Environ Monit Assess (2014) 186:72457257

DOI 10.1007/s10661-014-3924-1

Monitoring of trace amounts of heavy metals in different food

and water samples by flame atomic absorption
spectrophotometer after preconcentration
by amine-functionalized graphene nanosheet
Mohammad Behbahani & Nasim Akbari Ghareh Tapeh & Mojtaba Mahyari &
Ali Reza Pourali & Bahareh Golrokh Amin & Ahmad Shaabani

Received: 17 January 2014 / Accepted: 30 June 2014 / Published online: 15 July 2014
# Springer International Publishing Switzerland 2014

Abstract We are introducing graphene oxide modified

with amine groups as a new solid phase for extraction of
heavy metal ions including cadmium(II), copper(II),
nickel(II), zinc(II), and lead(II). Effects of pH value,
flow rates, type, concentration, and volume of the eluent, breakthrough volume, and the effect of potentially
interfering ions were studied. Under optimized conditions, the extraction efficiency is >97 %, the limit of
detections are 0.03, 0.05, 0.2, 0.1, and 1 g L1 for the
ions of cadmium, copper, nickel, zinc, and lead, respectively, and the adsorption capacities for these ions are
178, 142, 110, 125, and 210 mg g1. The aminofunctionalized graphene oxide was characterized by
thermogravimetric analysis, transmission electron microscopy, scanning electron microscopy, and Fourier
transform infrared spectrometry. The proposed method
was successfully applied in the analysis of environmental water and food samples. Good spiked recoveries over
the range of 95.8100.0 % were obtained. This work not
o n l y p r o p os e s a us e f u l m e t h o d f o r s am pl e
preconcentration but also reveals the great potential of
modified graphene as an excellent sorbent material in
analytical processes.
M. Behbahani (*) : M. Mahyari : B. G. Amin : A. Shaabani
Department of Chemistry, Shahid Beheshti University,
Evin, Tehran, Iran
e-mail: mohammadbehbahani89@yahoo.com
N. A. G. Tapeh : A. R. Pourali
Faculty of Chemistry, Damghan University,
Damghan 36715/364, Iran

Keywords Amine-functionalized graphene oxide .

Food and water samples . Solid phase extraction .
Preconcentration . Heavy metals

Introduction
The need for highly reliable methods and techniques for
the determination of trace heavy transition metals has
been recognized in analytical chemistry and environmental science. For that purpose, it is necessary to utilize
either very sensitive instrumental technique or
enrichment/separation methods for the quantification
of low concentrations of metals. Because of the higher
costs of the instrumental techniques like ICP-MS, generally, the preconcentration techniques are preferred
(Mir et al. 2004). Various separation techniques including coprecipitation (Krishna et al. 2004; Liang et al.
2004), solvent extraction (Saito et al. 1998; Tuzen
et al. 2002), membrane filtration (Soylak et al. 2004),
cloud point extraction (Wen et al. 2009), and ion exchange (Bazzi et al. 2005) have been used for the traces
of heavy metal ions in environmental samples.
Solid phase extraction has also emerged as a powerful tool for separation/enrichment of heavy metal ions
(Abdullin et al. 2000; Kobayashi 1994). Solid phase
extraction based on adsorption is a recently used method
that compensates solvent extraction disadvantages. The
solid phase extraction technique reduces organic solvent
usage and exposure, disposal costs, and extraction time
for sample preparation. Solid phase extraction allows

