Article

Cite This: J. Agric. Food Chem. XXXX, XXX, XXX−XXX pubs.acs.org/JAFC

Supercritical Fluid Chromatography and Gas Chromatography

Coupled to Tandem Mass Spectrometry for the Analysis of
Pyrethroids in Vegetable Matrices: A Comparative Study
María Murcia-Morales, Víctor Cutillas, and Amadeo R. Fernández-Alba*
Agrifood Campus of International Excellence (ceiA3), European Union Reference Laboratory for Pesticide Residues in Fruit and
Vegetables, Department of Hydrogeology and Analytical Chemistry, University of Almería, Carretera Sacramento s/n, La Cañada de
San Urbano, 04120 Almería, Spain
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ABSTRACT: This study describes a comprehensive comparison between supercritical fluid chromatography (SFC) and gas
chromatography (GC) coupled to mass spectrometry for the analysis of pyrethroids in vegetable matrices. The ionization
process used was electrospray ionization (ESI) in SFC and electron ionization in GC. In general, liquid chromatography
Downloaded via JAMES COOK UNIV on August 5, 2019 at 14:32:39 (UTC).

coupled to mass spectrometry with ESI sources provides poor results for pyrethroid detection, as described in previous
literature. A total of 14 pyrethroids were selected, together with 6 representative matrices. The differences in chromatographic
separation and ionization process were assessed. Similar results were obtained in terms of sensitivity (limits of quantification
close to 2 μg/kg, injecting the same amount of sample), matrix effect, and linearity. A total of 17 real samples were analyzed by
both systems, obtaining similar results. These data suggest that SFC offers a suitable alternative to GC in the analysis of
pyrethroids and allows for their inclusion in a wider multiresidue method.
KEYWORDS: supercritical fluid chromatography, gas chromatography, pyrethroids, vegetable matrices, electron ionization,
electrospray ionization

1. INTRODUCTION Pyrethroids are considered less toxic and safer to use

Pyrethrins are natural compounds present in Chrysanthemum
cinerariaefolium flowers. They are considered one of the most
effective natural insecticides and have been used to control
pest insects since ancient times. Pyrethroids are a class of
synthetic insecticides as consequence of a modification of the
chemical structures of pyrethrins. These changes provide
increased stability to light and air exposure and also improve
their biological performance through a more selective toxicity.1
The mechanism of the insecticidal activity resides in an
alteration of the sodium channels, thus altering the nerve
action potential.2 As a result of their efficiency and low toxicity
compared to organophosphorus pesticides, pyrethroids are
pesticides that are applied regularly.
Most pyrethroids are chiral molecules, with only a few
exceptions, such as etofenprox. Therefore, each pyrethroid has
a number of stereoisomers, and their identification is in some
cases influenced by the number of chromatographic peaks and
their resolution. Some pyrethroids, such as cypermethrin,
cyhalothrin, or cyfluthrin, possess three chiral centers, resulting
in eight possible stereoisomers (even though only four pairs of
enantiomers can be resolved by chromatography without the
use of chiral columns). The existence of these isomers also
affects their legislation, as in some cases, only some of them are
approved. For example, in the European Union, cyhalothrin
(as the sum of four pairs of stereoisomers) is not approved,3
whereas λ-cyhalothrin (only two of these isomers) and γ-
cyhalothrin (one isomer) are approved for their use as plant
protection products.4−6 In other cases, such as deltamethrin,
only one enantiomer form is formulated and there is no
ambiguity in its residue definition.
compared to organophosphate insecticides. Their toxicity by
dermal exposure is low as a result of their limited capability to
be absorbed through the skin. However, the average acute oral
LD50 for vegetable oils is in the range of 50−500 mg/kg, which
is considered to be moderately toxic.7 As a result of their high
lipophilicity, they tend to remain in the organism, which leads
to bioaccumulation. Corcellas et al. reported the bioaccumu-
lation of 11 pyrethroids in human breast milk, even though
these pesticides were assumed to hydrolyze in mammals.8
Pyrethroids show very low polarity, with log Kow values
higher than 4. As a consequence, most of the analytical
methods described for the analysis of these compounds use gas
chromatography (GC), instead of liquid chromatography
(LC).9−11 In general terms, LC usually provides lower
sensitivity in the analysis of pyrethroids when reverse-phase
and electrospray ionization (ESI) sources are applied. Few
studies have used LC for the analysis of pyrethroids, and in
these cases, a very specific sample extraction method, including
several cleanup steps or pre-concentration stages, is often
applied to increase the sensitivity of the analysis.12−15 Some
derivatization processes have also been tested, and the general
metabolite 3-phenoxybenzoic acid (3-PBA) is more sensitive
than the original pyrethroids.16 The main inconvenience is that
a high number of pyrethroids can be converted to this acid. On
Special Issue: 55th North American Chemical Residue Workshop
Received: January 30, 2019
Revised: May 6, 2019
Accepted: May 7, 2019
Published: May 7, 2019

