Anal. Method Environ. Chem. J. 3 (4) (2020) 17-29
Research Article, Issue 4
Analytical Methods in Environmental Chemistry Journal
Journal home page: www.amecj.com/ir
AMECJ
Sulfamethizole functionalized graphene oxide for in-
vitro separation and determination lead in blood serum of
batterymanufactoriesworkersbysyringelter-dispersive-
micro solid phase extraction
Abhijit De
a, *
and S. Mojtaba Mostafavi
b
a
HITech institute of theoretical and computational chemistry, Shivakote, Hesaraghatta Hobli, Bengaluru, India
b
Department of Chemistry, Iranian-Australian Community of Science, Hobart, University of Tasmania, Australia
ABSTRACT
Thetoxiceffectoflead(Pb)causestoanemiaandirondeciencyin
human body. So, the lead determination in blood/serum samples is very
important. In this study, a novel adsorbent based on sulfamethizole
functionalized on nanographene oxide (C
3
H
10
N
4
O
2
S
2
-NGO; SM-
NGO) was used for extraction of Pb(II) from human blood, serum and
plasma samples in battery manufactories workers by syringe lter-
dispersive-micro solid phase extraction procedure (SF-D-µ-SPE). By
procedure, 25 mg of SM-NGO mixed with 10 mL of human blood/
serum or plasma samples and aspirated by 10 mL of syringe tube.
After sonication of samples for 5 min, the Pb ions adsorbed based on
sulfur of SM-NGO adsorbent at pH=6 and the solid phase separated
by syringe coupled to Millex-FG hydrophobic PTFE membrane
(0.2 µm). Then, the lead ions were back-extracted from SM-NGO/
PTFE by elution phase with 0.5 mL of nitric acid solution (0.5 M).
Finally, the concentrations of Pb(II) ions were determined by atom
trapameatomicabsorptionspectrometry(AT-FAAS)afterdilution
with DW up to 1 mL. After optimization, the linear range (LR), LOD
and enrichment factor (EF) for Pb ions was obtained, 10-500 µg
L
-1
, 2.5 µg L
-1
and 9.86, respectively. The validation of procedure
wasconrmedbyelecthermalatomicabsorptionspectrophotometer
(ET-AAS) and certiedreference material (NIST,CRM) inhuman
samples.
Keywords:
Lead,
Human blood/serum,
Sulfamethizole,
Nano graphene oxide,
Syringelter,
Dispersive-micro solid phase extraction
procedure
ARTICLE INFO:
Received 18 Aug 2020
Revised form 24 Oct 2020
Accepted 14 Nov 2020
Available online 29 Dec 2020
*Corresponding Author: Abhijit De
Email: abhijit@iitcc.org, ade@actrec.gov.in
https://doi.org/10.24200/amecj.v3.i04.124
------------------------
1. Introduction
Heavy metals intakes to human body from air,
water and foods. Heavy metals such as lead (Pb),
chromium (Cr
VI
) and mercury (Hg) cause to serious
problem in humans and depended on way entrance
(Skin, lungs and gastrointestinal system) and
concentrations [1-3]. The battery manufactories,
gasoline, wastewater, x-ray protection and paint
are main source of lead in environment [4,5]. The
battery factories are a main source of lead toxicity in
workers and cause to dysfunction in blood red cells,
the central nervous system tissues (CNST), bones
and renal tissues [6]. Also, the lead poisoning can
18
Anal. Method Environ. Chem. J. 3 (4) (2020) 17-29
be effected on human organs such as braine, renal,
liver and bone [7,8]. The lead complexes in human
body create by interaction of lead with proteins/
amino acids and cause anemia [9,10]. On the other
hand the lead exposure causes to decrease the
erythrocytes cells (RBC) and reducing biosynthesis
of hemoglobin. The occupational safety and
health administration (OSHA) has reported that
the exposure lead (Pb) more than 600 μg L
-1
is
toxicandworkersmust beofinefromworkand
returntoworkwhentheBLLisbelow300μgL
-1
.
Also, the food and drug administration (FDA) has
announced the normal range of Pb in human blood
is below 250-300 µg L
-1
[11]. The limit exposure
oflead(Pb)inairisconsideredabout50μgm
-3
by
NIOSH [12]. Due to lead toxicity in human, the
accurate technology need to use for determination
of lead ions in the human blood/ serum samples.
