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4 AUTHORS, INCLUDING:
Virgnia Cruz Fernandes
Valentina F Domingues
University of Porto
REQUIMTE
SEE PROFILE
SEE PROFILE
This article was downloaded by: [b-on: Biblioteca do conhecimento online IPP]
On: 16 October 2012, At: 01:09
Publisher: Taylor & Francis
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House,
37-41 Mortimer Street, London W1T 3JH, UK
a b
To cite this article: V.C. Fernandes, V.F. Domingues, N. Mateus & C. Delerue-Matos (2012): Analysing organochlorine
pesticides in strawberry jams using GC-ECD, GC-MS/MS and QuEChERS sample preparation, Food Additives & Contaminants:
Part A, 29:7, 1074-1084
To link to this article: http://dx.doi.org/10.1080/19440049.2012.682319
a
REQUIMTE, Instituto Superior de Engenharia, Instituto Politecnico do Porto, Rua Dr Antonio Bernardino de Almeida,
431, P-4200-072 Porto, Portugal; bCIQ Faculdade de Ciencia da Universidade do Porto, Departamento de Qumica e
Bioqumica, Rua do Campo Alegre, 687, P-4169-007 Porto, Portugal
Introduction
Pesticides are used globally for the protection of food,
and more generally for human health. However, their
excessive use/misuse, especially in developing countries, volatility and long-distance transport eventually
results in widespread environmental contamination
(Kaushik et al. 2009; Perez-Serradilla et al. 2010).
Organochlorine pesticides (OCP) are a large class of
very persistent pollutants. They stay in the environment after being applied for long periods. Many OCPs
are endocrine-disrupting chemicals, meaning they have
subtle toxic effects on the bodys hormonal systems,
contributing to adverse health effects (Mansilha
et al. 2010).
The use of OCPs has been highly restricted in
European countries, but they have been reported in
several food samples (Correia et al. 2000; BarriadaPereira et al. 2003, 2007; Da Silva et al. 2010; Dubois
et al. 2010; Rashid et al. 2010; Fernandes et al. 2011a)
in the last decade. It is, therefore, not surprising that
residues of OCP in food have given rise to major
concerns. OCPs have been found in both raw and
processed foodstuffs (Keikotlhaile et al. 2010).
A reduction of pesticide residue levels occurred due
1075
Samples
Nine commercially available strawberry jams from
different types of production were obtained from local
shops and supermarkets in Porto, Portugal. One of
them was produced by organic farming strawberries
and the other eight from conventional farming (five
jams were produced by traditional methods and the
other four by industrial procedure).
Sample preparation
Nine bottles of strawberry jam were directly blended
and stored in a deep freezer (20 C). The samples were
homogenised in a grinder (Krups). A modified
QuEChERS extraction based on Standard Method
EN 15662 (Anastassiades 2008) citrate buffering was
developed. The homogenised jam samples (10 g) were
weighed into 50 ml polypropylene centrifuge tubes.
A known pesticide concentration mixture and the IS
were spiked into the samples, which were allowed to
stand for 1 h at room temperature to allow solvent
evaporation and then shaken vigorously. A total of
10 ml of acetonitrile (ACN) was added to the samples
and mixed using an automatic vortex for 5 min to swell
the matrix and extract the samples. The citrate
1076
GC-MS/MS analysis
A Trace GC Ultra gas chromatograph Polaris Q
coupled with an ion-trap mass spectrometer (Thermo
Fisher Scientific) operated in electron impact ionisation (EI) mode at 70 eV controlled by Xcalibur 1.3
software was used for the determination of residues.
Confirmation of residues was carried out by GC-MS/
MS using a Supelco column fitted with a SLBTM-5MS
(30 m 0.25 mm, 0.25 mm film thickness). The injector
operating in the splitless mode and an ultrapure grade
helium (Linde Sogas; purity 99.999%) were used as
the carrier gas at 1.3 ml min1 flow. The GC oven
temperature was programmed from an initial
temperature of 40 C (2 min hold), ramped at
30 C min1 to 220 C (5 min hold) and finally at
10 C min1 to 270 C with holding for 1 min. This
programme resulted in a total run time of 21 min.
The other optimised parameters included a transfer
line temperature of 250 C and an ion source of 250 C.
The MS/MS conditions were fixed for each compound,
trying to select as the precursor ion the one with the
highest m/z ratio and abundance (Table 1).