© XXXX American Chemical Society A DOI: 10.1021/acs.jafc.9b00732

J. Agric. Food Chem. XXXX, XXX, XXX−XXX
Journal of Agricultural and Food Chemistry
the other hand, as a result of their low polarity, GC is the most
widely used technique for the analysis of pyrethroids. Using
standard extraction methods, such as QuEChERS, an adequate
analysis of pyrethroids by GC can be achieved in multiple
matrices.17,18
Supercritical fluid chromatography (SFC) literature focused
on pyrethroid analysis is not very extensive. Most of the papers
emphasize the use of this type of chromatography for the
separation of enantiomers,19 and when it is used for the
analysis of pyrethroids, usually it is not coupled to mass
spectrometry (MS). However, there are numerous studies of
the use of supercritical fluid extraction (SFE) coupled or not to
SFC for the analysis of pyrethroids.20,21 One of the main
advantages of SFC regarding the ionization point of view
resides in the absence of water in the system.22 Some authors
achieve an increase in the sensitivity of pyrethroids by LC
when analyzing them using an isocratic flow with a very low
percentage of water in the mobile phase.23 These methods are
useful if the scope is mainly focused on pyrethroids but could
be impractical when considering a multiresidue method. SFC
provides high sensitivity for both polar and nonpolar
compounds and, therefore, allows the introduction of
pyrethroids in a broad multiresidue method.
The main objective of the present work is to highlight the
advantages that SFC with ESI sources can provide for the
analysis of pyrethroids, allowing for the enlargement of the
common electrospray scope compounds with this group of
pesticides. The capabilities of this new approach facilitate the
inclusion of pyrethroids in a wide-scope SFC multiresidue
method. For the comparative study, six different vegetable
matrices were investigated (tomato, pear, zucchini, orange,
onion, and tea). These samples represent a wide difficulty
range expected in an analysis laboratory, including complex
matrices, such as onion, orange, and tea. The nature of the
samples includes high-water matrices (tomato, pear, zucchini,
and onion), acid matrix (orange), and dry matrix (tea). The
QuEChERS standard extraction method was used in all cases,
together with common chromatographic conditions for each
separation technique.
2. MATERIALS AND METHODS
2.1. Reagents and Materials. Pesticide standards were acquired
from two different manufacturers: LGC (Teddington, U.K.) and
Sigma-Aldrich (Steinheim, Germany). The purity of these standards
was higher than 96% in all cases, except for permethrin (94.5%) and
flucythrinate (87.5%). The standards were stored at −30 °C. Prior to
the preparation of the mixed solution with all pyrethroids, individual
stock solutions were prepared with a concentration around 1000 mg
L−1 for each pyrethroid.
Reagents employed in the citrate QuEChERS extraction method
(anhydrous magnesium sulfate, sodium hydrogenocitrate sesquihy-
drate, sodium chloride, and sodium citrate tribasic dihydrate) as well
as formic acid and ammonium formate (for the mobile-phase
preparation) were obtained from Sigma-Aldrich (Steinheim, Ger-
many). All gases used by the chromatographic systems (CO2, N2, and
He) have been supplied by Air Liquide (Madrid, Spain).
2.2. Sample Preparation. Five representative fruits and
vegetables with different matrix compositions were purchased from
a local market in Almeriá (Spain). These matrices (tomato, zucchini,
pear, orange, onion, and tea) were extracted and analyzed in a first
step to ensure the absence of pyrethroids. The citrate buffer
QuEChERS method with primary/secondary amine (PSA) dispersive
solid-phase extraction (dSPE) cleanup was applied. The procedure
followed for the extraction did not have any modification, and its
B DOI: 10.1021/acs.jafc.9b00732
Article
parameters have been previously described.22,24 The final extract
resulting from the extraction method contained 1 g of matrix/mL.
