Bull Environ Contam Toxicol (2011) 86:1822

DOI 10.1007/s00128-010-0176-9

Evaluation of the QuEChERS Method and Gas Chromatography

Mass Spectrometry for the Analysis Pesticide Residues in Water
and Sediment
S. H. G. Brondi A. N. de Macedo
G. H. L. Vicente A. R. A. Nogueira

Received: 5 August 2010 / Accepted: 2 December 2010 / Published online: 17 December 2010
The Author(s) 2010. This article is published with open access at Springerlink.com

Abstract A method for the determination of pesticide

residues in water and sediment was developed using the
QuEChERS method followed by gas chromatography
mass spectrometry. The method was validated in terms of
accuracy, specificity, linearity, detection and quantification
limits. The recovery percentages obtained for the pesticides
in water at different concentrations ranged from 63 to
116%, with relative standard deviations below 12%. The
corresponding results from the sediment ranged from 48 to
115% with relative standard deviations below 16%. The
limits of detection for the pesticides in water and sediment
were below 0.003 mg L-1 and 0.02 mg kg-1, respectively.
Keywords
Sediment

Pesticides
QuEChERS method
Water

In recent years there has been growing environmental

concern, especially regarding the use and discharge of toxic
substances. Hence there is a growing need to monitor
organic micropollutants in environmental studies and to
control the quality of food. Pollution has been very damaging to aquatic ecosystems, and may consist of agricultural, urban, and industrial wastes containing contaminants
that have proven to be very damaging to aquatic habitats
and species (EPA 2009).

S. H. G. Brondi (&)
A. R. A. Nogueira

Embrapa Pecuaria Sudeste, Rodovia Washington Luiz,
km 234, Sao Carlos Zip Code: 13560-970, Brazil
e-mail: [email protected]
A. N. de Macedo
G. H. L. Vicente
Universidade de Sao Paulo, Avenida do Trabalhador
Sao-carlense, 400, Sao Carlos 13566-590, Brazil

123

Contamination of water resources by pesticide residues

is one of the major challenges for the preservation and
sustainability of the environment (Kustera et al. 2009).
However, these chemicals must be used properly,
respecting the law, the environment and human health,
since they are toxic and can cause contamination (Sanches
et al. 2003).
The determination of the residues of pesticides in food
and environmental samples is very important due to the
riscs that these compounds offer to human health, besides
their persistence in the environment and their tendency to
bioaccumulation (Prestes et al. 2009). The aquatic environment is one of the most affected by pesticide residues. It
is important to monitor the water quality, especially in
regions close to agricultural areas and in places where there
are primary sources of drinking water (Sanches et al.
2003). These pollutants present in water resources at low
concentrations can affect ecosystems and impact drinking
water supplies (Gervais et al. 2008).
The analysis of sediments should be included in environmental studies because they are the result of the integration of all processes (biological, physical and chemical)
that occur in an aquatic ecosystem, influencing the
metabolism of the whole system. Sediments are very different in composition, forms and processes and can provide
valuable information about water quality (Tundisi 2003). In
relation of reservoirs, sediments are considered to be
compartments of accumulation of pollutants, brought by
the water column, that have their origin in the different
uses of soil of the drainage areas basin (Bonai et al. 2009).
Materials carried from the terrestrial environment can
affect water bodies, among then sand, clay, contaminants,
fertilizers, etc. If they are insoluble or poorly soluble and
denser than water, they will be deposited as sediments,
remaining there or being released into the water column

Bull Environ Contam Toxicol (2011) 86:1822

under certain circumstances and in any event potentially

affecting the aquatic food chain.
Trace analysis of organic contaminants such as pesticides in food, biotic and environmental samples typically
consist of following consecutive steps: isolation of analytes
from the sample matrix, removal of bulk co-extracts from
crude extract, identification and quantification of target
analytes and examination the make sure there have been no
false positive results (Hajslova and Zrostlkova 2003).
Many innovations have occurred in analytical methods
for the extraction of organic compounds from different
matrices (e.g. food, biological, and environmental) that
reduce the analysis time, minimize the number of analytical steps, use fewer reagents in smaller amounts and provide high recovery. Recently, Anastassiades et al. (2003)
developed an approach called quick, easy, cheap, effective, rugged, and safe (QuEChERS), which involves
extraction with MeCN partitioned from the aqueous matrix
using anhydrous MgSO4 and NaCl followed by a dispersive-SPE cleanup with MgSO4 and primary secondary
amine (PSA). The QuEChERS method commonly uses
GCMS and LCMS/MS to cover the wide range of pesticides for analysis (Cunha et al. 2007).
Most applications in the analysis of pesticide residues
are based on chromatographic determination, both by gas
chromatography (GC) and high performance liquid chromatography (HPLC), but it is often necessary to use other
techniques such as mass spectrometry to identify the analytes separated by the chromatographic technique. GCMS
has become a standard laboratory instrument and can
provide qualitative and quantitative information for
essentially any GC-amenable analyte in a single injection.
The most common MS technique uses a very rugged and
practical quadrupole design, along with ionization is by
electron impact (EI) (Lehotay 2005).
Method validation is an important requirement in the
practice of chemical analysis. The analyst must generate
information to show that a method intended for these
purposes is capable of providing adequate specificity,
accuracy and precision, at relevant analyte concentrations
and in appropriate matrices (Hill and Reynolds 1999).
The aim of this study was to evaluate the QuEChERS
method in combination with gas chromatography-mass
spectrometry (GCMS) for the determination of atrazine,
fipronil and endosulfan in sediment and water samples.