1077
Pesticide
Molecular
weight
(g mol1)
Structure
Segment
Precursor
ions
(m/z)
Product
ions
for MS/MS
method (m/z)
1.0
0.1
1.0
6.5
5.00
219
109, 146
Solubility
in water
(mg l1)
at 20 C
-HCH
-HCH
-HCH (lindane)
-HCH
290.8
HCB
284.8
0.04
7.50
142
141
Aldrin
364.9
0.02
9.54
263
191
-Endosulfan
406.9
0.15
11.20
195
170
p,p0 -DDE
318.0
0.04
11.90
318
176, 211
Dieldrin
380.9
0.17
243
206, 211
Endrin
380.9
0.23
12.75
245
211
-Endosulfan
406.9
0.15
13.22
195
170
p,p0 -DDD
320.0
0.05
235
165, 199
o,p0 -DDT
354.5
0.003
235
165, 199
Methoxychlor
345.7
0.1
227
152, 169
8.10
15.00
The major analytical problem was due to the complexity of the matrix (Fugel et al. 2005) combined with
potential interferences from the presence of coextractives such as pigments and added pectin in the
jams. Consequently, the determination of pesticides in
jams must involve a sample preparation step including
an extraction process followed by a clean-up.
Recoveries (%)
Vortexed jam
Homogenised jam
1
-endosulfan
endrin
p-p' DDE
Dieldrin
-endosulfan
aldrin
lindane
HCB
-HCH
-HCH
95
94
92
87
82
76
77
77
73
69
65
62
66
69
58 62
60 62
59
4855 60 4657 59 48
56 59
49
48
43
47
132
127
115
101
91
77
131
98
95
67
methoxychlor
o-p' - DDT
The analytical method was validated as per the singlelaboratory validation approach. The performance of
the method was evaluated considering the following
validation parameters.
The calibration curves of the compounds in the
matrix were obtained by plotting the peak area against
the concentration at seven calibration levels ranging
from 5 to 100 mg kg1.
Limits of detection (LOD) and quantification
(LOQ) were determined by considering the slope of
the calibration line and the residual standard deviation
of a regression line (ICH 1996). The recovery experiments were carried out on strawberry jam by fortifying
the samples (10 g) in three replicates with the pesticide
mixture separately at five concentration levels of 0.005,
0.010, 0.030, 0.090 and 0.180 mg kg1, extracting by the
method described above. The quantification of recovery
samples was done by using the peak area of the analyte
and calculating the concentration by preparing a
calibration curve. The optimised method was applied
to the screening of nine strawberry jams samples.
-HCH
Method validation
p-p' - DDD
1078
Ultrasonic bath
1079
Chromatographic analysis
The GC-ECD demonstrated high performance compared with GC-MS/MS for the detection and quantification of OCP (Table 2). The chromatograms
obtained by these two different techniques show
a better chromatographic resolution by GC-ECD at
a concentration of 50 mg l1 when compared with the
Validation
The most common way to avoid matrix effects is to use
matrix-matched calibration standards, which was done
2
7.0
6
6.0
5.0
14
4.0
3.0
10
11
12 13
2.0
3
1.0
15
0.0
20.0
22.0
25.0
27.5
30.0
32.5
35.0
1
37.0
40.0
42.5
45.0
47.5
50.0 min
Figure 3. GC-ECD chromatogram of mixture OCP (50 mg l ). Peak identification in order of increasing retention time is as
follows: 1, -HCH; 2, HCB; 3, -HCH; 4, -HCH; 5, -HCH; 6, aldrin; 7, IS; 8, -endosulfan; 9, dieldrin; 10, p,p0 -DDE; 11,
endrin; 12, -endosulfan; 13, p,p0 -DDD; 14, o,p0 -DDT; and 15, methoxychlor.
1080
Conclusions
The proposed multi-residue methodology for the
determination of 14 OCPs in strawberry jam samples
by GC-ECD and GC-MS/MS using QuEChERS
coupled with a clean-up process is considered appropriate, except for -, -, -HCH, HCB and -HCH.
