Anal. Methods Environ. Chem. J. 4 (2) (2021) 47-59

Research Article, Issue 2

Analytical Methods in Environmental Chemistry Journal

Journal home page: www.amecj.com/ir

AMECJ

Evaluation and determination of occupational and

environmental exposure of lead in workplace air and human

workers based dispersive ionic liquid solid phase micro

extraction in battery manufacturing factories from Iran

Somayeh Mirza

a,*

and Azadeh Yahya Meymandi

Ph.D in Environment Chemistry and Management, Department of Environmental Management, Faculty

of Natural resources and Environment, Science and Research branch, Islamic Azad University, Tehran, Iran.

Faculty of Science, Department of chemistry, University of Birjand, Birjand, Iran

ABSTRACT

The exposure of lead in workplace air and human workers of battery

manufacturing factory was evaluated determined by nanotechnology

since 2019-2020. Human whole blood (HWB) for subject and healthy

peoples (25-55, Men, 40 N) and workplace air (40N) was prepared

based on NIOSH sampling. 10 mL of HWB samples added to 20

mg of mixture ionic liquid/ ligand ([HMIM][PF

]/APDC) modied

on graphene oxide nanostructures(GONs) at pH=6. After sonication,

the lead ions separated/extracted by dispersive ionic liquid solid

phase micro extraction (DIL-SPME) and determined by ame atomic

absorption spectrometry (F-AAS). All air samples in workplace were

analyzed based on NIOSH process. The results showed us the negative

correlation between Pb concentration in human blood subject and

healthy peoples (r=0.24). The range concentrations of lead in human

subject, healthy peoples and workplace air were obtained 193.4-

543.7 µg L

-1

, 85.6-175.9 µgL

-1

and 44.7-81.5 µgm

-3

, respectively. The

LOD, linear rang, enrichment factor(EF) and RSD% were achieved

1.25 µg L

-1

, 5.0- 310 µg L

-1

19.6 and less than 5% by procedure.

The method was validated by standard reference material (SRM), the

electrothermal atomic absorption spectrometry (ET-AAS) and ICP-

MS analyzer for human samples.

Keywords:

Lead,

Human whole blood,

Workplace air,

Nanotechnology,

Dispersive ionic liquid solid phase

micro extraction,

Battery manufacturing factories

ARTICLE INFO:

Received 5 Mar 2021

Revised form 19 May 2021

Accepted 3 Jun 2021

Available online 29 Jun 2021

*Corresponding Author: Somayeh Mirza

Email: somayeh.mirza@gmail.com

https://doi.org/10.24200/amecj.v4.i02.143

------------------------

1. Introduction

Heavy metals have toxic effects in environmental and

human health which it causes main problem in many

human organs such as brain. Lead (Pb) with dangerous

properties in human consider as a hazardous chemical

[1]. The battery and chemical factories use the lead

in their products. The lead enters in environment

by various sources such as, water, soil, the medical

industries, air dust, water and gas pipes, paint

factories and chemical products [2-5]. Lead causes

many problem in human organs such as, the nervous

central system (insomnia, delirium, cognitive

decits and tremor), kidney, liver, gastrointestinal

and bone disorders [6,7]. Also, an acute poisoning

of lead cause to neurological defect include, pain,

muscle weakness and numbness. In biochemistry,

the lead can be seen in various of proteins and

amino acids. Also, lead bonded to sulfur groups of

aminoacids such as cysteine (Cys) and a homoleptic

and hemidirectic (SR)3 complex [8,9]. Moreover,

Anal. Methods Environ. Chem. J. 4 (2) (2021) 47-59

lead can be complexed to copper/ zinc and caused

brain problems. Therefore, the lead determination

in the human biological samples (blood, serum and

urine) with accurate and precise method must be

considered. The environmental protection agency

(EPA), NIOSH , OSHA, the National toxicology

program (NTP) and food and drug administration

(FDA) reported that the lead concentration in water,

air and human blood samples are between 0.01-0.1

mg L

-1

, less than 50 μg m

-3

and 250 µg L

-1

, respectively

[10-12]. Recently, many researchers reported the

different lead analysis in water and human blood

samples [13]. The various techniques such as ame

atomic absorption spectrometry (F-AAS) [14,15],

the electrothermal atomic absorption spectrometry

(ET-AAS) [16], the inductively coupled plasma

mass spectrometry(ICP-MS), the microwave plasma

atomic emission spectrometry(MP-AES) [17, 18],

the gas chromatography mass spectrometry GC-

MS [19] and laser-induced breakdown spectroscopy

(LIBS)[20] were used for lead determination in

different matrixes. Due to difculty of matrixes and

interferences, the extraction/preconcentration method

for lead determination in blood samples is used.