Recently, the various methods were reported for lead
determination in human biological samples. The
instrumental techniques such as the electrothermal
atomic absorption spectrometry (ET-AAS) [13,14],
the inductively coupled plasma - atomic emission
spectroscopy or mass spectrometry (ICP-AES/MS)
[16], the high-performance liquid chromatographic
(HPLC) and the isotope dilution inductively coupled
plasma mass spectrometry (ID-ICP-MS) were used
for lead determination in different matrixes[17].
Moreover, the sample preparation method is required
for extracting of lead ions in different biological
samples before determination by spectrometry
techniques. Various sample preparation method
such as liquid-phase microextraction or dispersive
liquid-liquid microextraction (DLLME) [18,19],
hydrophobic deep eutectic solvents based on
microextraction techniques[20], the headspace
solid-phase microextraction [21], the carrier-
mediatedhollowberliquidphasemicroextraction
[22], the dispersive solid-phase extraction (DSPE)
combined with ultrasound-assisted emulsication
microextraction based on the solidication of
oating organic drop (UAEME-SFO)[23], the
microextraction based on precipitation[24], the
magnetic solid-phase extraction (MSPE) [25], and
dispersive-micro solid phase extraction procedure
(D-µ-SPE) [26], were used for lead determination
in different human matrixes. Between them, D-µ-
SPE procedure is used in different matrixes by
researchers. In addition, the characterizations of
sorbents are important factor for lead extraction
by the SPE method. For examples, the adsorbents
such as, the silica aerogel nanoadsorbent [27], the
magnetic phosphorus-containing polymer [28 ], the
magnetic metal organic frameworks MMOF[29],
the carboxylated graphene [30], the graphene
Oxide Sheets [31],andmodied-carbonnanotubes
with NiFe
2
O
4
[32] were reported by chemistry and
nanochemistry scientists. The previously works
showed that the different pretreatment techniques
based on metal nanoparticles, the drugs, the
functionalization of CNTs or GO, the magnetic
nanoparticles can be increased the efcient
extraction of heavy metals in human samples.
Recently, some drugs use for in-vitro and in-vivo
extraction of heavy metal in human samples and
depend on structure and function groups. The
antibiotic of sulfamethizole (SM) is a drug and use
as an insistent inhibitor of bacterial enzyme. The
SM can be complexed with metals in human body
and depended on pH and covalence bonding.
In this study, a new SM-NGO adsorbent
was used for determination lead in blood/serum
samples by syringe lter-dispersive-micro solid
phase extraction procedure (SF-D-µ-SPE) at
pH=6.0. The high recovery and absorption capacity
for lead extraction was obtained in optimized
conditions. The lead concentrations in blood/serum
samples were determined by AT-FAAS after sample
pretreatment.
2. Experimental
2.1. Instrumental
Lead in human blood and serum samples
determinedbytheGBC906atomtrapameatomic
absorption spectrophotometer (AT-FAAS, AUS)
after sample preparation. The atom trap accessory
put on burner and the fuel gas (air-acetylene), lamp
position, slit and light line were manually tuned.
Other parameters such as current and silt adjusted
by Avanta Software. The minimum sample volume
19
Lead extraction by SM-NGO adsorbent Abhijit Dea et al
(0.5-1 mL) vacuumed by nebulizer of AT-FAAS
after adjusting ow rate of sample (mL min
-1
).
The LOD for Pb determination with AT-FAAS
and F-AAS in standard solutions was obtained
0.025 mg L
-1
and 0.07 mg L
-1
, respectively. The
linear ranges of 0.08-6.0 mg L
-1
and 0.25-6.0 mg
L
-1
were obtained for AT-FAAS and F-AAS at
wavelength of 283.3 nm (5 mA). All samples were
injected with an auto-sampler injector from 100
μL to 1000 μL. The Inductively coupled plasma
mass spectrometry (ICP-MS, Perkin Elmer, Main
PlasmaGas:Arow~15Lmin
-1
,1.2s/m,USA)as
high sensitive analyzer was used for determining
of Pb in real human samples after microwave
digestion. ICP-MS atomizes the liquid sample and
generate atomic ions (M → M
+
+ e
−
), which are
then detected by quadrupole-based mass analyzer
system (mass-to-charge ratio (m/z), ICP-Q-MS).
