Food Control 42 (2014) 9e17

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Food Control
journal homepage: www.elsevier.com/locate/foodcont

Determination of thiamethoxam, triadimenol and deltamethrin in

pineapple using SLE-LTP extraction and gas chromatography
Elisa Helena da Costa Morais*, Alessandra Aparecida Zinato Rodrigues,
Maria Eliana Lopes Ribeiro de Queiroz, Antnio Augusto Neves,
Paulo Henrique Damasceno Morais
Department of Chemistry, Federal University of Viosa, 36570-000 Viosa, MG, Brazil

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 4 September 2013
Received in revised form
20 December 2013
Accepted 15 January 2014
Available online 31 January 2014

In this work, a simple and low cost method based on solideliquid extraction with low temperature
partitioning (SLEeLTP) was optimized and validated for the residue analyses of the pesticides thiamethoxam, triadimenol and deltamethrin in pineapple samples using gas chromatography with electron
capture detector (GC-ECD). The SLE-LTP technique was proven satisfactory with recoveries up to 90% and
linear chromatographic response for the analytes, ranging from 0.04 to 1.30 mg g
1 with correlation
coefcients above 0.99. The limits of quantication for the pesticides were lower than the allowed
maximum limits of residues established for these pesticides in pineapple by the European Union.
2014 Elsevier Ltd. All rights reserved.

Keywords:
Pesticide
Pineapple
Solideliquid extraction with low
temperature partitioning
Gas chromatography

1. Introduction
Pineapple, Ananas comosus (L.) Merrill., belonging to the family
Bromeliaceae, is originated in tropical and subtropical America,
according to naturalists and historians (Medina et al., 1987).
The consumption of this fruit is of paramount importance due to
its proteolytic activity, which acts as an adjunct in the digestion of
food (Medina et al., 1987; Souto, Durigan, Souza, Donadon, &
Menegucci, 2004). Currently, the world production of pineapple is
approximately 12.6 million tons. Thailand is the largest producer,
accounting for 16% of world production, followed by the Philippines
(12%) and Brazil (10%), while the variety Prola the most produced (ISHS, 2013). In Brazil, the cultivation of pineapple is
concentrated in the states of Minas Gerais and Paraba, which
represent more than half of Brazilian production (Morgado, Aquino,
& Terra, 2004).
Pineapple production, although very protable, demands
attention. The owering process is uneven, which affects the regularity of production, resulting in fruits not included in standard
commercialization. There are also diseases, weeds and pests, which
markedly affect the development of the pineapple crop, requiring,
therefore, the use of pesticides (Poel, Ceusters, & Proft, 2009).
* Corresponding author. Tel.: 55 31 3899 1430.
E-mail address: elisahelenabq@yahoo.com.br (E.H.C. Morais).
0956-7135/$ e see front matter 2014 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodcont.2014.01.024

About 85 different species of organisms have been found in

pineapples. For Brazilian conditions the following require attention: pineapple mealybug (Dysmicoccus brevipes), fruit borer
(Strymon basalides) and black rot caused by fungus (Ceratocystis
paradoxa). The use of the insecticides thiamethoxam, deltamethrin
and triadimenol fungicide is recommended for combating these
insects and fungus, respectively (AGROFIT, 2013; CEINFO, 2013).
The application of pesticides conscientiously can signicantly
reduce problems related to intoxication of humans, contamination
of groundwater, surface water, soil and food (Vercruysse, Drieghe,
Steurbaut, & Dejonckheere, 1999). However, what is observed is
that pesticides are often being used inappropriately and indiscriminately in attempts to maintain productivity and control diseases, weeds and pests. Due to negligence and the consequent
exposure of the public to the pesticides, there is increased demand
for food without these pesticides or food containing pesticide
residues below the maximum allowed limits. This has instigated
research to monitor and evaluate the risk of consuming contaminated food.
Several studies in the literature report the determination of
pesticide residues in fruits using the techniques of sample preparation solid-phase extraction (SPE) (Cherta, Beltran, Pitarch, &
Hernndez, 2013; Juan-Garca, Maes, Font, & Pic, 2004; Sanagi
et al., 2013), solid-phase microextraction (SPME) (Abdulrauf &
Tan, 2013; Blasco, Font, Maes, & Pic, 2003; Cherta et al., 2013)