2.3. Vial Preparation. To prepare the vials for the injection in the
system, the extracts were spiked with the 14 pyrethroids as follows:
each blank extract [50 μL for GC−tandem mass spectrometry (MS/
MS) and 100 μL for SFC−MS/MS] was evaporated under a gentle
stream of nitrogen and reconstituted with the same volume of an
organic solvent (ethyl acetate for GC−MS/MS and acetonitrile for
SFC−MS/MS) containing the mixture of the analyzed pyrethroids at
2, 5, 10, 20, or 100 μg/L.
2.4. SFC−MS/MS Analysis. The SFC analysis was performed
using a Nexera UC (Shimadzu Corporation, Kyoto, Japan). In
addition to the common devices of a liquid chromatograph, this
system is equipped with a CO2 pump and a back-pressure regulator
(BPR) splitless device just before the MS source.
Methanol with 1 mM ammonium formate was used as a modifier
and mixed with CO2 to build the method gradient. The composition
of the makeup was methanol with 0.1% formic acid and 5 mM
ammonium formate. The makeup solvent was introduced in the
system isocratically at 0.080 mL min−1. The SFC separation was
performed on a C18 stationary phase column Shimpack UC-X RP (3
μm, 250 × 2.1 mm). The oven temperature was set at 40 °C. The
BPR pressure and temperature were established at 150 bar and 50 °C,
respectively. A total mobile phase flow of 1.3 mL/min was used. The
gradient started with an isocratic flow of 1% of modifier that was kept
for 2 min. The modifier percentage increased linearly to 5% after 5
min and 40% after 8 min. This condition was kept for 2 min. The
modifier percentage was then reduced from 40 to 1% to recover initial
conditions and maintained over 3 min. The autosampler temperature
was set at 10 °C, and 2 μL was established as the injection volume.
The SFC system is coupled to a triple quadrupole mass
spectrometer LCMS 8060 (Shimadzu Corporation, Kyoto, Japan).
The study was carried out employing an ESI source operating with 5
ms of switching polarity time. The interface temperature was set at
300 °C, with 250 °C for the desolvation line (DL) and 400 °C in the
case of the heat block. The interface voltage used was 4 kV. With
regard to the nebulizer, heating and drying gas flows: 3, 10, and 10 L
min−1 were used, respectively.
2.5. Optimization of the SFC−MS/MS Parameters. A
Shimadzu extended multireaction monitoring (MRM) library was
used for the creation of the multiresidue method. This feature shows
many transitions for each pesticide; three of them were selected
following the sensitivity rank. Individual standard solutions of the
pesticides were injected to confirm the transition with a higher signal
(quantifier) and the second most sensitive (qualifier). Some
compounds, such as internal standards (dimethoate-d6, carbenda-
zim-d3, malathion-d10, and dichlorvos-d6), were not present in the
library and must be manually optimized using a precursor ion search.
For a proper identification, two transitions must be detected with an
ion ratio difference less than 30% and a retention time shift under 0.1
min. Acquisition windows of ±0.35 min were established for each
pesticide in the multiresidue method.
2.6. GC−MS/MS Analysis. The analyses of pyrethroids by GC
were performed with Agilent Intuvo 9000 GC coupled to an Agilent
7010 GC−MS/MS triple quadrupole and equipped with an Agilent
7693 autosampler. The samples were injected in splitless mode using
a multimode injector through ultra inert inlet liners with glass wool
(Agilent). A temperature ramp (80 °C for 0.1 min and then increased
to 300 °C at 600 °C min−1) was used in the injector, and the total
injection volume was 1 μL.
The run time was 12.4 min, followed by 2.1 min for backflush (310
°C). The temperature program of the oven started at 60 °C (0.5 min),
and then it was increased to 170 °C (80 °C min−1) and finally to 310
°C (20 °C min−1). The flows were kept constant during the analyses
(1.28 mL/min for the first column and 1.48 mL/min for the second
column). Helium was employed as the carrier gas. The collision gas
(nitrogen) flow was 1.5 mL/min, and the quenching gas (helium)
