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Contents lists available at ScienceDirect

Journal of Food Composition and Analysis

journal homepage: www.elsevier.com/locate/jfca
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Original Research Article

Determination of pyrethroid pesticides residues in vegetable oils using

liquidliquid extraction and dispersive liquidliquid microextraction
followed by gas chromatographyame ionization detection
Q1 Mir
a
b
c

Ali Farajzadeh a,*, Leila Khoshmaram b, Ali Akbar Alizadeh Nabil c

Department of Analytical Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz, Iran

Department of Chemistry, Faculty of Science, Azarbaijan Shahid Madani University, Tabriz, Iran
Toxicology Laboratory, Food Control Laboratory, Food and Drug Department, Tabriz University of Medical Science, Tabriz, Iran

A R T I C L E I N F O

A B S T R A C T

Article history:
Received 16 July 2013
Received in revised form 9 March 2014
Accepted 9 March 2014

A simple, fast and economical method has been developed for the determination of fenpropathrin,
sumithrin, cyhalothrin, permethrin and deltamethrin in vegetable oils. The method is based on a
dispersive liquidliquid microextraction (DLLME) technique after a preliminary liquidliquid extraction
step, which is followed by a gas chromatographyame ionization detection or gas chromatography
mass spectrometry determination. Initially oil samples were partitioned in a dimethylformamide
(DMF)hexane mixture and then DMF was removed and used as a disperser solvent in the following
DLLME procedure in which mL-level of 1,1,2-trichloroethane was used as an extraction solvent. The
detection limits ranged from 0.02 to 0.16 mg kg1 and the relative standard deviations (RSD %) for four
repeated experiments (C = 0.2 and 0.6 mg kg1 of each pesticide) varied from 2% to 13%, and enrichment
factors were in the ranges of 4070. In p value of 0.9, neither signicant matrix effects nor interferences
were observed by employing the proposed treatment in analysis of oil samples, and the extraction
recoveries obtained for the samples spiked with 0.2, 0.6 and 1.1 mg kg1 levels varied from 109  3% to
85  5%. The method was successfully applied in determination of target analytes in vegetable oils.
2014 Published by Elsevier Inc.

Keywords:
Dispersive liquidliquid microextraction
Sample preparation
Gas chromatography
Flame ionization detector
Mass spectrometry
Dimethylformamide
Pyrethroid pesticides
Vegetable oils
Pesticide residue
Food-chain contamination
Food safety
Food composition
Food analysis

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1. Introduction

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Pyrethroids are the synthetic analogs of pyrethrins which were

developed as pesticides from the extracts of dried and powdered

Abbreviations: DLLME,
dispersive
liquidliquid
microextraction;
DMF,
dimethylformamide; DMSO, dimethyl sulfoxide; DSPE, dispersive solid phase
extraction; ECD, electron capture detector; EF, enrichment factor; ER, extraction
recovery; FID, ame ionization detection; GC, gas chromatography; GPC, gel
permeation chromatography; LLE, liquidliquid extraction; LOD, limit of detection;
LOQ, limit of quantication; MS, mass spectrometry; MS2, tandem mass
spectrometry; MSPE, matrix solid-phase dispersion; RSD, relative standard
deviation; SEC, size-exclusion chromatography; SFE, supercritical uid extraction;
SPE, solid phase extraction; SPME, solid phase microextraction.
* Corresponding author. Tel.: +98 411 3393084; fax: +98 4113340191.
E-mail addresses: mafarajzadeh@yahoo.com, mafarajzadeh@tabrizu.ac.ir
(M.A. Farajzadeh).

ower heads of Chrysanthemum cinerariaefolium (Anadon et al., 16

2009). Pyrethroid pesticides are widely used in agriculture on a 17
variety of fruits and vegetables, to protect stored commodities, and 18
for the control of household, industrial and veterinary pests. 19
Pyrethroid pesticides are lipophilic compounds and their extrac- 20
tion from fatty matrices such as foods of animal origin, oils, cereals 21
and oil seeds, is accompanied by the simultaneous extraction of 22
considerable amounts of fatty material (Muccio et al., 1999).
Q223
Methods for fatty food analysis are usually laborious and not 24
fully effective in cleaning-up the sample if the compounds happen 25
to be evaluated below a few mg kg1 of lipid weight. To determine 26
the traces of lipophilic compounds, such as pyrethroid pesticides, 27
their separation from bulk of fatty material should be achieved 28
(Kodba and Voncina, 2007). Several sample preparation methods 29
have been developed for the analysis of lipophilic compounds in oil 30
samples, such as solid phase extraction (SPE) (Amvrazi and Albanis, Q331

http://dx.doi.org/10.1016/j.jfca.2014.03.004
0889-1575/ 2014 Published by Elsevier Inc.

