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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem
Analytical Methods
a r t i c l e
i n f o
Article history:
Received 2 October 2012
Received in revised form 6 November 2013
Accepted 7 March 2014
Available online 17 March 2014
Keywords:
Pyrethroids
Dispersive liquidliquid microextraction
(DLLME)
QuEChERS
Gas chromatographyelectron capture
detection (GCECD)
Fruit juices
Gas chromatographymass spectrometry
(GCMS)
a b s t r a c t
Dispersive liquidliquid microextraction (DLLME) sample preparation and the quick, easy, cheap, effective, rugged and safe (QuEChERS) method combined with DLLME were developed and compared for
the analysis of ten pyrethroids in various fruit juices using gas chromatography-electron capture detection (GCECD). QuEChERSDLLME method has found its widespread applications to all the fruit juices
including those samples with more complex matrices (orange, lemon, kiwi and mango) while DLLME
was conned to the fruit juices with simpler matrices (apple, pear, grape and peach). The two methods
provided acceptable recoveries and repeatability. In addition, the applicabilities of two methods were
demonstrated with the real samples and further conrmed by gas chromatographymass spectrometry
(GCMS).
2014 Published by Elsevier Ltd.
1. Introduction
Pyrethroids, a new-type of insecticide, have gained extensive
applications to prevent and treat insects in modern agriculture
due to their broad-spectrum insecticidal capacity and high effectiveness (Ye, Xie, Wu, & Lin, 2006). However, pyrethroid residues
are considered to be one of the most important sources of pollution
in agricultural production, and may be a potential threat to public
health (Kolaczinski & Curtis, 2004). Therefore, it is necessary to develop sensitive and selective methods for the analysis of pyrethroid
residues usually present in trace amounts. Potential analytical
methods include high performance liquid chromatography (HPLC)
(Boonchiangma, Ngeontae, & Srijaranai, 2012), capillary electrophoresis (CE) (Ye et al., 2006), gas chromatography (GC) (Du,
Yan, She, Liu, & Yang, 2010; Matsadiq et al., 2011), gas chromatographymass spectrometry (GCMS) (Cunha, Fernandes, & Oliveira,
Corresponding author at: Citrus Research Institute, Southwest University,
Chongqing 400712, China. Tel./fax: +86 23 68349046.
E-mail address: bljiao@tom.com (B. Jiao).
http://dx.doi.org/10.1016/j.foodchem.2014.03.028
0308-8146/ 2014 Published by Elsevier Ltd.
2009; Kok, Hiemstra, & Bodegraven, 2005), and gas chromatographytandem mass spectrometry (GCMS/MS) (Pay et al., 2007).
Quick and effective sample preparation coupled with a reliable
analytical method is imperative. Liquidliquid extraction (LLE) (Rezaee et al., 2006) and solid-phase extraction (SPE) (Sharif, Man, Hamid, & Keat, 2006) are the most common sample preparation
methods widely used for residue analysis. Recently, a growing
number of studies have focused on two kinds of micro-extractions
termed as liquid-phase microextraction (LPME) and solid-phase
microextraction (SPME), based on miniaturisation of conventional
LLE and SPE, respectively (Paillakis & Kalogerakis, 2003; Zambonin,
Cilenti, & Palmisano, 2002). In 2006, Rezaee et al. developed a novel liquidliquid microextraction method, dispersive liquidliquid
microextraction (DLLME) (Rezaee et al., 2006). The new method
has been widely recognised due to its simplicity, low cost and high
enrichment, which made it available to most analytical laboratories. Unfortunately, the lack of purication for samples with more
complex matrices, such as fruit and vegetable, has caused this
method to be limited to those with simpler matrices, specically
water and a few fruit juices.