19

sorbents included PSA, obtained from Varian (USA), and

C18 (50 lm), obtained from J.T.Baker (USA). NaCl was
obtained from Synth (Brazil) and MgSO4 from Mallinckrodt Baker (USA). The water and sediment samples used
for the blank and fortified studies were obtained from an
uncontaminated reservoir.
In this study QuEChERS method was applied according
to Anastassiades et al. (2003) and Lehotay et al. (2005). In
developing the method, the analytical procedure consisted
of the following steps: (a) placing a sample of 10 g of water
or dry sediment into a centrifuge tube; (b) adding atrazine,
fipronil and endosulfan in the required concentrations; (c)
adding 10 mL of MeCN, 4 g of MgSO4 and 1 g of NaCl in
each tube, and centrifuging it at 3,000 rpm for 1 min; (d)
transferring 5 mL of MeCN extract to a commercial SPE
cartridge containing 330 mg PSA, 330 mg C18 and a 1 cm
layer of MgSO4 activated with 3 mL of MeCN. Then, in the
column SPE, (e) the extract was passed and collected; (f)
1.0 mL of the extract was transferred to an autosampler vial
(Shimadzu AOC-20i autoinjector Kyoto, Japan) for
analysis by GCMS. The volume analyzed was 1 lL.
The analyte concentrations were determined through gas
chromatography combined with mass spectrometry using a
QP 2010 GCMS from Shimadzu, equipped with a mass
selective detector (MSD) and a 30-m DB-5 capillary column,
with an internal diameter of 0.25 mm and film thickness of
0.1 lm.
The analytical conditions were injector temperature: 250C,
interface temperature: 250C, the carrier gas with a flow rate of
0.75 mL min-1. The injections were done in the splitless
mode with an oven temperature program of 120C for 3 min,
18C min-1220C and 20C min-1270C for 5 min.
The mass spectrometer was operated initially in the
SCAN mode, sweeping between m/z 45 and m/z 475,
which allowed the detection of analytes in solvent to the
limit 1.0 mg L-1. Lower concentration could be detected
in the SIM mode by selecting the characteristic ions (m/z)
atrazine: 173, 200, 215; fipronil: 215, 351, 367; a-endosulfan: 161, 195, 241; b-endosulfan: 195, 239, 281.
To check the credibility of the data in the quantitative
analyses, analytical validation of the method was performed. The factors considered in the validation included
recovery, precision (relative standard deviation), determination coefficient (R2), linearity, detection (LOD) and
quantification (LOQ) limits (Lancas 2004; Hill and Reynolds 1999; Ribani et al. 2004; Aysal et al. 2007).

Materials and Methods

Results and Discussion
The standard pesticides were obtained from SigmaAldrich
(Germany) with 96.7% purity. The stock and working
solutions were prepared in MeCN solvent, HPLC grade,
obtained from Mallinckrodt (USA). Dispersive SPE

Pesticide residues were determined by GCMS-SIM in

order to obtain the separation of atrazine, fipronil and
endosulfan (a, b) with adequate sensitivity.

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20

Bull Environ Contam Toxicol (2011) 86:1822

(Anastassiades et al. 2003; Aysal et al. 2007), rice (Koesukwiwat et al. 2008), milk, eggs and avocados (Lehotay
et al. 2005), olives and olive oil (Cunha et al. 2007), and
soil (Lesueur et al. 2008), but to our knowledge this is the
first application of the method to sediments.
The representative chromatograms obtained from
extracts of pesticide-fortified in sediment (0.1 mg kg-1)
and in water (0.1 mg L-1) after the QuEChERS method
was applied are shown in Fig. 1. The QuEChERS method
resulted in extracts that contained the target analyte, with
high recovery, and free from interferences in the region of
the chromatogram near the retention time of the pesticides.
Validation is an essential requirement to ensure quality
and reliability of the results for all analytical applications
(Paschoal et al. 2008).
Accuracy: We calculated the percentage of recovery and
reproducibility, expressed as relative standard deviations
(RSD) of the proposed method. The experiments were
performed by spiking the water and sediment samples with
the pesticides being studied. The recoveries obtained for all
pesticides in water at different concentrations ranged from
63% to 116%, with relative standard deviations below
12%, and in the sediment ranged from 48% to 115%, with
relative standard deviations below 16% (Table 1). These
values are within the range stipulated by the U.S. Environmental Protection Agency (Tolosa et al. 1996), which is
from 70% to 130%, with relative standard deviations below
30%, with the exception of endosulfan in water at a concentration below 0.05 mg L-1 and atrazine in sediment at a
concentration of 0.02 mg kg-1.
Specificity: The specificity of the method was determined by analyzing blank water and sediment samples. The
absence of background peaks, above a signal-to-noise ratio
of 3, at the retention times of the target pesticides, showed
that no interferences occurred.
Linearity: An external calibration plot was constructed
in triplicate (n = 3) for analysis of blank water and sediment samples fortified by the addition of standard solutions
of the pesticides, at levels of 0.01, 0.02, 0.05, 0.10, 0.20
and 0.5 mg L-1 and mg kg-1, respectively. The response
for all pesticides was linear in the concentration range