7246

adsorbing chemical species directly onto a solid phase

as an adsorbent provides an effective separation. The
relatively high concentration factor and the ability of
treating large volume samples compared to the other
separationpreconcentration techniques are other advantages of the solid phase extraction. The sorbent
material used, which is the core of solid phase extraction
(SPE), determines the selectivity and sensitivity of the
method. Although numerous substances have been applied as SPE sorbents, such as C18 silica (Jak et al.
2004), polymeric sorbents (Rao et al. 2004), and polyurethane foam (Lemos et al. 2007), their applicability is
often limited only to a number of analytes. Reusability
of the SPE columns is also a problem. Thus, developing
new SPE adsorbents is of high value (Soylak et al. 1997,
2001; Soylak and Dogan 1996; Saracoglu et al. 2002).
Carbon materials are well known for their high adsorption capacity. They have been proven to possess
great potential as adsorbents for removing many kinds
of environmental pollutants: Some examples are activated carbon (Chakrapani et al. 2001), fullerenes
(Vallant et al. 2007), carbon nanotubes (CNTs)
(Shamspur and Mostafavi 2009), carbon nanohorn
(Zhu et al. 2009), and carbon nanocones/disks
(Jimenez-Soto et al. 2009). Graphene, which is considered as the basic building block of all graphitic forms
(including CNTs, graphite, and fullerene C60), is a
single-atom-thick, two-dimensional carbon material
(Rao et al. 2009). Compared with other graphitic forms,
graphene possesses extraordinary electronic, thermal,
and mechanical properties such as ultrahigh specific
surface area, good thermal conductivity, fast mobility
of charge carriers, and high values of Youngs modulus
and fracture strength (Stoller et al. 2008; Lee et al. 2008;
Balandin et al. 2008; Bolotin et al. 2008). To date, the
unique planar structure of graphene provides tremendous potential applications in many fields. For example,
graphene served as filler for the enhancement of mechanical and electrical properties in composite materials
(Watcharotone et al. 2007). Graphene-based materials
were also used as sensors for the sensitive and selective
detection of biomolecules (Lu et al. 2009; Chang et al.
2010; Shan et al. 2009), and graphene/polymer composite is a candidate for super capacitors because of their
high specific capacitance and good cycling stability
(Zhang et al. 2010). The exceptional properties of
graphene make it a superior candidate as an adsorbent
for SPE. Firstly, graphene has a large specific surface
area (theoretical value 2,630 m2 g1 (Stoller et al.

Environ Monit Assess (2014) 186:72457257

2008)), suggesting a high sorption capacity.

Specifically, both sides of the planar sheets of graphene
are available for molecule adsorption, whereas for CNTs
and fullerenes, steric hindrance may exist when molecules access their inner walls. Secondly, graphene can
be easily modified with functional groups, especially via
graphene oxide (GO) which has many reactive groups
(Park and Ruoff 2009). Functionalization may further
enhance the selectivity of SPE. Thirdly, CNTs usually
contain trace amounts of metallic impurities that come
from the metal catalysts used in their synthesis. These
impurities may have negative effects on the applications
of CNTs (Banks et al. 2006; Pumera and Miyahara
2009). Graphene, on the other hand, can be synthesized
from graphite without the use of metal catalysts, thus
obtaining pure material. Fourthly, the large delocalized
-electron system of graphene can form strong stacking interaction with the benzene ring (Allen et al.
2010; Dreyer et al. 2010), which might make graphene a
good choice as an adsorbent for the extraction of
benzenoid-form compounds. In fact, the SPE technique
using graphene as a novel adsorbent has been developed
by Liu et al. (2011) for the determination of eight
chlorophenols in aqueous samples. They demonstrated
that the preconcentration technique is an efficient, simple, and nonexpensive extraction procedure for HPLC
analysis. We extend its application to inorganic analysis
and have obtained the consistent conclusion.
In this work, graphene oxide has been modified with
amine groups and used as a novel sorbent for heavy
metal removal and these trace analysis. The effects of
pH, flow rates, type, concentration, and volume of eluent for elution of cadmium, copper, nickel, zinc, and
lead ions, break through volume and effect of potentially
interfering ions on the separation, and determination of
these heavy metals were investigated. The developed
method was applied for determination of cadmium,
copper, nickel, zinc, and lead ions in several real samples, and the accuracy of the method was confirmed by
certified reference material.

Experimental
Reagents and solutions
All reagents were of analytical grade. Glasswares were
cleaned by 10 % (v/v) HCl and then rinsed with deionized water. Graphite powder (325 mesh), 3-

