Anne Trégouët
a
, Masoud Khaleghi Abbasabadi
b
and Pooya Gholami
c,*
a
Department of Chemistry, University Paris-Saclay, Saint-Aubin, Paris, France.
b
Department of Chemistry, Iran University of Science and Technology, Tehran, Iran
c
Nano Technology Center, Research Institute of Petroleum Industry (RIPI), P.O. Box 14665-1998, Tehran, Iran
Research Article, Issue 1
Analytical Methods in Environmental Chemistry Journal
Journal home page: www.amecj.com/ir
AMECJ
------------------------
and humans[2]. Nickel is known to bind to specific
proteins and/or amino acids in the blood serum and
the placenta. Orally absorbed nickel is distributed
to the kidneys, followed by the liver, brain, and
heart The harmful health effects of nickel lead to
possible symptoms includes, chronic bronchitis,
lung dysfunction, cancer in lung and nasal sinus[3].
Other target organs include the cardiovascular
system, immune system, and the blood. In large
doses (>0.5 g), some forms of nickel may be acutely
toxic to humans when taken orally. Oral LD
50
values
for rats range from 67-9000 mg Ni per kg (ATSDR).
Nickel separation from human blood samples based on amine
and amide functionalized magnetic graphene oxide nano
structure by dispersive sonication micro solid phase extraction
1. Introduction
Nickel (Ni) is one of the toxic compounds in water
contamination and caused to acute and chorionic
effect in human bodies[1]. Ni(II) are released into
environment from waste of different chemical
factories such as battery manufacturing, mining
and electroplating. The air of near factories contain
the significant amount of heavy metals such as
nickel and caused to adverse effects in environment
*Corresponding Author: Pooya Gholami
Email: pooya1989gh@gmail.com
DOI: ttps://doi.org/10.24200/amecj
Nickel separation in blood by Fe
3
O
4
@A/A-GO Anne Trégouët, et al
A R T I C L E I N F O:
Received 18 Dec 2019
Revised form 6 Feb 2020
Accepted 27 Feb 2020
Available online 25 Mar 2020
Keywords:
Nickel,
Human blood,
Fe
3
O
4
-supported Amine
/Amide-functionalized graphene oxide,
Dispersive sonication micro solid phase
extraction
A B S T R A C T
Nickel (Ni) is toxic effect on human body and must be determined
in human blood samples. In this study, Ni ions separated
and preconcentrated from blood samples based on magnetic
Fe
3
O
4
-supported amine/amide-functionalized graphene oxide
(Fe
3
O
4
@A/A-GO) nanoparticles by dispersive sonication micro
solid phase extraction (DS-μ-SPE). By procedure, 10 mg of
Fe
3
O
4
@A/A-GO was dispersed in 10 mL of human blood samples
with sonication for 5.0 min and then separated from liquid phase
with magnetic accessory. The Ni ions was extracted based on
amine/amide covalence bonding of Fe
3
O
4
@A/A-GO sorbent
(Ni---: NH
2
). Then, the Ni ions back-extracted from Fe
3
O
4
@A/
A-GO in low pH with nitric acid (0.2 mL, 0.3 M) which was
diluted with DW up to 0.5 mL and finally, was determined by
ET-AAS (peak area). The LOD, linear range (LR), enrichment
factor (EF) and absorption capacity (AC) were obtained 35 ng
L
-1
, 0.15 -7.2 μg L
-1
, 19.8 and 131.6 mg g
-1
, respectively. The
method was validated by spiking samples.
