Anopheles
s tephensi larvae at 24h after application (Fig. 5).
3.1. Discussion
Tropical regions are more vulnerable to parasitic
diseases and risk contracting diseases due to climate
change and increased globalization. Mosquitoes
Fig. 4. Dose-response curve for Thymus carmanicus Jalas essential oil after 24h
Fig. 5. Thymus carmanicus
Jalas essential oil again s t An. s tephensi
Anal. Methods Environ. Chem. J. 6 (1) (2023) 69-78
75
are the mo s t important public health insects in
tropical and subtropical regions because they carry
important parasites and pathogens worldwide that
cause death, poverty, and social impairment [25].
Makizadeh Tafti et al. (2010) s tated that carvacrol
con s tituted the large s t part of the essential oil in
all thyme ecotypes in Kerman Province, followed
by thymol, para cymene, and gamma-terpinene.
It is worth noting that the content of each element
underwent changes in this research that could be
or even in-vitro conditions [10]. Ebrahimi et al.
contents in GC-MS device inve s tigations [26]. The
contents observed in the s tudy by Makizadeh Tafti
showed that carvacrol con s tituted about 80% of
reduced this amount to 60%. Eftekhar et al. (2010)
found 68% carvacrol in Thymus caramanicus.
This s tudy showed that the superiority of carvacrol
content compared to other con s tituents was around
62% [27]. In the research by Mazandarani and
Rezaei (2005) on Thymus caramanicus grown
in Mazandaran Province, it was observed that
pulegone (26%) was the mo s t frequent element in
the essential oil due to climate change, and carvacrol
content decreased to 8% [28]. It was observed that
Thymus caramanicus
The lethal property of Thymus caramanicus
essential oil was extremely high, so lethality
reached 50% and 90% at concentrations of 20 and
41, respectively. Damtie and Mekonnen (2021)
prevent Anopheles larvae proliferation and growth
at concentrations of 20-50 and showed desirable
resi s tance to adult insects at lower concentrations.
with observations [29]. Dargahi et al. (2014) found
that Thymus transcaspicus essential oil exhibited
s trong insecticidal activity again s t An. s tephensi,
which could be due to its con s tituent compounds,
especially carvacrol and thymol phenols [30].
These compounds were present in abundance in
Thymus caramanicus. Thymus transcaspicus could
154 and 248 µg L-1, respectively (P<0.05). There
plants in terms of concentration, which could be
attributed to the high concentration of carvacrol
and thymol in Thymus caramanicus compared to
Thymus transcaspicus.
Gupta et al. (2022) s tated that the phenolic
increase the larval population of various disease-
carrying mosquitoes (P<0.05). The results showed
LC50 and LC90 values of this plant for An. s tephensi,
Ae. aegypti and tritaeniorhynchus larvae were
equal to 56 and 124 µg L-1, 58 and 270 µg L-1,
and 22.58 and 193 µg L-1 [31]. In the current
s tudy, LC50 and LC90 values were decreased due
to changes in the phenolic compounds present in
Thymus caramanicus, which could be attributed to
the greater toxicity of these compounds. Kelidari
lipid nanoparticles containing Zataria multiora
essential oil and found that these particles could
Anopheles
s tephensi larvae [32]. Firooziyan et al. (2022)
Myrtus nanoemulsion
and found that this plant could eliminate 50% and
90% of An. s tephensi larvae at concentrations of
26 and 46 µg L-1 [33]. Zarenzhad et al. (2021)
chitosan nanoparticles containing Laurus nobilis
and Trachyspermum ammi essential oils again s t
An. s tephensi and s tated that the essential oil of
[34].
Similarly, in the present s tudy, the Thyme essential
An. s tephensi
larvae.
4. Conclusion
This s tudy indicated that T. Carmanicus has a rich
source of eco-friendly bioactive compounds for use
as a mosquito larvicide. Its considerable capability
might be the high percentage of Carvacrol, which
can be used as a larvicidal agent for mosquito
Chemical Analysis of Essential Oils by GC-MS Nazanin Sadat Mousavi et al
76
a possible way for further s tudies to determine the
active molecule. Carvacrol with 61.92% was the
highe s t compound of Thyme essential oil. LC90
of Thyme essential oil at 24h after the application
was 41.38 mg L-1. A concentration of 80% of this
essential oil killed 100% of larvae at 24 hours.
However, further inve s tigations mu s t be conducted
to describe the mode of action of each con s tituent
s
organisms.
5. Acknowledgements
Zahra Mahdavi and Miss Pakravanan for helping
in the essential oil preparation and interpretation
of GC-MS results. This article is part of Nazanin
supported by and conducted at Kerman University
of Medical Sciences (Project Number 400001144).
This s tudy was approved by the Ethics Committee
of Kerman University of Medical Sciences (IR.
KMU.REC1401.059).
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Anal. Methods Environ. Chem. J. 6 (1) (2023) 69-78
Anal. Methods Environ. Chem. J. 6 (1) (2023) 79-99
Research Article, Issue 1
Analytical Methods in Environmental Chemis try Journal
Journal home page: www.amecj.com/ir
AMECJ
modeling the competitive biosorption of pentachlorophenol
and 2,4,6-trichlorophenol to Canna indica L and analyzed by
UV-Vis spectrometry in Aquaponia
Chri s tian Ebere Enyoh a,c,*, Prosper Eguono Ovuorayeb, Beniah Obinna Isiukuc,
and Chinenye Adaobi Igwegbed
aGraduate School of Science and Engineering, Saitama University, Saitama, Japan
bDepartment of Chemical Engineering, Federal University of Petroleum Resource, P.M.B 1221 Eurun, Nigeria
cDepartment of Chemi s try, Faculty of Physical Sciences, Imo State University, Imo State, Nigeria
dDepartment of Chemical Engineering Nnamdi Azikiwe University, P. M. B. 5025, Awka, Nigeria
ABSTRACT
The continuous exposure of the environment to carcinogenic wa s tes
and toxic chlorophenols such as pentachlorophenol (PCP) and
2,4,6-trichlorophenol (TCP) resulting from indu s trial production
activities has become a great concern to research scienti s ts and
eco-friendly approach to the phytoremediation of water will
guarantee su s tainability. The present research concerns the co s t-
biosorption of PCP and TCP from aqueous solution to Cana indica.
neural network (ANN) model, and UV-Vis Spectrometry. The
predictive performances of the ANN model and the RSM were
compared based on their s tati s tical metrics. The antagoni s tic and
concentration, and exposure time) on biosorption were s tudied at
optimum conditions corresponding to predominantly acidic pH (4),
required initial concentration of 50 mg L-1, and exposure time of 25
days in aquaponia. The optimized output transcends to PCP and TCP
e s tablished that at the optimum conditions, the co s t of operating
the removal of TCP from the aqueous solution would save $ 7.72
at-a-time (OFAT) methodologies reported in previous research.
Keywords:
Phytoremediation,
UV-Vis spectrometry,
Chlorophenol biosorption,
Canna Indica L plant,
ANN modeling,
RSM optimization
ARTICLE INFO:
Received 11 Nov 2022
Revised form 21 Jan 2023
Accepted 16 Feb 2023
Available online 30 Mar 2023
*Corresponding Author: Chris tian Ebere Enyoh
Email: cenyoh@gmail.com
https://doi.org/10.24200/amecj.v6.i01.228
------------------------
1. Introduction
Chlorinated phenols, such as Pentachlorophenol
(PCP) and 2,4,6-Trichlorophenol (TCP), have
been used since the 1930s in a variety of indu s tries
including wood preservation, pe s t control, and
herbicide production [1,2]. As a result, wa s tewater
from these indu s tries can contain high levels of
80 Anal. Methods Environ. Chem. J. 6 (1) (2023) 79-99
these chemicals, leading to the pollution of water
resources and potential harm to ecosy s tems [3].
contributor to the contamination of water
resources with chlorinated compounds. s tudies
[4,5,6].
to their high toxicity, carcinogenic potential, and
environmental persi s tence [1,2]. These chemicals
can cause serious health issues such as respiratory
problems, cardiovascular disease, ga s trointe s tinal
issues, and cancer in humans and have been linked
to an increased risk of lymphomas, leukemia, and
liver cancer in animal s tudies [1,2]. Removing these
chemicals from water resources or wa s tewater
before they are released into the environment is
essential to prevent potential harm to humans
and ecosy s tems. Various methods are available
for removing PCP and TCP from water, including
biological and physicochemical approaches such
as photochemi s try, air s tripping, incineration,
and adsorption technologies using activated clay
and plant-based carbons [7]. While some of these
implement. Using aquatic plants for wa s tewater
treatment is a newer method for removing pollutants,
using con s tructed wetlands, pilot-scale sy s tems,
or hydroponic setups [7]
based treatment sy s tems can vary depending on
Nigeria and other tropical countries, various plants
used species for this purpose in Nigeria due to
their wide di s tribution and dominance in aquatic
environments. Additionally, canna lilies can
such as PCP and TCP through phytoremediation
[8-11] and can survive in polluted areas [12].