flow was 2.25 mL/min. The high-efficiency electron ionization (EI)
source and the transfer line were kept at 280 °C during the analyses,
and the quadrupoles were maintained at 150 °C.
J. Agric. Food Chem. XXXX, XXX, XXX−XXX
Journal of Agricultural and Food Chemistry
Figure 1. (a) Molecular formulas of permethrin, cyfluthrin, and cypermethrin and one common transition (163 > 127). (b) Chromatogram of
cyfluthrin obtained by GC showing an isobaric interference of permethrin and cypermethrin in the quantifier transition (163 > 127).
2.7. Optimization of the GC−MS/MS Parameters. The
optimization of the GC−MS/MS parameters was performed
according to a previously published paper.25 The individual
pyrethroids were first analyzed in full-scan mode to select the
precursor ion(s) and the retention time. Then, each pyrethroid was
reanalyzed with a product ion method, and the fragmentation patterns
were obtained at different collision energies (5, 10, 15, 20, 25, and 30
eV). The quantifier and qualifier transitions, together with their
corresponding collision energies, were selected.
3. RESULTS
Acrinathrin, bifenthrin, cyfluthrin, cypermethrin, deltamethrin,
etofenprox, fenpropathrin, fenvalerate, flucythrinate, λ-cyhalo-
thrin, permethrin, phenothrin, τ-fluvalinate, and tetramethrin
were the 14 pyrethroids selected to carry out this study. These
pesticides were analyzed using SFC and GC, both coupled to
MS/MS. The comparative study of these techniques was
carried out in terms of limits of quantification (LOQs),
linearity, and matrix effect. The differences in the chromato-
graphic separation and ionization process were also assessed.
3.1. Ionization. The ionization processes applied differ
considerably between the studied techniques as a result of the
type of ion source used in both equipment. In SFC−MS/MS,
an ESI source was used, producing soft ionization. Con-
sequently, the most intense ion of each compound
corresponded to the protonated molecular ion or ammonium
adduct. Except for phenothrin, τ-fluvalinate, and tetramethrin,
all pyrethorids formed an ammonium adduct, instead of the
usual protonation of the molecular ion. This could be related
to their proton affinity and the functional groups present in
C DOI: 10.1021/acs.jafc.9b00732
Article
these compounds, which share similar structures in most
cases.26,27
On the other hand, stronger ionizations are obtained when
using an EI source in GC−MS/MS. The parent ion of each
transition corresponds in most cases to a fragment of the
original molecule, and the molecular ion is not present. In the
case of pyrethroids whose chemical structures are similar, this
may affect the method selectivity, because less selective
transitions are formed with smaller ions. Therefore, it may
be necessary to consider co-elutions of other pyrethroids that
could result in isobaric interferences and interfere with their
quantification. This happened, for instance, in the case of
permethrin (retention time of 9.58 min), cyfluthrin (retention
time of 10.609 min), and cypermethrin (retention time of
10.862 min), which share the transition 163 > 127 (Figure 1a).
These three compounds are very close to each other in the
chromatographic window, and each one shows two (per-
methrin) or three (cyfluthrin and cypermethrin) separated
peaks as a result of the presence of stereoisomers (Figure 1b).
Therefore, the qualifier transition plays an essential role in
their proper identification. Softer ionizations can be also
achieved in GC with the use of chemical ionization (CI)
instead of EI.
3.2. Chromatographic Considerations. Very short run
times were achieved with both techniques, being 13 min for
SFC and 12.4 min for GC. In SFC, the compressibility
properties of CO2 make it possible to apply high flow rates (1.3
mL min−1), which are necessary to improve the chromato-
graphic performance of some compounds included in the
multiresidue method. Therefore, a very short chromatogram
J. Agric. Food Chem. XXXX, XXX, XXX−XXX
Journal of Agricultural and Food Chemistry Article