Please cite this article in press as: Farajzadeh, M.A., et al., Determination of pyrethroid pesticides residues in vegetable oils using liquid
liquid extraction and dispersive liquidliquid microextraction followed by gas chromatographyame ionization detection. J. Food
Compos. Anal. (2014), http://dx.doi.org/10.1016/j.jfca.2014.03.004

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2006; Bakas et al., 2013; Esteve-Turrillas et al., 2005; Ibrahim et al.,

2014; Lopez-Feria et al., 2009; Yague et al., 2005), dispersive solidphase extraction (Deme et al., 2014; Sobhanzadeh et al., 2012; Su
et al., 2011; Polgar et al., 2012), gel permeation chromatography
(GPC) (Guardia-Rubio et al., 2006; Patel et al., 2005; Wang et al.,
2014), solid phase microextraction (SPME) (Beltran et al., 2000;
Kataoka et al., 2000; Tsoutsi et al., 2006), matrix solid phase
dispersion (MSPD) (Li et al., 2013; Sobhanzadeh et al., 2011)
liquidliquid microextraction (Becerra-Herrera et al., 2014) and
supercritical uid extraction (SFE) (Hopper, 1999; Motohashi et al.,
2000; Pihlstrom et al., 2002).
Liquidliquid extraction (LLE) is known as a procedure which can
extract a wide variety of lipophilic compounds without or with
limited enrichment. On the other hand, dispersive liquidliquid
microextraction (DLLME) can be used as an efcient alternative to
the extraction and clean-up steps. It was introduced by Assadi and
coworkers in 2006 as a extraction technique with high enrichment
factor (EF) (Rezaee et al., 2006). It is based on the ternary
components solvent system such as homogeneous LLE and cloud
point extraction, including extraction solvent, dispersive solvent
and aqueous sample containing analytes of interest. The method is
attracting more and more attention due to its superior advantages of
high EF, high recovery, low cost, simplicity, rapidity and easy
operation (Rezaee et al., 2010). Also, DLLME has become a very
popular environmentally benign sample preparation technique,
because it consumes low volumes of organic solvent. DLLME has
been applied for the analysis of different compounds such as
opium alkaloids in human plasma (Ahmadi-Jouibari et al., 2013),
cyclic and linear siloxanes (Cortada et al., 2014), silver, lead,
copper and cadmium (Ramos and Borges, 2014), phthalate esters
in alcoholic beverages ( Fan et al., 2013), pesticide residues (Chen
et al., 2013), cobalt and nickel (Sorouraddin and Khoshmaram,
2010), amphetamines (Ahmadi-Jouibari et al., 2014), organophosphate (Ho et al., 2013) and bisphenol A (Liu et al., 2013) in
different samples.
In this study for the rst time the combination of LLE-DLLME is
proposed. The goal of this combination is to develop a simple and
rapid method for isolation, enrichment and determination of
pyrethroid pesticides (fenpropathrin, sumithrin, cyhalothrin,
permethrin and deltamethrin) in vegetable oils. It should be
noted that these ve pyrethroids were selected since they are used
more than others in Iran. This method consists of two steps: (i)
extraction of pesticides from vegetable oils by LLE; and (ii)
enrichment of the pesticides with DLLME. In the rst step,
pesticides are extracted into dimethylformamide (DMF) which is
used as a disperser in DLLME in the second step. The factors which
affect both LLE and DLLME were investigated and the optimal
conditions were selected. Figures of merit for the proposed
approach are reported using a gas chromatographyame ionization detection (GCFID) analysis and conrmation of the presence
of the studied pyrethroids in positive oil samples was done by GC
mass spectrometry (MS) analysis.

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2. Materials and methods

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2.1. Chemicals and solutions

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Fenpropathrin, sumithrin, cyhalothrin, permethrin (cis and trans

isomers mixture at a ratio of40:60), and deltamethrin were gifted by
GYAH Corporation (Karaj, Iran). Organic solvents such as dimethylformamide (DMF), acetonitrile, dimethyl sulfoxide (DMSO), 1,1,2,2tetrachloroethane, methanol, n-hexane, 1,2-dibromoethane, chloroform, and carbon tetrachloride were purchased from Merck
(Darmstadt, Germany). 1,1,2-Trichloroethane from Janssen Chimica
(Beerse, Belgium) was employed. De-ionized water (Ghazi Company, Tabriz, Iran) was used for the preparation of aqueous solutions.