Before mass separation, a beam of positive ions
has to be extracted from the plasma and focused
into the mass-analyzer. The pH meter was used
for determining of pH in liquid samples (Metrohm
E-744). The vortex mixer (Thermo, USA) with
100-500 rpm, ultrasonic heating (Iran) and Falcon
centrifuging (1000-4500rpm) prepared for this
study.
2.2. Reagents and Materials
The lead standard solution (Pb; 1000 mg L
-1
in 2 %
nitric acid) was purchased from Sigma (Germany).
All standard of lead (0.08, 0.1, 0.2, 0.5, 1, 2, 5
mg L
-1
) prepared by dilution of the stock lead
solution (1000 mg L
-1
)withultrapurewater(UPW,
Millipore,USA).Also, thesub-ppb concentration
(10-500 µg L
-1
) prepared by dilution of stock lead
standard solution (1 mg L
-1
). The pure reagents such
as, the polyoxyethylene octyl phenyl ether (TX-
100), nitric acid (HNO
3
), HCl, acetone, and ethanol
were purchased from Sigma Aldrich, Germany.
Sulfamethizole (4-Amino-N-(5-methyl-1,3,4-
thiadiazol-2-yl) enzenesulfonamide; C
9
H
10
N
4
O
2
S
2
;
CASN:144-82-1) and the hydrophobic ionic
liquid of 1-methyl-3-octylimidazolium
hexauorophosphate.([OMIM]PF6;C
12
H
23
F
6
N
2
P;
CASN: 304680-36-2) was purchased from Sigma
Aldrich, Germany. The GO and GO-COOH
was prepared from chemistry department, India.
The pH was adjusted to 6 by sodium phosphate
buffer solution (Na
2
HPO
4
/NaH
2
PO
4
) from Merck
(Germany).
2.3. Sample preparations
All glasses such as vials, volumetric, dishes and
beakers cleaned with HNO
3
and H
2
SO
4
solution
(1:1, 2 M) for at least 12 h. After cleaning, all
of glasses washed for 10 times with DW. The
normal Pb values in blood samples have less than
250 µg L
-1
or >25µg dL
-1
and more than 500 µg
L
-1
is toxic. By procedure, 10 mL of the blood or
serum samples were prepared from 50 workers of
lead–acid batteries factories in India (Men, 20-55
age). The sterilized syringes were used for blood
samples. The pure heparin based on free of Pb was
prepared and injected to blood. The blood samples
were maintained at –5 °C. In this work, all blood
samples prepared based on the world medical
association declaration of Helsinki for physicians
in human and all blood samples prepared from
worker with agreement forms.
2.4. Synthesis of GO@ Sulfamethizole
2.4.1.Preparation of GO
The graphene oxide (GO) was prepared with pure
graphite(G)bymodiedhummersprocedure[33].
The 0.2 g of graphite powder was added into mixing
solution (H
2
SO
4
/ H
3
PO
4
; 9:1) and then potassium
permanganate (KMnO
4
, 1.3 g) was slowly added
by stirring solution up to became black green.
The H
2
O
2
slowly added to solution and stirred for
removal of excess of KMnO4. After cooling, the
hydrochloric acid (HCl:10 mL in 30 mL DW) was
added and centrifuged. Then, the powder product
of GO was washed with HCl and DW for 5 times
and dried at 90 °C for 1 day.
2.4.2.Synthesis of GO@Cl
Chlorinated graphene oxide was prepared due to
the Liu procedure [34]. First, 1.0 g of GO, 20 ml
of benzene, and 100 ml of SOCl
2
were added in a
100-ml round ask and stirred(60-70°C).Then,
20
Anal. Method Environ. Chem. J. 3 (4) (2020) 17-29
the excess of SOCl
2
was exit out from mixture by
distillation process based on vacuum condition.
The solid product put in acetone and the suspension
in acetone was ltered with Watman lter (200
nm). Finally, the product washed with acetone for 3
times and dried at 60 °C.
2.4.3.Synthesis of Sulfamethizole functionalized
on nanographene oxide (SM@NGO)
1 g of sulfamethizole (SM) solved in 20 mL
H
2
O with 5 mL of NaOH and then 1 mL of SM
solution were mixed in 60 mL ethanol (99%) by
an ultrasonic bath (25 min) in a 100 mL round
bottom ask. Then, 0.1 mL of triethylamine
was added to mixture, and the mixture was
reuxed at 70 °C for 5 hours. The obtained
product was separated from the mixture by a
hydrophobic PTFE Membrane Filters (poly-
tetrauoroethylene)forltering SM@NGO. The
SM@NGO product washed with ethanol for many
timesandnallydriedundervacuumat95°C.