7246

adsorbing chemical species directly onto a solid phase

as an adsorbent provides an effective separation. The
relatively high concentration factor and the ability of
treating large volume samples compared to the other
separationpreconcentration techniques are other advantages of the solid phase extraction. The sorbent
material used, which is the core of solid phase extraction
(SPE), determines the selectivity and sensitivity of the
method. Although numerous substances have been applied as SPE sorbents, such as C18 silica (Jak et al.
2004), polymeric sorbents (Rao et al. 2004), and polyurethane foam (Lemos et al. 2007), their applicability is
often limited only to a number of analytes. Reusability
of the SPE columns is also a problem. Thus, developing
new SPE adsorbents is of high value (Soylak et al. 1997,
2001; Soylak and Dogan 1996; Saracoglu et al. 2002).
Carbon materials are well known for their high adsorption capacity. They have been proven to possess
great potential as adsorbents for removing many kinds
of environmental pollutants: Some examples are activated carbon (Chakrapani et al. 2001), fullerenes
(Vallant et al. 2007), carbon nanotubes (CNTs)
(Shamspur and Mostafavi 2009), carbon nanohorn
(Zhu et al. 2009), and carbon nanocones/disks
(Jimenez-Soto et al. 2009). Graphene, which is considered as the basic building block of all graphitic forms
(including CNTs, graphite, and fullerene C60), is a
single-atom-thick, two-dimensional carbon material
(Rao et al. 2009). Compared with other graphitic forms,
graphene possesses extraordinary electronic, thermal,
and mechanical properties such as ultrahigh specific
surface area, good thermal conductivity, fast mobility
of charge carriers, and high values of Youngs modulus
and fracture strength (Stoller et al. 2008; Lee et al. 2008;
Balandin et al. 2008; Bolotin et al. 2008). To date, the
unique planar structure of graphene provides tremendous potential applications in many fields. For example,
graphene served as filler for the enhancement of mechanical and electrical properties in composite materials
(Watcharotone et al. 2007). Graphene-based materials
were also used as sensors for the sensitive and selective
detection of biomolecules (Lu et al. 2009; Chang et al.
2010; Shan et al. 2009), and graphene/polymer composite is a candidate for super capacitors because of their
high specific capacitance and good cycling stability
(Zhang et al. 2010). The exceptional properties of
graphene make it a superior candidate as an adsorbent
for SPE. Firstly, graphene has a large specific surface
area (theoretical value 2,630 m2 g1 (Stoller et al.

Environ Monit Assess (2014) 186:72457257

2008)), suggesting a high sorption capacity.

Specifically, both sides of the planar sheets of graphene
are available for molecule adsorption, whereas for CNTs
and fullerenes, steric hindrance may exist when molecules access their inner walls. Secondly, graphene can
be easily modified with functional groups, especially via
graphene oxide (GO) which has many reactive groups
(Park and Ruoff 2009). Functionalization may further
enhance the selectivity of SPE. Thirdly, CNTs usually
contain trace amounts of metallic impurities that come
from the metal catalysts used in their synthesis. These
impurities may have negative effects on the applications
of CNTs (Banks et al. 2006; Pumera and Miyahara
2009). Graphene, on the other hand, can be synthesized
from graphite without the use of metal catalysts, thus
obtaining pure material. Fourthly, the large delocalized
-electron system of graphene can form strong stacking interaction with the benzene ring (Allen et al.
2010; Dreyer et al. 2010), which might make graphene a
good choice as an adsorbent for the extraction of
benzenoid-form compounds. In fact, the SPE technique
using graphene as a novel adsorbent has been developed
by Liu et al. (2011) for the determination of eight
chlorophenols in aqueous samples. They demonstrated
that the preconcentration technique is an efficient, simple, and nonexpensive extraction procedure for HPLC
analysis. We extend its application to inorganic analysis
and have obtained the consistent conclusion.
In this work, graphene oxide has been modified with
amine groups and used as a novel sorbent for heavy
metal removal and these trace analysis. The effects of
pH, flow rates, type, concentration, and volume of eluent for elution of cadmium, copper, nickel, zinc, and
lead ions, break through volume and effect of potentially
interfering ions on the separation, and determination of
these heavy metals were investigated. The developed
method was applied for determination of cadmium,
copper, nickel, zinc, and lead ions in several real samples, and the accuracy of the method was confirmed by
certified reference material.

Experimental
Reagents and solutions
All reagents were of analytical grade. Glasswares were
cleaned by 10 % (v/v) HCl and then rinsed with deionized water. Graphite powder (325 mesh), 3-

Environ Monit Assess (2014) 186:72457257

aminopropyltriethoxysilane, potassium permanganate

(KMnO4), concentrated sulfuric acid (H2SO4), hydrogen peroxide (H2O2), hydrochloric acid (HCl), nitric
acid (HNO3), acetic acid (HOAC), sodium nitrate
(NaNO3), and ethanol (EtOH) were purchased from
Merck (Darmstadt, Germany, www.merck.de). Double
distilled water from a Milli-Q purification system
(Millipore, Bedford, MA, USA) was used for the preparation of solutions. A stock solution (100.0 mg L1) of
cadmium, nickel, copper, zinc, and lead ions was prepared by dissolving an appropriate amount of corresponding nitrate salts in double distilled water.
Working standard solutions were prepared daily through
serial dilutions of the stock solution with deionized
water prior to analysis. Ore polymetallic gold
Zidarovo-PMZrZ (206 BG 326) from Bulgaria was used
as the reference material.