Various methodology such as, solid-phase extraction

(SPE)[21], the magnetic-SPE [22], the dispersive

liquid–liquid microextraction (DLLME) [23], the

cloud point extraction (CPE) [24], the dispersive solid

phase extraction (D-SPE), the ultrasound-assisted

dispersive micro solid phase extraction (USA-μ-

SPE) [25], and the emulsication microextraction

using a ionic liquid (IL-EME) [26] were reported.

Recently, the dispersive ionic liquid solid phase

micro extraction (DIL-SPME) for separation/

determination of heavy metals in liquid phases was

used. The DIL-SPME method has many advantages

such as high efciency/recovery and easy to use in

short time. The various sorbents include, the carbon

nanotubes (CNTs) [27], the silica gel functionalized

with thiosalicylic acid [28], and graphene/graphene

oxide [29] was used for extraction/removal of heavy

metals from solutions by scientists. The Ionic liquids

were used for the separation of heavy metals from

samples. Currently, safety, health and environmental

assessments in factories can help manage pollutants

in industry and pollutants are reduced in the long run

after analysis [30].

In this study, the mixture of ionic liquid/ ligand

([HMIM][PF

]/APDC) modied on graphene

oxide(NGO) was used for lead extraction from

blood samples at pH=6. The speciation of lead

was achieved based on IL/APDC/NGO adsorbent

before the DIL-SPME procedure. The method was

validated in blood and water samples by spiking

samples and ICP-MS analyzer.

2. Experimental

2.1. Apparatus

The GBC atomic absorption spectrophotometer

equipped with ame and graphite tube ( electrical

furnace) were used for the determination of lead

in blood and serum samples (F-AAS, ET-AAS,

GBC 932 plus, Australia). The Pal GF3000 as

auto sampler accessory for ET-AAS was used as

a low volume of samples for lead determination

by injecting 1-100 µL of sample to graphite tube

(wavelength 283.3 nm, slit 0.5 nm, lamp current

5.0 mA). Also, the auto sampler accessory for

ame technique was used (0.5-5 mL). After

atomization process in ame or graphite tube, the

ppm and ppb concentration of lead was determined

in liquid samples, respectively (Table 1). The ICP-

MS analyzer with high sensitivity was used for

as ultra-trace lead determination in human blood

samples (Perkin Elmer, USA, 1100 W; 14 L min

-1

;

1.2 sec per mass; auxiliary gas 1.1 L min

-1

). The pH

meter was used for measuring pH in blood samples

(Metrohm, E-744, Switzerland). The shacking of

samples was achieved by 300 rpm speeds by vortex

mixer (Thermo, USA) and samples centrifuged

with Falcon accessory (4000 rpm, 5-30 mL of

polypropylene conical tubes, USA). An ultrasonic

bath was used for dispersing of solid phase in blood

samples with the temperature controlling accessory

between 10-100

C (HB120, USA). The X-ray

diffraction (XRD) based on a Panalytical X’Pert

PRO X-ray diffractometer was used. The scanning

electron microscopy (SEM) images were obtained

using a Tescan Mira, Field Emission Scanning

Electron Microscope (FEG-SEM).

2.2. Reagents

The standard solution of lead (Pb

) was purchased

from Merck with a concentration of 1000 mg L

-1

in 1 % HNO

(CAS N: 119776, 1 Li, Germany).

The calibration standard of lead between 0.5-

60 µg L

-1

was daily prepared by diluting of lead

stock. Ammonium pyrrolidinedithiocarbamate

as ligand (APDC, CAS N.: 169209-63-6) was

prepared from Merck. Germany. Ultrapure water

was purchased from Millipore Company (USA) for

dilution of solutions or standards. The hydrophobic

ionic liquid of 1-Hexyl-3-methylimidazolium

hexauorophosphate was prepared from Sigma

Aldrich (CAS N: 304680-35). The pH of 6 was

adjusted by sodium phosphate buffer solution

(Merck, Germany, (Na

HPO

/NaH

). The

polyoxyethylene octyl phenyl ether (TX-100),

HNO

, HCl, acetone, and butanol were purchased

from Sigma Aldrich, Germany.

2.3. Human and air sample

For preparation of blood samples, the glass mixed

in nitric acid (1 M) for 24 h and washed with DW

for 8 times. The low concentrations of lead in blood

samples (<250 µg L

-1

), caused to the effect on

accurate results. So, the process of blood sampling

was carefully done based on standard methods. The

5 mL of blood samples were prepared from battery

workers from Iran (40 Men, 25-55 age) due to

world medical association declaration of Helsinki

and dilution with DW up to 10 mL. Clean tubes and

syringes with plastic needles were purchased for

Merck, Germany for blood sampling. The 10 μL

heparin (pure metals) was added to human blood

sample. The blood samples were maintained frozen

in refrigerator below -4°C.