Central composite design and response surface
methodology are s tati s tical approaches used in
pollutant removal s tudies to design experiments
and optimize treatment conditions [13]. They
conditions through s tati s tical software, reducing
the number of co s tly experiments and trials needed
[14]. These methods have been applied to various
processes, including coagulation to remove dyes
from wa s tewater [15,16]. They can be helpful in
predicting the behavior and outcomes of treatment
neural networks (ANN), which utilize learning
algorithms to evaluate the relationships between
input and output variables, can also be used to model
and predict the behavior of water management
processes [17, 18]
adaptable, and can produce real-time predictions
[18]. Both response surface methodology and
their predictive capabilities for various processes
[17]
using C. indica L (CiL-plant), an aquatic plant, to
remove PCP and TCP from water using response
The use of aquatic plants for wa s tewater treatment,
chlorophenols, is a relatively new method known
as aquatic phytoremediation. This s tudy aims to
optimize the ability of C. indica to remove PCP
and TCP from water using a hydroponic sy s tem,
of its kind. Furthermore, the removal behavior
of C. indica for PCP and TCP has been predicted
evaluation of the phytoremediation process was also
the suitability of the CiL-plant for Aquaponia.
2. Materials and methods
2.1. Preparation of plant material and pe s ticides
solutions
We discussed how the plant material and pe s ticide
solutions were manufactured in our earlier papers
[8-10]
s tate, Nigeria, Canna indica L. seeds and soil for
81
Analysis and Biosorption of P&T chlorophenol in Cana indica. L Plant Chris tian Ebere Enyoh et al
growing the plant were collected. The plant was
raised in nurseries with natural environmental
conditions. To conduct the s tudy, properly harve s ted
and TCP (analytical grade, 99.5%) was used after
being acquired from FinLab in Owerri. Di s tilled
water and ethanol were used to make the solutions
for this experiment. An ethanol-water solution (10%
v/v ethanol/di s tilled water) was used to dissolve
1.0 g of PCP/TCP per liter of solution in a 1.0 liter-
s tock had a 1000 mg L-1 equivalent. By dilutions
with di s tilled water, working solutions of 50, 100,
150, 200, and 250 mg L-1 were produced from the
s tock solution and labeled accordingly. Working
solutions were made, and absorbance was measured
at 220 nm for PCP and 296 nm for TCP using a
UV spectrophotometer. The calibration curve
(concentration vs. absorbance) was created using
the recorded absorbance, and it was then utilized
to calculate the amounts of PCP and TCP. With
absorbance for PCP and TCP s tarting concentrations
rose as the initial concentration increased. This
indicates s trong linearity of the regression line with
good correlation, consequently, and satisfaction of
the in s trument calibration.
2.2. Batch s tudies
Uptake of PCP and TCP by C. indica L. in
pe s ticide-contaminated water was s tudied in batch
culture experiment using hydroponic, cylindrical
(pots) containers with dimensions 18 cm in length,
37 cm in diameter (external) and 19 cm depth
[9, 10]
working solutions. Then the plant was introduced
into the solution and allowed to s tand. This was
durations (i.e., 10 days, 15 days, 20 days, and 25
days). In total, 5 pots were prepared and at each
interval of 5 days a plant was removed and the
residue was analyzed by UV-vis spectrophotometer
at 220 nm for PCP and 296 nm for TCP [9, 10]. The
C.
indica was determined in 500 mL of te s t solutions
containing 100 mg L-1
pH (4- 9). 1 M nitric acid (HNO3) and 1 M sodium
hydroxide (NaOH) were used for pH adju s tments.
The pH of each solution was measured with a digital
pH meter (Model Jenway 3510). The initial and
determined on a UV–visible spectrophotometer
(Spectrum Lab 23A) at its maximum absorbance
wavelength of 220 nm and 296 nm, respectively.
All set-ups were conducted in triplicate (total pots
were 80 for batch s tudies, including control and
placed randomly with position shifted once a week.
After one week, all set-ups were supplemented
with N.P.K. fertilizers (1%, i.e., 5 ml: 500 ml).
For each treatment method mentioned, there was a
corresponding control group that only consi s ted of
the nutrient needed for plant growth in water was
provided.
2.3. Response surface design of Experiment
The Central Composite Design (CCD) is an
empirical model used for multi-objective
optimization of the adsorption or bio-sorption of
micropla s tics from an aqueous solution [19,20].
The CCD optimization is based on the Response
Surface Methodology (RSM) [15]. It is used to
quadratic, cubic, or polynomial model [21]. The
e s tablish an optimal model equation and describe
the antagoni s tic or synergetic interactions and
relationship of experimental variables and their
range s tudied [13]. In this s tudy, the CCD matrices
consi s ted of 20 experimental runs. The modeling
of the bio-sorption of PCP and TCP to CiL-plant
(Canna indica L.) in terms of actual values is shown
the prediction of the optimum conditions for bio-
sorption (pH, initial concentration, and time)
for the removal of PCP and TCP is described by
Equation 1.
82
(Eq.1)
Where xij
independent variables (pH, time, and concentration),
is the model intercept, and Y is the response (PCP
and TCP removal rate). The optimization modeling
of the biosorption of TCP and PCP to the CIL
plant was executed using Design Expert software
v12.0. The experimental variables contact time (A)
(days), initial concentration (B) (mg L-1), and pH
(C) shown in Table 1 were varied to 3-Levels with
5 replications. The toxicity was modeled following
the CCD matrix. The initial concentration of the
biosorbent and the contact time was varied to
5-Levels at an experimentally determined pH of 4.
2.4. Articial Neural Network
Aside from RSM modeling, data modeling via
neural network (ANN) was implemented in this
s tudy to create a better under s tanding of the model
validation of the bioremediation process. The
neural network tool in MatLab 2018a was used to
model the CiL-plant biosorption process. As input
data, the experimental data set obtained from the
experimental design supplied by CCD space (Table
2) via the RSM was employed. The network was
trained using the Multi-Layer Perceptron (MLP)
Levenberg-Marquardt (LM) method (trainlm
the inputs and targets. The network was made up
parameters: time, concentration, and pH), neurons
(the hidden layer), and the output layer (which
expressed in %) (Fig. 1). The input data with 20
samples were divided randomly (dividrand) into
a training set (75%-14 points), validation (15%-
3 points) and te s ting sets (15%-3 points). Based
on R2 and mean square error values, the ideal
number of hidden layer neurons was determined
by trial and error. More data for training decreases
performance. The training was s topped when the
network generalization was improved indicated
by the increase in MSE error of the validation
samples. To eliminate network error, the input and
output variables were normalized between 0 and
1 [17].
2.5. Co s t e s timation theory for the biosorption
process
The techno-economic evaluation of the CiL-
plant-driven bioremediation of the comparative
removal of PCP and TCP from an aqueous solution
was determined following the e s tablished co s t-
[15].
alternative co s t models were used to describe the
feasibility of CiL-plant biosorption of PCP and
model equation described in Equations 2-5. The
total co s t for the biosorption of PCP, and TCP
from 1.0 L of the aqueous solution to CiL-plant at
optimum operating conditions was evaluated using
Table 1. Showing experimental factors in terms of coded values
Factor Name Units Minimum Maximum Coded Low Coded High Mean Std. Dev.