2.5. Extraction Procedure
By the SF-D-µ-SPE procedure, 10 mL of serum and
plasma samples were used for determination of lead
ions. Firstly, 25 mg of SM@NGO added to human
blood/serum and lead standard solution (10-500 µg
L
-1
) at pH=6.0 and aspirated by 10 mL of syringe
tube. The syringe tube placed on shaking tube for
5.0 min, after shaking, the lead was extracted by
the sulfur group of SM@NGO in optimized pH
(Pb
2+
→: SR@GO). The SM@NGO separated
from liquid phase by syringe hydrophobic PTFE
membrane. Then, the pb
2+
ions back-extracted from
SM@NGO-PTFE membrane with elution of nitric
acidic (0.5 mL, 0.5 M) and remain solution was
determined by AT-FAAS after dilution with DW up
to 1 mL (Fig.1).
The procedure run for 10 blank
solutions without any lead. The calibration curve
of lead based on SF-D-µ-SPE/AT-FAAS procedure
(10-500 µg L
-1
) and standard method by AT-FAAS
(0.1- 5 mg L
-1
) were done. The enrichment factor
(EF)calculatedbycurvettingrules(Tga=m
1
/m
2
).
3. Results and discussion
3.1. Mechanism of Extraction by SM@NGO
The pure sulfamethizole drug was solved in
H
2
O/NaOH solutions and then mixed with
ethanol (99%) by an ultrasonic bath. Then, the
triethylaminewereaddedtomixture.Afterreux,
the SM@NGO product was separated from the
mixture by PTFE membrane by washing and
drying. Finally, the sulfamethizole functionalized
on nanographene oxide caused to make favorite
adsorbent based on sulfur group (dative bond) for
lead extraction in blood, serum and plasma samples
Fig. 1. The SF-D-µ-SPE procedure for extraction of lead in human samples
21
Lead extraction by SM-NGO adsorbent Abhijit Dea et al
with high recovery. Therefore, the lead adsorption/
extraction with SM@NGO nanostructure caused
to increase the recovery up to 98.5 % as compared
to NGO adsorbent (26.4 %). The results showed,
the mechanism of lead extraction in serum, blood
and plasma samples depended on sulfur group
of sulfamethizole in SM@NGO adsorbent at
optimized pH (Fig.2).
3.2. SEM, TEM and FTIR of SM@NGO
The NGO adsorbent was synthesized from graphite
powder according to the modied Hummer›s
method from RIPI Company (Iran) [33] and used
for synthesis of SM@NGO. The TEM and SEM
of SM@NGO showed the minimum size between
30-100 nm which were shown in Figures 3 and 4,
respectively. The FT-IR spectrum of NGO-COOH
Fig.2. The mechanism of lead extraction in blood/serum samples based on SM@NGO
by the SF-D-µ-SPE procedure
Fig.3. The TEM of SM@NGO adsorbent Fig.4. The SEM of SM@NGO adsorbent
22
Anal. Method Environ. Chem. J. 3 (4) (2020) 17-29
and SM@NGO adsorbent was shown in Figure 5.
Based on FT-IR spectrum, the C=N double bond
is about twice as strong as a C-N single bond,
andtheC≡Ntriplebondissimilarlystrongerthan
double bond. The infrared stretching frequencies of
these groups vary in the same order, ranging from
1100 cm
-1
for C-N, to 1660 cm
-1
for C=N. Also,
symmetric and a symmetric traction for sulfur bond
(S-C) ranging from 1135-1165 cm
-1
and1310-1360
cm
-1
were seen. The O-H bond is about 3600 cm
-1
,
S-H bond in 2570 cm
-1
, C-H bond in 3000 cm
-1
and
N-H bond in 3400 cm
-1
observed.