The air sampling for lead was prepared based on

Filter (0.8 µm cellulose ester membrane) by 7082

NIOSH method. The owrate adjusted 1 to 4 L min

and ashing process with 6 mL of HNO

, 1 mL of

(30%) at 140

C was achieved. The working

standards covering the range 0.25 -20 µg mL

-1

Pb used for calibration method (2.5-200 µg lead per

sample). The lead concentration in workplace air

was determined by F-AAS and D

or H

continuum

or Zeeman background correction used to control

ame or molecular absorption. The working range

between 0.05 -1 mg m

-3

for a 200 L of air sample

was selected. In addition, the ow rate between 1.0

- 4.0 L min

-1

for 8 h was used by personal pump.

2.4. Synthesis of IL/GONPs

The mixture ionic liquid with ligand ([HMIM]

[PF

]/APDC) modied on graphene oxide

nanostructures(GONs) and used as adsorbent for

lead extraction. The synthesis of graphene oxide

(GO) was followed by the modied Hummers

method [31-33].

First, 10 g of graphite powder was

mixed with 500 mL of H

and stirred for one

day. Then, KMnO

(48 g) was mixed to the above

mixture at 55 °C. Next, the mixture was moved into

a beaker with 1000 mL of ice. Also, 100 mL of H

in 1000 mL of DW was added to the mixture up to

create a yellow color. The product was washed with

deionized water and HCl for many times and dried

at 70

C. Then, 0.1 g of APDC mixed with 0.25 g

Table 1. The AT-F-AAS and ET-AAS conditions for lead (Pb) determination

Features AT-F-AAS ET-AAS

Linear range 0.1-6.2 (mg L

-1

) 5-145 (µg L

-1)

Wavelength 283.3 nm 283.3 nm

Lamp current 5.0 mA 5.0 mA

Slit 0.5 nm 0.5 nm

Mode Peak Area Peak Area

Atomization Air-Acetylene Electrical

Auto Sampler 0.5-5.0(mL) 1-100(µL)

LOD 0.03(mg L

-1

) 1.2(µg L

-1

)

0.9998 0.9995

Lead analysis in air and human workers by IL/APDC/NGO Somayeh Mirza et al

Anal. Methods Environ. Chem. J. 4 (2) (2021) 34-59

of IL in presence of 2 mL of acetone and 0.2 g of

GO. After increasing temperature up to 50

C, the

mixture of IL/APDC were uniformly modied on

Go (Fig.1). Finally, 20 mg of adsorbent selected for

lead extraction in blood samples.

2.5. Extraction Procedure

In the DIL-SPME method, 10 mL of blood samples

were used for extraction and determination of lead

ions at pH 6.0. By procedure, 20 mg of IL/APDC

modied on GO were injected to human blood

samples and standard solution (5 - 310 µg L

−1

) at

pH=6.0 (Fig.2). After sonication of samples for 3.0

min, the lead separated and extracted with the APDC

on adsorbent as sulfur coordination bonding in liquid

phase (Pb

→: APDC/IL-GONPs). By centrifuging

(5 min; 4000 rpm), the solid phase trapped in [HMIM]

[PF

] phase and collected from the samples in the end

of the conical tube. The liquid phase was set aside

and the lead ions back-extracted from ([HMIM][PF

APDC)/NGO adsorbent in pH=2 (HNO

, 0.5 M, 200

μL) and diluted with 300 μL of DW before determined

by F-AAS. In addition, On the other hand, the 10 mL

of standard solution was used based on the DIL-SPME

procedure by the same conditions at pH=6.0. A blank

run without any lead ions was used for ten times. The

calibration curve for lead in standards solutions was

prepared. The analytical procedure was showed in

Table 2. Validation was followed by CRM of lead

and real samples, which was analyzed by ETAAS

and ICP-MS analyzer. The concentration lead in

work place air was achieved based on 7082 NIOSH

method for 40 workers and the mean concentration in

air were obtained 65.6 µgm

-3

for TWA measurements.

The recovery of lead procedure in blood samples

based on APDC/IL-GONPs was obtained by the

equation 1. The C

is the primary concentrations and

is the secondary concentration of Pb(II), which

was determined by APDC/IL-GONPs coupled DIL-

SPME procedure (n=8).