Anal. Method Environ. Chem. J. 3 (1) (2020) 5-16
6
Anal. Method Environ. Chem. J. 3 (1) (2020) 5-16
Toxic effects of oral exposure to nickel usually
involve the kidneys with some evidence from
animal studies showing a possible developmental/
reproductive toxicity effect (ATSDR). Normal
range for Ni in healthy peoples is 0.2 µgL
-1
in
serum and less than 3.0 µgL
-1
in human urine. A
national health and nutritional examination survey
(NHNES) of hair found mean nickel levels of 0.39
mg L
-1
, with 10% of the population having levels
less than1.5 mg L
-1
[4,5]. The vary techniques was
used for measurement of nickel (Ni
2+)
in human
bodies such as serum and blood samples. Due to
previous studies, the flame atomic absorption
spectrometry (F-AAS) and electrothermal atomic
absorption spectrometry (ET-AAS) were reported
for analysis of heavy metals such nickel (Ni)
which was suitable for the determination in
biological matrixes [6-9] . Occasionally, the atom
trapping flame atomic absorption spectrometry
(AT-FAAS) and fluorescence spectrometry (XRF)
also was used for heavy metal determaintion such
as nickel in biological samples [10, 11]. Also,
different methods such as, polarography [12],
inductively coupled plasma-optical emission
spectrometry (ICP OES, inductively coupled
plasma-mass spectrometry (ICP-MS) [13, 14],
and spectrophotometry [15] were used for nickel
analysis in human samples [16]. But as difficulty
matrixes in human biological samples such as
blood or serum, the sample preparation is required
to separation nickel ions from liquid phases. The
different sample preparation technology exist for
nickel extraction from blood samples such as liquid–
liquid extraction (LLE) [17], solid-phase extraction
(SPE) and functionalized magnetic-SPE [18],
dispersive solvent by liquid–liquid microextraction
method (DLLME) [19], phase extraction based
on cloud point (CPE) [20], dispersive solid phase
microextraction (D-SPME) [21], ultrasound-
assisted solid phase extraction (USA-SPE) [22].
Recently, the dispersive sorbent in the liquid
phase was presented as micro SPE (D-μ-SPE) for
separation/determination of nickel in water and
biological samples [23]. Some advantages such as
high extraction efficiency, simple usability, fast and
low time caused to select as a favorite technique for
metal extraction. The sorbent characterizations are
a main factor which was effected on heavy metal
extraction by the D-μ-SPE procedure. The selective
of favorite nanosturactures improved recovery. The
different nanosorbent such as, carbon nanotubes
(CNTs), graphene / graphene oxide sheets (NG,
NGO) and silica (MSN) were used for extraction
of Ni in blood samples [24-26]. Between them,
the NGO was reported as efficient sorbents for
metal extraction due to their surface properties.
In this study, a novel sorbent based on Fe
3
O
4
-
supported amine/amide-functionalized graphene
oxide (Fe
3
O
4
@A/A-GO) was used for separation/
speciation Ni(II) from human blood samples by
dispersive sonication micro solid phase extraction
(DS-μ-SPE) at pH=8.0. The method was developed
in blood samples by ET-AAS.
2. Experimental
2.1. Instrumental
The graphite furnace atomic absorption
spectrophotometer (GF-AAS, 932 GBC, Aus)
was used for nickel determination in blood
samples with the Avanta software. The linear
range of 3.0-150 µg L
-1
(peak Area, Abs=1.91) was
selected for Ni in optimized light. The current and
wavelength of HCL lamp was adjusted 3.0 mA
and 228.8 nm, respectively. The auto-sampler of
spectrophotometer (Pal 3000) was used for micro
injection (1µL) of sample volumes to furnace
tube after adjusted injector. The pH of the sample
was digitally calculated by Metrohm pH meter
(Swiss). Fourier transform infrared (FT‐IR) spectra
were recorded from KBr pellets using a Perkin
Elmer Spectrum 65 FT‐IR spectrophotometer.
Powder X-ray diffraction (XRD) was conducted
on a Panalytical X’Pert PRO X-ray diffractometer.