The X-ray diffraction (XRD) of NGO and SM@
NGO were shown in Figure 6. XRD of graphene
Fig.6. The X-ray diffraction (XRD) for NGO and SM@NGO adsorbents
Fig.5. The FT-IR spectrum of NGO-COOH and SM@NGO adsorbent
23
Lead extraction by SM-NGO adsorbent Abhijit Dea et al
oxideshowstheonemaindiffractionpeakat2θ=12°
related to the oxygen groups which are intercalated
betweengraphenesheets.Thepeaksat2θ=12°and
42.60
o
are related to the diffraction planes of (002)
and (100) respectively, which can be showed in both
of the XRD of GO and SM@NGO.
3.3. Optimization of extraction parameters
The SF-D-µ-SPE procedure based on new SM@
NGO adsorbent was used for the extraction and
determination Pb(II) ions in serum and plasma
samples.Forefcientleadextraction,theanalytical
parameters such as pH, sample volume and amount
of SM@NGO adsorbent were optimized.
3.3.1.Inuence of pH
The efcient extraction of lead in blood serum of
battery workers was depended to pH. So, the pH of
samples must be examined and optimized. The pH
can be affected on increasing /decreasing extraction
processes of lead in serum samples. The buffer
solutions help us to improved extraction recovery by
adjusting pH. Therefore, the different value of pH
between 2-10 was evaluated by lead concentration
from 10 to 500 µg L
-1
. The results showed, the
high extraction of lead ions in human biological
samples was obtained at pH of 6.0 by SM@NGO
adsorbent. On the other hand, the recoveries can be
decreased at acidic or basic pH. So, the pH point of
6 was used for extraction Pb in real samples (Fig.7).
Based on extraction mechanism, the coordination
of dative bond of sulfur as negative charge in SM@
NGO with the positively charged of Pb
2+
were
happened in optimized pH. At low pH (less than
5.5), the SM@NGO adsorbent protonated and
surface of adsorbent got the positively charged. Due
to the electrostatic repulsion, the recoveries of lead
extraction were decreased. Moreover, in optimized
pH(6), the surface of SM@NGO have negatively
charged and high recovery was obtained for Pb
2+
ions
by the SF-D-µ-SPE procedure (more than 96%).
3.3.2.Inuence of SM@NGO amount
The amount of SM@NGO nanoparticles for lead
extraction in human samples has studied with lead
concentration between 10-500 µg L
-1
as a low and
high LOQ ranges. For this proposed, 1-50 mg of
SM@NGO adsorbent were used for lead extraction
in serum, blood, plasma and standard solutions
by the SF-D-µ-SPE method. The obtained results
showed that the maximum of lead extraction in
human samples was achieved for 22 mg of SM@
NGO nanostructure. So, 25 mg of SM@NGO
adsorbent was used for further work (Fig. 8). The
extra mass of 25 mg had no effect on recovery of
lead extraction in human samples.
Fig.7.TheinuenceofpHonleadextractionbasedonSM@NGOadsorbent
by the SF-D-µ-SPE procedure
24
Anal. Method Environ. Chem. J. 3 (4) (2020) 17-29
3.3.3.Inuence of sample volume
Sample volume is an effective parameter for
extraction lead ions in human biological samples
such as serum, blood and plasma. So, the lead
extraction for sample volume between 1-30 mL was
studied and optimized with the low and high LOQ
concentration ranges (10-500 µg L
-1
) by the SF-D-
µ-SPE method (Fig. 9). The experimental results
conrmed that the best extraction was obtained
for less than 15 mL of human samples. Therefore,
10 mL was selected as optimum volume of human
samples for lead extraction with 25 mg of SM@NGO
adsorbent at pH=6. The recovery was decreased by
adding sample volume in optimized conditions.
Fig.8.TheinuenceofamountofSM@NGOonleadextraction
by the SF-D-µ-SPE procedure
Fig.9.TheinuenceofsamplevolumeonleadextractionbasedonSM@NGOadsorbent
by the SF-D-µ-SPE procedure
25
Lead extraction by SM-NGO adsorbent Abhijit Dea et al
3.3.4.Inuence of volume and concentration of
eluents
The inuence of volume and concentration
of eluents such as HCl, HNO
3
, NaOH and
CH
3
COOH for back-extraction of lead ions from
solid phase were studied and optimized by SF-D-
µ-SPE procedure. The syringe coupled to Millex-
FG hydrophobic PTFE membrane (0.2 µm) was
used for back-extraction lead from adsorbent by
different volume of eluents between 0.5-2 mL
(0.2-1 M). After pushing the plunger of syringe,
the elution of SM@NGO/hydrophobic PTFE
membrane was performed by different eluents.