Fig.1. Synthesis of ([HMIM][PF

]/APDC) modied on graphene oxide

R% = (C

-C

)/C

×100 (Eq.1)

For workplace air, after determination lead in

lter, the concentrations (C

, µg mL

-1

) of lead in

the sample air was achieved by NIOSH method

(The average blank, C

). The concentration in

air calculated based on equation 2, the volumes

(mL) of the sample and media blanks is V

and

, respectively. Finally, the C (mg m

-3

), of lead

showed the concentration lead in air where the air

volume sampled was V (L).

(Eq. 2)

Fig.2. Lead extraction based on APDC/IL/NGO coupled to DIL-SPME procedure

Table 2. The analytical features for determination lead in human blood samples

by DIL-SPME procedure

Features value

pH 6.0

Amount of APDC/IL modied on GONPs (mg) 20.0

Sample volume of blood, serum (mL) 10 .0

Volume of sample injection 0.5 mL

Linear range for blood

Mean RSD %, n=8

5-310 μg L

-1

4.6

LOD for blood 1.25 μg L

-1

Enrichment factor for blood 19.6

Volume and concentration of HNO

0.2 mL,0.5 M

Shaking/Centrifuging time 3.0 min, 5 min

Correlation coefcient R

= 0.9995

Lead analysis in air and human workers by IL/APDC/NGO Somayeh Mirza et al

Anal. Methods Environ. Chem. J. 4 (2) (2021) 34-59

3. Results and discussion

3.1. Characterization

XRD patterns of GO and APDC/IL/GO are

similar and shown in Figure 3. Graphene oxide

exhibits the sole main diffraction peak at 2θ = 12°

corresponding to the oxygen functional groups

which are intercalated between graphene sheets in

the course of oxidation.

The peaks at 2θ = 12° and

42.58

are related to the diffraction planes of (002)

and (100) respectively, which can be observed in

the XRD patterns of both GO and APDC/IL/GO.

However, existence of this peak at about 2θ = 12

agrees well with the modication of graphene

oxide with APDC and IL.

The morphology of GO and APDC/IL/GO are

evaluated by scanning electron microscope (SEM)

(Fig. 4). According to the SEM images (Fig. 4I and

4II), Comparison between the SEM images of GO

and APDC/IL/GO revealed that, the modication

of GO had no prominent effect on the morphology

of the graphene sheets.

3.2. Optimization of proposed procedure

The DIL-SPME procedure with new IL/APDC/

NGO was applied for the extraction and separation

of lead ions in human blood samples. The high

recovery for lead extraction was obtained by

optimizing of parameters.

3.2.1.The pH optimization

The pH is the main factor for lead extraction in

human blood samples by APDC ligand which was

mixed with IL and modied on NGO. The various

pH for lead extraction in blood samples was used

and studied between 2-12. The results showed,

the IL/APDC/NGO had highly lead extraction at

pH of 5.5-6.5 (<95%). The complexation of lead

with APDC were decreased at pH less than 5 and

Fig.3. XRD of NGO and IL/APDC/NGO

pH more than 6.5. So, the pH=6 was selected as

optimum pH for lead extraction in blood samples

(Fig. 5). The mechanism of lead extraction

depended on sulfur group in APDC which was

created a dative bond with Pb ions in liquid phase

(Pb

→:S….APDC/IL/NGO-GO). At lower pH

the surface of adsorbent had positive-charge

(S+) and extraction decreased due to similarity

charges law. At pH of 6.0, the surface of APDC/

IL/NGO have negatively charged (S─) and Pb (+)

complexed by sulfur groups. Moreover, the lead

participated in liquid phase (Pb(OH)

) and the

extraction efciency was decreased at more than

pH of 7. Therefore, the pH=6 used for further

studies in blood samples.

3.2.2. Optimization of APDC/IL/NGO mass

The high extraction was achieved based on APDC/

IL/NGO mass which was depended to sulfur

group of APDC as complexation. So, the mass of

APDC/IL/NGO must be optimized conditions.

The amounts of APDC/IL/NGO between 2-50 mg

in human blood samples and standard solution

were studied for lead extraction by DIL-SPME

procedure. As Figure 6, the maximum recoveries

were achieved for Pb extraction by 20 mg of APDC/

IL/NGO in human blood samples. So, 20 mg of

APDC/IL/NGO was selected for experimental run

by DIL-SPME procedure. The extraction efciency

of lead had constant rate for more than 20 mg of

APDC/IL/NGO in blood samples.

