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Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 120 (2014) 8896

Contents lists available at ScienceDirect

Spectrochimica Acta Part A: Molecular and


Biomolecular Spectroscopy
journal homepage: www.elsevier.com/locate/saa

Cloud-point extraction, preconcentration and spectrophotometric


determination of trace quantities of copper in food, water
and biological samples
Ayman A. Gouda a,, Alaa S. Amin b
a
b

Chemistry Department, Faculty of Science, Zagazig University, Zagazig, Egypt


Chemistry Department, Faculty of Science, Benha University, Benha, Egypt

h i g h l i g h t s

g r a p h i c a l a b s t r a c t

 New sensitive cloud point extraction

method for determination of Cu(II) in


various samples.
 Beers law is obeyed in the
concentration range 4.0115 ng mL1.
1
 The detection limit of 1.20 ng mL
with preconcentration factor of 125.

a r t i c l e

i n f o

Article history:
Received 5 August 2013
Received in revised form 14 September 2013
Accepted 29 September 2013
Available online 8 October 2013
Keywords:
Cloud point extraction
Copper
Spectrophotometry
Triton X-114
Food, water and biological samples

a b s t r a c t
A new, simple and sensitive cloud point extraction procedure was presented for the preconcentration and
determination of copper(II) ion in food, water and biological samples. The analyte was complexed with a
new synthesized reagent, 2-amino-4-(m-tolylazo)pyridine-3-ol (ATAP) as a new complexing agent and
Triton X-114 as the surfactant. After centrifugation, dilution of the surfactant-rich phase with 0.4 mL
of ethanol acidied with 1.0 M HNO3 was performed after phase separation, and the copper contents
were measured by spectrophotometry at kmax 608 nm. The inuence of analytical parameters including
concentration of complexing agent, Triton X-114, pH, equilibration temperature and time, centrifuge rate
and time were optimized. The analytical characteristics of the method (e.g. linear range, molar absorptivity, Sandell sensitivity, optimum Ringbom concentration ranges limits of detection and quantication,
preconcentration factor, and improvement factors) were obtained. Linearity was obeyed in the range
of 4.0115 ng mL1 of Cu(II) ion. The detection and quantication limits of the method were 1.20 and
3.94 ng mL1 of Cu(II) ion, respectively. The interference effect of some anions and cations was also
tested. The method was applied for determination of copper in food, water and biological samples.
Published by Elsevier B.V.

Introduction
In recent years, the toxicity and the effect of trace elements on
human health and the environment are receiving increasing
attention in pollution and nutritional studies. Copper is one of
Corresponding author. Tel.: +20 552420204; fax: +20 552308213.
E-mail address: aymangouda77@gmail.com (A.A. Gouda).
1386-1425/$ - see front matter Published by Elsevier B.V.
http://dx.doi.org/10.1016/j.saa.2013.09.146

the most widespread heavy metal contaminants in environment.


It is an important element for most life forms as a micronutrient,
but is also toxic at high concentrations [1]. Copper has a biological
action at low doses and a toxic effect when ingested in larger quantities. A concentration more than 1.0 lg mL1 of copper can impart
a bitter taste to water. Large oral doses can cause vomiting and
may eventually cause liver damage. Copper concentration in potable water is usually very low (620 lg L1) [2]. A high concentration

A.A. Gouda, A.S. Amin / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 120 (2014) 8896

of copper in foods originates mainly from fungicide residues containing this metal used in agriculture and from water plumbing.
The determination of trace levels of copper is important because
it could catalyze oxidation of fatty acid chains, exerting a deleterious inuence on shelf life and nutritional value [3].
Therefore, sensitive, reproducible and accurate analytical
methods are required for the determination of trace copper in
environmental biological and water samples. Although inductively
coupled plasma atomic emission spectrometry (ICP-AES) and
atomic absorption spectrometry (AAS) [4,5] are among the most
widely used methods for trace metal determination, they are
usually insufcient due to the matrix interferences and the very
low concentration of metal ions. For these reasons, an efcient separation and preconcentration procedure is often required prior to
the measurement step.
Currently, the most widely used preconcentration methods are
liquid phase microextraction [6], liquidliquid extraction [7], ionexchange [8], resins chelation [9], ber chelation [10], solid-phase
extraction (SPE) [11,12], electrochemical deposition [13] and cloud
point extraction (CPE) [14]. CPE technique has become increasingly
popular in comparison with the classical liquidliquid extraction
method because of its advantages of high enrichment factor, high
recovery, rapid phase separation, low cost, low consumption of
organic solvents and the ability of combination with different
detection.
Micelles and other organized amphiphilic assemblies are
increasingly utilized in analytical chemistry especially in separation and preconcentration procedures. Their unique micro heterogeneous structures capable of selective interaction with different
solute molecules can strongly modify solubility, chemical equilibrium, kinetics and the spectroscopic properties of analytes and
reagents [15,16]. Compared with other extraction methods such
as liquidliquid extraction, cloud point extraction (CPE) exhibits
much more environmentally friendly properties, and it is safer
because small volumes of noxious surfactants are used instead of
toxic organic solvents. CPE is an attractive technique that reduces
the consumption and exposures to a solvent and it also reduces
disposal costs and extraction time that has been used for preconcentration of metal ions after the formation of sparingly watersoluble complexes [1719]. Triton X-114 was chosen as the nonionic surfactant because of its low cloud point temperature and
high density of the surfactant-rich phase as well as its low cost
commercially and lower toxicity. The efciency of the cloud point
extraction depends on the hydrophobicity of the ligand and of the
complex formed, on the apparent equilibrium constants in the
micellar medium and on the formation kinetics of the complex
and on the transference between the phases.
In the present work, a new reagent, 2-amino-4-(m-tolylazo)
pyridine-3-ol (ATAP) was synthesized, characterized and used for
preconcentration and determination of copper(II) by spectrophotometry based on cloud-point extraction (CPE) of the copper(II)
complexed with ATAP using Triton X-114 as surfactant. The proposed method was also applied to the determination of copper in
food, water and biological samples.