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E.H.C. Morais et al. / Food Control 42 (2014) 9e17

and QuEChERS (Cherta et al., 2013; Lehotay et al., 2010), such as

pineapple (Castillo, Rodriguez-Valenciano, Moreno, & Blanch, 2012;
Cherta et al., 2013; Corts-Aguado, Snches-Morito, Arrebola, Frenich, & Vidal, 2008; Fytianos, Raikos, Theodoridis, Velinova, &
Tsoukali, 2006; Gilbert-Lpez et al., 2010; Ravelo-Prez, Hernndez-Borges, & Rodriguz-Delgado, 2008). However, techniques for
the determination of pesticide residues in complex samples have
evolved signicantly in terms of simplication, improvement of
extraction and purication of samples, with the principal objectives
of minimizing solvent and reagents consumption and optimizing
clean-up. The extraction with low temperature partitioning has
these advantages and has shown good results for the extraction of
pesticides from different matrices, due to the possibility of simultaneous performing by extraction of the pesticides and the cleanup of the extracts (Goulart, Queiroz, Neves, & Queiroz, 2008;
Pinho, Neves, Queiroz, & Silvrio, 2010).
Among modern analytical methods for quantication of pesticides in different matrices stand out the gas chromatography (GC)
(Abdulrauf & Tan, 2013; Cherta et al., 2013), the high performance
liquid chromatography (HPLC) (Sanagi et al., 2013; Wang, Chow, &
Leung, 2010) and ultra high performance (UHPLC) (Pic, Farr,
Soler, & Barcel, 2007; Wang et al., 2010), and the capillary electrophoresis (CE) (Font, Ruiz, Fernndez, & Pic, 2008), a function of
its facilities in effecting separations, and quantify the species present in a sample.
Based on this, the objective of this study was to optimize and
validate a method for the determination of the pesticide residues
thiamethoxam, deltamethrin and triadimenol in pineapples, used
in the control of common diseases in pineapple, caused by agents
shown above. In this determination was used the solideliquid
extraction with low temperature partitioning technique (SLEeLTP)
and gas chromatography.
2. Materials and methods
2.1. Reagents and solvents
The pesticides thiamethoxam (99.7% w/w) was obtained from
SigmaeAldrich, triadimenol (99.06% w/w) from Bayer, deltamethrin (99.0% w/w) from Chem Service, and bifenthrin (92.2% w/w),
used as an internal standard in the GC analysis, was from FMC.
The solvents acetonitrile (chromatographic grade e Vetec) and
ethyl acetate (analytical grade e Vetec) were employed as solvent
extractors. Anhydrous sodium sulfate (Vetec) was employed to dry
the extracts. The sorbents Florisil (SigmaeAldrich), octadecyl e C18
(Supelco) and silica (Merck), and sodium chloride (analytical grade
e ECIBRA), used for evaluation of ionic strength, were used as
received.
2.2. Stock solutions of pesticides
Stock solutions of each pesticide and the internal standard, at
concentration of 1000.0 mg L
1 were prepared by solubilization of
thiamethoxam, triadimenol, deltamethrin and bifenthrin standards
in acetonitrile.
Working solutions containing the pesticide triadimenol at a
concentration of 50.0 mg L
1 and thiamethoxam, bifenthrin and
deltamethrin at concentrations of 10.0 mg L
1 were prepared by
diluting the stock solutions in acetonitrile. These solutions were
stored at a temperature of around 4  C.
2.3. Equipments and instruments
The qualitative and quantitative analyses were performed on a
Shimadzu gas chromatograph (GC-2014) with electron capture