In acidic or basic pH, the lead (Pb) released
from SM@NGO/PTFF to elution phase and
due to participate of lead hydroxyl in basic pH,
the acidic pH was selected. Therefore, the acid
solution can be used for back-extraction of lead
from solid phase. The results demonstrated that
the lead ions were simply back-extracted from
SM@NGO adsorbent by nitric acid solution more
than 0.4 mol L
-1
. Therefore, 0.5 mol L
-1
of nitric
acid (HNO
3
) was used as optimum concentration
of eluent for this study. Also, The results showed,
the lead was efciently back-extracted from
SM@NGO by 0.5 mL of HNO
3
. Finally, the
lead concentration in remained solution was
determined with AT-FAAS after dilution with
DW up to 1 mL.
3.3.5.Membrane lter reusing
The reusability of the SM@NGO packed Millex-
FG hydrophobic PTFE membrane was evaluated
by several extraction and elution cycles under
optimized conditions. It was found that the
membrane lter could be reused after back-
extraction processes and then rinsed by 5 mL
of DW.The membrane lter was used for over
14adsorption–elutioncycles without signicant
decrease in extraction recoveries of lead ions.
As collection adsorbent on membrane, the ow
rateofeluentorsamplesthroughmembranelter
was hardly done and so, the extraction recovery
decreased more than 18 adsorption–elution
cycles.
3.3.6.Adsorption capacity
The main parameter for evaluating of lead
extraction with the SM@NGO packed Millex-
FG hydrophobic PTFE membrane is adsorption
capacity. So, the adsorption capacity of 25 mg
of SM@NGO adsorbent for Pb (II) in 10 mL of
human sample containing 5 mg L
−1
Pb (II) ions was
investigated at pH 6 in static system. After adjusting
pH with favorite buffer solution, the mixture was
shaked for 15 min and ltered with Millex-FG
hydrophobic PTFE membrane (0.2 μm). Finally
the residual concentrations of lead in the Millex-
FG hydrophobic PTFE membrane were determined
using AT-FAAS. The adsorption capacities of GO
and SM@NGO for Pb (II) ions were found to be
57.5 mg g
−1
and 162.1 mg g
−1
, respectively.
3.3.7.inuence of Interference ions
The inuence of some coexisting ions for lead
extraction in human samples was examined by
the SF-D-µ-SPE procedure. For investigation of
the effect of different interference ions on lead
extraction, the various ions (0.5-2 mg L
-1
) added to
10mLofleadsampleswith500μgL
-1
at pH 6.0. So,
the most of the concomitant ions in human blood
such as Cu, Zn, Mn, Mg, Se, Li, F, NO
3
, HCO
3
,
Ca, Na an K were considered for lead extraction
by SM@NGO adsorbent. The experimental results
showed, the interference coexisting ions had no
effect on lead extraction by proposed method.
(Table1). Therefore, the SM@NGO adsorbent
was efciently extracted lead ions from human
biological samples in the present of the interference
coexisting ions.
3.3.8.Determination of lead in real samples
The SM@NGO adsorbent was used for extraction
and determination of lead ions in human samples
by the SF-D-µ-SPE procedure. For validation of
the results, the human blood, serum and plasma
samples were spiked with lead standard solution
based on SM@NGO nanostructure before
determination by AT-FAAS. The high extraction
of spiked samples demonstrated that the SM@
NGO adsorbent had satisfactory results for lead
26
Anal. Method Environ. Chem. J. 3 (4) (2020) 17-29
extraction and determination in human samples
at pH=6.0 (Table 2). Also, 10 mL of the blood or
serum samples were prepared from 50
workers of lead–acid batteries factories in India
(Men, 20-55 age) by the SF-D-µ-SPE procedure
and compared to healthy peoples (Table 3).