Experimental
Apparatus
All Absorption spectra and absorbance measurements were
recorded and measured with a Perkin-Elmer k3B double beam
UVVIS spectrophotometer with 5.0 mm quartz cuvettes. An Orion
research model 601A/digital ionalyzer tted with a combined
glasscalomel electrode was used for pH adjustment and checking
the pH value of buffer solutions. An inductively coupled plasma

89

(ICP) model Varian Liberty 150AX Turbo was used for copper concentration determination. A Hettich, EBA 21 model centrifuge with
50 mL calibrated centrifuge tubes were used to achieve and accelerate the phase separation process, respectively. A thermostated
water bath with a good temperature control within 1.0 C was
used for cloud point temperature experiments.
In order to characterize the synthesized reagent, the IR
spectrum was recorded as KBr discs using Matson FTIR spectrophotometer in the 4000200 cm1 range. The 1H-NMR spectrum of the
reagent was performed using a varian EM 390-90 NMR spectrometer in d6-DMSO as solvent using tetramethylsilane (TME) as an
internal standard. The microanalysis of C, H and N of this
compound was performed in the Microanalytical Center of Cairo
University.
Chemicals and reagents
High-purity ethanol, methanol, isopentyl alcohol, chloroform,
nitric acid, hydrochloric acid, and ammonia solution all from Merck
(Darmstadt, Germany) were used without any further purication.
Analytical-grade cupric nitrate, mercuric chloride, sodium molybdate, sodium tungstate, sodium acetate, hydroxylamine hydrochloride and nitrate salts of sodium, potassium, lithium, calcium,
barium, beryllium, magnesium, manganese, strontium, cobalt, iron,
lead, and nickel all from Merck were of the highest purity available
and used without any further purication. Doubly distilled and
deionized water was used throughout.
A standard stock copper solution (1000 mg L1) was prepared
by dissolving 0.3802 g of Cu(NO3)23H2O, (Merck) in water and
diluting to 100 mL. This solution was standardized by iodimetric
titration. Working solutions were prepared by appropriate dilution
of the stock solution. A 2.0% (v/v) Triton X-114 from Merck was
prepared by dissolving 2.0 mL of Triton X-114 in bidistilled water
in 100 mL volumetric ask with stirring. Cloud point of Triton
X-114 in aqueous solution is 24 C [20].
Thiel buffer solutions of pH 2.012 were prepared by mixing
different proportions of two successive solutions in the following
list to obtain the required pH-value:
(a) 0.05 M oxalic acid +0.20 M boric acid;
(b) 0.20 M boric acid +0.05 M succinic acid +0.05 M sodium
sulphate;
(c) 0.05 M sodium tetraborate;
(d) 0.05 M sodium bicarbonate; resulting in a + b (pH 1.52.5),
b + c (pH 3.09.0), and c + d (pH 9.512) [21].
Synthesis of 2-amino-4-(m-tolylazo)pyridine-3-ol (ATAP) reagent
The azo dye under investigation was prepared by the common
way used for preparing azo dye derivatives of aromatic amines. A
0.01 mole of p-toluidine was converted to the hydrochloric form
by adding the least amount of 1:1 HCl then diluting with water
and cooling at 2.0 C. A cooled solution of NaNO2 (0.01 mole) is
added gradually with continuous stirring to the amine salt. The
resulting diazonium salt solution is allowed to stand in ice bath
for 15 min with stirring at 2.0 C and added gradually to a solution of 0.01 mole of 2-amino-3-hydroxypyridine dissolved in 10%
NaOH which cooled at 2.0 C. The resulting solution is allowed
to stand for 15 min with constant stirring until the azo dye completely formed. The obtained azo compound is ltered off, dried
and recrystallized in ethanol. The purity of the resulting azo compound is checked by measuring the melting point constancy. The
chemical structure of the synthesized reagent (ATAP) (Fig. 1) was
detected by elemental analysis (C, H, N), IR and 1H-NMR spectra.
A 5.0  104 M solution of the ATAP reagent was prepared by
dissolving an appropriate weight of reagent in 10 mL ethanol and

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A.A. Gouda, A.S. Amin / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 120 (2014) 8896
H2N

OH

CH 3

Fig. 1. The structural formula of 2-amino-4-(m-tolylazo)pyridine-3-ol (ATAP).

then completed to the mark in 100 mL calibrated ask with the


same solvent.
General procedure
For the cloud point extraction, 7.5 mL of thiel buffer solution
(pH 4.5), 6.0 mL of 5.0  104 M ATAP solution and 5.0 mL of
1.0% (v/v) Triton X-114 solution and an aliquot of 10 mL of a solution containing Cu(II) ion (0.25.75 lg), were kept for 10 min in
the thermostatic bath at 50 C and then completed with deionized
water to 50 mL in a 50 mL measuring ask. Separation of two
phases was achieved by centrifugation for 10 min at 4000 rpm.
The mixture was cooled in an ice bath to increase the viscosity of
the surfactant-rich phase, and the aqueous phase was easily decanted by simply inverting the tube. The micellar extract from this
procedure was diluted with 0.4 mL of methanol acidied with
1.0 M HNO3 and transferred into 5.0 mm quartz cell to measure
its absorbance at 608 nm.
Pretreatment of samples
Analysis of water samples (tap, river and waste water) for
determination of copper ions content was performed as following:
400 mL of sample was poured in a beaker and 8.0 mL concentrated
HNO3 and 3.0 mL of 30% (v/v) H2O2 for elimination and decomposition of organic compound were added. The samples, while stirring was heated to one tenth volume. After adjustment of
samples pH to desired value, the complexation and CPE were performed according to described general procedure.
Vegetable sample (Spinach) was purchased from Benha, Egypt.
Afterwards, it was taken in small mesh. For the digestion of the
sample, the procedure given by Ghaedi [16,22] was applied. A
40 g of spinach sample was heated in silica crucible for 3.0 h on a
hot plate and the charred material was transferred to furnace for
overnight heating at 650 C. The residue was cooled, treated with
10 mL concentrated nitric acid and 3.0 mL 30% H2O2 again kept
in furnace for 2.0 h at the same temperature so that no organic
compound traces are left. The nal residue was treated with
3.0 mL concentrated hydrochloric acid and 2.04.0 mL 70% perchloric acid and evaporated to fumes, so that all the metals change
to respective ions. The solid residue was dissolved in bidistilled
water, ltered and by keeping the pH at 4.5 made up to 25 mL
by addition of thiel buffer. The dissolved solution was suitably diluted and presented procedure was applied.
A 20 g of tablet chocolate sample was ashed in silica crucible for
4.0 h on a hot plate and the charred material was transferred to
furnace for overnight heating at 450 C. The residue was cooled
and treated with 10 ml concentrated nitric acid and 3.0 ml 30%
H2O2 again kept in furnace for 2.0 h at the same temperature so
that no organic compound traces are left. The nal residue was