detector equipped with an AOC-20i autosampler. The capillary

column used in separating the compounds was a HP-5 from Agilent
Technologies, with a stationary phase consisting of 5% phenylmethylsiloxane and 95% dimethylsiloxane, 30 m length, 0.25 mm
internal diameter and 0.1 mm lm thickness. The analytical conditions set for the simultaneous analysis of pesticides were: injector
and detector temperature equal to 280  C and 300  C respectively,
initial column temperature 150  C, kept at this temperature for
2 min, with heating rates of 20  C/min to 210  C and 10  C/min to
290  C, kept at this temperature for 5 min. The total analysis time
was equal to 18 min; the injected volume was equal to 1 mL; the gas
ow drag (N2) was equal to 1.2 mL min
1; and the split was 1:5.
An analytical balance (Sartorius BP 2215), with an accuracy of
1  10
4 g, was used to measure mass. A shaker (Tecnal TE e 420),
an ultrasonic bath (Unique) (150 W and 25 kHz) and a vortex
(Certomat) were used in the extraction process.
2.4. Samples preparation and fortication of pineapple
Pineapple samples free of pesticide residues of the variety
Prola were chopped into pieces and crushed in a home multiprocessor to complete homogenization. Into glass vials of 22.0 mL
capacity were added 3.0 g of pineapple pulp spiked with 100.0 mL of
working solution containing all four pesticides. Samples fortied
with 1.67 mg g
1 of triadimenol and 0.33 mg g
1 of thiamethoxam
and of deltamethrin were allowed to stand for about 3 h in an open
container and then subjected to pesticide extraction procedures.
2.5. SLE-LTP of pesticides in pineapple samples
Into the glass vials containing the crushed and fortied pineapple samples ware added 10.0 mL of an extraction mixture consisting of 7.0 mL of acetonitrile, 1.5 mL of water and 1.5 mL of ethyl
acetate. After mechanical stirring for 10 min at 25  C and 200 oscillations per minute, the mixture was cooled at
20  C for
approximately 12 h. After separation of the phases by freezing of
pineapple pulp along with the aqueous phase, the supernatant was
passed through an ordinary lter paper containing 1.5 g of anhydrous sodium sulfate and 1.0 g of Florisil placed successively in
layers. To this collected extract were added 100.0 mL of bifenthrin
solution at a concentration of 10.0 mg L
1 and the volume
completed to 10.0 mL with acetonitrile. This solution was stored in
a freezer until the time of analysis with a GC-ECD.
2.6. Optimization of the SLEeLTP using pineapple samples
The solideliquid extraction with low temperature partitioning
technique for determination of the pesticides thiamethoxam, deltamethrin and triadimenol in samples of pineapple was optimized
based on the procedures used for extraction of pyrethroids from
milk (Vieira, Neves, & Queiroz, 2007) and chlorpyrifos, cypermethrin, l-cyhalothrin and deltamethrin from tomato (Pinho,
2007). In the process of optimization factors that inuence the
percentage recovery of pesticides were evaluated. These are shown
in Table 1.
2.7. Validation of the analytical method
The analytical selectivity, limit of detection (LOD), limit of
quantication (LOQ), linearity, accuracy and precision of the
method were evaluated using the guidelines of the National Agency
for Sanitary Vigilance (ANVISA, 2003), the National Institute of
Metrology, Standardization and Industrial Quality (INMETRO, 2010)
and related publications (Ribani, Bottoli, Collins, Jardim, & Melo,
2004; Lanas, 2004a).