Apparatus
Cadmium, copper, nickel, zinc, and lead concentrations
were determined by an AA-680 Shimadzu (Kyoto,
Japan) flame atomic absorption spectrometer (FAAS)
in an air-acetylene flame, according to the users manual, provided by the manufacturer. Cadmium, copper,
nickel, zinc, and lead hollow cathode lamps (HCL) were
used as the radiation source with wavelengths of 228.8,
324.8, 232, 213.9, and 283 nm, respectively. The pH
was measured at 251 C with a digital WTW Metrohm
827 Ion analyzer (Herisau, Switzerland) equipped with a
combined glass-calomel electrode. A peristaltic pump
was obtained from Leybold (Cologne, Germany), and
an adjustable vacuum gauge and controller were obtained from Analytichem International (Harber City, CA).
The adjustable vacuum gauge allowed the control of the
flow rate during extraction. The CHN analysis was
performed on a Thermo Finnigan Flash EA112 elemental analyzer (Okehampton, UK). IR spectra were

Fig. 1 A scheme for synthesis of amine-functionalized graphene

7247

recorded on a Bruker IFS-66 Fourier transform infrared

(FTIR) spectrophotometer. The transmission electron
microscopy (TEM) images were taken on a JEOL
JEM-2100 F field emission transmission electron microscope. X-ray diffraction patterns were obtained on a
Philips-PW 17C diffractometer with Cu K radiation.
Thermogeravimetric analysis (TGA/DSC) was carried
out on a Bahr STA-503 instrument under air atmosphere. Scanning electron microscopy (SEM) was performed by gently distributing the powder sample on the
stainless steel stubs, using SEM (Philips, XL-30,
Almelo, the Netherlands) instrument.
Preparation of graphene oxide
GO was prepared using Hummers method (Hummers
and Offeman 1958). Graphite powder (1.00 g, 325 mesh)
and H2SO4 (95 %) (23 mL) were added to a 250-mL
conical flask, and the mixture was stirred with a magnetic bar. Sodium nitrate (0.50 g) was added, and the
resulting mixture was cooled to 0 C. Under vigorous
agitation, KMnO4 (3.00 g) was added slowly, and the
mixture was stirred for 1 h, while the temperature was
kept below 35 C. Then, H2O (45 mL) was added
slowly to the reaction mixture, and the solution was
stirred for 30 min at 90 C. Next, H2O2 (10 mL of a
30 % solution) and deionized water (140 mL) were
added to the mixture. Then, resulting precipitate was
centrifuged and washed repeatedly with HCl (5 %, 3
15 mL) and EtOH. The mixture was then vacuum dried
at 60 C. The GO was obtained as a brown powder.
Preparation of amine-functionalized graphene oxide
In a typical run, GO (0.05 g) and DMF (5.0 mL) in a
round flask were subjected to ultrasonic pulse for
30 min, and then 3-aminopropyltriethoxysilane was
added to the solution under nitrogen atmosphere. The

7248

reaction mixture was heated to 80110 C and stirred for

40 h. For coupling reaction, 20 L of triethylamine was
typically added to the reaction mixture to promote the
graft reaction. Then, the mixture was subjected to

Environ Monit Assess (2014) 186:72457257

centrifugation and thoroughly washed with toluene

and THF for at least five times. The product was carefully collected and dried under vacuum at 60 C until
constant weight. Amine-functionalized graphene was

Fig. 2 A schematic illustration of extraction procedure of heavy metals by amine-functionalized graphene

Environ Monit Assess (2014) 186:72457257

obtained as a black powder. Figure 1 provides a scheme

to illustrate the synthesis of amine-functionalized
graphene oxide.
Column method
One hundred milligrams of amine-functionalized
graphene was packed in a glass column with dimensions
of 120 mm in length and 20 mm in diameter and blocked
by two polypropylene filters at the ends to prevent the
loss of the material during sample loading. Prior to
extraction, the column was preconditioned successively
with 5 mL of absolute ethanol, 5 mL of toluene, and
5 mL of absolute ethanol and then washed with double
distilled water until it became free of organic solvents.
Twenty-five milliliters of standard solution containing
2 mg L1 cadmium, copper, nickel, zinc, and lead ions,
after adjusting the pH of sample to 6.0 by adding HNO3
or NaOH solution, was passed through the column at a
flow rate of 18 mL min1 using a peristaltic pump. The
sorbed cadmium, copper, nickel, zinc, and lead ions were
eluted from the column with HCl (1 mol L1) at a flow
rate of 3 mL min1. Afterward, the analytes in the eluent
were determined by FAAS. Figure 2 provides a scheme
for illustration of the mentioned extraction procedure.
Sample pretreatment for metal analysis
Food samples
Fish The fish samples were bought from four local
supermarkets in four different sites randomly in
Tehran, Iran. After collection, four samples of fish from
each supermarket were randomly selected, mixed, and