H 2N

CH 3

OH

treated with 0.5 mL concentrated hydrochloric acid and 1.0


2.0 mL 70% perchloric acid and evaporated to fumes, so that all
the copper metal changes to copper ions. The solid residue was dissolved in water and ltered and by keeping the pH at 4.5 made up
to 25 mL by addition of thiel buffer. Then the procedure was applied. Copper levels in the nal solutions were determined by
ICP-AES and the above general procedures.
A 50 g of liver and meat samples was taken and dried for 48 h in
an oven at 120 C to remove the water content and to obtain a constant weight (about 68% water). Dried liver sample transferred into
a glass ask. For the digestion of the sample, the procedure given
by Ghaedi et al. [23] was applied and the samples were treated
according to previous publication [24]. A concentrated acid mixture of 3.0 mL H2SO4, 15 mL HClO4 and 15 mL HNO3 were added
and left to stand over night. The solution was kept in an oil bath
at 50 C until the foaming stopped. Then the temperature was increased to 150 C and heating was continued until the evolution
of brown fumes of nitrogen oxides ceased. When a dark brown in
mixture was appeared, the ask was cooled for about 2.0 min then
a 5.0 mL of nitric acid had to be added. Heating was continued until
nitrogen oxides fumes were longer given off. Appearance of white
fume of perchloric acid in 1.0 mL solution is an indication of
complete digestion. The solid residue was dissolved in water and
ltered and by keeping the pH at 4.5 made up to 25 mL by addition
of thiel buffer. Then the procedure was applied. Copper levels in
the nal solutions were determined by ICP-AES and the above general procedures.
Homogenized soil sample 20 g or blood sample 20 mL was taken accurately and in a 250 mL beaker was digested in the presence of an oxidizing agent with addition of 10 mL concentrated
HNO3 and 2.0 mL HClO4 70% was added and heated for 1.0 h. The
content of beaker was ltered through a Whatman No. 40 lter paper into a 250 mL calibrated ask and its pH was adjusted to desired value and diluted to mark with de-ionized water. Then the
general procedure given at Section 2.3 was applied. Copper levels
in the nal solutions were determined by ICP-AES and the above
general procedures.
Results and discussion
Characteristic elemental analysis, infrared and 1H-NMR spectra of
ATAP
The elemental analysis of the synthesized ATAP (C12H12N4O;
M.Wt. 228.24), (%): C, 53.15; H, 5.29; N, 24.55 and Found: C,
52.85; H, 4.91; N, 24.81 with melting point 186 C.
For ATAP reagent the mOH band stretching vibration appears as
sharp absorption with medium intensity at frequency 3477 cm1.
The mNH band appears at 3328 cm1. The mCH band for aromatic system appears at 3010 cm1 and another mCH band appears at
2920 cm1. The band at 1587 cm1 is due to mC=C vibrations. The
mC=N band is observed at 1509 cm1 while the mN=N is observed at
1453 cm1. The appearance of the mC=N band for the reagent gives
an indication that hydroxy azo M hydrazo equilibrium is taken
place which conrmed by the appearance of weak band at 3261
3196 cm1of the stretching vibration of NH group resulting from
hydroxy azo M hydrazo equilibrium and this can be represented
as follow:

H 2N

H
N

CH 3

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A.A. Gouda, A.S. Amin / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 120 (2014) 8896

2
OH

3
CH 3

0.3

0.2

0.1

0
500

540

580

620

660

700

Wavelength (nm)
Fig. 2. Absorption spectra for 75 ngAbsorption spectra for 75mL1 and lg mL1
Cu(II) complexed with 3.0  105 M ATAP using 0.1% (v/v) Triton X-114 at pH 4.5,
with and without CPE, respectively.

0.6

CH3

0.4

II

The different types of signals for hydrogens which are expected


for ATAP reagent and the chemical shift of different types of protons are detected. All signals observed take the integration value
which gives evidence and helps to assign the signals. The signal
lying at very downeld side 2.48 ppm is due to the hydrogen of
OH group that are attached to pyridine ring. The values of magnetic
resonance of hydrogen of OH group are shifted to downeld because the presence of NH2 group increase the electron density
around the hydrogen of OH group and causes shielding on hydrogen. Also the position of this signal at downeld can be taken as an
argument for the contribution of the OH group in an interamolecular hydrogen bond taking place as follow:
H 2N

0.5

0.5
N

The signal lying at very downeld side 3.84 ppm are due to the
hydrogens of NH2 group. A new multiple signals at 1.23 ppm is due
to the protons of CH3 group. The multi signals appear at 7.327.07
and 6.586.45 ppm are due to the protons of rings I and II
respectively.
Absorption spectra
ATAP shows maximum absorbance at 496 nm at pH 4.5; Cu(II)
in buffer medium of pH 4.5 reacts with ATAP in absence (kmax
575 nm) and in the presence of Triton X-114 and the absorbance
of solution shift to bathochromic at 608 nm. Therefore the ternary
complex of Cu(II)ATAPTriton X-114 can be extracted by CPE
method. After separation of surfactant-rich phase, the absorbance
was measured at 608 nm against a reagent blank (Fig. 2). Therefore, the method is suitable for preconcentration and spectrophotometric determination of Cu(II).
Optimization of the system
To take full advantage of the procedure, the reagent concentrations and reaction conditions must be optimized. Various experimental parameters were studied in order to obtain optimized
system. These parameters were optimized by setting all parameters to be constant and optimizing one each time.
The effect of pH
The formation of metalcomplex and its chemical stability are
the two important inuence factors for CPE. The pH plays an unique role on metalcomplex formation and subsequent extraction,