Scanning electron microscopy (SEM) images were
obtained using a Tescan Mira-3 Field Emission Gun
Scanning Electron Microscope (SEM). Magnetic
7
Nickel separation in blood by Fe3O4@A/A-GO Anne Trégouët, et al
property of the catalyst sample was measured with
a vibrating sample magnetometer (VSM) model
LBKFB, Meghnatis Daghigh Kavir Co, Iran, at room
temperature
2.2. Reagents
Chemicals including natural flake graphite
(325 mesh, 99.95%), were purchased from Merck
chemical company and used as received. The
standard stock solutions (1000 mg L
-1
) of Ni (II),
acetone, acetate, HNO
3
, NaOH, HCl and other
reagents were purchased from Merck (Darmstadt,
Germany). Ultra-pure deionized water (DW)
purchased from Milli-Q plus water purification
system (USA). The standard and experimental
solutions of Ni
2+
(0.1, 0.2.0.5, 1.0, 3, 5.9 and 7.0
μg L
-1
) were prepared daily by appropriate dilution
of the stock solutions with DW. The pH of samples
were used with appropriate buffer solutions
including sodium acetate for pH 3.5–5.6, sodium
phosphate for pH of 5.8–8.0, and ammonium
chloride for pH 8–10. All the laboratory glassware
and plastic tubes were cleaned by 5% (v/v) HNO
3
for at least 12 h and then washed many times with
DW and dried in oven prior to use. All reagents
for synthesis of Fe
3
O
4
@A/A-GO prepared by RIPI
Company.
2.3. Synthesis of graphene oxide (GO)
Graphite black powder was oxidized to GO
following the modified Hummers method in
the several steps [27-29].
Fore pre oxidation of
graphite powder, 250 mL of H
2
SO
4
was added to
5 g of graphite powder and the resulting mixture
was stirred for 24 h. After 24 h, 30 g of KMnO
4
was added to the mixture stirring for 72 h at 50 °C.
Next, a solution of 45 mL of H
2
O
2
(30%) in 400
mL of deionized water was added to the mixture
after which the brown color of the mixture turned
into bright yellow. The GO dark solution was
centrifuged and washed with deionized water and
10% HCl solution, and dried at 65
°
C.
2.4. Preparation of nanomagnetic Fe
3
O
4
-
supported Amine/Amide-functionalized graphene
oxide (Fe
3
O
4
@A/A-GO)
2.4.1. Acyl-chlorination of graphene oxide
In the first step, thionyl chloride 60 mL was added
GO (0.3 g) and stirred in at 70°C for 24 h. After
acyl-chlorination reaction, the Acyl-chlorination
of GO was washed with THF four times and the
precipitated dried at 65°C [30].
2.4.1 Functionalization of graphene oxide with
Ammonia
A total of 1 g of Ammonia was added to Acyl-
chlorinated GO (0.3 g). Then, the mixture was
refluxed for 72 h at 120°C under argon condition.
Finally, the mixture reaction washed with DI water.
A dark powder of Amine/Amide-functionalized
graphene oxide was obtained [31].
2.4.2 Nano magnetization of Amine/Amide-funcf-
tionalized graphene oxide
The Fe
3
O
4
-supported Amine/Amide-functionalized
graphene oxide (Fe
3
O
4
@A/A-GO) nanoparticles
were synthesized by co-precipitation of
FeCl
2
·4H
2
O and FeCl
3
·6H
2
O, in the presence
of Amine/Amide-grafted graphene oxide. First,
a solution of FeCl
2
·4H
2
O and FeCl
3
·6H
2
O was
prepared with a molar ratio of 2:1. The weight ratio
of GO to FeCl
3
in the nano composite was 1 per
20 (mGO: mFeCl
3
= 1:20). Afterward, 10 mg of
Amine/Amide-grafted graphene oxide in 15 mL of
DI water was ultrasonicated for 20 min. 12.5 mL
solution of FeCl
2
·4H
2
O (125 mg) and FeCl
3
·6H
2
O
(200 mg) in deionized water (10 mL) was added
to the reaction mixture at room temperature. In
order to raise pH value to 11, an aqua solution of
30% ammonia was added in the reaction mixture at
70
°
C. Then, the reaction mixture was allowed to
cool to room temperature and Fe
3
O
4
@A/A-GO
washed five times with DI H
2
O dried at 70
°
C [32].
2.5 Analytical Procedure
In proposed procedure, 10 mL of blood and
Nickel separation in blood by Fe
3
O
4
@A/A-GO Anne Trégouët, et al
8
Anal. Method Environ. Chem. J. 3 (1) (2020) 5-16
standard solutions was prepared by buffer solution.