368
At present, quick, easy, cheap, effective, rugged and safe (QuEChERS) sample preparation, is the most common technique for
multi-residue pesticides analysis in food, especially fruit and vegetable (Anastassiades, Lehotay, Stajnbaher, & Schenck, 2003). However, the major disadvantage of this technique is the poor
enrichment factor, which can lead to higher detection limits, i.e.
lower sensitivity, compared with other techniques. Researchers
proposed a new method comprised of DLLME pre-concentration
after QuEChERS extraction (Cunha & Fernandes, 2011; Zhao, Zhao,
Han, Jiang, & Zhou, 2007). Coupling these techniques takes advantages of the benets of both methods whilst reducing some of their
drawback. Moreover, QuEChERSDLLME sample preparation widens the use of DLLME to those samples with more complex matrices. To the best of our knowledge, there are a few reports on the
extraction and enrichment of pyrethroid residues in fruit juices
using QuEChERSDLLME method, although several studies on the
analysis of pyrethroids in fruit juices using only DLLME have been
reported (Boonchiangma et al., 2012; Cunha et al., 2009; Du et al.,
2010; Matsadiq et al., 2011).
In the paper, two sample preparation including DLLME and
QuEChERSDLLME followed by gas chromatography-electron capture detection (GCECD) were developed and compared for the
analysis of ten pyrethroids (tetramethrin, bifenthrin, lambdacyhalothrin, permethrin, cyuthrin, cypermethrin, ucythrinate,
fenvalerate, tau-uvalinate and deltamethrin) in various fruit
juices. Compared with the existing reports on the analysis of pyrethroid residues in fruit juices, more pyrethroids and fruit juicestypes were analysed in this study.
Enrichment factor (EF), dened as the ratio of the analyte concentration after DLLME (Csed) to the initial analyte concentration
(C0), was calculated as:
EF
ER
C sed
C0
C sed V sed
100
C 0 V aq
where Vsed and Vaq are the volumes of the sedimented phase and the
sample, respectively.
2.7. Samples
Eight types of fruit juices were purchased from local supermarkets. Samples were ltered using Buchner funnel before extraction
to remove the sediments. All fruit pulps (2 kg each), excluding lemon, were prepared from peeled fruit. The whole fruit was used to
prepare lemon pulp. A representative portion of these samples
(200 g each) was chopped and homogenised in a food chopper.
2.8. DLLME procedure
2.2. Apparatus
An Agilent 7890A gas chromatograph (Palo Alto, USA) was used
to perform all GC analysis. Ten pyrethroid residues were separated
in a capillary column (DB-1, 30 m 0.25 mm id 0.25 lm lm).
An Agilent 5795C/7890A GC/MS (Palo Alto, USA) was used to perform the conrmation. A HP-5MS (30 m 0.25 mm id 0.25 lm
lm) capillary column was used.
The DLLME procedure described below was followed for enrichment. 5 g of deionized water was placed in a centrifuge tube. 1 mL
of the extract and 60 lL of CCl4 were rapidly injected into 5 g of
deionized water with a syringe to form cloudy solution. The samples were vortexed for 5 min and then centrifuged at 4000 rpm
for 5 min. Finally, the sedimented phase was transferred to a trace
intubation in a sample vial.
3. Results and discussion
3.1. Optimisation of DLLME
Factors likely to impact DLLME and QuEChERSDLLME such as
types and volumes of extraction solvents and dispersive solvents,
ultrasonic time, salt concentration, vortex and centrifugation time
were investigated in detailed.
3.1.1. Selection of extraction solvent and its volume
The correct solvents are vital for the success of DLLME. In our
study, four common halogenated solvents including CHCl3,
C6H5Cl, C2Cl4 and CCl4 were selected for extraction. Extraction efciency was evaluated by comparing the recoveries of the analytes.
Fig. 1A showed that carbon tetrachloride was most efcient and it
was therefore used in subsequent experiments.
To investigate the volume effect of CCl4, different quantities of
CCl4 (3070 lL) were used whilst the dispersant was maintained
at 1 mL. Observably, the extraction efciency was improved with
369
the increase of volume (Fig. 1B). When CCl4 was increased from
30 to 60 lL, the recovery of ten pyrethroids was increased from
4368% to 61103%. However, those peaks of some non-analytes
including interfering substances appeared more obviously with
volumes greater than 60 lL although the recovery of the pyrethroids continued to increase. Thus, 60 lL of CCl4 was selected as
the optimum volume.
Fig. 1. Effects (A) of extraction solvents, (B) of the volumes of extraction solvents, (C) of dispersive solvents, and (D) of the volumes of dispersive solvents on the recoveries of
analytes in DLLME. 5.00 g of spiked apple juices at 0.01 mg kg1.