0.4

Absorbance

1
H 2N

with CPE

without CPE
0.6

Absorbance

It is worthy to mention that the stretching vibration of the C@O


group resulted from hydroxy azo M hydrazo equilibrium, taking
place in azo compound ATAP which must be observed in this region is masked by the intense bands due to C@C ring vibrations.
A further support for the conclusion obtained from elemental
analysis and IR spectrum for the reagent under investigation is
gained by a consideration of its 1H-NMR spectrum. The different
types of hydrogen protons which are expected for the reagent under investigation can be formulated as follow:

0.3

0.2

0.1

10

11

pH
Fig. 3. Effect of pH of thiel buffer on the absorbance of the Cu(II)ATAP complex.
Conditions: 75 ng mL1 Cu(II), 3.0  105 M ATAP, 0.1% (v/v) Triton X-114, temperature 50 C.

and is proved to be a main parameter for CPE. Extraction yield


depends on the pH at which complex formation is carried out.
The effect of pH on the absorbance at a constant concentration of
complex in surfactant-rich phase was investigated in the range of
2.511. Different types of buffers (acetate, borate, phosphate, thiel,
and universal) were examined. The optimum one is thiel buffer and
the best values were between pH 4.05.0. As Fig. 3 shows, the pH
4.5 was selected for all further studies, since the results are higher
concordant at this value. The complexation reaction at pH values
lower than 3.0 is incomplete due to protonation of ATAP and complexation reaction is incomplete. The decrease in absorbance at pH
values higher than 8.0 could be due to the hydrolysis of Cu(II).
Moreover the optimum volume of pH 4.5 per 50 mL sample was
found to be 7.5 mL, since the results are highly concordance at this
volume.

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A.A. Gouda, A.S. Amin / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 120 (2014) 8896

The effect of reagent concentration


The effect of concentration of the complexing agent on the analytical responses was subsequently studied. The effect of ATAP concentration on the extraction and determination of Cu(II) was
investigated in the range (0.55.0)  105 M. The sensitivity of
the method increased by increasing ATAP concentration up to
3.0  105 M and remained constant at higher concentrations.
Therefore, 3.0  105 M of ATAP was used in further works (Fig. 4).
The effect of temperature and time
It was desirable to employ the shortest equilibration time and
the lowest possible equilibration temperature as a compromise between completion of extraction and efcient separation of phases.
The dependence of extraction efciency upon equilibration temperature and time above the cloud point in the range of 3060 C
and 5.025 min were thoroughly optimized, respectively. The results showed that an equilibration temperature of 50 C and an
equilibration time of 10 min were adequate to achieve quantitative
extraction. It was found that 50 C is adequate for these analyses.
An equilibration time of 10 min was chosen as the optimal to
achieve maximum sensitivity. Unreasonably high temperatures
are not suitable in the proposed analytical method since they could
create stability problems for complexes and complexing agents,
while at higher temperature, critical micelle concentration (CMC)
of non-ionic surfactants decreases [25]. The results indicate that
the experiment in the optimized reagent concentration after heating for 10 min at 50 C and centrifuging by 10 min in 4000 rpm and
cooling in 10 min in ice-bath lead to high recovery of copper(II) ion
in short time. Moreover, non-ionic surfactants appear relatively
more hydrophobic at higher temperatures, due to an equilibrium
shift that favors dehydration of the ether oxygens [26]. This leads
to an increase in the number concentration of micelles. As reported
previously [26], the solubilization capability of the micellar solution increases with temperature leading to an increase in the
extraction.

Fig. 5. Effect of Triton X-114 concentration on the absorbance of the Cu(II)ATAP


complex. Conditions: 75 ng mL1 Cu(II), 3.0  105 M ATAP, pH 4.5, temperature
50 C.

Table 1
Analytical characteristics of the proposed method.

The effect of surfactant concentration


The concentration of surfactant that is used in CPE is critical factor. To obtain the optimal concentration of Triton X-114, the effect
of surfactant concentration on the increase in the absorbance of the
system was investigated in the range 0.020.20% (v/v). As Fig. 5

Parameters

After CPE

Before CPE

Amount of ethanol
pH
Optimum [ATAP]
Reaction time (min)
Stirring time (min)
kmax (nm)
Beers range (ng mL1)
Ringbom range (ng mL1)
Molar absorptivity (L mol1 cm1)
Sandell sensitivity (ng cm2)
Regression equation a
Slope
Intercept
Correlation coefcient (r)
RSD a (%)
Detection limits (ng mL1)
Quantication limits (ng mL1)
Preconcentration factor
Enrichment factor
Consumptive index (mL)

0.4
4.5
3.0  105
10
10
608
4.0115
10110
4.37  105
0.0145

4.5
3.0  105
20

575
5000250,000
7500225,000
1.40  103
45

6.87
0.005
0.9996
1.25
1.20
3.94
125
3122
0.016

0.0022
0.009
0.9985
2.90
1500
4950

A = a + bC, where C is the concentration of Cu(II) in lg mL1.

shows, the absorbance of the solutions increased by increasing


the Triton X-114 concentration up to 0.08% (v/v) and remained
constant at higher concentrations. The increase in absorbance by
increasing surfactant concentration can be due to both more efcient extraction of the complex and medium effect. Therefore,
0.1% (v/v) Triton X-114 was used in further studies.
As the surfactant-rich phase was very viscous, ethanol was
added to the surfactant-rich phase after CPE to facilitate its transfer
into spectrophotometric cell. The amount of 0.4 mL methanol acidied with 1.0 M HNO3 was chosen to have an appropriate amount
of sample for transferring and measuring the sample absorbance.
Therefore, a preconcentration factor of 125 was archived using
the proposed method.
Stoichiometric ratio
Fig. 4. Effect of ATAP concentration on the absorbance of the Cu(II)ATAP complex.
Conditions: 75 ng mL1 Cu(II), pH 4.5, 0.1% (v/v) Triton X-114, temperature: 50 C.