Fig.4. I (SEM of NGO II) SEM of IL/APDC/NGO

Fig.5. The effect of pH on lead extraction based on APDC/IL/NGO by DIL-SPME procedure

Lead analysis in air and human workers by IL/APDC/NGO Somayeh Mirza et al

3.2.3.Optimization of sample volume and eluent

The volume and concentration of eluents such

as acid or bases is important for back extraction

Pb ions from APDC/IL/NGO were studied. The

low/ high pH caused to dissociate the sulfur-Pb

covalent bonds and released the Pb ions from

adsorbent. So, the different reagents (HCl, HNO

COOH, H

, H

) were used for the

back-extraction of lead from APDC/IL/NGO to

liquid solutions. The 0.1-1.0 molar of various

eluents with different volumes from 0.1 to 0.7

mL was studied. Due to Figure 7, the efcient

recovery obtained by 0.2 mL of HNO

(0.5 M).

In addition, the volume of blood and standard

solutions for lead extraction based on APDC/IL/

NGO adsorbent was optimized from 1.0 mL to

20.0 mL for LLOQ and ULLOQ concentration of

Pb(II) (5.0-310 µg L

-1

). The results showed that

the best recovery was achieved less than 10 mL

for in blood samples at pH=6. therefore, the 10

mL of blood or standard solution was used for

further work (Fig. 8).

Fig.6. The effect of amount of APDC/IL/NGO on lead extraction based

on APDC/IL/NGO by DIL-SPME procedure

Fig.7. The effect of eluents on lead extraction based

on APDC/IL/NGO by DIL-SPME procedure

Anal. Methods Environ. Chem. J. 4 (2) (2021) 47-59

3.2.4.Optimization of time, reusability and

absorption capacity

The time dispersion of APDC/IL/NGO adsorbent

in blood or standard samples had a main factor for

efcient extraction of lead. Moreover, the mass-

transference between APDC/IL/NGO and Pb ions

was occurred at pH=6 by DIL-SPME procedure.

In optimized conditions, the sonication time was

evaluated between 1 - 8 min. the results showed

that the sonication time of 3.0 min has maximum

extraction of lead in blood samples. In addition, the

centrifuging time between 1-10 min (4000 rpm)

was used for collecting sorbent in end of conical

tube. Based on results, 5 min of centrifuge time is

enough time for efcient extraction of lead. Finally,

the lead ions were back-extracted from APDC/IL/

NGO by changing pH and determined by F-AAS.

The reusability of APDC/IL/NGO was obtained

with many times extraction process by DIL-SPME

procedure. The results showed, the DIL-SPME can

be used for less than 7 cycles for lead extraction at

optimized pH in room temperature. The absorption

capacities for lead depended on chemical and

physical properties of APDC/IL/NGO in bath

system. The absorption capacities of APDC/IL/

NGO for Pb ions examined with 50 mg L

-1

optimized conditions. The adsorption capacity of

APDC/IL/NGO and NGO for Pb ions was achieved

154.7 mg g

-1

and 28.9 mg g

-1

, respectively.

3.2.5.Interference cations and anions

For extraction lead in blood samples, the effect of

interference of ions was studied by DIL-SPME

procedure. So, the concentrations of cations and

anions in human blood such as Cu, Zn, Mn, Mg, Ca,

Fe and etc. (1-2 ppm) added to 10 mL of standard

solution or human blood samples in presence of

300 μg L

-1

of lead at pH=6. The recovery of lead

extraction in presence of the concomitant cations/

anions couldn’t decreased at pH=6. Therefore, the

Pb ions can be extracted based on APDC/IL/NGO

in blood samples in presence of some coexisting

ions (Table 3).

3.2.6.Real sample analysis

The concentration of lead in human blood and

standard samples was determined by DIL-SPME

procedure. The lead was efciently extracted based

on APDC/IL/NGO adsorbent from human blood

samples and obtained results were validated by

electrothermal atomic absorption spectrometry

(ET-AAS) and ICP-MS techniques (Table 4). In

this study, the validation based on spiking real

Fig.8. The effect of sample volume on lead extraction based on APDC/IL/NGO

by DIL-SPME procedure

Lead analysis in air and human workers by IL/APDC/NGO Somayeh Mirza et al

samples with a standard solution were achieved

in human blood and serum samples by APDC/IL/

NGO adsorbent (Table 5). The results demonstrated

the efcient recovery for lead ions in difculty

matrixes in blood samples. The spiked samples

showed a accurate results for separation/extraction/

determination /preconcentration of human blood

samples by APDC/IL/NGO adsorbent. As intra-day

and inter-day analysis, the 40 human blood workers

of battery manufacturing factories determined by

same procedure ((25-55, Men, Iran) and compared

to ofce people as control markers (Table 6).