7249

transferred into an ice bag and transported to the laboratory and stored at 20 C prior to analysis. The muscle
tissue of the fish sample was taken out quickly and was
air dried in an oven at 70 C for 48 h (Behbahani et al.
2013a, b). About 0.5 g of the sample was digested with
5 mL of concentrated HNO3 for 4 h at 100 C. The
resulted solution was filtered and transferred into a beaker and diluted to 250 mL before adjusting the pH to 6.0.
Tomato, mushroom, and apple Food products, including tomato, mushroom, and apple, were collected from
an agricultural land in south of Tehran, Iran. All of the
samples were stored in plastic bags and brought to the
laboratory for preparation and treatment. After washing
the samples with distilled water, 5.0 g of each sample
was grounded, homogenized, and dried at 80 C and
triturated in porcelain mortar. After fractionation of
samples by sieving, sizes less than 20 m were dissolved in 10 mL of 3 mol L1 HNO3 solution and diluted
with distilled water to a final volume of 250 mL, and
then the pH was adjusted to 6.0 for further experiments
(Behbahani et al. 2013a, b).
Water samples
Distilled, tap, and river water The sorbent was successfully used for determination of cadmium, copper, nickel,
zinc, and lead in distilled, tap, and river water samples
(from Shahrood River and Derka River (Ghaemshahr,
Iran)). The polyethylene bottles filled with the samples
were cleaned with detergent, water, diluted nitric acid,
and water in sequence. The samples were immediately
filtered through a cellulose filter membrane (pore size
0.45 m) and were acidified to pH of 2.0 for storage.

Fig. 3 The FTIR of GO (blue line) and amine-functionalized graphene (red line)

7250

Before analysis, the water samples (250 mL) were adjusted to pH of 6.0 according to optimized experimental
conditions.

Results and discussion

Sorbent characterization
The synthesized amine-functionalized graphene oxide
was characterized by FTIR spectroscopy, TEM, SEM,
Fig. 4 The scanning electron
microscopy (a) and transmission
electron microscopy (b) graph of
modified graphene

Environ Monit Assess (2014) 186:72457257

and TGA and elemental analysis (CHN). The FTIR

spectra of amine-functionalized graphene and GO are
shown in Fig. 3. FTIR spectroscopy is one of the important tools for characterization of GO (blue line). The
weak bands at 3,410 cm1, 1,610 cm1, and 1,059 cm1
show stretching vibrations ofCOOH groups. After the
modification of GO sheets with amine groups (red line),
the appearance of a relatively strong band at 2,900 cm1
(CH band of 3-aminopropyltriethoxysilane) proves a
successful modification of graphene oxide with amine
groups. SEM image of the suspended modified

(a)

(b)

Environ Monit Assess (2014) 186:72457257

7251

Fig. 5 The TG plot of graphene

oxide (A) and aminefunctionalized graphene oxide (B)

Heavy metal adsorptiondesorption studies

graphene sheets is shown in Fig. 4a. The exfoliated

graphene sheets with lateral dimensions of several micrometers were observed by SEM image. TEM (Fig. 4b)
is one of the most powerful techniques used to investigate single layers and the level of the dispersion of
graphene sheets in aqueous solution. The thermal stability of GO and amine-functionalized graphene
oxide has been investigated by thermal analysis.
According to these results in Fig. 5, the difference
between weight loss of amine-functionalized
graphene oxide and GO proves the modification
of GO with amine groups.

100

Retention (%)

80

Optimization of the adsorption steps

Among the tested variables, pH was found to be the
most critical parameter for the adsorption of metals on
the modified graphene. To evaluate the effect of pH on
the retention efficiency, the pH of 25 mL of sample
solution containing 2 mg L1 of cadmium, copper, nickel, zinc, and lead ions was adjusted to fit in the range of
29. As can be seen in Fig. 6, the quantitative adsorption
of cadmium, copper, nickel, zinc, and lead ions on the

Cadmium
Copper
Nickel
Zinc
Lead

60
40
20
0
2

pH
Fig. 6 The effect of pH of sample solutions on retention of lead ions by amine-functionalized graphene (conditions: sample concentration,
2 mg L1; sample volume, 25 mL)