Fig. 1 GC-MS chromatogram of a mixture of pesticides obtained

after QuEChERS method was applied, in the concentration of
0.1 mg kg-1 in sediment (a) and 0.1 mg L-1 in water (b) samples.
1 = atrazine (tr = 7.15 min); 2 = fipronil (tr = 9.00 min); 3 =
a-endosulfan (tr = 9.17 min); 4 = b-endosulfan (tr = 9.75 min)

The QuEChERS method was applied to sample preparation in this study, because it has several advantages over
most of the traditional extraction techniques, according to
Lehotay (2005): high recoveries for a wide polarity and
volatility range of pesticides; very accurate results; low
solvent usage and waste; and high sample throughput.
Besides these advantages, a single person can perform the
method without much training or technical skill, the
method is quite rugged, relatively inexpensive and few
materials and glassware are needed. This method is nowadays the most applied extraction method for the determination of pesticide residues in food samples, providing
acceptable recoveries for acidic, neutral and basic pesticides (Prestes et al. 2009), such as fruits and vegetables

Table 1 Recovery and precision (RSD) from fortified water and sediment samples using the QuEChERS method and GCMS
Pesticides

Recovery (%) RSD (%)

Water (mg L-1)
0.01

Sediment (mg kg-1)

0.05

0.5

0.02

0.05

0.5

Atrazine

111 8

72 10

99 4

48 15

91 4

Fipronil

105 7

72 5

116 8

80 16

115 13

101 7

67 6
68 12

77 1
87 2

80 2
69 9

76 4
74 3

72 4
76 3

a-Endosulfan
b-Endosulfan

123

64 11
63 12

78 12

Bull Environ Contam Toxicol (2011) 86:1822

21

Table 2 Determination coefficient, calibration equation, and values of detection and quantification limits for the investigated pesticides in water
and sediment samples
Pesticides

Water

Sediment

Determination Regression equation

coefficient
(y = ax b)

LOQ
Determination Regression equation
LOD
(y = ax b)
(mg L-1) (mg L-1) coefficient

LOD
LOQ
(mg kg-1) (mg kg-1)

Atrazine

0.9964

y = 654.9x - 11695.0 0.003

0.01

0.9853

y = 257.2x - 3463.7 0.02

Fipronil

0.9970

y = 179.3x - 2033.7

0.003

0.01

0.9835

y = 127.4x - 2048.5 0.003

0.01

a-Endosulfan 0.9969

y = 40.1x - 418.6

0.002

0.005

0.9993

y = 38.8x - 9.2

0.003

0.01

b-Endosulfan 0.9972

y = 23.1x - 209.1

0.002

0.005

0.9860

y = 15.0x - 276.2

0.003

0.01

evaluated, with determination coefficients [0.996 and

0.983 for water and sediment, respectively. The determination coefficient value and regression equation for the
pesticides are shown in Table 2.
Detection and Quantification Limits: The limits of
detection (LOD) and quantification (LOQ) for all pesticides were calculated by considering a value 3 and 10 times
the background noise obtained for blank samples, respectively (Lancas 2004). Table 2 presents the LOD and LOQ
values for atrazine, fipronil and endosulfan (a, b). The low
quantification limit achieved below 0.01 mg L-1 and
0.05 mg kg-1 in water and sediment samples, respectively
allow application of the QuEChERS method followed by
GCMS to monitor pesticide residues in environmental
studies.
After validating of the methods for pesticide residue
analysis in water and sediment, we applied them to determine the levels of atrazine, fipronil and endosulfan in a
river located near a sugarcane plantation. The samples
were analyzed by the QuEChERS method, followed by gas
chromatography (GC) and mass spectrometry (MS). None
of the pesticides analysed were registered, considering the
limit of detection of the method for each compound.
Acknowledgments We are grateful to the Sao Paulo State Research
Foundation (FAPESP) and the National Research Council (CNPq) for
the financial support given to this study.
Open Access This article is distributed under the terms of the
Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any
medium, provided the original author(s) and source are credited.

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