Moreover,thecertiedreferencematerials(NIST;
CRM) and lead analysis with ICP-MS were used
for validating of methodology based on SM@NGO
by the proposed procedure (Table 4)
Table 1. Theinuenceofinterferencescoexistingionsonleadextractioninhumanbloodsamples
by the SF-D-µ-SPE procedure
Interfering Ions
Mean ratio
(C
I
/C
Pb(II)
)
Recovery (%)
Pb(II) Pb(II)
Cr
3+
, Se
2+
, Mn
2+
, 600 97.4
Zn
2+
, Cu
2+
700 98.6
I
-
, Br
-
, F
-
, Cl
-
1200 97.2
Na
+
, K
+
, Li
+
1000 99.2
Ca
2+
, Mg
2+
800 97.7
Ni
2+
, Co
2+
450 98.4
CO
3
2-
, PO
4
3-
, HCO3
-,
NO
3
-
950 96.6
Table 2. Determination of lead based on SM@NGO adsorbent and spiking samples
by the SF-D-µ-SPE procedure coupled to AT-FAAS
*Sample
Added
(μg L
-1
)
*
Found (μg L
-1
)
Extraction efciency (%)
Blood
--- 9.7 ± 224.2 ---
200 17.8 ± 418.6 97.2
Plasma
--- 2.8 ± 55.4 ---
50 4.8 ± 106.7 102.6
Serum
--- 7.6 ± 168.9 ---
150 14.3 ± 311.6 95.1
Blood
--- 8.2 ± 178.8 ---
150 13.8 ± 322.3 95.6
Serum
--- 9.4 ± 192.3 ---
200 19.3 ± 399.5 103.6
--- 4.2 ± 88.5 ---
Plasma 100 8.4 ± 186.8 98.3
*Mean of three determinations of samples ± SD (P = 0.95, n =10)
27
Lead extraction by SM-NGO adsorbent Abhijit Dea et al
Table 3. Determination of lead in serum, blood and plasma by the SF-D-µ-SPE procedure coupled
to AT-FAAS (intra –day and inter day, n=50, µgL
-1
)
*
Sample
b
Workers (n=50)
b
Healthy peoples (n=50)
a
Workers
Intra-day Inter day Intra-day Inter day r P value
Blood 352.8 ± 15.7 360.2 ± 16.2 33.1 ± 1.4 29.5 ± 1.3 0.092 <0.001
Serum 276.7 ± 12.6 281.2 ± 13.9 24.6 ± 1.1 28.2 ± 1.2 0.112 <0.001
Plasma 147.5 ± 6.8 152.4 ± 7.3 12.8 ± 0.6 14.6 ± 0.7
0.086 <0.001
a
CorrelationsarebasedonPearsoncoefcients(r).StatisticalsignicancewillbeobservedifP<0.05
b
Meanofthreedeterminationsofsamples±condenceinterval(P=0.95,n=10)
*
50 workers of lead–acid batteries factories in India (Men, 20-55 age)
Table 4. Validation of methodologyforleaddeterminationbycertiedreferencematerials(CRM,NIST)
Recovery (%)Found
*
( μg L
-1
)AddedCRM*( μg L
-1
)Sample
-----135.8 ± 6.5-----139.5 ± 0.8Caprine blood, level 2
97.8233.6± 11.4100
-----272.6 ± 13.1-----277.6 ± 1.6Caprine blood, level 3
96.5465.7± 21.8200
-----84.2 ± 4.4-----86.4 ± 2.1Serum by ICP-MS
94.6131.5 ± 4.450
*
Mean of three determinations of samples ± SD (P = 0.95, n =10)
CRM955c,caprineblood,level2,139.5±0.8μgL
-1
CRM955c,caprineblood,level3,277.6±1.6μgL
-1
4. Conclusions
A novel SM@NGO adsorbent was used for lead
separation/extraction in human blood, serum and
plasma samples by the SF-D-µ-SPE coupled to
AT-FAAS. The Millex-FG hydrophobic PTFE
membrane (0.2 µm) was used as a lter for
separating solid phased from liquid phase. By
the SF-D-µ-SPE method, the high extraction, the
low cost, the fast separation and simple method
in short time was obtained in optimized pH. The
trace amount of SM@NGO as a solid-phase caused
to extract the lead ions from the human biological
samples without any ligand. The developed SF-D-µ-
SPE method presented a analytical nanotechnology
for lead separation/extraction/preconcentration in
human blood samples with low interference ions,
good reusability and simple sample preparation
in difculty matrixes. Therefore, the SM@NGO
nanostructure was used as a perfect adsorbent for
determination and extraction of lead in blood,
serum and plasma samples by AT-FAAS.
5. Acknowledgments
The authors wish to thank from Department of
Chemistry, Hi Tech institute of theoretical and
computational chemistry, India
28
Anal. Method Environ. Chem. J. 3 (4) (2020) 17-29
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