E.H.C. Morais et al. / Food Control 42 (2014) 9e17

Table 1
Optimization of parameters using the SLEeLTP technique of pesticides in pineapple
samples.
Parameters

Levels

Sorbent
Amount of sorbent (g)
Extraction mixture

Florisil, octadecilesilica
1.0e2.0
Acetonitrile/water/ethyl acetate
(7.0 mL/1.5 mL/1.5 mL)
(5.5 mL/3.0 mL/1.5 mL)
(7.0 mL/3.0 mL/0.0 mL)
Shaking table (10e20 min)
Ultrasound (5e10 min)
Vortex (30 se1 min)
1.5 mL of water
1.5 mL of aqueous NaCl
solution (0.2 mol L
1)

Mode and time of extraction

Ionic strength

11

Table 2
Regression equations and coefcients of determination of the analytical curves of
the pesticides studied.
Pesticide

Concentration
range (mg L
1)

Equation

r2

Thiamethoxam
Triadimenol
Deltamethrin

10.0e600.0
50.0e600.0
10.0e100.0

y 0.0062x 0.0075
y 0.0011x 0.0013
y 0.0102x
0.0288

0.9936
0.9961
0.9980

*r2: Coefcient of determination; x: concentration of the pesticide in solvent; y: peak

area of the pesticide/peak area of internal standard.

500.0 mg L
1 for triadimenol, and 10.0 and 100.0 mg L
1 for deltamethrin and thiamethoxam.
2.9. Application of the SLE-LTP method to pineapple samples

The LOD of the method was considered as the concentration of

each pesticide in the matrix whose signal was equal to three times
the baseline signal (noise) in an area obtained by injection and
analysis by GC-ECD of pineapple extract free of the analytes. The
LOQ was determined considering a signal ten times greater than
the noise. The response linearity of the method was evaluated by
injection and subsequent analysis of extracts obtained from samples of spiked pineapple using seven concentrations of the active
ingredients thiamethoxam (0.04e0.40 mg g
1), triadimenol (0.13e
1.30 mg g
1) and deltamethrin (0.04e0.40 mg g
1) in triplicate and
submitted to the optimized extraction procedure. The recoveries of
pesticides resulting from fortication of pineapple samples in three
concentration levels (LOQ, 2  LOQ and 10  LOQ), in triplicate,
were used to assess the accuracy of the method, and relative
standard deviations (RSD) corresponding to recoveries of analytes
at concentrations equal to 2  LOQ, to ascertain the accuracy and
repeatability through in seven replicates. Intermediate precision
was determined by analyses on three different days (1st, 7th and
28th days).

The optimized and validated method was applied for determination of the pesticides thiamethoxam, deltamethrin and triadimenol in three samples of pineapple from the cities Maratazes
(ES), Miracema (TO) and Monte Alegre de Minas (MG).
3. Results and discussions
3.1. Chromatographic analysis
Fig. 1 shows a chromatogram referring to a standard solution
having the pesticide triadimenol at a concentration of 500.0 mg L
1
and the pesticides thiamethoxam and deltamethrin at concentrations of 100.0 mg L
1.
The presence of more than one peak for the pyrethroid deltamethrin (compound 4) is attributed to the conversion of this pyrethroid into isomers (Mastovsk & Lehotay, 2004). Quantication
was performed considering the sum of peak areas of the isomers in
the chromatograms relating to samples and standard solutions.
Quantication of pesticides was performed by the method of
internal standardization, using bifenthrin compound. This internal
standard was used since it is similar to the analytes, has a retention
time close to them, and different from the coextractives, do not react
with the matrix component, besides not being one of its constituents.
Standard curves were constructed for each compound. The
range of concentrations of pesticides, the regression equation for
each compound and the coefcients of determination (r2) are
shown in Table 2.

2.8. Evaluation of the matrix effect

The inuence of the components of pineapple on detector
response was evaluated by the analysis and comparison of analytical curves from injection of standard solutions of pesticides prepared in solvent and in an extract of a pesticide free matrix. The
concentration ranges of pesticides in these solutions were: 50.0e

uV x 10,000
5.0
4.5
4.0

3.5
3.0

2.5

2.0

1.5
1.0
0.5
0.0
2.5

5.0

7.5

10.0

12.5

15.0

min

Fig. 1. Chromatogram of a solution containing the pesticides studied, where: 1 (retention time 8.5 min) is thiamethoxam; 2 (retention time 8.9 min) is triadimenol; 3 (retention
time 11.7 min) is bifenthrin; and 4 (retention time 17.0 min) is deltamethrin.