The nature of the complex was established at the optimum


conditions described above using the molar ratio and continuous

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A.A. Gouda, A.S. Amin / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 120 (2014) 8896
Table 2
Comparative data from some recent studies on CPE preconcentrationseparation of Cu(II).
Reagent

Micellar system

SRP diluting agent

Detection

DL
(ng mL1)

PF/
EF

Sample

Ref.

Dithizone
HEPTS

Triton X-114
Triton X-114

Tetrahydrofuran
2.0 mL methanol

ST
ST

4.6
0.1

10
5

[29]
[30]

4-BPDC
Isoleucine
Diethyldithiocarbamate
Thiamine

Triton
Triton
Triton
Triton

X-114
X-100
X-100
X-114

NA
Up to 1.0 mL with methanol
3.0 mL ethanol
At 2.5 mL with ethanol/water (1:1)

ST
ST
ST
SF

1.6
5
0.4
0.29

NA
22
18
10

Me-BTABr

Triton X-114

FAAS

1.08

17

NDTT

Triton X-114

FAAS

0.22

22.4

Water

[36]

Cupron

Triton X-114

FAAS

0.04

88

Triton X-114

FAAS

1.5

14

River water and


seawater
Water

[37]

Me-BDBD
Alizarin Red S

Triton X-114

FAAS

1.07

21

Water

[39]

PDBDM (chrysoidine)

Triton X-114

200 lL methanol solution of 1.0 M


HNO3
200 ll of methanol solution
containing 0.1 M HNO3
800 lL methanol solution of 0.1 M
HNO3
200 lL methanol solution of 1.0 M
HNO3
1 mL methanol solution of 0.1 M
HNO3
0.5 mLof 1.0 M HNO3 in methanol

Liver
Water and saturated
saline
Natural water
Food and water
Water and food
Water and parenteral
solutions
Water samples

FAAS

0.6

15

[24]

PTU
PAN
DFID (thiophenytoin)

Triton X-114
OP-7
Triton X-114

0.5 mL 0.5 M HNO3 in methanol


NA
0.5 mL of 0.5 M HNO3 in methanol

FAAS
FAAS
FAAS

1.6
8.54
1.5

30
NA
39

[40]
[41]
[42]

N,N0 -bis(2-hydroxyacetophenone)1,2-propanediimine
Sulfathiazolylazo resorsin
Caproic acid and octylamine
N-phenyl benzohydroxamic acid
MPMP

Triton X-114

0.3 mL of 0.5 M HNO3 in methanol

FAAS

0.06

20

Environmental and
biological
Environmental
Waters
Water, blood,
vegetables
Waters

Triton X-100
OP-10
Triton X-114
Triton X-114

FAAS
FAAS
FI-FAAS
FI-FAAS

0.64
10
1
0.15

25
NA
45
81

Waters
Water
Water and hair
Rice our and water

[44]
[45]
[46]
[47]

DDTC

Triton X-114

FAAS

1.1

11

Tea, milk and water

[48]

ATAP

Triton X-114

0.8 mL of 0.1 M HNO3 in methanol


Distilled water
Ethanol with 1% HNO3
300 lL with 1.0 M solution of HNO3
in methanol
0.5 mL by HNO3 in
methanol(0.1 M)
0.4 mL of ethanol acidied with
1.0 M HNO3

ST

1.2

125

Food, water and


biological

This work

[31]
[32]
[33]
[34]
[35]

[38]

[43]

HEPTS: 4-ethyl-1-(pyridin-2-yl)thiosemicarbazide; 4-BPDC: (4-bencylpiperidine-ditiocarbamate; Me-BTABr: 2-[20 -(6-methyl-benzothiazolylazo)]-4-bromophenol; NDTT: 6(2-naphthyl)-2,3-dihydro-as-triazine-3-thione; Me-BDBD: 6-[20 -(60 -methyl-benzothiazolylazo)]-1,2-dihydroxy-3,5-benzenedisulfonic acid; Alizarin Red S: 1,2-dihydroxyanthraquinone-3-sulfonic acid sodium salt; PDBDM: 4-(Phenyl diazenyl) benzene-1,3-diamine; PTU: 4-hydroxy-2-mercapto-6-propylpyrimidine; PAN: 1-(2-pyridylazo)-2naphthol; MPMP: 2-[(2-mercaptophenylimino)-methyl]phenol; DDTC: sodium diethyldithiocarbamate; OP-10: Polyoxyethylated phenol; OP = 7: polyoxyethylated alkylphenol; Cupron: 1,5-diphenyl-benzoin; DFID (thiophenytoin): 5,5-diphenylimidazolidine-2-thione-,4-one; TritonX-114: octylphenoxypolyethoxyethanol; FAAS: ame
atomic absorption spectrometry, ST: spectrophotometry; SF: spectrouorimetry; CE: capillary electrophoresis; SRP surfactant-rich phase; PF preconcentration factor; EF:
enrichment factor; DL detection limit; NA: non-available data.

Table 3
Comparison of selected reagents for the spectrophotometric determination of Cu(II).
Reagent
2-(5-Bromo-2-pyridylazo)-5-diethylamino amino phenol, polyglycol octylphenyl ether
1,5-Bis(di-2-pyridylmethylene) thiocarbonohydrazide
Diethyldithiocarbamate, b-cyclodextrin
Bis(acetylacetone)ethylenediimine
Poly[allylamine-co-N-4-(8-aminoquinolyl-5-azo) benzylideneallylamine]
3,30 -(1,3-Propanediyldiimine)bis-[3-methyl-2-butanone]dioxime
1-[Pyridyl-(2)-azo]-naphthol-(2), TX-100, N,N0 -diphenylbenzamidine
S,S0 -bis(2-aminophenyl)oxalate
3-{2-[2-(2-Hydroxyimino-1-methyl-propylideneamino)-ethylamino]-ethyl-imino}-butan-2one oxime
Naphthazarin
1-Phenyl-1,2-propanedione-2-oxime thiosemi-carbazone
Thiomichlersketone, polyethylene octyl phenyl ether
Dimethylindodicarbocyanine
Bromosulphonazo III
4-(20 -benzothiazolylazo)-salicylic acid (BTAS)
ATAP

kmax
(nm)

e/

Linear range
(lg mL1)

Ref.