Figure 2. Chromatograms of permethrin (two transitions) obtained by (left) GC and (right) SFC.

Table 1. LOQs (μg L−1) of the 14 Pyrethroids in the Matrices Included in the Comparative Studies
tomato pear zuccini orange onion tea
SFC GC SFC GC SFC GC SFC GC SFC GC SFC GC
achrinathrin 2 2 2 2 2 2 2 2 2 2 2 2
bifenthrin 2 2 2 2 2 2 2 2 2 2 2 2
cyfluthrin 2 2 2 2 2 2 20 5 20 2 10 2
cypermethrin 2 5 2 5 2 5 2 10 2 5 5 10
deltamethrin 2 2 2 2 2 2 2 2 2 2 2 100
etofenprox 2 2 2 2 2 2 2 2 2 2 2 2
fenprotathrin 2 2 2 2 2 2 2 5 2 2 2 5
fenvalerate 2 2 2 2 2 2 5 5 2 2 5 5
flucythrinate 2 2 2 2 2 2 5 2 5 2 10 2
λ-cyhalothrin 2 2 2 2 2 2 2 2 2 2 5 2
permethrin 2 2 2 2 2 2 2 2 2 2 2 5
phenothrin 2 10 2 10 2 10 5 20 2 5 10 10
τ-fluvalinate 2 2 2 2 2 2 2 2 2 2 2 2
tetramethrin 2 2 2 2 2 2 2 2 2 2 2 2