Sodium chloride (analytical-reagent grade, Mojallali, Tehran, Iran)

was used to adjust the ionic strength of aqueous solutions.
Standard solution of pesticides (100 mg L1, each pesticide)
was prepared by dissolving an appropriate amount of each
pesticide into 1,1,2-trichloroethane. This solution was injected
into the separation system each day (three times) for quality
control and the obtained peak areas were used in the calculation of
EFs and recoveries. Also, a stock solution of pesticides was
prepared in n-hexane at a concentration of 50 mg L1 (each
pesticide). This solution was prepared weekly to spike the samples
or to obtain the optimum experimental conditions.

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2.2. Samples

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Four oil samples including corn (Famila Company, Golestan,

Iran), sunower and colza (Ladan Company, Tehran, Iran) and olive
oils (Roudbar Company, Gilan, Iran) were purchased from local
supermarkets in August 2012 (Tabriz, Iran). The sample size for
each oil was 1 L (ve samples were collected per oil type). They
were analyzed immediately after opening.

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2.3. Apparatus

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A gas chromatograph (GC-2014, Shimadzu, Japan) equipped with

a split/splitless injector system, and a ame ionization detector was
used for separation and determination of the selected pyrethroid
pesticides. Helium (99.999%, Gulf Cryo, United Arab Emirates) was
used as the carrier gas at a constant linear velocity of 30 cm s1.
Separation was carried out on an SPB-1 capillary column (100%
dimethyl siloxane, 30 m  0.25 mm i.d., and lm thickness
0.25 mm) (Supelco, Bellefonte, USA). The oven temperature was
programmed as follows: initial temperature 125 8C (held 1 min), and
ramped to 220 8C at a rate of 25 8C min1, then to 280 8C at a rate of
5 8C min1 and held for 3 min. The total time for one GC run was
about 20 min. The injector and FID temperatures were maintained at
300 8C. Hydrogen gas was generated with a hydrogen generator
(OPGU-1500S, Shimadzu, Japan) for FID at a ow rate of
40 mL min1. The ow rate of air for FID was 300 mL min1 and
that of make up gas (helium) was 30 mL min1. GCMS analysis was
carried out on an Agilent 6890N gas chromatograph with a 5973
mass-selective detector (Agilent Technologies, CA, USA). The
separation was carried out on an HP-5 MS capillary column
(30 m  0.25 mm i.d., and lm thickness 0.25 mm). Helium was
used as carrier gas at a ow rate of 1.0 mL min1. The temperature
program of the oven was the same as that of GCFID analysis
mentioned above. The ion source temperature of MS was maintained
at 250 8C, with an ionizing energy at 70 eV. A scan range of 50
500 m/z was used for full scan analysis of samples.

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2.4. Extraction/preconcentration procedure

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2.4.1. Liquidliquid extraction procedure

A 0.5 mL sample of different vegetable oils or pesticide-free
sunower oil was spiked with analytes at the concentration of
1.1 mg kg1 (each pesticide) into a 10-mL screw cap glass test tube.
Then, 4.5 mL n-hexane was added and the mixture was shaken
manually for 30 s to homogenize. After that 1 mL DMF was added
to the tube. After manual shaking (2 min), the mixture was
centrifuged at 4000 rpm for 2 min. After centrifugation, the
pesticides were extracted into lower phase (DMF). About 0.7 mL
DMF was collected. Finally, 0.6 mL from the lower organic phase
was removed and used in the following DLLME procedure.

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2.4.2. Dispersive liquidliquid microextraction procedure

First, 5 mL NaCl aqueous solution (15%, w/v) was placed into
a glass test tube with a conical bottom. An aliquot of 20 mL

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Please cite this article in press as: Farajzadeh, M.A., et al., Determination of pyrethroid pesticides residues in vegetable oils using liquid
liquid extraction and dispersive liquidliquid microextraction followed by gas chromatographyame ionization detection. J. Food
Compos. Anal. (2014), http://dx.doi.org/10.1016/j.jfca.2014.03.004

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1,1,2-trichloroethane was added to 0.6 mL of DMF extract obtained

from the previous extraction step and injected rapidly into NaCl
solution by a 1 mL-syringe. A cloudy solution, resulting from the
dispersion of the ne droplets of 1,1,2-trichloroethane into the
aqueous solution, was formed in the tube. The mixture was
centrifuged at 10 000 rpm for 5 min. The dispersed 1,1,2-trichloroethane droplets were sedimented (5 mL) at the bottom of the
conical test tube. Finally, 1 mL of the sedimented phase was injected
into the separation system.
It should be noted that permethrin was a mixture of cis and
trans isomers and therefore gave rise to split peaks in chromatograms. The preferred approach for the reporting of these isomeric
compounds was to integrate the peaks separately.