7252

Environ Monit Assess (2014) 186:72457257

amine-functionalized graphene was obtained in the pH

of 6.0. Due to hydrolysis of the heavy metals, pH above
9.0 was not tested.
To optimize the sample flow rate, 25 mL solutions of
2 mg L1 cadmium, copper, nickel, zinc, and lead ions
were adjusted to pH of 6.0 and then passed through the
column at flow rates in the range of 122 mL min1 with
a peristaltic pump. The obtained results demonstrated
that the sample flow rate variation in the ranges of 1
18 mL min1 had no effect on the simultaneous retention
of the heavy metals on amine-functionalized graphene
oxide.
Optimization of the elution conditions
For desorption of cadmium, copper, nickel, zinc, and
lead ions from amine-functionalized graphene, series of
selected eluents, such as HNO3, HCl, and CH3COOH at
different concentrations, were used. As shown in
Table 1, it was eventually found that HCl (1 mol L1)
provided an effective elution of cadmium, copper, nickel, zinc, and lead ions from amine-functionalized
graphene oxide. The effect of eluent volume on the
recovery of the heavy metals was also studied. As
Table 1 shows, quantitative recovery could be obtained
with 2.5 mL of HCl (1 mol L1). Therefore, volumes of
2.5 mL of eluent for desorption of cadmium, copper,
nickel, zinc, and lead ions were used in the remaining
experiments. The influence of the eluent flow rate on
metal recovery was also studied. The results demonstrated that the quantitative recoveries for cadmium, copper,

nickel, zinc, and lead ions were obtained at a flow rate

range of 0.53.0 mL min1 with HCl (1 mol L1).
Effect of the volume of sample solutions
Due to low concentrations of heavy metals,
preconcentration was performed on large volumes of
real samples. Therefore, the maximum volume of sample solution was investigated by increasing the volume
of metal ion solution with a constant amount of ions
(0.1 mg of cadmium, copper, nickel, zinc, and lead
ions). Sample solution volumes of 50, 100, 300, 500,
600, 700, 800, 900, 1,000, 1,100, and 1,200 mL containing cadmium, copper, nickel, zinc, and lead ions
were passed through the column. The results (Fig. 7)
demonstrated that the simultaneous quantitative recovery of cadmium, copper, nickel, zinc, and lead ions on
amine-functionalized graphene oxide can be obtained
for sample volume up to 800 mL.
Effect of the potentially interfering ions
To investigate the effect of various cations found in
natural samples, elements that are known as alkaline,
alkaline earth, and transition metals were added to
100 mL of solution containing 0.01 mg of cadmium,
0.01 mg of copper, 0.01 mg of nickel, 0.01 mg of zinc,
and 0.01 mg of lead ions. The degrees of tolerance for
some alkaline, alkaline earth, and transition metal ions
are presented in Table 2. From the tolerance data, it can
be seen that the potentially interfering ions have no

Table 1 The effect of type, concentration, and volume of eluent on extraction efficiency of the study heavy metals
Eluent

Concentration (mol L1)

Volume (mL)

R%S
Copper

Lead

Cadmium

Nickel

Zinc

HNO3

10

87.01.0

76.01.0

91.01.0

79.01.2

92.01.0

HCl

10

99.01.0

99.01.0

99.01.0

99.01.0

99.01.0

CH3COOH

10

37.01.4

42.01.2

38.01.3

49.01.2

52.01.3

HCl

10

99.01.0

99.01.0

99.01.0

99.01.0

99.01.0

HCl

10

99.01.0

99.01.0

99.01.0

99.01.0

99.01.0

HCl

0.5

10

72.01.2

67.01.4

82.01.3

71.01.5

62.01.3

HCl

99.00.8

99.01.0

99.01.0

99.01.0

99.01.0

HCl

99.01.0

99.01.0

99.01.0

99.01.0

99.01.0

HCl

2.5

99.00.8

99.01.0

99.01.0

99.01.0

99.01.0

HCl

80.01.3

78.01.5

71.01.3

69.01.3

54.01.0

R recovery, S standard deviation

Environ Monit Assess (2014) 186:72457257

110
100

Recovery (%)

Fig. 7 The effect of sample

volume on the recovery of the
heavy metals on aminefunctionalized graphene (0.1 mg
of cadmium, copper, nickel, zinc,
and lead ions)

7253

Cadmium
Copper
Nickel
Zinc
Lead

90
80
70
60
50
40
0

200

400

600

800

1000

1200

1400

Sample volume (mL)

significant effects on preconcentration of cadmium, copper, nickel, zinc, and lead ions at pH of 6.0.

by using a FAAS. The amount of adsorbed heavy metal

ions was calculated using the following equation:
Q C o C A  V =m

Adsorption capacity
The concentration of the metal ions in the aqueous
phases after desired treatment periods was measured

where Q is the amount of metal ions adsorbed onto unit

amount of the composites (mg g1), Co and CA are the
concentrations of metal ions in the initial solution and in