The whole blood samples diluted with DW (1:1)
before procedure. By DS-μ-SPE procedure, the
pH of the standard solution containing 0.2 -7.0 μg
L
−1
of nickel was adjusted up to 8 and then 0.01
g of Fe
3
O
4
@A/A-GO as adsorbent was added to
samples. The standard / blood samples was shaked
for 5.0 min at room temperature and Ni ions
physically adsorbed on surface of Fe
3
O
4
@A/A-
GO and chemically extracted by amine and amide
covalence bonding at pH=8. Then, the Fe
3
O
4
@A/
A-GO separated from liquid phase by magnetic
accessory. Finally, the Ni ions back-extraction with
nitric acid (0.2 M) from Fe
3
O
4
@A/A-GO and after
dilution up to 0.5 mL with DW was determined by
ET-AAS. In optimized conditions, the recovery of
physically and chemically adsorption by Fe
3
O
4
@A/
A-GO was almost obtained 25.3% and 97.8%,
respectively. The extraction efficiency of proposed
method based on Fe
3
O
4
@A/A-GO was calculated
by equation 1. The C
A
is the stock concentrations of
nickel and C
B
is the remain concentration of Ni(II)
after procedure (n=10, Eq. 1).
Recovery of extraction=(C
A
-C
B
)/C
A
×100 (Eq.1)
3. Results and Discussion
3.1. Characterization of the nano Fe
3
O
4
@A/A-GO
The Fe
3
O
4
-supported Amine/Amide-functionalized
graphene oxide nanomagnetic (Fe
3
O
4
@A/A-GO)
was synthesized according to the synthetic route
shown in Figure 1. Firstly, graphite was oxidized
to GO by the modified Hummers method [27-29].
Afterward, the GO was amination and amidation
with Ammonia by according to our previously
reported procedure [30, 31].
Finally, the resulted
Amine/Amide-functionalized graphene oxide
(Fe
3
O
4
@A/A-GO) was nano-magnetized by co‐
precipitation of ferrous (Fe
2+
) and ferric (Fe
3+
)
ions in the presence of A/A-GO to afford the target
adsorbent Fe
3
O
4
@A/A-GO [32].
Figure 2 shows the FT-IR spectra of GO and
Fe
3
O
4
@A/A-GO. The broad peak in the range
between 2600-3500 cm
-1
in the IR spectra of these
compounds is related to the (O-H stretching)
Fig 1. Synthetic route of Fe
3
O
4
@A/A-GO
Scheme 2. Synthetic route of Fe
3
O
4
@A/A-GO
2.5. Analytical Procedure
In proposed procedure, 10 mL of blood and standard solutions was prepared by buffer solution. The
whole blood samples diluted with DW (1:1) before procedure. By DS-μ-SPE procedure, the pH of the
standard solution containing 0.2 -7.0 μg L
−1
of nickel was adjusted up to 8 and then 0.01 g of
Fe
3
O
4
@A/A-GO as adsorbent was added to samples. The standard / blood samples was shacked for 5.0
min at room temperature and Ni ions physically adsorbed on surface of Fe
3
O
4
@A/A-GO and chemically
extracted by amine and amide covalence bonding at pH=8. Then, the Fe
3
O
4
@A/A-GO separated from
liquid phase by magnetic accessory. Finally, The Ni ions back-extraction with nitric acid (0.2 M) from
Fe
3
O
4
@A/A-GO and after dilution up to 0.5 mL with DW was determined by ET-AAS. In optimized
conditions, the recovery of physically and chemically adsorption by Fe
3
O
4
@A/A-GO was almost
obtained 25.3% and 97.8%, respectively. The extraction efficiency of proposed method based on
Fe
3
O
4
@A/A-GO was calculated by equation 1. The C
A
is the stock concentrations of nickel and C
B
is the
remain concentration of Ni(II) after procedure (n=10, Eq. 1).
Recovery of extraction = (C
A
-C
B
)/C
A
×100 (Eq.1)
3. Results and Discussion
3.1. Characterization of the nano Fe
3
O
4
@A/A-GO
The Fe
3
O
4
-supported Amine/Amide-functionalized graphene oxide nanomagnetic (Fe
3
O
4
@A/A-GO) was
synthesized according to the synthetic route shown in Scheme 2. Firstly, graphite was oxidized to GO by
the modified Hummers method [27-29].