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2.5. Calculation of enrichment factor and extraction recovery

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The EF was dened as the ratio between the analyte

concentration in the sedimented phase (Csed) and the initial
concentration of analyte (C0) in the sample, in Eq. (1):
EF

C sed
;
C0

(1)

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where Csed is obtained from comparing peak areas of the analytes

achieved in two cases, direct injection of standard solution of the
selected pesticides in extraction solvent, and injection of
sedimented phase having enriched analytes.
Extraction recovery (ER) is dened as the percentage of the total
analyte amount (n0) which is extracted into the sedimented phase
(nsed), in Eq. (2):




nsed
C sed  V sed
 100
 100
ER
C0  V 0
 n0 
;
(2)
V sed
 EF  100
ER
V0

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where Vsed and V0 are the volumes of the sedimented phase and oil
sample, respectively.

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3. Results and discussion

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In this study, initially the selected pyrethroid pesticides were

extracted from the oil sample into an organic solvent which acts
as an extractant for analytes in the rst step and as a disperser in
the second step (DLLME). In order to select the optimum
conditions for the determination of analytes by the proposed
method, it is required to optimize the different parameters that
affect the extraction of pesticides from oil sample and DLLME.
Some of these parameters are as follows: selection of suitable
extraction solvent for the extraction of analytes from the oil
sample (also disperser solvent for the following DLLME), the
volume of extraction solvent in the rst step, type of extraction
solvent in DLLME and the ionic strength of aqueous phase in
DLLME. It is important to optimize them in order to obtain high
EFs for analytes and to improve analytical signals. It should be
noted that in optimization of different steps, one sample was
analyzed in triplicate ( n = 3) and mean of data was used in
preparation of tables or plotting gures.

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3.1. Optimization of parameters in the extraction of pesticides from oil

sample

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3.1.1. Selection of extraction solvent

In this study, extraction solvent of pesticides from oil sample
will be used as the disperser solvent in the following DLLME step.
Therefore, the choice of an appropriate extraction solvent is very
important. The requirements for the selection of this solvent are:
(i) capability for the extraction of analytes from a lipophilic matrix

Fig. 1. Selection of extraction solvent type in extraction of the pesticides from oil
sample. Extraction conditions: sample, 0.5 mL oil (spiked with 110 mg kg1 of each
pesticide) diluted with 4.5 mL n-hexane; extraction solvent volume, 1 mL. The error
bars indicate the minimum and maximum of three determinations.

(oil sample); (ii) low solubility in n-hexane (diluent for oil

sample); and (iii) miscibility with aqueous phase and extraction
solvent used in DLLME step. So methanol, acetonitrile, DMF and
DMSO were tested for this purpose. A 0.5 mL of the analytes-free
oil sample spiked with 110 mg kg1 of each pyrethroid pesticide
along with 4.5 mL n -hexane were placed into a 10-mL screw cap
glass test tube and the mixture was homogenized. Then 1.0 mL
of the mentioned extraction solvents was added to the mixture.
After manual shaking and centrifugation at 4000 rpm for 2 min,
1 mL of the lower phase was injected into the GC. It should be
noted that each experiment was repeated three times. The
obtained results showed that methanol was not able to extract
any of the analytes. Also, comparison of the peak areas obtained
with other extraction solvents (Fig. 1) shows that DMF is the
most effective extraction solvent among the tested solvents and
gives the highest analytical signals for most analytes. Since
DMSO has a melting point near to room temperature (1619 8C),
its usage is difcult. On the other hand, in the cases of
acetonitrile and DMF, similar analytical signals were obtained
for sumithrin and permethrin (cis). However , by considering
that acetonitrile is relatively volatile (b.p. = 81 8C), DMF was
selected as the extraction solvent for the subsequent experiments.

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3.1.2. Study of DMF volume

To examine the effect of the extraction solvent volume,
different volumes of DMF (0.50, 0.75, 1.00, 1.25 and 1.50 mL)
were subjected to the extraction procedure described above. The
results (not shown) indicated that the highest analytical signals
are observed in the case of 1.00 mL DMF. It should be noted that
the sedimented DMF was about 0.7 mL. It seems that at volumes
less than 1.00 mL, extraction of the analytes is incomplete and at
volumes more than 1.00 mL, dilution of the extracted analytes in
DMF is occurred. So, 1.00 mL DMF was used for the further
studies.