A Time Days 5.00 25.00 14.50 8.87
B Conc mgL-1 50.00 250.00 122.50 67.81
C pH 4.00 9.00 5.85 2.06
Anal. Methods Environ. Chem. J. 6 (1) (2023) 79-99
83
Table 2. Design matrix in terms of actual and predicted values for the RSM and ANN optimization process
Factors
Response 1: Response 1:
%PCP Removal Eciency %TCP Removal Eciency
Std Run
A:
Time
(Days)
B:
Concentration
(mg L-1)
C:
pH Actual
RSM
predicted
values
ANN
predicted
values
Actual
RSM
predicted
values
ANN
predicted
values
11 1 25 250 4 78.31 77.91 78.04 78.74 78.70 78.76
14 2 15 100 6 52.70 54.81 52.64 66.19 64.97 61.62
18 3 5 100 9 7.56 12.20 7.56 4.81 8.34 5.78
8 4 5 100 6 50.37 33.52 50.32 32.33 21.30 31.17
16 5 25 100 4 82.00 77.11 81.96 85.71 81.89 85.75
13 6 15 100 9 36.33 27.88 36.33 52.29 33.60 48.70
1 7 5 50 4 34.64 35.65 34.6 9.04 9.15 9.07
17 8 25 100 9 49.26 48..85 49.19 53.00 53.81 52.85
10 9 15 100 6 52.70 54.81 52.64 66.19 64.97 61.62
12 10 25 100 6 73.00 81.40 76.81 81.05 83.45 80.53
311 15 100 6 52.70 54.81 52.64 52.70 64.97 61.62
15 12 5 250 4 13.24 13.58 12.71 3.85 3.89 3.81
2 13 25 50 4 90.00 87.99 85.37 82.09 81.87 82.18
7 14 5 100 9 7.56 12.20 7.56 4.81 8.31 5.78
9 15 5 100 4 16.86 21.75 27.52 5.00 8.73 2.12
6 16 15 100 6 52.70 54.81 52.64 66.19 64.97 61.62
4 17 5 250 4 13.24 13.58 12.71 3.85 3.89 3.81
19 18 25 100 9 49.26 48..85 49.19 53.00 53.81 52.85
5 19 5 50 4 34.64 35.65 34.60 9.04 9.049.15 9.07
20 20 25 250 4 78.31 77.97 78.04 78.74 78.70 78.76
Fig. 1. ANN network of the PCP and TCP optimization sequence
the expression shown in Equation 3. The energy
consumption (EC) was evaluated using Equation 2
[15, 23]. and given by:
(Eq.2)
Where PC is the power consumption by the device
(kW), f is the load factor. In a full mode, f =1, t is
the time of usage of the device (hour), and C is the
energy e s timated co s t ($) per (KWh) in Nigeria as
of the month of April 9, 2021.
Total co s t is a function of all co s ts, including
Analysis and Biosorption of P&T chlorophenol in Cana indica. L Plant Chris tian Ebere Enyoh et al
84
biosorbent production, labour, and energy. Cm is the
co s ts incurred from transportation, and renting [24].
(Eq.3)
(Eq.4)
Where FO is the return on the selected and forgone
performance of CiL-plant for the bioremediation of
aqueous medium and CO is the return on chosen option
from PCP versus TCP, and CB is the opportunity co s t
based derived based on environmental impact and
regulatory risk (Eq. 4). In this case, the return on
as a function of direct and indirect co s t [15]. The
parameter F0
model Equation 5, expressed as:
(Eq.5)
3. Result and Discussion
In our previous s tudies [8-10], the results for the
removal of PCP and TCP have been presented. This
current s tudy is a s tep further in which removal
processes are optimized and predicted using RSM
and ANN, respectively, to determine the optimum
operating variable for modeling the performance
CiL-plant-driven bioremediation process.
analysis for the method was evaluated in the current
s tudy to ascertain the feasibility of the CiL-driven
bioremediation of PCP and TCP in aquaponia
3.1. Central composite design modeling of the
CiL-driven biosorption process
following the biosorption of PCP and TCP to CIL-
plant from an aqueous solution follow a second-
order quadratic model shown in the ANOVA (Tables
3 and 4). Tables 3 and 4 showed that the selected
quadratic model recorded consi s tent outputs
from the CCD that adequately describes the CiL-
plant-driven biosorption of PCP and TCP from
an aqueous solution. It was observed that model
f-values PCP (30.55) and TCP (62.75) obtained a
0.05. This s tati s tical output indicates that there
is only a 0.01% chance that f-values this large
could occur in the optimization modeling of the
phytoremediation process variables due to noise
[19, 24]
space sugge s ts that the quadratic model terms and
subsequent assumptions on the phytoremediation
The s tati s tical output also sugge s ts that the quadratic
driven biosorption of PCP and TCP from an aqueous
medium [21]
the removal of PCP from the aqueous medium
e s tablished that the predicted R² (0.8322), adju s ted
R2 (0.9256), is in reasonable agreement with the
the central composite design space. Similarly, the
model predicted R² (0.9329) was also in reasonable
agreement with the adju s ted R² (0.9630) reported for
the CiL-plant biosorption of TCP from an aqueous
solution. These r-squared values are close to unity
that the selected quadratic model description of
the CiL-plant-driven phytoremediation process is
[19, 21, 22].
However, where the adequacy of precision output
>4 is desirable [15], the selected quadratic model
recorded adequacy of precision (16.11) value
measures the signal-to-noise ratio (16.11), and the
model f-value (5.36) can be used to navigate design
space for modeling the PCP removal rate [24].
The adequacy of precision (19.41), and signal ratio
(19.41) recorded for the TCP biosorption modeling
were
following the design space adequately describes
the modeling of the biosorption of PCP and TCP
reported with the central composite design space,
not counting those required to support hierarchy.
Anal. Methods Environ. Chem. J. 6 (1) (2023) 79-99
85
pH, and initial concentration, and the second-order
outcome indicated that contact time (A), initial
concentration of CiL-plant (B), and pH (C) all have
biosorption of PCP from aqueous solution. The
impact on removing PCP from aqueous solution
compliance to CiL-plant at p-values <0.100
[15]. The s tati s tical outcome sugge s ts model
of model factors A*B, and A*C which transcends
to contact time*concentration (A*B), and contact
the biosorption of PCP to Canna indica. L (CiL-
plant) in the combined sy s tem. Comparatively,
the quadratic model s tati s tics and assumptions
describing the CiL-plant-driven TCP removal rate
e s tablished that contact time (A) and pH (C) of
on the TCP removal from the aqueous solution.
Also, the model assumption e s tablished that a
higher order degree of contact time (A²) and pH (C²)
to Canna indica L. (CiL-plant). This translates to
second-order impact on TCP biosorption to CiL-
plant. The contact time-initial concentration (A*B)
and PCP to Canna indica L. Consequently, the
contact time and optimized initial concentration
of samples are consequential to the overall
performance of CiL-plant in Aquaponia. The results
e s timate representing the expected change in TCP
[21]
for the CiL-plant driven biosorption of PCP from
aqueous solution. This range of VIFs output (1.22
of TCP from an aqueous solution. The model-
e s tablished VIFs outputs fell within the range
orthogonal design [17]
are adju s tments around that average based on the
plant. The factors are orthogonal when the VIFs
collinearity at VIFs outputs >1 [17]. The moderate
VIFs recorded with the CCD indicate a negligible
level of severity of the correlation of factors [21].
Consequently, the VIFs <10 recorded for PCP and
TCP are tolerable. The summary based on the VIFs
obtained via the e s tablished quadratic model, the
subsequent model hierarchy based on the level
the biosorption of PCP, and TCP to Canna indica L.
(CiL-plant) from an aqueous solution follows the
Table order.
The e s tablished quadratic model equations
describing the biosorption of PCP, and TCP to
Canna indica L were obtained from the CCD
optimization outputs. The outcome showed that the
given by Equation 6, and the TCP removal rate is
described by Equation 7.
PCP Contact time > Initial Concentration > pH > Time*pH > Time*Initial Concentration
TCP Contact time > Initial Concentration > pH > Time*pH > Time*Initial Concentration
Analysis and Biosorption of P&T chlorophenol in Cana indica. L Plant Chris tian Ebere Enyoh et al
86
Table 3. ANOVA for the Quadratic modeling of PCP biosorption to CiL-plant
Source Sum of Squares df Mean Square F-value p-value
Model 12028.39 8 1503.55 30.55 < 0.0001 Signicant
A-Time 6707.47 1 6707.47 136.28 < 0.0001
B-Concentration 421.62 1 421.62 8.57 0.0138
C-pH 497.35 1 497.35 10.11 0.0088
AB 66.62 1 66.62 1.35 0.2693
AC 215.55 1 215.55 4.38 0.0604
A² 14.93 1 14.93 0.3034 0.5928
B² 97.68 1 97.68 1.98 0.1865
C² 450.18 1 450.18 9.15 0.0116
Residual 541.40 11 49.22
Lack of Fit 541.40 3 180.47 Not signicant
Pure Error 0.0000 8 0.0000
Cor Total 12569.79 19
Pred R2 2 2
Table 4. Showing ANOVA for Quadratic modeling of the TCP biosorption to CiL-plant
Source Sum of Squares df Mean Square F-value p-value
Model 18743.53 8 2342.94 62.75 < 0.0001 Signicant
A-Time 10015.46 1 10015.46 268.24 < 0.0001
B-Concentration 29.10 1 29.10 0.7794 0.3962
C-pH 284.22 1 284.22 7.61 0.0186
AB 2.02 1 2.02 0.0540 0.8205
AC 473.96 1 473.96 12.69 0.0045
A² 338.64 1 338.64 9.07 0.0118
B² 1.12 1 1.12 0.0301 0.8654
C² 300.46 1 300.46 8.05 0.0162
Residual 410.71 11 37.34
Lack of Fit 274.23 3 91.41 5.36 0.0257 Not signicant
Pure Error 136.49 8 17.06
Cor Total 19154.25 19
Pred R2 2 2
3.2 Optimization outputs following the CiL-plant-
driven biosorption process
The interpretation of the CiL-plant-driven biosorption
of PCP, and TCP from an aqueous solution follows
from the e s tablished model Equations 6-7. The results
based on the interpretation of the optimization ramp
(Fig. 2)
conditions describing the be s t performance of the
CiL-plant-driven biosorption of PCP and TCP from
an aqueous solution correspond to pH (4), initial
concentration (50 mg L-1), and contact time (25
days). The predicted optimum based on the quadratic
ramp shown in Figure 2. The predicted optimum
and 81.87 % for the biosorption of PCP, and TCP to
CiL-plant as indicated in the optimization ramp in
Figure 2. The optimum points transcend to a s tandard
deviation of PCP (7.01), and TCP (6.80) from the
actual observations practicable. The outcome is
plots in Figure 3 (a-b)
that the predicted optimum points are located within
the CCD design space [17, 24], and maintained
within the range of the experimental values under
inve s tigation.