12

E.H.C. Morais et al. / Food Control 42 (2014) 9e17

Coefcients of determination of the analytical curves greater

than 0.9900, residues random distributed along the line of
regression and relative errors less than 20.0% indicate that there is a
linear response of the electron capture detector within the concentration range of the analytes studied.
3.2. Optimization of the SLE-LTP technique with pineapple samples
The solideliquid extraction with low temperature partitioning,
similarly to the liquideliquid extraction with low temperature
partition (LLEeLTP) (Goulart et al., 2008; Vieira et al., 2007), appears as an alternative technique for the extraction of analytes in
complex matrices such as pineapple, because of its low cost, due to
low consumption of reagents, compared to the sample preparation
techniques like QuEChERS. Moreover, it is simple, the extraction of
analytes is efcient and ensures precision and accuracy of results,
often not observed when the techniques SPE and SPME are used.
According to studies by Pinho (2007) and Vieira et al. (2007), for
the determination of pesticide residues in tomato and milk,
respectively, the mixture containing matrix and extractor solvent
should remain in freezer, approximately
20  C for at least 12 h, to
freezing the pineapple pulp along with the aqueous phase and
efcient extraction of the analytes.
3.2.1. Evaluation of the efciency of the sorbent for the clean-up of
the extracts
Due to yellowish pigments present in the extracts obtained from
samples of pineapple, and the presence of other coextractants, the
sorbents Florisil, silica and C18 were compared for clean-up of the
extracts. Recoveries obtained from the SLE-LTP of the pesticides
using these sorbents are shown in Fig. 2a.
The use of C18 sorbent did not provide clearer extracts.
Furthermore, shown recoveries below 70.0% were found for the
pesticide triadimenol. Silica had a positive inuence on the cleanup of the extracts; however, it provided low percent recoveries
for this same compound. The Florisil sorbent proved to be the most
efcient in removing unwanted components of the matrix and
showed higher percent recoveries of the pesticide triadimenol,
compared to the other two sorbents, with recoveries between 70.0
and 120.0% (range of acceptable values for determining residues
pesticides) (Ribani et al., 2004) for the compound thiamethoxam.
Recoveries above 120.0% were observed for the pesticides
thiamethoxam and deltamethrin, probably due to the increase of
the chromatographic response of these compounds when present
in the extracts. This effect was more pronounced when using C18 or
silica. This indicates that Florisil is more efcient in removing some
of the matrix components, such as sugars and avonoids, which
interfere with the chromatographic analysis. Thus it was decided to
use Florisil as sorbent in the clean-up step.
3.2.2. Determination of the amount of sorbent needed for clean-up
of the extracts
The percent recoveries of the pesticides using SLE-LTP technique, followed by clean-up are shown in Fig. 2b.
We observed a signicant decrease in the recovery of the pesticides thiamethoxam and deltamethrin when performing the
clean-up of the extracts. This shows that although both 1.0 and 2.0 g
of Florisil are effective in removing some coextractives of the matrix
that interfere with the chromatographic analysis, use of an
increased mass of Florisil induced a reduction in the percent recoveries of the pesticides.
For these reasons, we chose to use the mass of 1.0 g of Florisil for
the clean-up of the extracts.