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3.2. Optimization of parameters in dispersive liquidliquid

microextraction step

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3.2.1. Selection of extraction solvent type

Five organic solvents namely 1,2-dibromoethane, 1,1,2,2tetrachloroethane, 1,1,2-trichloroethane, carbon tetrachloride
and chloroform were chosen as extraction solvents in DLLME,
because they have the properties such as higher density than
water, extraction capability for analytes, low solubility in water
and good chromatographic behavior. For this purpose, after the
extraction of pyrethroid pesticides from oil sample into DMF,

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Please cite this article in press as: Farajzadeh, M.A., et al., Determination of pyrethroid pesticides residues in vegetable oils using liquid
liquid extraction and dispersive liquidliquid microextraction followed by gas chromatographyame ionization detection. J. Food
Compos. Anal. (2014), http://dx.doi.org/10.1016/j.jfca.2014.03.004

G Model

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Fig. 2. Selection of extraction solvent type in DLLME. Extraction conditions: sample, 0.5 mL oil sample spiked with 1.1 mg kg1 of each pesticide diluted with 4.5 mL n-hexane;
extraction solvent in LLE, 1.00 mL DMF; extraction solvent in DLLME, 1,2-dibromoethane (50 mL), 1,1,2,2-tetrachloroethane (55 mL), 1,1,2-trichloroethane (60 mL), carbon
tetrachloride (75 mL) and chloroform (100 mL); centrifuge rate, 10 000 rpm; and centrifuge time, 5 min. The error bars indicate the minimum and maximum of three
determinations.

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0.6 mL DMF was removed, mixed with different volumes of

extraction solvents at microlitre level and injected rapidly into
5.0 mL deionized water by a 1 mL-syringe to achieve 5.0 mL of the
sedimented phase volume. As solubility of various solvents in
aqueous phase is different, if the same volume of the extraction
solvents had been used, different volumes of the sedimented phase
would have been obtained. In this case, besides the extraction
solvent type, the difference in volumes of the sedimented phase
would also affect the peak area. So in order to obtain the same
volume of sedimented phase (5.0 mL), different volumes of
extraction solvents were chosen. Thereby 50, 55, 60, 75 and
100 mL of 1,2-dibromoethane, 1,1,2,2-tetrachloroethane, 1,1,2trichloroethane, carbon tetrachloride and chloroform were used,
respectively. The results (Fig. 2) reveal that 1,1,2-trichloroethane
has the highest extraction efciency in comparison with the other
solvents. Therefore it was selected as the extraction solvent in
DLLME procedure.

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3.2.2. Inuence of the extraction solvent volume

In DLLME method, the volume of extraction solvent should be
optimized, because this factor can affect the extraction efciency.
For this purpose, after the extraction of pyrethroid pesticides from
oil sample into DMF, 0.6 mL DMF was removed, mixed with
different volumes of 1,1,2-trichloroethane (70200 mL) and
subjected to the same DLLME procedure. The results (not shown)
revealed that, by increasing the volume of 1,1,2-trichloroethane
from 75 to 200 mL, the analytical signals decreased rapidly. It
should be noted that the volume of sedimented phase increased
with increasing volume of 1,1,2-trichloroethane, and EFs decreased synchronously. Therefore, 75 mL of 1,1,2-trichloroethane
was selected as the volume of extraction solvent in DLLME. In the
case of 70 mL 1,1,2-trichloroethane, the volume of sedimented
phase was about 3 mL and so its removal was difcult and the
repeatability of the method decreased.

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3.2.3. Inuence of salt addition

The salting-out effect is commonly used for the enhancement of
extraction efciency. Generally, the addition of salt can decrease
the solubility of analytes in the aqueous phase (and can also reduce
the solubility of organic solvents in water) while enhancing their
partitioning into the organic phase. Salting-out effect on the
performance of DLLME step was evaluated by adding sodium
chloride into aqueous phase in the range of 025% (w/v). For this
purpose, different volumes of 1,1,2-trichloroethane were tested to
achieve 5 mL of sedimented phase volume after DLLME (75, 35, 25,

20, 17 and 15 mL for 0, 5, 10, 15, 20 and 25%, w/v, NaCl

concentration, respectively). Plots of peak areas against amount (%)
of NaCl are presented in Fig. 3. It was found that extraction
efciency increased up to 10% (w/v) NaCl concentration and after
that remained constant. Therefore, 15% (w/v) NaCl concentration
was selected in the further experiments in order to have higher
extraction recoveries and relatively lower extraction solvent
volume (20 mL).