The predicted optimum indicates that the be s t
biosorption of PCP, and TCP occurred via an active
Anal. Methods Environ. Chem. J. 6 (1) (2023) 79-99
87
initial concentration of 50 mg L-1 of Canna indica
L. plant. The phytoremediation progressed in a
predominantly acidic medium (pH 4), and requires
biosorption of PCP, and TCP to lower residual that
guarantees su s tainability. The optimization results
e s tablished that under similar optimum operating
conditions, the comparative biosorption rate of
PCP to CiL-plant was mo s t favorable compared to
3.3 Articial neural network performance
validation of the CiL-driven biosorption process
The validation performance in Figures 4 (a-b)
shows how the number of epochs varied with the
MSE for the optimal neural network. The be s t
validation performance was 4.8753 and 2.8482 at
epoch 3 for PCP and TCP biosorption. The scatter
plots depicting the linearity of the output values of
the network with the target values for the training,
te s ting, validation, and overall data (as generated
Fig 2. Optimization ramp for CiL-plant driven biosorption of PCP and TCP
Fig 3: 3D surfaces plot of CCD optimization
of the CiL-plant driven biosorption of: (A) TCP, and (B) PCP
Fig. 3.
Analysis and Biosorption of P&T chlorophenol in Cana indica. L Plant Chris tian Ebere Enyoh et al
88
by the MLP platform) are illu s trated in Figures 5 (a, b).
The predicted R2 values were used to indicate the
linearity – with the training network having the
highe s t value of 0.9999 and 0.9945 for the optimal
neural network for PCP and TCP biosorption,
respectively. The outputs design matrix in
Table 2 (presented in section 2.4 above)
displayed the anticipated responses at various
experimental setups. It can be concluded from the
outline of Figure 4 that the summary of the s tati s tical
and evaluation metrics from the ANN indicates an
increase in model errors as the number of epochs
increased. This outcome reasonably agrees with the
optimization modeling procedure reported in the
literature [16, 27]
minimized [27].
The performance of each model (ANN and RSM)
was validated by evaluating their prediction
accuracy using s tati s tical tools (MSE, RSME, X2
and SSE) [27, 28]. The mathematical equations
representing the s tati s tical tools are summarized in
Table 5. A better predictive model has a high R2
value (almo s t 1) and low s tati s tical errors (close
to 0) [11, 12]. The high R2 values and s tati s tical
errors proved a good correlation with the actual
observations practicable than the RSM. When
compared to the RSM, the ANN outputs yielded
error (R22
The low s tati s tical error indicates reliable
adequacy of precession [25, 26], sugge s ting
minimal error due to noise [27]. However, the
s tati s tical outcome from both optimization tools
is in reasonable agreement with the actual values
obtained from the experimentation with the RSM
ANN output. The RSM performance evaluation
parameters and their interactions on the response
[18]. The s tati s tical model assumptions of the RSM
have been ascertained for reliability, and the design
space (CCD) has been te s ted based on the design
of experiments (DoE). As a result, the predicted
optimum reported for the RSM was employed to
further optimize the CiL-plant driven biosorption
process.
Table 5.
Error factor Equation RSM ANN
MSE 0.0080 0.0036
RMSE 0.0894 0.0604
X22.8478 0.0001
SSE
Predicted R2
0.1600
0.9329
0.0729
0.9954
Where yi, yi*, and ym s tand for the experimental, predicted, and mean value of the actual responses, N represents the number
2: Chi-square and SSE: sum of
squares errors
Anal. Methods Environ. Chem. J. 6 (1) (2023) 79-99
89
012345678
8 Epochs
10
0
10
1
10
2
10
3
10
4
Mean Squared Error (mse)
Best Validation Performance is 2.8482 at epoch 3
Train
Validation
Test
Best
0 1 2 3 4 5 6
6 Epochs
10
-25
10
-20
10
-15
10
-10
10
-5
10
0
Mean Squared Error (mse)
Best Validation Performance is 4.8753 at epoch 3
Train
Validation
Test
Best
Fig. 4.
for the (a) PCP, and (b) TCP biosorption processes
Analysis and Biosorption of P&T chlorophenol in Cana indica. L Plant Chris tian Ebere Enyoh et al
90
Fig. 5(a).
20 40 60 80
Target
10
20
30
40
50
60
70
80
Output ~= 1*Target + -0.11
Training: R=0.99998
Data
Fit
Y = T
40 50 60 70
Target
35
40
45
50
55
60
65
70
75
Output ~= 1*Target + -0.93
Validation: R=0.99622
Data
Fit
Y = T
20 40 60 80
Target
20
30
40
50
60
70
80
90
Output ~= 0.88*Target + 6.8
Test: R=0.98843
Data
Fit
Y = T
20 40 60 80
Target
10
20
30
40
50
60
70
80
90
Output ~= 0.97*Target + 1.9
All: R=0.99433
Data
Fit
Y = T
Anal. Methods Environ. Chem. J. 6 (1) (2023) 79-99
91
Fig. 5(b).
20 40 60 80
Target
10
20
30
40
50
60
70
80
Output ~= 0.99*Target + 0.34
Training: R=0.99448
Data
Fit
Y = T
20 40 60 80
Target
10
20
30
40
50
60
70
80
Output ~= 1*Target + -1.5
Validation: R=0.99949
Data
Fit
Y = T
10 20 30 40 50 60
Target
10
20
30
40
50
60
Output ~= 0.93*Target + 1.7
Test: R=0.99826
Data
Fit
Y = T
20 40 60 80
Target
10
20
30
40
50
60
70
80
Output ~= 0.99*Target + -0.054
All: R=0.9958
Data
Fit
Y = T
Analysis and Biosorption of P&T chlorophenol in Cana indica. L Plant Chris tian Ebere Enyoh et al
92
3.4 Eects of experimental variables on the
overall performance of the CiL-plant
on the CiL-plant driven biosorption of PCP, and
TCP from an aqueous solution were based on the
CCD s tati s tics (VIFs), and model hierarchy model
parameters shown in section 3.1. The relative
contact time (A) on the biosorption of PCP, and
TCP aqueous solution compliance to CiL-plant
in the single and interactive sy s tem when the
in Figures 6 and 7.