Afterward, the GO was amination and amidation with Ammonia
9
Nickel separation in blood by Fe3O4@A/A-GO Anne Trégouët, et al
vibration of carboxylic and enolic functionalities
[35].
The peaks at 3420, 1719, 1621, and 1060
cm
-1
shown in the spectra of GO and Fe
3
O
4
@A/A-
GO are ascribed to the (C-O stretching), (C=C
stretching), (C=O stretching), and (O-H stretching)
respectively [33, 34]. Also, The absorption bands
at 3360, 3181, 1630, and 1234 cm
-1
shown in the
spectra of GO and amination GO are ascribed to
the stretching bands ν(O-H), ν(N-H), ν(C=O), and
ν(C-N) respectively.
In the spectrum of Fe
3
O
4
@A/
A-GO, the peaks observed at around 628 and 583
cm
-1
are related to the Fe–O stretching vibration
[37-39].
These results prove that the successful
Amination and amidation of GO and synthesis of
Fe
3
O
4
@A/A-GO.
The XRD patterns of GO, and Fe
3
O
4
@A/A-GO
are demonstrated in Figure 3 (a,b). GO has the two
main peak at 2θ = 11.5°, 42.58
o
are related to the
diffraction planes of (002) and (100) respectively,
which can be observed in the XRD patterns of
both GO and Fe
3
O
4
@A/A-GO [40-42]. As shown
in Figure 3 (b),
the peaks at 2θ = 24° and 42.58
o
It
is evident from the XRD pattern of Fe
3
O
4
@A/A-
GO that was proved the presence of GO [42].
The
main peaks at 2θ° = 30.12, 35.45, 43.07, 56.97,
62.47 in XRD pattern of Fe
3
O
4
@A/A-GO are in
good accordance with the standard XRD data of
magnetite Fe
3
O
4
. The main diffraction peaks at
by according to our previously reported procedure [30, 31].
Finally, the resulted Amine/Amide-
functionalized graphene oxide (Fe
3
O
4
@A/A-GO) was nano-magnetized by co‐precipitation of ferrous
(Fe
2+
) and ferric (Fe
3+
) ions in the presence of A/A-GO to afford the target adsorbent Fe
3
O
4
@A/A-GO
[32].
Fig. 2 shows the FT-IR spectra of GO and Fe
3
O
4
@A/A-GO. The broad peak in the range between 2600-
3500 cm
-1
in the IR spectra of these compounds is related to the (O-H stretching) vibration of carboxylic
and enolic functionalities [35].
The peaks at 3420, 1719, 1621, and 1060 cm
-1
shown in the spectra of GO
and Fe
3
O
4
@A/A-GO are ascribed to the (C-O stretching), (C=C stretching), (C=O stretching), and (O-H
stretching) respectively [33, 34]. Also, The absorption bands at 3360, 3181, 1630, and 1234 cm-1 shown
in the spectra of GO and amination GO are ascribed to the stretching bands ν(O-H), ν(N-H), ν(C=O), and
ν(C-N) respectively.
In the spectrum of Fe
3
O
4
@A/A-GO, the peaks observed at around 628 and 583 cm
-1
are related to the Fe–O stretching vibration [37-39].
These results prove that the successful Amination and
amidation of GO and synthesis of Fe
3
O
4
@A/A-GO.
Fig. 2. FT-IR spectra of (a) GO, and (b) Fe
3
O
4
@A/A-GO
The XRD patterns of GO, and Fe
3
O
4
@A/A-GO are demonstrated in Fig. 4. GO has the two main peak at
2θ = 11.5°, 42.58
o
are related to the diffraction planes of (002) and (100) respectively, which can be
observed in the XRD patterns of both GO and Fe
3
O
4
@A/A-GO [40-42]. As shown in Fig. 4 (b),
the peaks
at 2θ = 24° and 42.58
o
It is evident from the XRD pattern of Fe
3
O
4
@A/A-GO that was proved the
presence of GO [42].