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3.2.4. Optimization of centrifuge rate and time

Centrifugation is a mandatory process in DLLME, and hence
optimization from the views of rate and time is necessary. The
effect of centrifuging speed was examined in the range of 2000
10 000 rpm for 5 min. The results showed that, by increasing the
speed of centrifuging from 6000 to 10 000 rpm, the volume of
sedimented phase increased from 3 to 5 mL and the repeatability of
method increased. It should be noted that in the case of 5000 rpm
or less, no sedimented phase was collected. So, 10 000 rpm was
selected as optimum speed of centrifuging.
The effect of centrifuging time was examined in the range of
110 min at 10 000 rpm. The results showed that, in the range of
410 min the volume of sedimented phase was 5 mL and peak
areas were nearly constant. It should be noted that in the case of
3 min or less, the volume of sedimented phase was 3 mL or less, and

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Fig. 3. Effect of NaCl addition on the method performance. Extraction conditions:

NaCl concentration, 0%, 5%, 10%, 15%, 20%, and 25% (w/v) for 1,1,2-trichloroethane
volume 75 mL, 35 mL, 25 mL, 20 mL, 17 mL and 15 mL, respectively. Other conditions
are the same as Fig. 2. The error bars indicate the minimum and maximum of three
determinations.

Please cite this article in press as: Farajzadeh, M.A., et al., Determination of pyrethroid pesticides residues in vegetable oils using liquid
liquid extraction and dispersive liquidliquid microextraction followed by gas chromatographyame ionization detection. J. Food
Compos. Anal. (2014), http://dx.doi.org/10.1016/j.jfca.2014.03.004

G Model

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Table 1
Recoveries obtained by the proposed method in oil samples spiked at 0.20, 0.60 and 1.10 mg kg1 and analyzed by GCFID.
Mean recovery (%)  relative standard deviation (n = 3)a

Sample

Spiked concentration (mg kg1)

Cyhalothrin

cis-Permethrin

trans-Permethrin

Deltamethrin

Sunower oilb

0.20
0.60
1.10

93  4
95  4
94  3

88  5
90  5
92  5

109  3
104  3
102  4

108  2
107  2
98  3

89  2
100  3
99  3

<LOQc
89  5
98  2

Corn oild

0.20
0.60
1.10

91  6
93  6
92  4

93  5
92  4
98  2

89  5
107  5
99  2

87  5
98  5
95  4

92  3
90  3
94  4

<LOQ
85  5
88  3

Colza oilb

0.20
0.60
1.10

89  3
95  3
102  4

102  7
95  7
97  3

99  4
93  4
98  2

93  4
87  4
93  4

89  6
88  6
101  2

<LOQ
95  4
103  5

Olive oile

0.20
0.60
1.10

103  4
101  2
99  4

108  2
106  2
104  3

101  2
102  3
98  4

101  5
103  2
102  6

97  5
95  6
96  5

<LOQ
95  6
96  4

Fenpropathrin

a
b
c
d
e

Sumithrin

Analytes contents of samples were subtracted.

From Ladan company (Tehran, Iran).
LOQ for deltametrin is 0.555 mg kg1.
From Famila company (Golestan, Iran).
From Roudbar company (Gilan, Iran)

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319

so its removal was difcult. Hence, 5 min was selected as the

optimum time for centrifuging.

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3.2.5. Investigation of the matrix effect on recovery rates

To evaluate the matrix effect in different vegetable oils, the
added-found method was used. The relative recoveries for
analytes in oil sample in comparison with the results obtained
for 5 mL n -hexane spiked at the same concentrations were
calculated after performing the proposed method on them.
According to the obtained results, relatively strong matrix effect
was observed when 1 mL vegetable oil was applied in partitioning
DMFn-hexane step. But by applying 0.5 mL vegetable oil in this
step, there was no signicant effect on the performance of the
presented technique. The obtained results are summarized in
Table 1. So, 0.5 mL vegetable oil was used in the further
experiments.

333

3.3. Analytical features of the proposed method

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Quantitative characteristics of the proposed method were

obtained under the optimized conditions. Some quantitative
parameters including linear ranges of calibration graphs,
correlation coefcients ( r), limits of detection (LOD), limits of
quantication (LOQ), relative standard deviations (RSD), enrichment factors (EF) and extraction recoveries (ER) were calculated

and summarized in Table 2. Good linearities were obtained

for the calibration graphs with square of correlation coefcients
higher than 0.990. Comparing with the direct injection, the
EFs of fenpropathrin, sumithrin, cyhalothrin, cis-permethrin,
trans-permethrin and deltamethrin were calculated as 70,
56, 69, 42, 66 and 40, respectively. ERs were in the range of
4070%.
Precision of the method was evaluated from the relative
standard deviation of four analyses performed on the samples
spiked with either 0.2 or 0.6 mg kg1 of each analyte and the
obtained RSDs were from 2% to 13%. LODs and LOQs were calculated
at a signal-to-noise ratio (S/N) of 3 and 10, respectively. LODs ranged
from 0.02 to 0.16 mg kg1 (0.019 mg kg1, 0.020 mg kg1,
0.023 mg kg1, 0.045 mg kg1, 0.046 mg kg1 and 0.166 mg kg1
for cyhalothrin fenpropathrin, sumithrin, trans-permethrin, cispermethrin and deltamethrin, respectively) and LOQs ranged from
0.06 to 0.55 mg kg1, which are completely low for a GC-FID method
and sample volume 0.5 mL. Low LODs and LOQs, relatively high ERs
and good repeatability are the major advantages of the proposed
method.