3.4.1 Antagoni s tic eect of contact time on the
Phytoremediation process
Figure 6
contact time on the biosorption of PCP, and TCP
to CiL-plant at the optimum concentration (50 mg
L-1). The graph showed that the CiL-plant-driven
biosorption of PCP, and TCP from an aqueous
increased in days. The overall performance under
maximum PCP removal rate (90%) recorded in
25 days and was consi s tent with the maximum
removal rate recorded for TCP (87.99%). At
contact time
less than the outcome sugge s ts biosorption
of the chemical species (TCP) was slow on the
CiL-plant surface or had not yet occurred for
PCP removal from the aqueous solution compared
with the performance of CiL-plant biosorbent on
the removal of TCP at maximum contact for 25
biosorption of the contaminants (TCP and PCP) on
CiL-plant from the solution to reach equilibrium
[29]. The outline of the red and blue bars indicated
that biosorption of TCP reached equilibrium fa s ter
level of tolerance of the CiL-plant index to PCP-
contaminated medium [8]. Overall performance
of the CiL-plant as an active biosorbent for the
su s tainable removal of PCP and TCP to guarantee a
tolerable residual contaminant level. The outcomes
time on the removal of PCP and TCP from an
aqueous solution recorded at a p-value of 0.0001 at
Fig 6.
at an optimum initial concentration (50 mg L-1) and pH 4
Days
Removal Rate (%)
0 5 10 15 20 25
0
20
40
60
80
100
TCP Removal rate (%) PCP Removal rate (%)
Anal. Methods Environ. Chem. J. 6 (1) (2023) 79-99
93
3.4.2 Antagoni s tic impact of pH on the
Phytoremediation process
biosorption of TCP from an aqueous solution at an
initial concentration of 100 mg L-1 at pH 4 is shown
in Figure 7. The graph showed that the biosorption of
PCP and TCP to CiL-plant at varying pH decreased
rapidly in an alkaline solution. The performance
of the CiL-plant translates to a removal rate of
49.3% for PCP, and 53.2% for TCP at pH 9 and an
optimum of 25 days, respectively. The removal rate
medium transcending to 82% for PCP, and 85.7%
for TCP at pH 4. The protonated chlorophenols
were more absorbable [30], which accounted for
and TCP at the lower pH value. The analysis of the
the CiL-driven biosorption process favored TCP
removal from an aqueous medium in an acidic
medium compared to PCP at a p-value value of
was consi s tent with the optimum pH 2 reported for
the removal of PCP and TCP reported in the work
of Radhika and Palanivelu et al. [29]. The outline of
[8], sugge s ting a superior solubility of 2,4,6-TCP
in water than PCP at optimum pH 4 in aquaponia.
corresponding (82, 85.7%) e s tablished that contact
performance of CiL-plant-driven phytoremediation
for su s tainability.
3.4.3. Synergetic impact of Time-pH and
concentration-pH on the Biosorption process
pH*Time and Time*initial concentration on
the response PCP and TCP removal rate was
at a p-value less than 0.005 and 95% CI. The 3-D
surface plots in Figure 8 (a-d) were obtained from
the response surface design space to under s tand
better how the biosorption process works under
[5, 8]. The red color gradient corresponds to higher
yellow contour gradient corresponds to moderate
Fig 7.
at optimum Time (25days) and initial concentration of 100 mg L-1 at pH 4
pH
Removal rate (%)
4 5 6 7 8 9
40
50
60
70
80
90
TCP Removal rate (%) PCP Removal rate (%)
Analysis and Biosorption of P&T chlorophenol in Cana indica. L Plant Chris tian Ebere Enyoh et al
94
dominant greenish-blue contour orientation
can be traced to possible charge reversal from
surplus ions arising from the binary solution of
PCP and TCP under changing pH in the medium.
These excess negative charges contributed to the
building up of the concentrations of the PCP and
TCP molecules in an aqueous solution causing
the red hue gradient on the base of the surfaces
Figures 8 (a-b)
of the solution decreased from 9 to predominantly
pH 4, as the exposure time of CiL-plant increased
from 20 to 25 days. The phytoremediation
performance of the biosorbent in aquaponia
transcends to increase in PCP biosorption rate
from 70 to 90%, while the TCP removal rate
increased from 70 to 87.99%, as illu s trated by the
Figure
8 (a,b). The pH depression from 7 to 4 yielded
better performances that can be attributed to the
the acidic window and the prolonged exposure
time of 25 days. This outcome is indicated by the
curvatures of the dominant red contour deviation
from the yellowish-green contour lines shown
in Figure 8 (a,b). The basic tendency of CiL-
plant-driven biosorption of PCP and TCP can be
removal rate towards a dominantly acidic medium
of the superior pH on the overall biosorption of
PCP and TCP in aquaponia was attributed to the
dissociation of mo s t chlorophenol in the form of
a salt which loses its negative charge easily when
pH is increased [30]
e s tablished that CiL-plant is tolerant in a solution
of PCP, compared to TCP. This observation agrees
with previous research works reported in the
literature [8, 30]
initial concentration*contact time is illu s trated
in the outline the contour plot in Figure 8 (c,d).
The overall performance of the CiL-plant in the
phytoremediation remediation of aquaponia is
illu s trated by the deviation of the green color
contour from the dominant blue gradient on the
base of the surfaces in Figure 8c and Figure 8d,
respectively. The curvature of the light green
from the dominant blue color orientation is
attributed to areas of good performance of the
biosorbent morphology and adaptation of Canna
indica L for the removal of PCP and TCP. The
data points and orientation of the dominant
blue contour lines transcend to areas of poor
performance of the biosorbent on PCP and TCP
in solution. The intensity of the blue contour
gradients in Figure 8 (c,d) confirmed that the be s t
performance of the CiL-plant is adapted to an
initial concentration of less than 100 mg L-1. The
decreased as the initial concentration was
increased beyond 100 to 250 mg L-1. The output
<65% for TCP, as indicated by the curvature of
the blue contour gradient in Figure 8 (c,d) and
corresponding 3D surfaces in Figure 3. This
outcome indicates that the initial concentration
plant biosorption of PCP and TCP [30, 31]. In
contra s t, the contact time or exposure had a
reports on TCP biosorption [29]. The authors
reasoned that if the concentration was to be
increased slightly beyond 100 mg L-1, and a
reduction in equilibrium exposure time below 25
days would decrease mass transfer to the surface
from 90% to 40%. This outcome e s tablished
of initial concentration and exposure time on
the overall performance of the CiL-plant-driven
phytoremediation process in Aquaponia.
Anal. Methods Environ. Chem. J. 6 (1) (2023) 79-99
95
Fig. 8.
of Aquaponia (a) pH and Time on PCP, (b) pH-Time on TCP, (c) Conc-Time on PCP, (d) Conc-Time on TCP
Analysis and Biosorption of P&T chlorophenol in Cana indica. L Plant Chris tian Ebere Enyoh et al
96
3.5. Co s t analysis of the treatment process
the biosorption process as a decision-making tool
to te s t the feasibility of the active CiL-plant as a
biosorbent for removing PCP and TCP from an
aqueous solution. The co s t of the phytoremediation
operation is based on the performance of the active
CiL-plant, energy consumption, transportation to
the remediation site, labor and technology used to
remove contaminants [4, 8], and environmental and
regulatory risk. The techno-economic feasibility of
the biosorption process and phytotoxicity handling
necessitates using low-co s t materials (Canna
indica L.) with negligible environmental impact
and regulatory risk. The operating co s t of treating
1.0 L of the aqueous solution was calculated by
considering the co s t of preparing the 100 mg L-1
initial concentration of the aqueous solution for CiL-
plant as biosorbent. The labor co s t was determined
as a function of the number of working personnel
on board for the treatment operation. The power
consumption rate per unit of equipment utilized at
full scale (f=1), and the time spent following the
model report from previous research [15], were
evaluated following equations 2-5 presented in
section 2.5.
Analysis of Figure 9
co s t of energy consumption corresponds to $ 27.80
for PCP, and $ 21.28 for TCP, respectively. It can be
observed from Figure 9 that, the co s t of operating
the phytoremediation process for PCP removal was
slightly higher than the co s t of TCP removal in terms
of energy consumption by $ 6.52. The preparation
of 100 mg L-1 initial concentration of CiL-plant
for removal of PCP from the aqueous solution
co s t $ 177.4 again s t $ 176.2 for removal of TCP
from the aqueous solution under similar operating
conditions. The labor co s t was projected to be $
100.2 per annum, while the co s t of transportation of
materials and personnel on board to the remediation
site was $12.00, irrespective of operating with PCP
or TCP. It can be concluded from the analysis of
Figure 9 showed that the overall co s t for using
the biosorption of PCP from aqueous solution to
CiL-plant at optimum conditions was computed as
$ 321.20 and $ 313.48 for TCP, respectively. The
analysis of the phytoremediation process proved
that, at the e s tablished optimum condition, the
opportunity co s t of operating the biosorption of
TCP from aqueous solution to CiL-plant would
save $ 7.72 compared with PCP for su s tainability.
The authors reasoned that the outcome is largely
due to higher solubility and rapid biosorption of
TCP to the surface of the CiL-plant in aquaponia,
irrespective of the longer exposure time required
for the CiL-plant driven biosorption of PCP and
TCP to reach equilibrium.
Fig. 9. Co s t evaluation summary of the CiL-plant-driven biosorption treatment
Anal. Methods Environ. Chem. J. 6 (1) (2023) 79-99
97
3.6. Comparison of CiL-plant for the remediation
of PCP and TCP from solution
The performance comparison of the biosorption of
PCP, and TCP from aquaponia was analyzed and the
report is summarized in Table 6 below. The previous
research reports on the CiL-plant phytoremediation
analysis applied the one-factor-at-a-time (OFAT)
approach for determining the optimum PCP and
TCP removal rate. The current s tudy applied the
design of experiment (DoE) approach via the RSM
for the optimization modeling of the uptake of PCP
from the comparison of the results showed that with
the optimum reported via the OFAT approach. The
PCP, respectively.