Table 2 The tolerance limit of various potentially interfering ions on determination of the study heavy metals
Foreign ion

K+

Tolerable concentration
Ratio X/Cd, Cu, Ni, Zn, Pb

R%S
Cadmium

Copper

Nickel

Lead

Zinc

10,000

99.01.0

99.01.0

98.01.2

99.01.0

99.01.0

Na

10,000

99.01.0

98.01.5

98.01.4

99.01.0

99.01.0

Cs+

10,000

98.01.5

98.01.2

98.01.4

98.01.5

99.01.0

Ca2+

1,000

97.01.4

96.01.8

98.01.0

97.01.3

97.01.2

Mg2+

1,000

96.01.2

97.01.3

97.01.1

97.01.5

98.01.0

Al3+

700

97.01.0

96.01.0

96.01.5

96.01.5

97.01.0

3+

Cr

800

97.01.2

96.01.0

96.01.4

97.01.2

96.01.5

Fe2+

800

97.01.0

96.01.3

96.01.2

96.01.5

97.01.2

As3+

500

97.01.2

96.01.5

97.01.3

96.01.5

97.01.2

Sb3+

700

96.01.5

96.01.3

97.01.2

97.01.3

98.01.0

Hg2+

600

96.01.3

97.01.5

97.01.5

96.01.2

96.01.5

2+

Mn

700

97.01.2

96.01.4

97.01.6

96.01.2

96.01.3

CrO42

1,000

98.01.3

97.01.1

96.01.3

97.01.2

97.01.0

PO43

1,000

97.01.3

97.01.5

97.01.5

96.01.4

96.01.4

Conditions: sample pH, 6.0; sample volume, 100 mL, 0.01 mg of each metal; sample flow rate, 18 mL min1 ; eluent, 2.5 mL of 1 mol L1
HCl; eluent flow rate, 3 mL min1 . X indicates concentration of diverse ions
R recovery, S standard deviation

7254

Environ Monit Assess (2014) 186:72457257

Fig. 8 Effect of the study heavy

metal equilibrium concentrations
on the adsorption capacity of
amine-functionalized graphene
(adsorption isotherm)

250
Cadmium
Copper
Nickel
Zinc
Lead

Q (mg g-1)

200

150

100

50

0
0

50
100 150 200 250 300 350 400
Initial conecntration of heavy metals (mg L-1)

450

the aqueous phase after adsorption, respectively

(mg L1), V is the volume of the aqueous phase (L),
and m is the weight of the composites (g).
The heavy metal ion adsorption capacities of the
amine-functionalized graphene oxide are given as a
function of initial concentration of heavy metal ions
within the aqueous phase in Fig. 8. These adsorption
curves were obtained from experiments where adsorptions from the single heavy metal aqueous solutions
were studied. It was observed that the amount of adsorption was significantly increased with initial heavy
metal concentration in the studied concentration range.
The maximum adsorption capacities of the aminefunctionalized graphene oxide in the studied range at
optimum adsorption conditions are 178, 142, 110, 125,
and 210 mg g1 for cadmium, copper, nickel, zinc, and
lead, respectively.

defined as CLOD =3Sb /m, where Sb is the standard deviation of ten replicate blank signals and m is the slope of
the calibration curve after preconcentration, for a sample
volume of 250 mL, was found to be 0.03 g L1 for
cadmium, 0.05 g L1 for copper, 0.2 g L1 for nickel,
0.1 g L1 for zinc, and 1 g L1 for lead ions, respectively. The relative standard deviations for eight separate
column experiments for determination of 7.5 g of cadmium, copper, nickel, zinc, and lead ions in 250 mL of
water were 2.4, 2.9, 2.7, 3.1, and 2.4 %, respectively.

Analytical performance

Table 3 The analysis of cadmium, copper, nickel, zinc, and lead

in certified reference material

Under the optimized conditions, calibration curves were

sketched for the determination of cadmium, copper,
nickel, zinc, and lead ions according to the general
procedure. Linearity was maintained at 0.1100 g L1
for cadmium, 0.1100 g L 1 for copper, 0.9
170 g L1 for nickel, 0.5150 g L1 for zinc, and
1.5350 g L1 for lead ions in initial solution. The
correlation of determination (r2) was 0.992 for cadmium,
0.996 for copper, 0.997 for nickel, 0.997 for zinc, and
0.995 for lead ions. The limit of detection which is