104 L mol1 cm1

560
500
436
370
590
525
520
504
570

15
4.2
1.3

4.1
2.95
11.4
0.54
0.16

0.018
NA
06.0
Up to 80
0l.0
0.5350
To 0.6
0.4150
0.2225

[49]
[50]
[51]
[52]
[53]
[54]
[55]
[56]
[57]

330
465
500
650
616.8
485
608

1.84
0.56
5.7
12
33
2.35
43.7

Up to 4.5
0.387.63
00.6
0.01.2
01.024
0.635.04
0.0040.115

[58]
[59]
[60]
[61]
[62]
[63]
This work

NA: non-available data.

variation methods [27]. The plot of absorbance versus the molar


ratio of ATAP to Cu(II), obtained by varying the ATAP concentration, showed inection at molar ratio 2.0, indicating presence of

two ATAP molecules in the formed complex. Moreover, the Job


method showed a ratio of ATAP to Cu(II) = 2.0. Consequently, the
results indicated that the stoichiometric ratio was (2:1) [ATAP:

94

A.A. Gouda, A.S. Amin / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 120 (2014) 8896

Table 4
Tolerance ratio of diverse ions on the determination of 75 ng mL1 Cu(II) (relative
error 5.0%).
Ion added

Tolerance ratio (wion/wCu(II))

K+, Li+, Na+, succinate, acetate, tartaric acid

6000a
4500

Ag+, Ba2+, Ca2+, oxalic acid, C2 O2


4
Ba2+, Ca2+, Mg2+, S, Sr2+, I, Br, B(III)
3

Al3+, Cr3+, SO2
4 , PO4 , F
Sn(IV), V(V), Cl, NO
3
Co2+, Cd2+, borate, hydrazine
La3+, Y3+, Sc3+, Br, hydrazine

Fe2+, Hg2+, ClO3

Mn2+, Zn2+, SCN, SO2


3
Mo(V), Sb(III), Thiourea

3+
Fe , NO2
a

2500
1750
1000
750
400
250
125
70
45

Maximum limit tested.

Cu(II)]. The conditional formation constant (log K), calculated using


Harvey and Manning equation applying the data obtained from the
above two methods, was found to be 4.68, whereas the true constant was 4.55.
For ion associate complexes of CuATAPTriton X-114, the stoichiometric ratio as obtained from molar ratio indicated the formation of 1:1 for [Cu(ATAP)2]: Triton X-114; so we conjectured that
an ion association complex [Cu(ATAP)2]: [Triton X-114] is formed
in the system. The structure of which is probably as follows:

CuII 2ATAP  Cu  ATAP2 


Cu  ATAP2  Triton X  114fCu  ATAP2 Triton X  114g

Calibration curve and sensitivity


The calibration curve showed that the system obeys Beers
law in the concentration range of 4.0115 ng Cu(II) per mL in
the measured solution. For more accurate results, Ringbom optimum concentration ranges was found to be 10110 ng Cu(II) per
mL in the measured solution. The linear regression equation obtained was A = 6.87C + 0.005 (r = 0.9996). The molar absorptivity
was calculated to be 4.37  105 L mol1 cm1 at 608 nm, whereas
Sandell sensitivity was found as 0.145 ng cm2. Because the
amount of Cu(II) in 50 mL of sample solution is measured after
preconcentration by CPE in a nal volume of 0.4 mL, the maximum preconcentration factor of the solution is 125. Enrichment
factor, dened as the ratio of the slope of the calibration graph
for the CPE method to that of the calibration graph in aqueous
media without CPE, for Cu(II) was 3122. Other factors that characterize preconcentration systems, such as consumptive index
(CI), were also determined (Table 1). The consumptive index is
dened as the sample volume, in millilitres, consumed to reach
an unit of enrichment factor (EF): CI = Vs (mL)/EF, where Vs is
the sample volume.
The standard deviations of the absorbance measurements were
calculated from a series of 13 blank solutions. The limits of detection (K = 3) and of quantication (K = 10) of the method were
established [28] and recorded in Table 1, according to the IUPAC
denitions (C1 = KSo/s where C1 is the limit of detection, So is the
standard error of blank, s is the slope of the standard curve and
K is the constant related to the condence interval). The relative
standard deviation was 1.25% obtained from a series of 10 standards each containing 75 ng mL1 of Cu(II).

Table 5
Recovery studies of Cu(II) in real samples.
Sample

Added

Vegetablec

0.0
30
60
0.0
35
70
0.0
25
50
0.0
40
80
0.0
45
90
0.0
20
35
0.0
30
60
90
0.0
15
30
0.0
20
40
0.0
35
70
105

Liverc

Meatc

Lotus (tree)

Soilc

Blood

Tap waterd

Waste waterd

River waterd

Chocolate

a
b
c
d

Founda
Proposed

ICP-AES

28.9
59.2
88.5
29.1
64.2
99.5
30.6
55.3
80.2
17.7
57.5
98.3
18.0
63.5
107.3
77.7
98.2
112.4
6.2
36.3
66.5
95.8
75.6
91.0
105.1
63.2
83.0
102.5
0.2
35.3
70.1
105.4

28.8
59.0
89.2
29.0
64.5
99.7
30.5
55.8
81.0
17.8
57.6
88.0
18.1
63.6
108.0
77.5
98.4
113.1
6.3
36.5
66.0
96.6
75.5
91.2
105.9
63.0
83.6
104.0
0.2
35.4
70.3
105.3

RSD (%)

Recovery (%)

1.4
1.1

100.51
99.55

100.16
100.40

99.46
99.50

99.65
100.61

100.79
99.35

100.51
99.73

100.28
100.46
99.58

100.44
99.52

99.76
99.32

100.28
99.86
100.19

1.3
1.0
1.3
0.9
1.3
1.0
1.4
1.0
1.3
0.9
1.2
1.4
1.1
1.2
1.5
0.9
1.3
1.1
0.8
1.1
1.0
1.2
1.4
1.1
0.9

Average values of six determinations.