The use of an ultrasonic bath achieved better results
compared with the two versions previously tested. The
use of matrix-matched calibration standards avoids
matrix interference effects. The method yields recoveries between 62% and 129%, and the LOQs were, in all
cases, significantly lower than the MRL established for
strawberries (10 or 50 mg kg1 depending on the OCP)
in the European Union. The presence of organochlorines in several samples has been frequently reported in
-HCH
-HCHa
-HCHa
HCBb
Lindaneb
Aldrinc
-Endosulfand
p,p0 -DDEe
Dieldrinc
Endrinb
-Endosulfand
p,p0 -DDDe
o,p0 -DDTe
Methoxychlorb
ECD
MS/MS
ECD
MS/MS
ECD
MS/MS
0.9996
0.9994
0.9986
0.9970
0.9986
0.9972
0.9974
0.9980
0.9974
0.9977
0.9988
0.9987
0.9996
0.9995
0.9989
0.9991
0.9993
0.9994
0.9995
0.9989
0.9985
0.9991
0.9986
0.9991
0.9992
0.9994
0.9990
0.9989
0.8
1.1
1.5
7.6
1.5
7.3
7.0
6.2
7.1
6.7
4.7
5.1
2.9
3.1
0.8
0.9
0.9
6.1
5.6
7.9
7.3
8.9
6.7
8.5
6.3
6.0
6.3
7.7
2.7
3.5
5.1
25.4
5.1
24.4
23.4
20.6
23.7
22.3
15.7
16.9
9.6
10.3
2.5
2.9
3.0
20.4
18.6
26.3
24.5
29.5
22.3
28.3
21.1
20.3
21.1
25.7
0.010
mg kg1
66 11
69 10
67 10
63 8
65 11
92 10
95 6
94 5
74 9
97 9
110 11
124 9
120 13
121 12
0.005
mg kg1
66 10
67 11
65 7
64 8
62 13
96 8
90 9
96 11
70 12
90 12
115 9
121 11
122 8
121 8
Name
-HCH
-HCH
-HCH
HCB
Lindane
Aldrin
-Endosulfan
Dieldrin
p,p0 -DDE
Endrin
-Endosulfan
p,p0 -DDD
o,p0 -DDT
Methoxychlor
62 8
65 10
60 5
63 9
62 11
91 12
94 10
92 8
72 5
92 6
114 8
121 10
128 12
123 9
0.030
mg kg1
59 10
60 11
62 6
64 11
72 9
95 10
99 12
98 7
80 9
96 9
119 10
123 12
129 13
121 9
0.090
mg kg1
Commercial jam (n 3)
62 5
60 6
61 10
62 8
69 10
91 8
93 11
89 10
79 7
90 10
110 9
125 9
125 8
122 11
0.180
mg kg1
71 11
75 6
73 5
69 7
70 9
90 11
91 9
89 9
69 8
88 10
100 13
112 11
112 9
113 8
0.005
mg kg1
68 12
70 12
69 5
73 5
70 7
92 9
90 12
89 10
70 10
93 8
116 6
114 9
113 13
114 8
0.010
mg kg1
69 10
71 11
70 9
71 8
74 7
93 9
96 11
92 6
72 11
90 12
106 8
124 10
124 12
119 8
0.030
mg kg1
72 10
69 11
71 9
70 8
76 7
95 9
98 11
95 6
76 11
95 12
110 8
122 10
126 12
119 8
0.090
mg kg1
Traditional jam (n 3)
Recoveries (%)
Table 3. Influence of five spiked levels on the recovery efficiency using the version 3 procedure.
70 12
74 12
73 9
71 7
72 7
94 11
91 10
90 8
71 12
92 10
104 9
118 11
115 10
116 9
0.180
mg kg1
72 11
70 5
71 5
71 9
70 8
90 9
91 11
86 13
75 12
86 9
113 9
110 10
109 6
103 7
0.005
mg kg1
72 11
75 13
73 13
72 12
70 13
90 11
93 10
88 13
78 11
88 9
107 13
110 13
111 10
112 12
70 9
71 11
74 8
71 6
73 7
96 10
95 9
89 12
74 10
85 5
103 10
105 11
103 12
106 11
0.030
mg kg1
72 6
76 11
77 9
72 10
78 8
100 12
102 10
95 13
79 11
90 7
110 12
110 11
109 9
110 10
0.090
mg kg1
Organic jam (n 3)
0.010
mg kg1
70 9
72 11
75 7
70 9
75 8
98 11
97 12
89 10
78 11
89 9
105 15
107 10
105 7
113 8
0.180
mg kg1
1082
Relative Abundance
2
100
95
90
85
80
75
70
65
60
55
50
45
40
35
30
25
20
15
10
5
0
3
1
4
8
10
5
9
10
11
12
Time (min)
13
11 12
13
14
14
15
15
16
Figure 4. GC-MS/MS chromatogram of a mixture of OCP (100 mg l1). Peak identification in order of increasing retention time
is as follows: 1, -HCH; 2, HCB; 3, -HCH; 4, -HCH; 5, -HCH; 6, aldrin; 7, IS; 8, -endosulfan; 9, dieldrin; 10, p,p0 -DDE;
11, endrin; 12, -endosulfan; 13, p,p0 -DDD; 14, o,p0 -DDT; and 15, methoxychlor.
Acknowledgements
This research was supported by a PhD grant from the FCT
(Fundacao para a Ciencia e a Tecnologia BD/47200/2008)
and Grant Number PEst-C/EQB/LA0006/2011.
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