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3.4. Application of the proposed method to real samples

360

To evaluate performance of the presented method, extraction/

preconcentration and determination of the selected pyrethroid

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Table 2
Analytical parameters of the proposed method.
Analyte

Fenpropathrin
Sumithrin
Cyhalothrin
cis-Permethrin
trans-Permethrin
Deltamethrin
a
b
c
d
e
f
g

LRa

0.066
0.066
0.066
0.156
0.156
0.556

rb

0.990
0.991
0.998
0.999
0.995
0.990

LODc

0.02
0.02
0.02
0.05
0.04
0.17

LOQd

0.07
0.07
0.06
0.16
0.15
0.55

Precision (RSD %)e

0.20 mg kg1

0.60 mg kg1

5
6
2
8
8
<LOQ

5
5
2
7
6
13

EF  SDf

ER %  SDg

70  4
56  5
69  2
42  4
66  5
40  5

70  4
56  5
69  2
42  4
66  5
40  5

Linear range (mg kg1).

Correlation coefcient.
Limit of detection (S/N = 3) (mg kg1).
Limit of quantication (S/N = 10) (mg kg1).
Relative standard deviation (n = 4).
Enrichment factor  standard derivation (n = 3).
Extraction recovery  standard derivation (n = 3).

Please cite this article in press as: Farajzadeh, M.A., et al., Determination of pyrethroid pesticides residues in vegetable oils using liquid
liquid extraction and dispersive liquidliquid microextraction followed by gas chromatographyame ionization detection. J. Food
Compos. Anal. (2014), http://dx.doi.org/10.1016/j.jfca.2014.03.004

G Model

YJFCA 2406 18
M.A. Farajzadeh et al. / Journal of Food Composition and Analysis xxx (2014) xxxxxx

Fig. 4. GCFID chromatograms of (a) standard solution (100 mg L1 of each

pesticide in 1,1,2-trichloroethane) (direct injection), (b) corn oil spiked with
pyrethroid pesticides (0.13 mg kg1 of each pesticide), (c) corn oil, (d) colza oil, (e)
olive oil and (f) blank (n-hexane). In all cases except chromatogram (a), the
proposed method was performed and 1 mL of the sedimented phase was injected
into GC. Peaks identication: 1, fenpropathrin; 2, sumithrin; 3, cyhalothrin; 4, cispermethrin; 5, trans-permethrin; and 6, deltamethrin.

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365
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pesticides in four oil samples including corn oil, colza oil, olive
oil and sunower oil were carried out under the optimum
conditions established above. It should be noted that ve
samples of every oil type were analyzed. The obtained results
showed that there were three positive oil samples; however, in

sunower oil, the examined pesticides were in concentrations

below the LOD values. Fenpropathrin was found in the
concentrations of 0.0870.093 mg kg1 in corn oil. Cyhalothrin
was determined in colza oil (0.0770.083 mg kg1). Sumethrin
and cyhalothrin were found in the concentrations of 0.125
0.135 mg kg1 and 0.1160.124 mg kg1, respectively, in olive
oil. Fig. 4 shows the typical GC FID chromatograms of olive oil,
corn oil and colza oil, standard solution (100 mg L1 of each
pesticide in 1,1,2-trichloroethane), corn oil spiked with the
studied pesticides (0.13 mg kg1 of each pesticide) and blank n hexane.
The proposed extraction procedure followed by GCMS
detection was performed on samples in which some of analytes
were detected by GCFID to conrm the obtained results. A typical
total ions chromatogram (TIC) for corn oil along with mass spectra
for fenpropathrin and scan 780 (retention time 13.22 min)
conrmed the presence of fenpropathrin in this sample (Fig. 5).
Presence of cyhalothrin in colza oil and sumethrin and cyhalothrin
in olive oil was also veried by GCMS data (not shown).