3.2.3. Evaluation of the extraction mixture

To improve the extraction of these compounds three extraction
mixtures were evaluated. The results shown in Fig. 2c indicate that
decreases in the polarity of the extraction mixture, increases the
recoveries percent of the pesticides. That is, the less polar extraction mixture, besides promoting an increase in the percent of recovery of less polar compounds such as deltamethrin (logarithm of
octanolewater partition coefcient (log P) 4.6) and triadimenol
(log P 3.18), also caused an increase in the percent recovery of
thiamethoxam (log P
0.13), a more polar compound (IUPAC,
2013).
Therefore, the ideal extraction mixture is the less polar one
consisting of 7.0 mL of acetonitrile, 1.5 mL water and 1.5 mL of ethyl
acetate.
3.2.4. Evaluation of the mode and time of extraction
One of the most important steps in the extraction of pesticides is
the homogenization between the extraction mixture and the
sample.
According to the results shown in Fig. 2d and applying the
Students t-test, at 95.0% probability it can be stated that homogenization of the three modes exhibited statistically similar recovery
percentages for triadimenol.
Considering the pesticides thiamethoxam and deltamethrin, it
can be seen that the use of 5 and 10 min of ultrasound led to a
decrease in the percent recoveries when compared to 10 and
20 min in a shaker, and 30 s and 1 min in a vortex, are considered
statistically identical using the Students t-test. However, for the
vortex homogenization, the coefcients of variation were high,
which can be associated with possible adherence of the compounds
to the walls of the containers.
Therefore, 10 min using a shaker was chosen for homogenization between of the extraction mixture and sample, since it provided more accurate results and high recovery percentages.
3.2.5. Evaluation of the effect of the ionic strength
The addition of salt often causes an increase in recovery, mainly
for polar compounds since their solubility in the matrix decreases,
facilitating its extraction to the organic phase (Lanas, 2004b). The
results obtained after evaluating this variable showed that the
addition of salt did not change, at 95.0% probability by the Students
t-test, the recovery of the compounds. Therefore, it was decided not
to add salt to the extraction mixture.
3.3. Validation of the analytical method
3.3.1. Selectivity
The selectivity was evaluated by comparing chromatograms of
pesticide-free matrix extracts with chromatograms of fortied extracts (Fig. 3).
The extracts of the pineapple samples, despite having a complex
composition, showed no interfering substances at the same
retention times as the compounds tested, which makes the method
selective. The selectivity of the electron capture detector that detects only molecules that capture electrons (Collins, Braga, &
Bonato, 2006), also contributed to achieving these results.
3.3.2. Limit of detection (LOD) and limit of quantication (LOQ)
Extracts of matrix obtained from samples fortied with the
pesticides thiamethoxam, deltamethrin and triadimenol in
different concentrations were injected to determine the signal/
noise ratios at 3:1 and 10:1 to calculate detection and quantication
limits, respectively. These values are presented in Table 3.
It was found that the limit of detections obtained for the compounds thiamethoxam, triadimenol and deltamethrin using GC-

E.H.C. Morais et al. / Food Control 42 (2014) 9e17

Fig. 2. Inuence of parameters on the recovery of pesticides. (a) sorbent, (b) amount of sorbent, (c) extraction mixture, (d) mode and extraction time.

13

14

E.H.C. Morais et al. / Food Control 42 (2014) 9e17

uV x 10,000
5.0
4.5

4.0
3.5
3.0
2.5

8.5

2.0

8.9

17.0

11.7

1.5
1.0
0.5
0.0
2.5

5.0

7.5

10.0

12.5

15.0

min

uV x 10,000
5.0
4.5

4.0

11.7

3.5
3.0

8.5

2.5

17.0

8.9

2.0
1.5
1.0
0.5
0.0
2.5

5.0

7.5

10.0

12.5

15.0

min

Fig. 3. Chromatograms of extracts obtained from samples of pineapple. (a) without pesticides, (b) fortied and subject to SLE-LTP with 1.67 mg g
1 of triadimenol and 0.33 mg g
1 of
thiamethoxam and deltamethrin. Identication of peaks: retention time 8.5 min: thiamethoxam; retention time 8.9 min: triadimenol, retention time 11.7 min: bifenthrin,
retention time 17.0 min: deltamethrin.