was obtained with all of the compounds eluting during the first
7 min. This strong eluent flow hinders a proper separation of
the pyrethroid isomers, which, in some cases, are not fully
resolved. Figure 2 shows the chromatograms of permethrin in
the zucchini matrix at the concentration of 2 ppb obtained by
both SFC and GC. It can be observed that the peaks are only
resolved in GC. This could have been an inconvenience if the
pyrethroid residue definition required individual quantification
of a certain isomer. However, for the analyzed pyrethroids, the
maximum residue levels (MRLs) are defined for the sum of
isomers; therefore, a low chromatographic resolution is not a
problem for their quantitative analysis.
The main inconvenience of the short chromatograms
obtained in SFC−MS/MS consists generally of the presence
of isobaric interferences with the matrix. This effect is more
intense when working with complex matrices that possess a
large number of co-eluting compounds, and it is especially
troublesome at low concentration levels. It is sometimes
necessary to add more selective transitions to the affected
compounds to identify them.22 However, pyrethroids are a
pesticide group with molecular masses above the average,
ranging from 331.4 g mol−1 (tetramethrin) to 514.4 g mol−1
(acrinathrin) for the 14 pyrethroids included in this study.
These high molecular weights result in more selective
transitions, and therefore, the probability of finding isobaric
D DOI: 10.1021/acs.jafc.9b00732
interferences decreases dramatically. In the present study, only
cyfluthrin showed a transition affected by interferences that
modified the ion ratio in complex matrices (orange, onion, and
tea). The use of different transitions avoided the effect of this
interference, but their lower sensitivity did not allow for the
identification of this compound at the lowest concentration
levels.
This problem is less common in GC−MS/MS, because this
technique employs longer chromatographic columns that allow
for a better separation efficiency, and therefore, a lower
number of compounds co-elute with the analytes.
3.3. LOQs. The LOQs for the 14 pyrethroids were
evaluated by the use of matrix-matched standards with a
concentration range of 2−100 μg L−1. The results are detailed
in Table 1 as the lower limits of the instrumental concentration
range. Similar results were obtained with both techniques,
being slightly better in SFC (Figure 3). The high sensitivity
achieved in SFC is related to the absence of water in the
mobile phase and the low flows that reach the ESI source,
resulting in low LOQs.22 In GC, for its part, the good results
are associated with the high sensitivity of the instrument used.
The LOQ values achieved are in most cases lower than the
European MRLs for pyrethroids.
The following pesticides: acrinathrin, bifenthrin, etofenprox,
τ-fluvalinate, and tetramethrin, could be identified at the lowest
J. Agric. Food Chem. XXXX, XXX, XXX−XXX
Journal of Agricultural and Food Chemistry
Figure 3. Number of results with a LOQ of 2, 5, and 10 or higher in
the different matrices.
concentration level of 2 μg L−1 in all matrices in both SFC and
GC. Deltamethrin also showed LOQs of 2 μg L−1 in all
matrices, with the exception of tea in GC. In this case, an
interference modified the ion ratios and made it impossible to
identify the pesticide at concentration levels lower than 100 μg
L−1, even with the use of different qualifier transitions (Figure
4). Cyfluthrin showed the highest LOQs in SFC (10−20 μg
L−1) in tea, orange, and onion as a result of the use of less
sensitive transitions to avoid the effects of an isobaric
interference (see section 3.2). This pesticide showed LOQs
of 2 μg L−1 in most matrices with the use of GC. On the other
hand, phenothrin and cypermethrin showed higher LOQs in
all cases with GC than with SFC.
With regard to the matrices, LOQs in tea were in general
higher than the rest, with several pesticides reaching values of
5−10 μg L−1. In tomato, pear, and zucchini, all pesticides
showed LOQs of 2 μg L−1 by SFC and GC, with the exception
of cypermethrin and phenothrin in the latter technique.
The responses were linear across the whole concentration
range in all samples, with the lowest limit corresponding to the
LOQ of each pesticide in the matrix (Table 1). Coefficients of
determination (r2) were higher than 0.99 for all of the studied
cases.
3.4. Matrix Effect. Complex matrices, such as orange or
tea, tend to have a high number of interfering matrix
compounds. These compounds can co-elute with the analytes
Figure 4. Chromatogram of deltamethrin in tea by GC at (left) 10 μg/kg (ion ratio of 337.7%) and (right) 100 μg/kg (ion ratio of 106.7%).
E DOI: 10.1021/acs.jafc.9b00732
Article
in the source, producing signal suppression or signal
enhancement. Co-eluting matrix compounds produce signal
suppression in most cases when an ESI source is employed,28
because the competition for the available charges between the
analytes and the co-eluting matrix compounds produces a
decrease of the ionization efficiency in the interface. However,
the signal suppression using an ESI source performs differently
in SFC compared to LC. In SFC, the analytes reach the
ionization source together with the modifier and the makeup
flow. Methanol is commonly used in SFC as a co-solvent. This
solvent provides lower density and surface tension compared
to water, increasing the solvent evaporation rate and improving
the ionization efficiency. This fact, combined with the low flow
reaching the source and the small amount of sample injected
(2 mg per injection), produces a lower matrix effect compared
to LC.
On the other hand, using GC, matrix interferents usually
cause an enhancement of the analyte signal. This fact is due to
the occupation of some available sites in the liner by the co-
extractive matrix components producing the transfer of a larger
amount of analyte to the chromatographic column.29
The matrix effect obtained for the pyrethroids analyzed in
the six matrices studied is detailed in Table 2. Three different
ranges of the matrix effect can be defined depending upon the
percentage of signal suppression/enhancement. Between 0 and
20% is considered a low or non-existent matrix effect; however,
alterations of the signal between 20 and 50% and >50% are
considered medium and strong matrix effects, respectively. In
all cases, in SFC, the medium and strong matrix effects
correspond to signal suppression. Considering GC, the
situation was the opposite: all pyrethroids with a matrix effect
higher than 20% showed signal enhancement with one
exception, tetramethrin, which presented a signal suppression
of −38% in tea.
A low matrix effect was observed in pear and zucchini
matrices by both techniques. These are high water content
matrices and present lower co-extracted matrix compounds.
With regard to onion, four pesticides presented a medium
matrix effect (bifenthrin, fenpropathrin, fenvalerate, and
flucythrinate) and two pesticides showed a strong matrix
effect (etofenprox and deltamethrin) in SFC. In GC, there was
no matrix effect higher than 50% but most of the pesticides
presented a medium matrix effect, except for deltamethrin,
J. Agric. Food Chem. XXXX, XXX, XXX−XXX
Journal of Agricultural and Food Chemistry Article