The main peaks at 2θ° = 30.12, 35.45, 43.07, 56.97, 62.47 in XRD pattern of
Fe
3
O
4
@A/A-GO are in good accordance with the standard XRD data of magnetite Fe
3
O
4
. The main
diffraction peaks at 2θ° = 62.47, 56.97, 43.07, 35.45, 30.12 are related to the reflection planes of cubic
spinel crystal structure of Fe
3
O
4
at (440), (511), (400), (311), (220) respectively [43-50].
Fig. 2. FT-IR spectra of (a) GO, and (b) Fe
3
O
4
@A/A-GO
Fig. 3. XRD patterns of (a) GO, (b) Fe
3
O
4
@A/A-GO
Fig. 4. XRD patterns of (a) GO, (b) Fe
3
O
4
@A/A-GO
The SEM image of GO shows that graphene oxide nano sheets consists of randomly accumulated and
wrinkled thin sheets. Also, the SEM image of Fe
3
O
4
@A/A-GO shows in Fig. 5(b) that the average
diameter of Fe
3
O
4
nanoparticles was about 35.43 nm
and indicate a regularly spherical morphology on
A/A-GO [51-52].
Fig. 5. SEM images of (a) GO, (b) Fe
3
O
4
@A/A-GO
The Magnetic behavior of the Fe
3
O
4
@A/A-GO was recorded using a VSM. As shown in the Fig. 6a, the
saturation magnetization of Fe
3
O
4
@A/A-GO was found to be 42 emu g
-1
. This amount of a saturation
magnetization value, the magnetized nanocomposite is expected to have considerable paramagnetism to
make it magnetically separable from reaction mixture [32, 48].
Nickel separation in blood by Fe
3
O
4
@A/A-GO Anne Trégouët, et al
10
Anal. Method Environ. Chem. J. 3 (1) (2020) 5-16
2θ° = 62.47, 56.97, 43.07, 35.45, 30.12 are related
to the reflection planes of cubic spinel crystal
structure of Fe
3
O
4
at (440), (511), (400), (311),
(220) respectively [43-50].
The SEM image of GO shows that graphene
oxide nano sheets consists of randomly accumulated
and wrinkled thin sheets (Fig. 4a). Also, the SEM
image of Fe
3
O
4
@A/A-GO shows in Figure 4(b) that
the average diameter of Fe
3
O
4
nanoparticles was
about 35.43 nm
and indicate a regularly spherical
morphology on A/A-GO [51-52].
The Magnetic behavior of the Fe
3
O
4
@A/A-GO
was recorded using a VSM. As shown in the Figure
5, the saturation magnetization of Fe
3
O
4
@A/A-
GO was found to be 42 emu g
-1
. This amount of
a saturation magnetization value, the magnetized
nanocomposite is expected to have considerable
paramagnetism to make it magnetically separable
from reaction mixture [32, 48].
3.2. Optimization of extraction procedure
For efficient extraction of nickel ions in human
blood samples, the main parameters must be
studied. The effective features such as, pH, sample
volume, amount of Fe
3
O
4
@A/A-GO, shacking
time and interferences ions must be optimized.
Chemical bonding was strongly depended on the
pH solutions. By procedure, the effect of pH on
extraction of nickel through the amine functional
group of Fe
3
O
4
@A/A-GO was evaluated. For this
purpose, the different pH values from 1 to 11 with
nickel concentration of 0.2-7 μg L
-1
as LLOQ and
ULOQ was examined according to DS-μ-SPE
procedure. Obviously, the maximum of extraction
Fig. 4. XRD patterns of (a) GO, (b) Fe
3
O
4
@A/A-GO
The SEM image of GO shows that graphene oxide nano sheets consists of randomly accumulated and
wrinkled thin sheets. Also, the SEM image of Fe
3
O
4
@A/A-GO shows in Fig. 5(b) that the average
diameter of Fe
3
O
4
nanoparticles was about 35.43 nm
and indicate a regularly spherical morphology on
A/A-GO [51-52].