Table 3. The effect of interference cations and anions for lead extraction by the DIL-SPME procedure

Interfering Ions in blood (A)

Mean ratio

Pb(II)

)

Recovery (%)

Pb(II) Pb(II)

, Fe

550 99.3

, Cu

850 98.1

, Mo

600 96.5

, Br

, F

, Cl

1100 97.4

, K

1000 99.3

, Mg

1200 98.4

, PO

,NO

650 97.4

300 98.1

, V

, Cr

900 97.7

, Ag

120 98.2

Table 4. The validation of proposed method for lead extraction

in human matrixes by ICP-MS and ET-AAS (µgL

-1

)

Samples

ICP-MS DIL-SPME /F-AAS  ET-AAS

Serum 253.8 ± 7.2 244.9 ± 10.2 248.3 ± 12.7

Blood 302.7 ± 7.8 290.6 ± 11.6 295.1 ± 14.2

Plasma 142.5 ± 3.8 139.4 ± 5.5 145.6 ± 6.9

Mean of three determinations ± ccondence interval (P= 0.95, n=5),

As low LOD for ET-AAS and ICP-MS the samples diluted with DW before analysis

Table 5. Spiking real samples with a standard solution in human blood, serum

and plasma samples by APDC/IL/NGO adsorbent(µgL

-1

)

Sample Added

 Found

Recovery (%)

Serum

----- 123.2± 5.3 ----

50 170.9 ± 7.6 95.4

100 225.4± 9.8 102.2

Blood

----- 111.3± 4.9 -----

50 159.4 ± 6.1 96.2

100 209.8± 9.2 98.5

Plasma

----- 84.5 ± 4.2 -----

50 133.6 ± 5.8 98.2

100 183.7 ± 8.8 99.2

 Mean of three determinations ± ccondence interval (P= 0.95, n=5)

Anal. Methods Environ. Chem. J. 4 (2) (2021) 47-59

In addition, the blood samples were validated by

certied reference materials (CRM, NIST) by DIL-

SPME procedure (Table 7). Based on table 4-7, the

precision and accuracy of results was satisfactorily

conrmed in human blood samples.

4. Conclusions

An applied adsorbent based on APDC/IL/NGO

was used for separation, preconcentration and

determination of lead in blood samples by the

DIL-SPME procedure. The IL was helped to

separate the solid phase from liquid matrixes

in bottom of conical tube which was modied

on NGO. By DIL-SPME procedure, the simple,

perfect recovery and fast analysis was achieved

for lead at pH=6.0 by APDC/IL/NGO sorbent. The

modication of NGO surface with IL/APDC

helped

to enhance the lead extraction in blood samples.

Also, some advantages of proposed method such

as, fast separation, low cost, and high extraction

caused to comparable to ICP-MS analyzer. The

LOD and linear range(LR) has acceptable level

for lead analysis in human biological samples.

Therefore, the extraction and determination lead

with APDC/IL modied on NGO in human blood

were simply achieved based on sulfur dative

bond in optimized conditions by DIL-SPME

procedure before determined by F-AAS. The lead

analysis in workplace air was achieved based on

NIOSH method for 40 workers and range/mean

concentrations in air were obtained 44.7-81.5

µgm

-3

and 65.6 µgm

-3

, respectively for a 200 L of

air sample which was higher than standard of lead

in air by NIOSH (50 µgm

-3

5. Acknowledgements

The authors wish to thank from Faculty

of Natural resources and Environment, Science

and Research branch, Islamic Azad University,

Tehran, Iran and Birjand university, Birjand, Iran.

Table 6. Comparing of mean lead concentration in human blood

of battery workers with control peoples by DIL-SPME procedure (µgL

-1

)

Sample

Mean of Subjects (n=40) Mean of Controls (n=40)

Data Subject

Intra-day Inter day Intra-day Inter day r P value

Plasma 194.2± 8.7 191.5 ± 8.6 47.1 ± 2.3 48.4 ± 2.5 0.090 <0.001

Serum 401.2 ± 19.3 397.7± 18.9 94.4 ± 4.6 90.8 ± 4.4 0.088 <0.001

Blood 382.5 ± 18.4 386.3 ± 18.6 88.2 ± 4.1 85.9± 3.9 0.101 <0.001

Mean of three determinations of samples ± condence interval (P = 0.95, n =10)

Correlations are based on Pearson coefcients (r). Statistical signicance will be observed if P < 0.001

Subject and control groups belong to battery workers and control peoples

Table 7. The validation of methodology for lead extraction with certied reference material (CRM, ICP-MS)

by DIL-SPME procedure(µgL

-1

)

SRM Added

Found by μ-SPE



Recovery (%)

ICP-MS A -------

120.4 ± 5.7 -------

168.1 ± 7.3 95.4

100 218.5 ± 9.9 98.1

ICP-MS B -------

208.7 ± 9.8 -------

260.1 ± 14.2 102.8

100

305.4 ± 14.2 96.7



Mean value ± standard deviation based on three replicate measurements

ICP-MS: blood analysis for lead, A=122.5 ngL

-1

, B=210.3 (µgL

-1

)

Lead analysis in air and human workers by IL/APDC/NGO Somayeh Mirza et al

6. References

[1] J. Briffa, E. Sinagra, R, Blundell, Heavy

metal pollution in the environment and their

toxicological effects on humans, Heliyon, 6

(2020) e04691.