370
Fig. 3. Chromatograms (A) of a standard mixture at 0.01 mg L1, (B) of a spiked apple juice at 0.01 mg kg1 after DLLME, (C) of a spiked orange juice at 0.01 mg kg1 after
QuEChERSDLLME, and (D) a spiked orange juice at 0.01 mg kg1 after DLLME while the spiked sample is diluted to ve times: 1, tetramethrin; 2, bifenthrin; 3, lambdacyhalothrin; 4, permethrin; 5, cyuthrin; 6, cypermethrin; 7, ucythrinate; 8, fenvalerate; 9, tau-uvalinate; 10, deltamethrin.
371
Linear equation
Tetramethrin
Bifenthrin
Lambda-
210,000
0.510,000
Permethrin
Cyuthrin
Cypermethrin
Flucythrinate
Fenvalerate
Tauuvalinate
Deltamethrin
Pyrethroids
Correlation
coefcient (r2)
LOD
(lg L1)
LOQ
(lg L1)
Enrichment
factor of DLLME
Enrichment factor of
QuEChERSDDLME
Precision (% RSD)
Intraday
(n = 3)
Inter-day
(n = 3 3)
0.9998
0.9998
0.210,000
0.9996
0.9996
0.9995
0.9995
0.9990
0.9993
0.9992
2
0.5
5
2
110
65
25
18
2.9
2.2
4.1
4.9
210,000
110,000
210,000
110,000
0.510,000
0.510,000
y = 251.85x + 9849.36
y = 358.74x + 29175.27
cyhalothrin
y = 1570.54x + 74876.51
y = 205.10x + 8595.29
y = 2201.21x + 67955.49
y = 1460.15x + 83946.31
y = 201.19x + 29722.88
y = 564.36x + 33944.35
y = 1381.12x + 57236.53
0.2
2
1
2
1
0.5
0.5
1
5
4
5
4
2
2
64
104
78
86
104
107
93
19
24
18
20
24
16
26
3.9
2.1
2.8
2.5
2.3
2.4
2.8
6.3
4.0
6.4
4.5
5.3
5.8
6.2
0.510,000
y = 383.27x + 56526.79
0.9990
0.5
92
17
3.1
6.0
Table 2
Comparison of the proposed methods and some other methods for the determination of pyrethroids residues in fruit juices.
Extraction method
Instrument
detector
Analyte
Sample
LOD
Recovery (%)
EF
Ref.
DLLME
HPLCUV
6 Pyrethroids
25 lg L1
8494
6284
Boonchiangma, Ngeontae,
and Srijaranai (2012)
UA-DLLME
GCFID
2 Pyrethroids
23 lg kg1
92107
344351
DLLME
DLLME
GCECD
GCMS
4 Pyrethroids
24 Pesticides including 2
pyrethroids
10 Pyrethroids
73106
60105
4091089
4258
62108 and
63138
64110 and
1626
DDLME and
GCECD
QuEChERSDDLME
0.22 lg L1
372
Fig. 4. Chromatograms (A) of apple juice by GCECD after DLLME, (B) by of apple juice GCMS after DLLME, (C) of orange pulp by GCECD after QuEChERSDLLME, and (D) of
orange pulp by GCMS after QuEChERSDLLME: (1) lambda-cyhalothrin standard at 0.2 mg L1; (2) sample contaminated with lambda-cyhalothrin; (3) tau-uvalinate
standard at 0.2 mg L1; (4) sample contaminated with tau-uvalinate.
4. Conclusions
In this paper, two rapid and simple sample preparation methods DLLME and QuEChERSDLLME followed by GCECD were
developed and compared for ten pyrethroid residues in various
fruit juices. These methods offered acceptable recoveries and limited variation from day-to-day. DLLME was suitable for those fruit
juices with simper matrices and QuEChERSDLLME for all the fruit
juices. The proposed methods demonstrated pyrethroid residues in
some fruit juices in the Chinese market.
Acknowledgements
This work was supported by the National Key Technology R&D
Program (Nos. 2007BAD47B07 and 2009BADB7B04), China
Agriculture Research System (No. CARS-27), Natural Science
Foundation of Chongqing (Nos. cstc2013jcyjA0435 and
cstc2013jjB80009) and Fundamental Research Fund for the Central
Universities (No. XDJK2012C059).
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