ECD are between 0.012 and 0.050 mg g
1, while limit of quantications are between 0.040 and 0.17 mg g
1. The lowest values of LOD
and LOQ are observed for the pesticides thiamethoxam and
deltamethrin.
Taking into account the maximum residue limits (MRLs)
established by the European Union, equal to 0.050, 3.00, and
0.050 mg g
1, for thiamethoxam, deltamethrin and triadimenol
respectively, it can be observed that the LOQ values obtained are
equal to or lower than the values allowed for each pesticide in the
food type evaluated herein (EU, 2013).
Several papers on the injection of extracts of diverse samples in
the gas chromatograph are found in the literature, using the so
called splitless technique (Shuling, Xiaodong, & Chongjiu, 2007).
This procedure usually provides smaller values of LOD and LOQ, as a

Table 3
Limit of detections (LOD), limit of quantications (LOQ) and linearity of the method
for the pesticides studied.
Pesticide

LOD
LOQ
Concentration Equation
(mg g
1) (mg g
1) range (mg g
1)

Thiamethoxam 0.015
Triadimenol
0.051
Deltamethrin 0.012

0.05
0.17
0.04

0.04e0.40
0.13e1.30
0.04e0.40

r2

y 1.5205x 0.0329 0.9998

y 0.1805x 0.1431 0.9933
y 2.8434x 0.0305 0.9982

*LOD: limit of detection; LOQ: limit of quantication; x: concentration of the

pesticide in solvent; y: peak area of the pesticide/peak area of internal standard; r2:
coefcient of determination.

larger sample is introduced into the chromatographic column.

However, for complex samples such as pineapple, where the
presence of coextractives is high, this procedure is not convenient.
3.3.3. Linearity of the response of the method
After injection and chromatographic analysis of extracts obtained using the SLE-LTP procedure with samples that were fortied with the pesticides, analytical curves were constructed, using
the area ratios of the analytes and the internal standard, thus
obtaining regression equations and coefcients of determination.
These results are shown in Table 3.
The correlation coefcients of the calibration curves were
greater than 0.9900, featuring an optimal t of the data of the linear
regression and response linearity of the proposed method,
conrmed by the randomness of the distribution of residuals and
by relative errors less than 20.0% (Fig. 4).
3.3.4. Accuracy and precision
The accuracy was evaluated by recovery experiments on samples of pineapple, resulting from the fortication of these matrices
with pesticides at three concentration levels. Satisfactory recoveries between 70.0 and 120.0%, the acceptable range for the
determination of pesticide residues, were obtained for the pesticides (Table 4).
Precision was evaluated in terms of repeatability and intermediate precision, using seven pineapple samples fortied with pesticides on three different days and subjected to the proposed

E.H.C. Morais et al. / Food Control 42 (2014) 9e17

15

Fig. 4. Graph of relative error versus concentration corresponding to pesticides (a) thiamethoxam, (b) triadimenol, (c) deltamethrin.

Table 4
Recoveries and relative standard deviations obtained by the analysis of extracts of pineapple samples fortied with pesticides.
Pesticide

Thiamethoxam
Triadimenol
Deltamethrin

Accuracy

Repeatability

Recovery (%)

Relative standard deviations (%)

Intermediate precision

LOQ (mg g
1)

2  LOQ (mg g
1)

10  LOQ (mg g
1)

2  LOQ (mg g
1)

2  LOQ (mg g
1)

97.3
98.2
103.4

96.4
97.4
104.0

92.3
95.2
105.5

3.7
8.3
5.5

5.2
8.4
3.9

*LOD: limit of detection; LOQ: limit of quantication.

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E.H.C. Morais et al. / Food Control 42 (2014) 9e17

Table 5
Equations of regression and coefcients of determination of the analytical curves
obtained by analysing standard solutions in acetonitrile and pineapple extracts.
Pesticide

Thiamethoxam
Triadimenol
Deltamethrin

Equation; coefcient of determination (r2)

Acetonitrile

Extract of pineapple

y 0.0063x
0.0388;
r2 0.9998
y 0.0011x 0.0218;
r2 0,9936
y 0.0047x 0.0406;
r2 0.9970

y 0.0095x 0.1683;
r2 0,9949
y 0.0010x 0.0863;
r2 0,9988
y 0.0125x 0.0240;
r2 0.9976