Theoretical values for t-values and F-values at 95% condence level for ve degrees of freedom are 2.57 and 5.05, respectively.
All values are lg g1 after suitable dilution.
All values are ng mL1.

t-Testb

F-Valueb

0.87
1.11

2.12
2.60

0.78
1.19

1.93
2.92

0.96
1.10

2.13
2.54

1.23
1.15

3.18
2.78

1.24
0.90

3.19
2.02

1.17

2.85

0.82
1.09
1.34

1.99
2.47
3.41

1.11
1.25

2.69
3.13

0.94
1.18

2.17
2.90

0.89
0.98
1.08

2.12
2.33
2.41

A.A. Gouda, A.S. Amin / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 120 (2014) 8896

The separation occurred efciently, resulting in good enrichment factor and low LOD. A comparison of the proposed method
with other preconcentration procedures [24,2948] for Cu using
several reagents is given in Table 2. The proposed methodology offers a simple, rapid, inexpensive and nonpolluting alternative to
other preconcentration techniques. The detection limits of analytes
are superior to those of preconcentration techniques. The sensitivity expressed as molar absorptivity of the proposed method is
compared with those of published spectrophotometric methods
(Table 3). The proposed method is more sensitive than other methods [4963], that based on spectrophotometry (Table 3).
Interference
In order to study the selective separation and determination of
copper ions from its binary mixtures with diverse metal ions, an
aliquot of aqueous solutions (50 mL) containing 75 ng Cu(II) and
mg amounts of other cations was taken and the proposed procedure was followed. The tolerance limit was taken as the ratio of
foreign ions to the Cu(II) ion determined that causes maximum
limit tested 5.0% error in the absorbance value. The results are
summarized in Table 4. The result shows that most common ions
do not interfere with the determination suggesting the highly
selectivity of the proposed method.
Analytical applications
In order to conrm the applicability of the proposed method, it
has been applied to the determination of nanogram amounts of
Cu(II) in food, water and biological samples. The reliability of the
presented method was checked by spiking experiments and independent analysis. The results for this study are presented in Table 5.
An ICP-AES method was used as a reference method and the results
are also shown in Table 5. The recovery of spiked samples is satisfactorily reasonable and was conrmed using addition method,
which indicates the capability of the system in the determination
of ions. A good agreement was obtained between the added and
measured analyte amounts. The recovery values calculated for
the added standards were always higher than 95%, thus conrming
the accuracy of the procedure and its independence from the matrix effects.
The performance of the proposed method was assessed by
calculation of the t-value (for accuracy) and F-test (for precision)
compared with ICP-AES method. The mean values were obtained
in a Students t- and F-tests at 95% condence limits for ve
degrees of freedom [64]. The results showed that the calculated
values (Table 5) did not exceed the theoretical values. A wider
range of determination, higher accuracy, more stability and less
time consuming, shows the advantage of the proposed method
over other method.

Conclusions
Cloud point extraction was used to the preconcentration of
copper in different samples. The method, which is based on the
cloud point extraction of the complex of copper with ATAP, allows
the determination of copper as low as 1.2 ng mL1. The proposed
method requires inexpensive instrumentation and offers safety,
good selectivity, accuracy and precision that can be applied to
the determination of copper in real samples. The surfactant has
been used for preconcentration of copper in samples, and thus
toxic solvent extraction, has been avoided. The methodology offers
a simple, rapid, inexpensive and nonpolluting alternative to other
preconcentration techniques. The method is relatively rapid

95

comparing with previously reported procedures for the enrichment of analyte ions.