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381
382
383
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385
386

3.5. Comparison of the proposed method with relevant literature

387

Table 3 summarizes linear range (LR), LOD, LOQ and RSD of

some analytical methods along with those of the proposed method
for extraction and determination of the selected pesticides in
vegetable oils. In this study, GC along with FID was applied for
separation, detection and determination of the target pesticides
but the used detectors in other methods are expensive and
sensitive detectors such as electron capture detector (ECD) or MS.
However, LODs and LOQs for the proposed method were
comparable with those of other methods and RSDs of the proposed
method are better or comparable with other methods. Also LRs of
the proposed method are larger than other methods except sizeexclusion chromatography (SEC)GCMS method. These results
reveal that the presented technique is simple, inexpensive and
repeatable and it can be used successfully for extraction,
enrichment and determination of the pesticides residues from
fatty matrices.

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394
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397
398
399
400
401
402
403

Table 3
Comparison of the proposed LLEDLLMEGCFID method with other methods in determination of the selected pyrethroid pesticides in real samples.
Analyte

LRa

Method
e

LODb

LOQc

RSDd

Reference

0.03

0.0450.047

0.05
0.008
0.1500.156

12.512.9
613
26
67

Barrek et al. (2003)

Lentza-Rizos et al. (2001)
Nguyen et al. (2010)
This method

Permethrin

SECGCMS
LLESPEGCECDg
LLEDSPEGCMSi
LLEDLLMEGCFID

0.0670

0.156

Deltamethrin

MSPDGCMS2f
LLESPEGCECDg
MSPDGCMSh
LLEDSPEGCMSi
LLEDLLMEGCFID

0.010.5

0.0250.5

0.556

0.006

0.060

0.166

0.05

0.05
0.555

4
716

56
13

Garca-Reyes et al. (2007)

Lentza-Rizos et al. (2001)
Ferrer et al. (2005)
Nguyen et al. (2010)
This method

Cyhalothrin

LLESPEGCECDg
LLEDSPEGCMSi
LLEDLLMEGCFID

0.066

0.019

0.02
0.008
0.063

417
27
2

Lentza-Rizos et al. (2001)

Nguyen et al. (2010)
This method

Fenpropathrin

LLEDSPEGCMSi
LLEDLLMEGCFID

0.066

0.020

0.008
0.066

27
5

a
b
c
d
e
f
g
h
i

Nguyen et al. (2010)

This method

Linear range (the concentration unit is mg kg1, except for SECGCMS method which is mg L1).
Limit of detection (the concentration unit is mg kg1, except for SECGCMS method which is mg L1).
Limit of quantication (mg kg1).
Relative standard deviation (%).
Size-exclusion chromatographygas chromatographymass spectrometry.
Matrix solid-phase dispersiongas chromatography-tandem mass spectrometry.
Liquidliquid extractionsolid phase extractiongas chromatographyelectron-capture detector.
Matrix solid-phase dispersiongas chromatographymass spectrometry.
Liquidliquid extractiondispersive solid phase extractiongas chromatographymass spectrometry.

Please cite this article in press as: Farajzadeh, M.A., et al., Determination of pyrethroid pesticides residues in vegetable oils using liquid
liquid extraction and dispersive liquidliquid microextraction followed by gas chromatographyame ionization detection. J. Food
Compos. Anal. (2014), http://dx.doi.org/10.1016/j.jfca.2014.03.004

G Model

YJFCA 2406 18
M.A. Farajzadeh et al. / Journal of Food Composition and Analysis xxx (2014) xxxxxx

Fig. 5. Total ion chromatogram (TIC) of (a) corn oil after performing the proposed extraction technique; and mass spectra of (b) scan 780, retention time 13.22 min and (c)
fenpropathrin.

404

4. Conclusions

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In this study, an extraction/preconcentration method was

presented for pyrethroid pesticides from fatty matrices, vegetable
oils, followed by GCFID determination and GCMS conrmation.
Extraction of analytes was performed through LLE by DMF, which
acts as a disperser in the following preconcentration step, DLLME.
To the best of our knowledge, this is the rst report on coupling
LLEDLLME techniques in fatty samples. The results indicated that
the proposed method has some advantages such as low extraction
time and organic solvent consumption which makes it an
economical method. Hence, it seems possible to extend this
method to the extraction of interest analytes in other similar
samples by varying the extraction conditions.

417

Q4 Uncited references

418

Casado et al. (2014) and Xu et al. (2014).

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421

Acknowledgment
Q5
Q6

The authors thank the Research Council of Tabriz University for

the nancial support.

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Please cite this article in press as: Farajzadeh, M.A., et al., Determination of pyrethroid pesticides residues in vegetable oils using liquid
liquid extraction and dispersive liquidliquid microextraction followed by gas chromatographyame ionization detection. J. Food
Compos. Anal. (2014), http://dx.doi.org/10.1016/j.jfca.2014.03.004

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