4. Conclusion
The techno-economic evaluation and optimization
modeling of the competitive biosorption of PCP
and TCP from aqueous solution to the Cana indica
plant have been inve s tigated. The aqueous solution
of fertilizer contaminated with PCP and TCP was
prepared. The CiL-plant-driven phytoremediation
of the aqueous medium was s tudied at varying
pH, initial concentration, and con s tant time based
on the design of experiments. The optimization
modeling tools for ANN and RSM have yielded
good s tati s tical evaluation metrics for modeling
the CiL-plant-driven phytoremediation process.
2
adopted RSM optimization outputs have to te s t their
reliability based on DoE. The predicted optimum
corresponds to pH, concentration, and exposure
time of 4, 50 mg L-1, and 25 days guaranteed PCP
e s tablished optimum condition required $7.75 more
for su s tainable PCP removal than TCP.
5. Acknowledgement
Chemi s try Laboratory, Imo s tate University, for
their support during the experimental set-up and
analysis
6. Conict of intere s t
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Analysis and Biosorption of P&T chlorophenol in Cana indica. L Plant Chris tian Ebere Enyoh et al
Anal. Methods Environ. Chem. J. 6 (1) (2023) 100-114
Research Article, Issue 1
Analytical Methods in Environmental Chemis try Journal
Journal home page: www.amecj.com/ir
AMECJ
grown and greenhouse-grown tomatoes using liquid
chromatography-mass spectrometry
Fatemeh Norouzi a,b, Maryam Faraji a,b,*, Ramezan Sadeghi c, Ali Faghihi-Zarandi d,
and Farshid Shabani Boroujeni e
a Environmental Health Engineering Research Center, Kerman University of Medical Sciences, Kerman, Iran
b Department of Environmental Health Engineering, Faculty of Public Health,
Kerman University of Medical Sciences, Kerman, Iran
c Department of Environmental Health of Engineering, Faculty of Health,
Shahrekord University of Medical Sciences, Shahrekord, Iran
d Department of Occupational Health Engineering and Safety at Work, Faculty of Public Health,
Kerman University of Medical Sciences, Kerman, Iran
e Department of Plant Protection, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
ABSTRACT
greenhouse-grown tomatoes and homemade pas te based on the quick,
(QuEChERS) before determined by the liquid chromatography-mass
of deltamethrin (DLM) and acetamiprid (ACT) in raw tomatoes of
greenhouse-grown was obtained at 91.42 and 90.00%, respectively,
86.34%). Maximum reduction percentages of the DLM in pas te under
95.86% and 93.11%, respectively. The residual concentration of both
DLM (91.42%) and ACT (90.00%) in the greenhouse decreased more
reached below the MRL in a shorter time after spraying (2 days).
Considering the pre-harves t interval (PHI) period of deltamethrin
and abamectin can reach their residual concentration to the MRL in
both conditions, which were determined by LC-MS. According to
the results of the current s tudy, 7 and 5 days can be sugges ted as
tomatoes, respectively. Therefore, using pes ticides in the proper
dosage, considering appropriate PHI, and harves ting can reduce their
residues in agricultural products.
Keywords:
Liquid chromatography-mass
spectrometry,
Analysis,
QuEChERS method,
Pes ticide residues,
Field and Greenhouse-grown tomatoes
ARTICLE INFO:
Received 14 Nov 2022
Revised form 20 Jan 2023
Accepted 12 Feb 2023
Available online 29 Mar 2023
*Corresponding Author: Maryam Faraji
Email: m.faraji@kmu.ac.ir and m_faraji28@yahoo.com
https://doi.org/10.24200/amecj.v6.i01.234
------------------------
1. Introduction
Solanum
Lycopersicum
processed due to having high antioxidants such
as ascorbic acid, vitamins E and A, carotenoids,
of cardiovascular diseases and prevent diabetes and
cancer [1, 2]. Several pes ticides are used to maintain
agricultural products. Improper consumption of
pes ticides in farm products and non-compliance
with the pre-harves t interval (PHI) period can
101
pes ticides in the crops [3]. Therefore, the European
Union (EU) has set a maximum pes ticide residue
limit (MRL). The MRLs for s tudied pes ticides in the
present s tudy included acetamiprid, deltamethrin,
70, and 90, respectively, regardless of the growth
conditions [4, 5]
residual acetamiprid in okra showed that using 10 g
and 20 g of 20% acetamiprid per hectare led to the
residual of 2.034 and 4.044 mg kg-1, respectively
[6]
the residual pes ticides in the greenhouse tomatoes
during 2, 5, 7, 10, 14, 17, and 21 days after spraying
showed that the residual of acetamiprid, diazinon,
imidacloprid, and pirimicarb declined after the PHI
period approached [7]. Mohamed et al. reported that
imidacloprid decomposed fas ter than acetamiprid
in tomatoes grown under greenhouse conditions
[8]. Iran ranks seventh globally, accounting for
4.7% of the total world production of tomatoes,
with an annual production of 5.8 million tons
and an average yield of 38 tons per hectare [9].
According to the high production and consumption
of raw and processed tomatoes in Iran and the use
of high levels of pes ticides in their cultivation, this
s tudy aimed to determine deltamethrin, abamectin,
and acetamiprid residues in cultivated tomatoes
processed using QuEChERS (quick, easy, cheap,
residual pes ticides by liquid chromatography-mass
spectrometry (LC-MS) method.
In this s tudy, the residual concentrations of
pes ticides, including acetamiprid, deltamethrin,
and abamectin, were extracted and determined by
the QuEChERS procedure coupled to LC-MS. The
residual concentration of three high-consumption
pes ticides of Iran in raw and processed tomatoes
was determined and compared. Also, the residual
concentration of the three mentioned pes ticides
tomatoes were s tudied and compared together.
The current s tudy was innovative in comparing the
residual pes ticides.
2. Material and Methods
This s tudy was done in several s tages included
greenhouse and harves ting, preparation of the
samples through the QuEChERS method and their
analysis of samples via LC-MS and s tatis tical
analysis of the data. S tudy s tages are illus trated in
Schema 1.
Schema 1. S tudy s tages of sampling, the QuEChERS preparation method and determination by LC-MS
Determination and analysis of pes ticides by LC-MS Fatemeh Norouzi et al
102
2.1. Ins trumental
LC-MS is an accurate and precise method to
quality control analysis of compounds. It can
also be used in combination with other analytical
methods to further elucidate the components of
mixtures [17]. LC-MS (model: Waters Alliance
2695 (UK)) using a matrix-matched method was
used to analyze samples in the present s tudy. The
type of detector was Micromass Quattro Micro
API Triple Quadruple Mass Spectrometer (UK).
Column 100 Å, 150 mm × 2.1 mm × 1.5 µm. The
samples of 20 µL were injected into the device.
Chromatograms of the s tandard samples to provide
calibration curve have been illus trated in Schema 2.
2.2. Chemicals and reagents
S tandards of Acetamiprid (99.9%), abamectin
(95%), deltamethrin (98.5%), deltamethrin
(2.5%EC), acetamiprid (20%SP), abamectin
(1.8%EC) and other chemicals and reagents included
acetonitrile, anhydrous magnesium sulfate, the
internal s tandard of triphenyl phosphate, sodium
chloride, trisodium citrate dihydrate, disodium
hydrogen citrate, primary, secondary amine (PSA),
and carbon adsorbent (C18) were purchased from
Sigma Aldrich, Germany. Dilutions of 50, 100,
250, 500, and 1000 ng g-1 were used to plot the
calibration curve of the pes ticides using a matrix-
matched method. The limit of detection (LOD),
equation of calibration, and the MRL for the s tudied
pes ticides are mentioned in Table 1.
2.3. Planting and spraying of tomatoes in the
eld and greenhouse
considered for planting tomatoes outdoors and in
a greenhouse in the summer of 2020. The average
temperature in the s tudy period, namely the summer
dis tance between tomato plants was considered
to be 40 cm. Dis tances of 120 and 100 cm were
overlap and possibly dispersion of pes ticides
through the wind. The control samples were grown
on the unsprayed terrace. Randomized spraying
was performed with a 20 L calibrated rechargeable
back sprayer (model: IAC CODE: E2) according to
the doses recommended by the Iran Plant Protection
Organization, including 0.6 liters per hectare for
abamectin, 300 cc per hectare for deltamethrin, and
Schema 2.
and abamectin for calibration curve
Anal. Methods Environ. Chem. J. 6 (1) (2023) 100-114
103
0.5 kg per 1000 liters of water for acetamiprid. The
physical and chemical characteris tics of the s tudied
pes ticides [10-12] are reported in Table 2.