References
[1] J.X. Li, J. Hu, G.D. Sheng, G.X. Zhao, Q. Huang, Colloids Surf. A Physicochem. Eng.
Asp. 349 (2009) 195199.
2] Y. Yamini, J. Hassan, M.H. Karbasi, Microchim. Acta 148 (2004) 305309.
[3] J.C. Cypriano, M.A.C. Matos, R.C. Matos, Microchem. J. 90 (2008) 2630.
[4] M.R. Ganjali, M.R. Pourjavid, L.H. Babaei, M.S. Niasari, Qum. Nova 27 (2004)
213220.
[5] E.C. de Oliveira, M.I. Monteiro, F.V. Pontes, M.D. de Almeida, M.C. Carneiro, L.I.
da Silva, A. Alcover Neto, J. AOAC Int. 95 (2012) 560566.
[6] S. Dadfarnia, A.M.H. Shabani, Anal. Chim. Acta 658 (2010) 107111.
[7] M. Pesavento, G. Alberti, R. Biesuz, Anal. Chim. Acta 631 (2009) 129133.
[8] K. Pyrzynska, Microchim. Acta 153 (2006) 117120.
[9] D. Yang, X.J. Chang, Y.W. Liu, S. Wang, Microchim. Acta 147 (2004) 219222.
[10] R.S. Juang, P. Huang, J. Chem. Technol. Biotechnol. 75 (2000) 610616.
[11] M. Wongkaew, A. Imyim, P. Eamchan, J. Hazard. Mater 154 (2008) 739744.
[12] M.A. Taher, S.Z.M. Mobarakeh, A.R. Mohadesi, Turk. J. Chem. 29 (2005) 1724.
[13] Y.C. Sun, J. Mierzwa, C.R. Lan, Talanta 52 (2000) 417421.
[14] Y. Wei, Y. Li, X. Quan, W. Liao, Microchim. Acta 169 (2010) 297301.
[15] M. Ghaedi, A. Shokrollahi, F. Ahmadi, H.R. Rajabi, M. Soylak, J. Hazard. Mater.
150 (2008) 533540.
[16] M. Ghaedi, Spectrochim. Acta A 66 (2007) 295301.
[17] M.D. Bezerra, M.A.Z. Arruda, S.L.C. Ferreira, Appl. Spectrosc. Rev. 40 (2005)
269299.
[18] C.B. Ojeda, F.S. Rojas, Anal. Bioanal. Chem. 394 (2009) 759782.
[19] C.B. Ojeda, F.S. Rojas, Microchim. Acta 177 (2012) 117.
[20] M. Ghaedi, F. Ahmadi, M. Soylak, J. Hazard. Mater. 147 (2007) 226231.
[21] H.T.S. Britton, Hydrogen Ions, second ed., vol. 1, Longmans, London, 1952.
[22] M. Ghaedi, A. Shokrollahi, A.H. Kianfar, A.S. Mirsadeghi, A. Pourfarokhi, M.
Soylak, J. Hazard. Mater. 154 (2008) 128134.
[23] M. Ghaedi, F. Ahmadi, M.R. Fathi, F. Marahel, Fresenius. Environ. Bull. 14
(2005) 11581163.
[24] A. Shokrollahi, M. Ghaedi, M.R. Fathi, S. Gharaghani, M. Soylak, Quim. Nova 31
(2008) 7074.
[25] J.H. Clint, Surfactant Aggregation, Blackie, Glasgow, 1992. pp. 154.
[26] P.C. Hiemenz, R.H. Rajagopalan, Principles of Colloid and Surface Chemistry,
third ed., Marcel Dekker Inc., New York, 1997. pp. 377.
[27] P. Job, Advanced Physicochemical Experiments, second ed., Oliner and Boyd,
Edinburgh, 1964. pp. 54.
[28] International Conference on Harmonization of Technical Requirements for
Registration of Pharmaceuticals for Human Use (2005) ICH Harmonized
Tripartite Guideline, Validation of Analytical Procedures: Text and
Methodology, Q2(R 1), Complementary Guideline on Methodology dated 06
November 1996, ICH, London.
[29] L.J. Manzoori, G. Karim-Nezhad, Iran. J. Chem. Chem. Eng. 24 (2005) 4752.
[30] M.M. Hassanien, M.H. Abdel-Rhman, A.A. El-Asmy, Trans. Met. Chem. 32
(2007) 10251029.
[31] F. Shemirani, M.R. Jamali, R.R. Kozani, Chem. Anal. 52 (2007) 327336.
[32] P. Liang, J. Yang, J. Food Compos. Anal. 23 (2010) 9599.
[33] X. Wen, L. Ye, Q. Deng, L. Peng, Spectrochim. Acta A 83 (2011) 259264.
[34] A.B. Tabrizi, J. Hazard. Mater. 139 (2007) 260264.
[35] V.A. Lemos, J.S. Santos, P.X. Baliza, J. Braz. Chem. Soc. 17 (2006) 3035.
[36] P. Biparva, M.R. Hadjmohammadi, Acta Chim. Slov. 54 (2007) 805810.
[37] N. Goudarzi, J. Braz. Chem. Soc. 18 (2007) 13481352.
[38] V.A. Lemos, M.S. Santos, M.J.S. Santos, D.R. Vieira, C.G. Novaes, Microchim. Acta
157 (2007) 215222.
[39] N. Satiroglu, C. Arpa, Microchim. Acta 162 (2008) 107112.
[40] A. Shokrollahi, M. Ghaedi, O. Hossainia, N. Khanjaria, M. Soylak, J. Hazard.
Mater. 160 (2008) 435440.
[41] Y.D. Su, L.J. Zhang, Y.Y. Zhu, L.H. Gan, Yejin Fenxi/Metallurg. Anal. 28 (2008)
3638.
[42] F. Ahmadi, A. Khanmohammadi, Z. Tavakoli, E-J. Chem. 6 (2009) S33S40.
[43] S.A.M. Fathi, M.R. Yaftian, J. Colloid. Interface Sci. 334 (2009) 167170.
[44] E. Kk Yetimoglu, O. Aydin Urucu, Z. Yurtman Gndz, H. Filik, Anal. Lett. 43
(2010) 18461856.
[45] S.A. Kulichenko, V.O. Doroschuk, S.O. Lelyushok, Talanta 59 (2003) 767773.
[46] N. Javadi, N. Dalali, J. Iran. Chem. Soc. 8 (2011) 231239.
[47] N. Baghban, A.M.H. Shabani, S. Dadfarnia, A.A. Jafari, Croat. Chem. Acta 85
(2012) 8590.
[48] X. Guoqiang, W. Shengping, J. Xiuming, L. Xing, H. Lijun, Iran. J. Chem. Chem.
Eng. 30 (2011) 101107.
[49] S. Yi, Microchem. J. 36 (1987) 386389.
[50] C.B. Ojeda, A.G. de Torres, F.S. Rojas, J.M. Cano Pavon, Microchem. J. 35 (1987)
164170.
[51] S.H. Li, S.Q. Li, A. Chen, Talanta 40 (1993) 10851089.
[52] N. Chimpalee, D. Chimpalee, S. Lohwithee, L. Nakwatchara, D. Thorburn Burns,
Anal. Chim. Acta 329 (1996) 315318.
[53] H.-M. Ma, Y.-X. Huang, S.-C. Liang, Anal. Chim. Acta 334 (1996) 213218.
[54] S. Karabocek, S. Nohut, O. Dalman, S. Guner, Anal. Chim. Acta 408 (2000) 163
167.
[55] M. Thakur, M.K. Deb, Talanta 49 (1999) 561566.

96

A.A. Gouda, A.S. Amin / Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 120 (2014) 8896

[56] S. Nohut, S. Karabocek, S. Guner, Y. Gok, J. Pharm. Biomed. Anal. 20 (1999)


309314.
[57] O. Dalman, M. Tufekci, S. Nohut, S. Guner, S. Karabocek, J. Pharm. Biomed. Anal.
27 (2002) 183187.
[58] R. Chaisuksant, W. Palkawong-na-ayuthaya, K. Grudpan, Talanta 53 (2000)
579584.
[59] K.H. Reddy, N.B.L. Prasad, T.S. Reddy, Talanta 59 (2003) 425429.

[60]
[61]
[62]
[63]

D. Fu, D. Yuan, Spectrochim. Acta A 66 (2007) 434439.


I.S. Balogh, M. Ruschak, V. Andruch, Y. Bazel, Talanta 76 (2008) 111115.
L. Yuan, S.H. Huo, X.N. Ren, H. Chen, Chin. Chem. Lett. 19 (2008) 9294.
E.Y. Hashem, M.M. Seleim, A.M. El-Zohry, J. Appl. Spectrosc. 78 (2011) 586
593.
[64] J.N. Miller, J.C. Miller, Statistics and Chemometrics for Analytical Chemistry,
fth ed., Prentice Hall, England, 2005.

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