[2] A.L. Wani, A. Ara, J.A. Usmani, Lead toxicity:

A review, Int. Toxicol., 8 (2015) 55–64.

[3] Lead toxicity: what Is the biological Fate of Lead

in the Body, environmental health and medicine

education, Agency for toxic substances and

disease registry (ATSDR), 2017. https://www.

atsdr.cdc.gov/csem/csem.asp?csem=34&po=9

[4] M. Tamayo y Ortiz, M.M. Téllez-Rojo, H.

Hu, M. Hernandez-Avila, R. Wright, C.

Amarasiriwardena, N. Lupoli, A. Mercado-

Garcia, I. Pantic, H. Lamadrid-Figueroa, Lead

in candy consumed and blood lead levels of

children living in Mexico City, Environ. Res.,

147 (2016) 497–502

[5] J. Wieczorek, A. Baran, K. Urba ´nski, R.

Mazurek, A. Klimowicz-Pawlas, Assessment

of the pollution and ecological risk of lead and

cadmium in soils, Environ. Geochem. Health,

40 (2018) 2325–2342.

[6] J. Ahn, M.Y. Park, M.Y. Kang, I.S. Shin, S.

An, H.R. Kim, Occupational lead exposure

and brain tumors: systematic review and meta-

analysis, Int. J. Environ. Res. Public Health, 17

(2020) 3975.

[7] J. Sirivarasai, S. Kaojarern, S. Chanprasertyothin,

P. Panpunuan, K. Petchpoung, A.

Tatsaneeyapant, K. Yoovathaworn, T. Sura,

S. Kaojarern, P. Sritara, Environmental lead

exposure, catalase gene, and markers of

antioxidant and oxidative stress relation to

hypertension: An analysis based on the EGAT

study, Biomed. Res. Int., 2015 (2015) 856319.

[8] F. Aarabi, M. Kusajima, T. Tohge, T. Konishi,

T. Gigolashvili, M. Takamune, Y. Sasazaki, M.

Watanabe, H. Nakashita, A.R. Fernie, Sulfur

deciency-induced repressor proteins optimize

glucosinolate biosynthesis in plants, Sci. Adv.,

2 (2016) e1601087.

[9] O. P. Ajsuvakovaa, A. A. Tinkov, Sulfhydryl

groups as targets of mercury toxicity, Coord.

Chem. Rev., 15 (2020) 417. http://doi.

org/10.1016/j.ccr.2020.213343.

[10] Environmental Protection Agency (EPA), Basic

information about lead in drinking water, 2014.

[11] United States Food and Drug Administration

(FDA), Elemental impurities guidance for

industry, Department of Health and Human

Services, p. 41, 2017.

[12] US department of health and human services,

national institute for occupational safety and

health (NIOSH); Adult blood lead epidemiology

and surveillance (ABLES), 2017.

[13] S. Triantafyllidou, J. Burkhardt, Variability

and sampling of lead (Pb) in drinking water:

Assessing potential human exposure depends

on the sampling protocol, Environ. Int., 146

(2021) 106259.

[14] S. Kapitány, D. Nagy, J. Posta, A .Béni,

Determination of atmospheric sulphur dioxide

and sulphuric acid traces by indirect ame

atomic absorption method, Microchem. J., 157

( 2020) 104853.

[15] T. Amiri-Yazani, R. Zare-Dorabei, M. Rabbani,

A. Mollahosseini, Highly efcient ultrasonic-

assisted pre-concentration and simultaneous

determination of trace amounts of Pb (II)

and Cd (II) ions using modied magnetic

natural clinoptilolite zeolite: Response surface

methodology, Microchem. J., 146 (2019) 498-

508.

[16] P. Montoro-Leal, J.C. García-Mesa, M.T. Siles

Cordero, Magnetic dispersive solid phase

extraction for simultaneous enrichment of

cadmium and lead in environmental water

samples, Microchem. J., 155 (2019) 104796.

[17] J.S.F. Pereira, D. P. Moraes, Determination of

metals and metalloids in light and heavy crude

oil by ICP-MS after digestion by microwave-

induced combustion, Microchem. J., 96 (2010)

4-11.