2.4. Sample harves ting
PHI period for deltamethrin and abamectin was
in the case of deltamethrin and abamectin was
done in the sugges ted PHI period and before and
after that, 1, 2, 3, 4, and 5 days after spraying. The
acetamiprid. Thus, sample harves ting in the case of
acetamiprid was done according to similar s tudies
[13, 14]
9, and 11 days after spraying. After time elapsed, 2
mixed and, after coding, placed in a black bag and
maintained at 4 °C. Then, part of the samples was
homogenized after washing to measure the residual
pes ticide in the raw sample, and the other part was
used to prepare homemade tomato pas te. To make
tomato pas te, the washed tomatoes were chopped,
salted, and s tored at room temperature for 24 hours.
Then, the tomato juice was s trained and heated at
96 °C for one hour. After cooling, the samples were
packaged and coded separately.
Also, one sample of each treatment was taken
one hour after spraying to compare the amount of
pes ticide residues in washed and unwashed tomatoes.
one part was washed with tap water, and another part
was reserved unwashed. Finally, the samples were
maintained at -21 °C until experiments.
Table 1.
residue limit (MRL) for the s tudied pes ticides
Pes ticide Abamectin Deltamethrin Acetamiprid
Chemical s tructure
Chemical formula C95H142O28 C22H19Br2NO3C10H11ClN4
MW (g mol-1)1732.1 505.21 222.68
Water solubility 1.21 mg L-1 at 25 ºC <0.002 mg L-1 at 25 ºC 4.25 g L-1 at 25 ºC
Octanol/water partition
4.4 6.10 0.8
Chemical Family
Insecticide, a natural
fermentation product
of soil-dwelling
actinomycete,
S treptomyces avermitilis
Pyrethroid insecticide Neonicotinoid insecticide
Table 2. Physical and chemical characteris tics of the s tudied pes ticides
Pes ticide LOD
(mg kg-1)
LOQ
(mg kg-1)
Regression equation
of calibration R2*MRL
Abamectin 13.2 40 y=3.37761x +0.313794 0.9848 0.09
Deltamethrin 13.2 40 y=7.78742x-7.7343 0.9931 0.07
Acetamiprid 13.2 40 y=11.8763x-8.70884 0.9946 0.50
*MRL: maximum residue limit of the European Union
Determination and analysis of pes ticides by LC-MS Fatemeh Norouzi et al
104
2.5. Preparation and analysis of samples
The QuEChERS method, with its high sensitivity, is
used to extract the residual pes ticides in the products
in many reference laboratories [15]. To extract
pes ticides in the current s tudy by the QuEChERS
method, each sample was homogenized in a blender,
and 10 g of samples were transferred to the centrifuge
tube. Then, 10 mL of acetonitrile and 100 µL of the
internal s tandard of triphenyl phosphate were added
to each centrifuge tube at the concentration of 10
ppm. Next, 4g anhydrous magnesium sulfate, 1.0 g
sodium chloride, 1.0 g Trisodium citrate dehydrate,
and 0.5 g disodium hydrogen citrate were added to
each centrifuge tube after a vigorous shake for one
minute. Again, the mixture was vortexed for one
minute at 5000 rpm for 5 minutes at -10 °C. Then, 3
mL of the transparent top layer was transferred into
the tube containing 75 mg PSA, 450 mg anhydrous
magnesium sulfate, and 75 mg C18 adsorbent.
for one minute and re-centrifuged [16]. Residual
concentrations of pes ticides in the samples were
measured by the method of LC-MS. LC is an accurate
and precise method to separate, identify and analyze
adopted for quality control analysis of compounds. It
can also be combined with other analytical methods
to further elucidate the components of mixtures [17].
2.6. S tatis tical analysis
S tatis tical analysis was performed using R software
version 3.4.1. Results were reported as the mean
ANOVA. P-value < 0.05 was considered as the
3. Results and discussion
3.1. Deltamethrin
The EU has determined the deltamethrin
MRL in tomatoes as 70 µg kg-1. The residual
concentration of deltamethrin was reached less
(Fig. 1a). Therefore, considering the PHI period
its residual concentration met the MRL in both
conditions. Residual concentration and reduction
percentage of deltamethrin in raw tomato and
in Table 3. Comparison between the mean residual
times from 1 to 5 days with the MRL showed a
of percentage reduction in raw tomato between
residual concentration of deltamethrin in tomato
grown products showed a decreasing trend (Fig.1b).
The concentration of deltamethrin in the pas te from
day after spraying. While its removal was more
than 95% in the greenhouse products (Table 3).
3.2. Abamectin
The comparison of the residual concentration
tomatoes with the MRL of 90 µg kg-1 was shown in
Figure 2a. The residual concentration of abamectin
was less than MRL on the second day after spraying
in both growing conditions (58 µg kg-1 and 77 µg
kg-1, respectively). The PHI period for abamectin
PHI period for abamectin can reach its residual
concentration below the MRL in tomatoes grown
was observed in the residual concentration of
grown and greenhouse-grown products (Fig. 2b).
The concentration of abamectin was reduced to
more than 89% in the pas te made from crops in both
(Table 3). The residual
concentration of abamectin in the pas te can reach
below 40 µg kg-1-1 in the
greenhouse, considering the PHI period in tomato
(three days).
Anal. Methods Environ. Chem. J. 6 (1) (2023) 100-114
105
Fig. 1.
and greenhouse-grown tomatoes (a) and pas te (b)
Determination and analysis of pes ticides by LC-MS Fatemeh Norouzi et al
106
Fig. 2.
and greenhouse-grown tomatoes (a) and pas te (b) and the EU maximum residue limits
Anal. Methods Environ. Chem. J. 6 (1) (2023) 100-114
107
Table 3. Residual concentration and reduction percentage of deltamethrin, abamectin, and acetamiprid
Tomato pas teRaw tomatoes
Pes ticide
GreenhouseFieldGreenhouseField
PR
(%)
RC
(µg kg-1)
PR
(%)
RC
(µg kg-1)
PR
(%)
RC
(µg kg-1)
PR
(%)
RC
(µg kg-1)
Day
91.3683.5079.45198.7086.36131.9169.74292.621
Deltamethrin
93.5762.1088.91107.2089.9397.3081.92174.802
95.0847.9092.7769.9192.5171.6188.27113.403
>95.86<4094.5752.5093.8059.8291.5381.914
>95.86<4095.6941.6094.5052.5293.1166.605
-
-90.2893.9891.4282.6384.91145.87
Mean
-
----70-70
MRL
-
----0.43*0.18**0.14*
p-value
73.21101.6079.0679.4070.38112.3475.5892.601
Abamectin
81.7769.1187.4747.5079.7476.8084.7557.802
87.8446.12>89.45<4085.8953.5088.1844.813
>89.45<40>89.45<4088.2144.70>89.45<404
>89.45<40>89.45<40>89.45<40>89.45<405
--------
Mean
-------90
MRL
--------
p-value
91.27328.1187.32476.3385.67538.4177.76835.613
93.53242.9190.93340.7188.85418.9184.63577.6225
94.25215.8192.61277.4090.42359.7187.69462.337
Acetamipride
94.85193.2193.77234.1191.97301.6290.11371.619
95.41172.5194.39210.6193.08259.8191.49319.5211
93.86230.5191.80307.8390.00375.6986.34513.34
Mean
-----500-500
MRL
--0.19**--0.06*0.22**0.89*
p-value
*Comparison between mean concentration and maximum residue limit (MRL),
RC: Residual concentration
PR: Percentage reduction
Determination and analysis of pes ticides by LC-MS Fatemeh Norouzi et al
108
3.3. Acetamiprid
-1 as
the MRL level for acetamiprid. The mean residual
and MRL level is compared in Figure 3a. The results
showed that the residual concentration of acetamiprid
µg kg-1
(419 µg kg-1) reached below the MRL by LC-MS.
acetamiprid. Therefore, the acetamiprid PHI period
greenhouse-grown tomato can be sugges ted based
in the mean reduction percentage of acetamiprid in
Comparing the mean residual concentration of
acetamiprid in raw tomato and the MRL (500 µg
kg-1
(p=0.89) and greenhouse (p=0.06). The residual
concentration of the acetamiprid in the tomato pas te
products followed a decreasing trend (Fig. 3b). The
concentration of acetamiprid was approximately
reduced to 95% in the pas te made from crops in
both conditions after 11 days (Table 3). The mean
percentage reduction of acetamiprid in the pas te
Elbashir et al. measured the residual concentrations
that the pes ticide residues of fenpropathrin after 27
after three days immediately after washing reached
[18].
organochlorine, pyrothyroid, and dicarboximide
in greenhouse-grown tomatoes, the residual
concentration of deltamethrin was reported in the
range of 1-0.01 mg kg-1. The residual concentration of
pes ticides in the two s tudied samples was higher than
the MRL [19]
of deltamethrin in greenhouse-grown cucumber
showed that the residual concentration of pes ticide
reached the allowable limit (0.2 mg kg-1
day after spraying and was not measurable on the
seventh day after it [20]
residual concentrations of abamectin, acetamiprid,
abamectin and acetamiprid were reported one hour
after spraying as 5.80 and 1.10 mg kg-1, respectively.