Fig. 5. SEM images of (a) GO, (b) Fe
3
O
4
@A/A-GO
The Magnetic behavior of the Fe
3
O
4
@A/A-GO was recorded using a VSM. As shown in the Fig. 6a, the
saturation magnetization of Fe
3
O
4
@A/A-GO was found to be 42 emu g
-1
. This amount of a saturation
magnetization value, the magnetized nanocomposite is expected to have considerable paramagnetism to
make it magnetically separable from reaction mixture [32, 48].
Fig. 5. VSM curve of Fe
3
O
4
@A/A-GO
Fig. 4. SEM images of (a) GO, (b) Fe
3
O
4
@A/A-GO
11
Nickel separation in blood by Fe3O4@A/A-GO Anne Trégouët, et al
efficiencies for Ni (II) ions based on Fe
3
O
4
@A/A-
GO were obtained at pH range of 8.2, and then
the recoveries were decreased by increasing or
decreasing of pH (Fig. 6). In optimized conditions,
the effect of sample volume of blood on nickel
extraction was studied between 2-20 mL. The results
showed, the optimum extraction was achieved for
12 mL of blood sample and 15 mL of standard
samples, So, 10 mL of sample volume was used for
further studies. Also, the amount of Fe
3
O
4
@A/A-
GO for nickel extraction was evaluated by DS-μ-
SPE procedure. Based on experimental results, 10
mg of Fe
3
O
4
@A/A-GO was selected as optimum
point. The sonication time for extraction of Ni(II)
was studied in optimized pH. Based on previous
research, kind and size of adsorbents are the most
important factors for extraction and sonication time.
Therefore, the effect of sonication in blood samples
was examined by DS-μ-SPE procedure. The results
showed, the maximum efficiency was achieved at
5 min. The concentration of Interfering ions such
as Na
+
, K
+
, Mg
2+
, Ca
2+
, Cu
2+
, Zn
2+
, Co
2+
, Al
3+
, Hg
2+
,
SO
3
2-
, I
-
, NO
3
-
, Cl
-
and F
-
caused less than ± 5%
deviation in the recovery of Ni(II) as the tolerance
limit. So, the interfering ions has no effect on the
recovery efficiencies of Ni(II) in blood samples.
3.3 . Validation methodology
Many methods was used for validation of
methodology by SPE [53-55]. The analytical
results of the developed DS-μ-SPE procedure
were shown method at optimum conditions (Table
1). After sample preparation, Ni concentration in
human blood and standard samples was determined
by ET-AAS. The Human blood, serum and plasma
as a real sample was used for determination of Ni
by DS-μ-SPE procedure. The results was verified
by analyzing the spiked samples with standard
concentration of Ni (II) in human samples (Table
2). Based on results, a high and favorite recovery
was obtained by spiking samples which confirms
the accuracy of results in difficulty matrix. The
Fig. 6. The effect of pH on nickel extraction by DS-μ-SPE procedure
Fig.7. The effect of pH on nickel extraction by DS-μ-SPE procedure
3.3 . Validation methodology
Many methods was used for validation of methodology by SPE [ 53-55]. The analytical results of the
developed DS-μ-SPE procedure were shown method at optimum conditions (Table 1). After sample
preparation, Ni concentration in human blood and standard samples was determined by ET-AAS. The
Human blood, serum and plasma as a real sample was used for determination of Ni by DS-μ-SPE
procedure. The results was verified by analyzing the spiked samples with standard concentration of Ni (II)
in human samples (Tables 2). Based on results, a high and favorite recovery was obtained by spiking
samples which confirms the accuracy of results in difficulty matrix. The recoveries of spiked samples
demonstrated that the results was satisfactory for Ni analysis by DS-μ-SPE. In order to validate the
method, the extraction efficiency for intra-day and inter day analysis was evaluated by spiking samples
(Table 3).