Sample

Validation of the method

The concentration of cadmium, copper, nickel, zinc, and
lead ions obtained by amine-functionalized graphene
oxide was compared to the standard material. For this

Element Concentration
(mg kg1)

Relative
error
(%)

Certified Found
Ore Polymetallic gold
Zidarovo PMZrZ(206
BG 326)

210.20 0.8

Cd

212.0

Pb

5.47

5.35

2.2

Ni

12.0

11.70

2.5

Cu

0.51

0.49

3.9

Zn

4.06

3.90

3.9

Environ Monit Assess (2014) 186:72457257

7255

Table 4 Analysis of cadmium, copper, nickel, zinc, and lead in different food (a) and water (b) samples
Sample

Element

Real sample (g kg1)

Added (g kg1)

Found (g kg1)

Cadmium

5.3

10.0

15.1

98.7

Copper

78.3

10.0

86.5

98.0

Recovery (%)

A
Fish

Tomato

Mushroom

Apple

Nickel

52.2

10.0

61.3

98.5

Lead

38.5

10.0

46.7

96.3

Zinc

81.7

10.0

89.5

97.6

Cadmium

10.0

9.8

98.0

Copper

8.1

10.0

17.8

98.3

Nickel

5.1

10.0

14.7

97.4

Lead

7.2

10.0

16.9

98.2
98.5

Zinc

9.8

10.0

19.5

Cadmium

3.5

10.0

12.9

95.5

Copper

19.1

10.0

28.5

97.9

Nickel

5.4

10.0

15.1

98.0

Lead

6.1

10.0

15.8

98.1

Zinc

18.7

10.0

27.5

95.8

Cadmium

10.0

9.7

97.0

Copper

17.6

10.0

17.2

97.7

Nickel

6.1

10.0

15.8

98.1

Lead

4.2

10.0

13.9

97.9

Zinc

16.5

10.0

26.1

98.5

B
Distilled water

Tap water

Shahrood river water

Derka river water

Cadmium

10.0

9.9

99.0

Copper

10.0

10.0

100.0

Nickel

10.0

9.9

99.0

Lead

10.0

9.9

99.0

Zinc

10.0

10.0

100.0

Cadmium

10.0

9.8

98.0

Copper

10.1

10.0

10.0

99.0
96.7

Nickel

2.3

10.0

11.9

Lead

10.0

9.9

99.0

Zinc

6.8

10.0

16.3

97.0

Cadmium

2.9

10.0

12.6

97.7

Copper

21.1

10.0

30.5

98.1

Nickel

45.1

10.0

54.2

98.4

Lead

10.2

10.0

19.8

98.0

Zinc

19.2

10.0

28.5

97.6

Cadmium

3.3

10.0

12.9

97.0

Copper

18.9

10.0

28.5

98.6

Nickel

34.2

10.0

42.5

96.2

Lead

10.3

10.0

19.9

98.0

Zinc

20.5

10.0

29.8

97.7

Conditions: sample pH, 6.0; sample volume, 250 mL; sample flow rate, 18 mL min1 ; eluent, 2.5 mL of 1 mol L1 HCl; eluent flow rate,
3 mL min1

7256

reason, the concentration of the heavy metal ions was

determined at optimum conditions in standard reference
material (Ore Polymetallic gold Zidarovo-PMZrZ (206
BG 326)). As it can be seen in Table 3, good correlation
was achieved between estimated content by the present
method and reference materials. Therefore, aminefunctionalized graphene oxide can be used as a reliable
solid phase for extraction and determination of cadmium, copper, nickel, zinc, and lead ions in real samples.
Determination of cadmium, copper, nickel, zinc,
and lead in real samples
Since natural samples have complex matrices, nonspecific background absorption was caused by interfering
species of the sample matrix. To reduce this undesirable
effect, modified graphene oxide was applied for selective extraction of cadmium, copper, nickel, zinc, and
lead ions in pH of 6.0. Table 4 shows the cadmium,
copper, nickel, zinc, and lead ion recoveries in various
food and water samples.

Conclusions
The proposed SPE method is easy, safe, and economic
for the preconcentration and determination of trace
amounts of toxic heavy metals in different samples.
Due to a relatively high preconcentration factor, toxic
heavy metals at trace level can be determined accurately.
The amine-functionalized graphene oxide showed high
tolerance to interferences from the matrix ions, and in a
nutshell, the important features of the proposed method
were its high adsorption capacity, good preconcentration
factor, and low detection limit which are distinctive
among other SPE methods. This work not only proposes
a useful method for environmental water and food samples pretreatment but also reveals the great potential of
graphene as an excellent sorbent material in analytical
processes.

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