The results of this s tudy showed that ten days after
spraying with abamectin and one day after spraying
with acetamiprid, the residual pes ticides reached
below the EU MRL [21]. The s tudy of Fujita et al
on the residual amount of acetamiprid, azoxys trobin,
greenhouse-grown lettuce showed that the residual
concentrations of pes ticides in the greenhouse crop
dinotefuran, the residual pes ticides in the greenhouse
[22]. According
acetamiprid and imidacloprid in greenhouse-grown
tomatoes reached below the Europe MRL within
[8]
concentration of propamocarb in greenhouse-grown
of propamocarb in the greenhouse crop was higher
[23].
3.4. Comparison of dierent condition
In comparison between mean reduction percentages
in the present s tudy, it can be s tated that residual
concentration of both deltametrin (91.42%) and
acetamiprid (90.00%) in the greenhouse was
respectively) by LC-MS. Abamectin reached below
the MRL in a shorter time after spraying (2 days)
compared to other pes ticides. The extent of pes ticide
residues in the agricultural products depends on
several factors such as the properties of pes ticide,
its formulation and applied concentration, light,
temperature, plant morphology and plant growth
factors [24].
Anal. Methods Environ. Chem. J. 6 (1) (2023) 100-114
109
Fig. 3.
and greenhouse-grown tomatoes (a) and pas te (b)
Determination and analysis of pes ticides by LC-MS Fatemeh Norouzi et al
110
In comparison between raw tomato and tomato pas te
in both grow condition, it was found that processing
of the raw tomato through cooking could decrease
the concentration of pes ticides in all experiments
pes ticides in the raw and processed products was
reduction was not observed with the processing
of cooking on the residual pes ticides deltamethrin,
penconazole, cresoxime methyl, cyproconazole,
epoxiconazole, and azoxys trobin in rice, the results
reported the reduction of pes ticides as 20.73% to
57.72% for home cooking, 32.74% to 70.39% for
washing with excess water, and 68.87% to 87.50% for
soaking rice before cooking, respectively [25]. The
eggplant showed that washing 24.73%, boiling
reduction of its residual one day after spraying with
the recommended dose [26]
examined the reduction of non-sys temic and low-
sys temic (indoxacarb, chlorfenapyr, and fenarimol)
and sys temic (acetamiprid) pes ticides in okra after
the cooking process. The residual acetamiprid was
reduced up to 90% using cooking methods, indicating
that the tissues of the okra disintegrated during
cooking, so the internal remnants of acetamiprid
were exposed to water dissolution and thermal
decomposition [27]. The reduction percentage of
pes ticides in washed and unwashed tomato samples
residual pes ticides of abamectin, deltamethrin, and
acetamiprid was observed after washing with tap
water. Rinsing with tap water reduced the residual
concentrations of acetamiprid, abamectin, and
deltamethrin in the crops harves ted during one hour
after spraying up to 66.85%, 51.62%, and 50.52%,
respectively (Table 4). Acetamiprid, as a sys temic
pes ticide, with the highes t solubility in water (4250
mg L-1), had the highes t reduction percentage after
washing compared to the other pes ticides. Washing
processing methods. Many residual pes ticides can
be removed by washing them with tap water. Various
including the location of the pes ticide in the crop
(on the surface or in the tissue), washing method,
soaking time, physicochemical properties of the plant
and pes ticide, and the type of pes ticide. Pes ticides
with high water solubility can be more easily
eliminated, probably due to their reduced tendency
to enter the inner layers [24, 28, 29]
washing with an acetic acid solution on the residual
chlorpyrifos, cypermethrin, and fenvalerate) and
one herbicide (2, 4-dichloro phenoxy acetic acid) in
tomato showed that both washing methods reduced
the concentration of pes ticides by a maximum of
63.08% [30]
washing with tap water and acetic acid (1%) could
decrease the residual concentrations of abamectin
and buprofezin in eggplant and pepper plants two
hours after spraying up to 21.86% for washing with
water and 41.68% with acetic acid [31]
for chlorfenapyr and acetamiprid was reported to be
7.5 mg kg-1 and 0.8 mg kg-1, respectively, which after
washing the okra with water, the residual reduction
percentage was reported to be 90% for chlorfenapyr
the water solubility of two s tudied pes ticides [27].
in outdoor-grown tomatoes were measured over 30
days. The results showed that the residual pes ticides
days, and deltamethrin after three days in unwashed
samples reached below the MRL set by the Codex
and the EU. This amount immediately after washing
reached below the MRL in the washed samples
[18]. Moreover, some methods such as ultrasound-
assis ted dispersive micro solid-phase extraction,
micro-column solid-phase extraction, adsorption
used for extraction process [33-38]. The results
of similar s tudies were compared with proposed
methods in Table 5.
Anal. Methods Environ. Chem. J. 6 (1) (2023) 100-114
111
4. Conclusion
The present s tudy aimed to inves tigate the residual
concentrations of pes ticides deltamethrin, abamectin
grown tomatoes as raw and processed in the form
of homemade tomato pas te by LC-MS. The rank
of reduction percentage of pes ticides at the end of
the harves t period in the raw and pas te products
under both conditions followed as deltamethrin,
acetamipride and abamectin. Considering the PHI
period for deltamethrin and abamectin (3 days) can
reach their residual concentration to the MRL in both
conditions. According to results of the current s tudy,
the times of 7 days and 5 days can be sugges ted as
greenhouse-grown tomato, respectively. According
to the data obtained from the current s tudy and
the reduction percentage of the residual amount of
Table 5. Comparison of proposed method based on LC-MS technique with the published similar s tudies
Pes ticide Ins trument Product Condition Pes ticide Residues Ref.
Acetamiprid HPLC Tomato Greenhouse
Acetamiprid residues were below the already
es tablished European maximum residue
limits (EU MRLs) (0.5 mg/kg) 3 days after
application.
[8]
Abamectin HPLC Tomato Field
The maximum residues level (MRL) values
set by EU for abamectin are 0.02 mg/kg (EU,
2005). Based on these MRL values, PHIs
were 7 d.
[21]
Acetamiprid HPLC Tomato Greenhouse
The residual amount of acetamiprid
pes ticides in tomatoes is decreasing as the
PHI approaches.
[32]
Acetamiprid LC-MS/MS Lettuce Field and
Greenhouse
conditions was observed. [22]
Acetamiprid
LC-MS Tomato Field and
Greenhouse
The reduction rate of acetamiprid residue in
tomato was fas ter in greenhouse conditions
Deltamethrin
The reduction rate of delthamethrin residue
in tomato was fas ter in greenhouse conditions
This
Work
Abamectin
The reduction rate of abamectin residue in
greenhouse.
Table 4. Comparison of reduction percentage of deltamethrin, abamectin, and acetamiprid in unwashed
and washed tomato
Pes ticide Unwashed Washed Reduction (%)
Acetamiprid 3758.40 1245.80 66.85
Abamectin 967.10 467.80 51.62
Deltamethrin 379.20 187.60 50.52
Determination and analysis of pes ticides by LC-MS Fatemeh Norouzi et al
112
pes ticide from raw product to processed product
processing product. The general conclusion that can
be inferred from this s tudy was that the highes t and
mos t remarkable reduction in the residual amounts
of pes ticide was related to the washing s tep, which
can reduce the residual pes ticide up to 66% which
analyzed by LC-MS. It can be sugges ted to s tudy
the initial residues in unwashed, washed, and
processed samples, and the residual concentration of
pes ticides in the soil during the harves t period, and
5. Acknowledgment
The authors would like to acknowledge the Vice-
Chancellor for Research and Technology of Kerman
supports. The datasets analyzed during the current
s tudy are available from the corresponding author on
reasonable reques t. FN performed executive part of
FSB analyzed data, and MF designed and managed
the s tudy.
6. Funding
This work was extracted from the Mas ter of Science
thesis supported by the Vice-Chancellor for Research
and Technology of Kerman University of Medical
Sciences under grant number 99000482.
7. Conicts of interes t
The authors declare that they have no known
reported in this paper. This work was supported by
the Vice-Chancellor for Research and Technology
of Kerman University of Medical Sciences under
REC.1399.600.
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