Table 1. Analytical results for Ni(II) extraction based on Fe
3
O
4
@A/A-GO by DS-μ-SPE
Element
a
SV
b
LR
R
2
c
LOD (n = 10)
d
RSD
b
(%)
F
EF
Ni(II)
10
0.15- 7.2
0.9997
0.035
2.8%
19.8
a
sample volume (mL),
b
Linear rang (g L
−1
) ,
C
Limit of detection (g L
−1
) ,
d
Relative standard deviation,
F
enrichment factor(EF),
0
20
40
60
80
100
120
2 3 4 5 6 7 7.5 8 8.5 9 10
Recovery (%)
pH
Table 1. Analytical results for Ni(II) extraction based on Fe
3
O
4
@A/A-GO by DS-μ-SPE
Element
a
SV
b
LR R
2 c
LOD (n = 10)
d
RSD
b
(%)
F
EF
Ni(II) 10 0.15- 7.2 0.9997 0.035 2.8% 19.8
a
sample volume (mL),
b
Linear rang (𝜇g L
−1
) ,
C
Limit of detection (𝜇g L
−1
) ,
d
Relative standard deviation,
F
enrichment
factor(EF),
Nickel separation in blood by Fe
3
O
4
@A/A-GO Anne Trégouët, et al
12
Anal. Method Environ. Chem. J. 3 (1) (2020) 5-16
recoveries of spiked samples demonstrated that the
results was satisfactory for Ni analysis by DS-μ-
SPE. In order to validate the method, the extraction
efficiency for intra-day and inter day analysis was
evaluated by spiking samples (Table 3).
4. Conclusions
A fast and efficient method based on Fe
3
O
4
@A/A-
GO as adsorbent was used for preconcentration,
separation of trace Ni (II) in human blood samples
by DS-μ-SPE procedure. The mechanism of
extraction was achieved by interaction between
negative charge (-) of amine group of Fe
3
O
4
@A/
A-GO with positive charge (+) of nickel ions in
favorite pH (Ni
2+
…..
NH
2
). After extraction, the
concentration of nickel was determined by ET-
AAS technique. Finally, the developed method
has low ion interference, simple usage with low
LOD, favorite RSD(%) values and good LR with
high recoveries for Ni extraction in blood samples
(>95%). Therefore, the proposed method can be
considered as applied techniques for Ni separation
and determination in blood samples by DS- μ-SPE
coupled to ET-AAS.
5. Acknowledgements
The authors wish to thank the Research Institute of
Petroleum Industry (RIPI) for financial support to
Table 3. Validation of methodology for Ni(II) extraction from human samples based on Fe
3
O
4
@A/A-GO by DS-
μ-SPE procedure
Sample*
Added
(μg L
-1
)
*
Found (μg L
-1
)
Recovery (%)
Blood A --- 2.61 ± 0.13 ---
2.5 5.07 ± 0.24 98.4
Blood B --- 1.73 ± 0.08 ---
2.0 3.76 ± 0.18 101.5
Serum C --- 3.06 ± 0.15 ---
3.0 5.98 ± 0.33 97.3
Serum D --- 2.72 ± 0.14 ---
3.0 5.68 ± 0.28 98.6
Plasma E --- 1.46 ± 0.11 ---
1.5 3.02 ± 0.16 104.6
Plasma F --- 0.25 ± 0.02 ---
0.25 0.49 ± 0.03 96.0
*Mean of three determinations of samples ± confidence interval (P = 0.95, n =10)
Table 2. Determination of nickel in blood, serum and plasma solutions by DS- μ-SPE procedure (mean intra –day
and inter day for 10 samples)
Sample
Added(μg L
−1
) * DS-μ-SPE (μg L
−1
)
Recovery (%)Intra-day Inter day Intra-day Inter day
blood ------ ------ 3.65 ± 0.18 3.48 ± 0.15 ------
3.0 ------ 6.57± 0.28 ------ 97.3
------ 3.0 ------ 6.51± 0.32 101.0
Plasma ------ ------ 1.45 ± 0.06 1.57 ± 0.07 ------
1.5 ------ 2.88 ± 0.13 ------ 95.3
------ 1.5 ------ 2.99 ± 0.14 94.6
Serum ------ ------ 2.92 ± 0.14 2.86 ± 0.12 ------
3.0 ------ 6.02± 0.27 ------ 103.3
------ 3.0 ------ 5.82± 0.25 98.6
*Mean of three determinations of samples ± confidence interval (P = 0.95, n =10)
13
Nickel separation in blood by Fe3O4@A/A-GO Anne Trégouët, et al
carry out this research.
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