20.0% (Ribani et al., 2004) either of the two forms of quantication were suitable.
3.5. Application of the SLEeLTP technique to pineapple samples
The optimized and validated SLE-LTP technique was applied to
samples of pineapples from the cities of Maratazes (ES), Miracema
(TO) and Monte Alegre de Minas (MG). The presence of any residues
of pesticides studied was not observed.
4. Conclusions

method. The results are expressed in terms of relative standard

deviations and are presented in Table 4, which shows accurate results, since the relative standard deviations were lower than 20.0%.
3.4. Evaluation of the matrix effect
The matrix effect, also known as enhancement or suppression
of chromatographic response induced by the matrix, is one of the
factors responsible for the overestimation and the low accuracy of
results (Menkissoglu-Spiroudi & Fotopoulou, 2004).
The liner of the injector is often responsible for matrix effects
when using gas chromatography as a technique for the determination of analytes, since active sites on this can cause adsorption or
thermal degradation of analytes. When standard solutions are
prepared in solvent, injected and analyzed by gas chromatography,
the active sites of the liner are available to interact with the pesticides. However, when standard solutions are prepared in the
matrix extract, injected and analyzed in a gas chromatograph,
pesticides compete with matrix components for the active sites of
the liner. Depending on the characteristics of the matrix and analyte, matrix components may be adsorbed on the liner and more
pesticide may be introduced into the chromatographic column,
giving a greater response for the analyte as compared to the
response when using pure solvent (Hajslov & Zrostlkov, 2003).
During the development of this work it was observed that the
adsorption of components from pineapple in the chromatographic
system led to an increased response of the compounds thiamethoxam, triadimenol and deltamethrin. This increase is evidenced
by comparing analytical curves resulting from injection and analysis of solutions prepared in pesticide-free matrix extracts and in
solvent, and by the ratio of the slopes being above 1.0 (thiamethoxam:1.5; deltamethrin:2.6) as shown in Table 5 (Pinho, Neves,
Queiroz, & Silvrio, 2009).
Considering aspects related to the matrix effect the best way to
quantify residues of pesticides thiamethoxam, deltamethrin and
triadimenol in samples of pineapple when employing the SLE-LTP
was evaluated. The pineapple samples were spiked with pesticides and subjected to the extraction process. Quantication of
pesticides was performed using the standard curves prepared in
acetonitrile and in pineapple extract. The recoveries percent of the
pesticides thiamethoxam, deltamethrin and triadimenol obtained
using the analytical curve in acetonitrile and in pineapple extract
were equal to 96.3, 89.3 and 102.6%, and 77.5, 92.4 and 86,9%,
respectively, with relative standard deviations lower than 5.0%.
Applying the Students t-test, with 95.0% probability, it is observed
that there was no signicant difference in the quantication of the
pesticide triadimenol. For the pesticides thiamethoxam and deltamethrin, a reduction in the recovery percent was found when
using standard prepared in the matrix extract, which was to be
expected, since they suffer greater interference due to the presence
of coextractives from the matrix.
In general, in the analysis of pesticide residues recoveries are in
the range of 70.0e120.0% are permitted, with accuracies of up to

The optimization of the SLEeLTP technique for the extraction of

the pesticides thiamethoxam, deltamethrin and triadimenol from
pineapple, using determinations by GC-ECD, resulted in an efcient
and low cost procedure, with high percent recoveries. With this
technique, the adsorbent, extraction mixture and homogenization
mode were evaluated to determine the best conditions for analysis
of the compounds of interest. The stage of clean-up with Florisil
was necessary to remove coextractives presents in extracts from
pineapple. The change in the polarity of the extraction phase consisting of acetonitrile, by adding small amount of ethyl acetate
improved the extraction of the pesticides. It was observed that the
matrix components inuenced the measurement of the pesticides
by gas chromatography. However, analytical curves prepared in
pure solvent and in the matrix extract were both suitable for
quantication of these pesticides by the proposed method.
Acknowledgments
The authors acknowledge nancial support and fellowships
from the Conselho Nacional de Desenvolvimento Cientco e Tecnolgico (CNPq) and also C. H. Collins for helpful discussions and
suggestions.
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