Table 2. Matrix Effect (%) of the 14 Pyrethroids in the Matrices Studied

pear zuccini orange onion tea
SFC GC SFC GC SFC GC SFC GC SFC GC
achrinathrin 5 10 5 7 0 94 0 44 −22 52
bifenthrin 10 4 8 8 −7 33 −24 28 −3 5
cyfluthrin 10 0 15 −1 −4 40 −2 23 −57 −5
cypermethrin −2 1 4 −2 −8 43 −17 26 −32 3
deltamethrin 1 11 3 −3 −14 23 −63 15 −18 -
etofenprox 0 1 −1 0 −8 21 −65 15 −10 −12
fenprotathrin 18 5 20 4 −7 41 −25 30 −1 13
fenvalerate 0 −1 4 10 −6 24 −40 12 −32 1
flucythrinate −5 6 −5 1 −8 52 −37 34 −86 16
λ-cyhalothrin −7 2 −6 2 −9 43 −18 25 −19 18
permethrin 2 3 8 5 2 48 −11 28 −10 2
phenothrin 6 9 11 12 −8 53 −17 41 −75 18
τ-fluvalinate 6 9 10 −4 −6 62 −12 22 −50 35
tetramethrin −3 4 3 6 −4 64 −2 35 −11 −38

etofenprox, and fenvalerate. Green tea was the most complex
matrix in SFC: four compounds showed a matrix effect
between 20 and 50% (acrinathrin, cypermethrin, fenvalerate,
and τ-fluvalinate) and three pesticides presented a matrix effect
higher than 50% (cyfluthrin, flucythrinate, and phenothrin).
Better results were obtained in GC when analyzing green tea.
Using this technique, only two pesticides were affected, τ-
fluvalinate and acrinathrin, which showed medium and strong
matrix effects, respectively. No matrix effect was obtained
analyzing orange by SFC; however, this matrix produced a
remarkable enhancement of the signal for all of the compounds
in GC, except for fenvalerate.
With focus on each pesticide, the matrix effect performance
was different depending upon the type of chromatography
used. For example, tetramethrin did not present any matrix
effect in SFC for the matrices studied; however, in GC,
medium and strong matrix effects were observed in orange,
onion, and green tea. A similar situation takes place with λ-
cyhalothrin and permethrin.
only one pesticide each. Cypermethrin was the only pyrethroid
present in more than one sample (pepper and potato). With
regard to quantitation, the concentrations obtained using both
instruments were similar. With the concentrations obtained by
SFC taken as reference, the difference from those of GC is
lower than 25% in all cases.
■ AUTHOR INFORMATION
Corresponding Author
*Telephone: +34-950-015-034. E-mail: amadeo@ual.es.
ORCID
Amadeo R. Fernández-Alba: 0000-0002-9715-3489
Funding
The authors acknowledge support from the European
Commission, DG SANTE (Document SANTE/11813/
2017), and the European Union Reference Laboratory for
Fruits and Vegetables (EURL-FV).
Notes
The authors declare no competing financial interest.

3.5. Real Samples. A total of 17 real samples of fruits and
vegetables were analyzed by both systems to prove the
similarity and reliability of these two types of chromatog-
raphies regarding pyrethroid quantitation. A variety of matrices
were acquired from local markets in Almeriá (apple, aubergine,
banana, broccoli, carrot, green beans, kiwi, leek, lemon,
mandarina, pear, pepper, potato, pumpkin, spinach, and
zucchini). A total of 7 of the 14 pyrethroids validated in the
method were detected in six different samples (Table 3). The
■
ABBREVIATIONS USED
BPR, back-pressure regulator; CI, chemical ionization; DL,
desolvation line; EI, electron ionization; ESI, electrospray
ionization; GC, gas chromatography; LC, liquid chromatog-
raphy; LOQ, limit of quantification; MRL, maximum residue
limit; MRM, multireaction monitoring; MS, mass spectrome-
try; PSA, primary/secondary amine; SFC, supercritical fluid
chromatography; SFE, supercritical fluid extraction

■
pepper sample showed 3 pyrethroids (acrinathrin, cypermeth-
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