[18] V. Balaram, Microwave plasma atomic

emission spectrometry (MP-AES) and its

applications – A critical review, Microchem. J.,

159 (2020)105483.

Anal. Methods Environ. Chem. J. 4 (2) (2021) 47-59

[19] S.M. S. Khademi, A. Salemi, Development

and comparison of direct immersion solid

phase micro extraction Arrow-GC-MS for the

determination of selected pesticides in water,

Microchem. J., 164 (2021) 106006.

[20] V. CâmaraCosta, M. Limade Mello, Calibration

strategies for determination of Pb content in

recycled polypropylene from car batteries using

laser-induced breakdown spectroscopy (LIBS),

Microchem. J., 159 (2020) 105558.

[21] Y. Song, Q. Ma, H. Cheng, J. Liu, Y. Wang,

Simultaneous enrichment of inorganic and

organic species of lead and mercury in

pg L

−1

levels by solid phase extraction online

combined with high performance liquid

chromatography and inductively coupled

plasma mass spectrometry, Anal. Chim. Acta,

1157 ( 2021) 338388.

[22] Y.-K. Li, W. Li, X. Liu, T. Yang, M.L.

Chen, J.H. Wang, Functionalized magnetic

composites based on the aptamer serve as

novel bio-adsorbent for the separation and

preconcentration of trace lead, Talanta, 203

(2019) 210-219.

[23] V. de Jesus Ferreira, J. S. Almeida, Determination

of Cu, Ni, Mn, and Pb in diesel oil samples

using reversed-phase vortex-assisted liquid-

liquid microextraction associated with energy

dispersive X-ray uorescence spectrometry,

Talanta, 222 (2021) 121514.

[24] A. H. Kame, A.Ei-Galil, E.Amr, Pre-

concentration based on cloud point extraction

for ultra-trace monitoring of lead (II) using

ame atomic absorption spectrometry, Appl.

Sci., 22 (2019) 4752. https://doi.org/10.3390/

app9224752

[25] M. Ghazaghi, H. Shirkhanloo, H.Z. Mousavi,

A.M. Rashidi, Ultrasound-assisted dispersive

solid phase extraction of cadmium(II)

and lead(II) using a hybrid nanoadsorbent

composed of graphene and the zeolite

clinoptilolite, Microchim. Acta, 182

(2015)1263-1272.

[26] M. Shokri, A. Beiraghi, S. Seidi, In

situ emulsication microextraction using a

dicationic ionic liquid followed by magnetic

assisted physisorption for determination of

lead prior to micro-sampling ame atomic

absorption spectrometry, Anal. Chim. Acta, 889

(2015) 123-129.

[27] M. Vesali-Naseh, M. RezaVesali Naseh, P.

Amer, Adsorption of Pb (II) ions from aqueous

solutions using carbon nanotubes: A systematic

review, J. Clean. Prod., 291 (2021) 125917.

[28] N. Rahman, M. Nasir, P.Varshney, Efcient

removal of Pb(II) from water using silica gel

functionalized with thiosalicylic acid: Response

surface methodology for optimization, J. King

Saud. Un. Sci., 33 (2021) 101232.

[29] N. Manousi, E. Rosenberg, E. A. Deliyanni,

Sample preparation using graphene-oxide-

derived nanomaterials for the extraction of

metals, Molecules, 25 ( 2020) 2411. https://doi.

org/10.3390/molecules25102411

[30] S. Mirza, N. Mansouri, R. Arjmandi, R.

Azizinejad, Evaluation and comparison of

health, safety and environmental management

system in oil and petrochemical downstream

industries (Case Study of Textile Factories), J.

Health safe. Work, 1132-35 (2021) .

[31] W.S.J. Hummers, R.E. Offeman, Preparation of

Graphitic Oxide, J. Am. Chem. Soc., 80 (1958)

1339.

[32] M.K. Abbasabadi, A. Rashidi, S. Khodabakhshi,

Benzenesulfonic acid-grafted graphene as a new

and green nanoadsorbent in hydrogen sulde

removal, J. Nat. Gas Sci. Eng., 28 (2016) 87-94.

[33] M. Khaleghi-Abbasabadi, D. Azarifar, Magnetic

Fe3O4-supported sulfonic acid-functionalized

graphene oxide (Fe3O4@GO-naphthalene-

SO3H): a novel and recyclable nanocatalyst

for green one-pot synthesis of 5-oxo-

dihydropyrano[3,2-c]chromenes and 2-amino-

3-cyano-1,4,5,6-tetrahydropyrano[3,2-c]

quinolin-5-ones, Res. Chem. Intermed. 45

(2019) 2095-2118.

Lead analysis in air and human workers by IL/APDC/NGO Somayeh Mirza et al