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Talanta
journal homepage: www.elsevier.com/locate/talanta

Multi-residue method for determination of 58 pesticides,

pharmaceuticals and personal care products in water using solvent
demulsication dispersive liquidliquid microextraction combined
with liquid chromatography-tandem mass spectrometry
Sergiane Souza Caldas, Caroline Rombaldi, Jean Lucas de Oliveira Arias,
Liziane Cardoso Marube, Ednei Gilberto Primel n
Escola de Qumica e Alimentos, Universidade Federal do Rio Grande-FURG, 96203-900 Rio Grande, RS, Brazil

art ic l e i nf o a b s t r a c t

Article history: A rapid and efcient sample pretreatment using solvent-based de-emulsication dispersive liquidliquid
Received 2 March 2015 microextraction (SD-DLLME) coupled with liquid chromatography-tandem mass spectrometry (LCMS/
Received in revised form MS) was studied for the extraction of 58 pharmaceuticals and personal care products (PPCPs) and pes-
11 June 2015
ticides from water samples. Type and volume of extraction and disperser solvents, pH, salt addition,
Accepted 17 June 2015
amount of salt and type of demulsication solvent were evaluated. Limits of quantication (LOQ) in the
range from 0.0125 to 1.25 mg L
1 were reached, and linearity was in the range from the LOQ of each
Keywords: compound to 25 g L
1. Recoveries ranged from 60% to 120% for 84% of the compounds, with relative
Dispersive liquidliquid microextraction standard deviations lower than 29%. The proposed method demonstrated, for the rst time, that sample
Pharmaceuticals
preparation by SD-DLLME with determination by LCMS/MS can be successfully used for the simulta-
Personal care products
neous extraction of 32 pesticides and 26 PPCPs from water samples. The entire procedure, including the
Pesticides
Liquid chromatography extraction of 58 organic compounds from the aqueous sample solution and the breaking up of the
Mass spectrometry emulsion after extraction with water, rather than with an organic solvent, was environmentally friendly.
Water samples In addition, this technique was less expensive and faster than traditional techniques. Finally, the ana-
lytical method under study was successfully applied to the analysis of all 58 pesticides and PPCPs in
surface water samples.
& 2015 Elsevier B.V. All rights reserved.

1. Introduction treatment processes [3]. In addition, approximately 3000 different

A large number of synthetic organic compounds, such as
pharmaceuticals and personal care products (PPCPs), as well as
pesticides, has been found in the aquatic environment [1].
The important role that pesticides play in food protection has
led to their massive global use in agriculture and consequent de-
tection in the environment. Pesticides in water sources have been
a topic of considerable interest due to the detection of an in-
creasing number of pesticides in the environment, a fact that re-
quires the establishment of strict regulations to minimize their
impact [2].
PPCPs have become emerging contaminants because of the
threat they pose to drinking water, their effects on human life and
wildlife, besides their incomplete removal in wastewater
n
Corresponding author.
E-mail address: eprimelfurg@gmail.com (E.G. Primel).
substances seem to be used as pharmaceutical ingredients, such as
pharmaceuticals used by humans and those that aim at veterinary
use for livestock, poultry and sh farming [4].
To quantitatively evaluate the fate of these chemicals and en-
sure the quality of drinking water, effective analytical methods are
highly desirable. Because the concentrations of pesticides and
PPCPs in water are usually very low (ng L
1 or lower), it is ne-
cessary to incorporate a concentration step into the analytical
procedure prior to gas chromatographic or liquid chromatographic
determinations. Solid phase extraction (SPE) is the most com-
monly used extraction method to extract multiresidue compounds
from water samples [5]; however, new extraction techniques,
aiming at reducing the overall analytical time and solvent con-
sumption, have been recently proposed.
Multiresidue analytical methods are preferred to single group
analysis because the former provide broader knowledge about the
occurrence, removal, partition and fate of pollutants in the

http://dx.doi.org/10.1016/j.talanta.2015.06.047
0039-9140/& 2015 Elsevier B.V. All rights reserved.

Please cite this article as: S.S. Caldas, et al., Talanta (2015), http://dx.doi.org/10.1016/j.talanta.2015.06.047i
2 S.S. Caldas et al. / Talanta ()
environment [6]. However, the biggest challenge comes from the
fact that groups of contaminants have a broad spectrum of che-
mical and physical properties, thus requiring a solvent which is
capable of extracting this group of diverse compounds.
Since the development of Dispersive LiquidLiquid Micro-
extraction (DLLME) by Rezaee et al. [7] in 2006, the technique has
been very popular among analytical chemists. Since then, DLLME
has undergone many changes, mainly related to the requirements
of the extraction and disperser solvent. Most changes attempted to
use solvents which were lighter than water [8], solvents with low
toxicity and more convenient practical procedures [9], such as the
DLLME based on the solidication of a oating organic drop
(DLLME-SFO) and the solvent-terminated dispersive liquidliquid
microextraction (ST-DLLME) [10].
ST-DLLME, also known as solvent-based de-emulsication
DLLME (SD-DLLME), was rst introduced by Chen et al. [10]. This
change in DLLME was studied to employ low-density extraction
solvents to DLLME and remove the centrifugation step. Because
the oil/water (O/W) emulsion that was formed after the injection
of the dispersion and extraction solvent into the sample was
thermodynamically unstable, solvents-usually used as disperser
solvents-were introduced as chemical demulsiers to break up the
dispersed system, considering their characteristics of low surface
tension and high surface activity. After this injection, the emulsion
cleared quickly into two phases. Therefore, separation of the or-
ganic phase from the aqueous bulk was achieved without the use
of centrifugation.
After its development, the SD-DLLME technique has been em-
ployed to the extraction of organic compounds, such as polycyclic
aromatic hydrocarbons [11,12], organophosphorus [13], and or-
ganochlorine pesticides [14], carbamates [10], s-triazine herbicides
[15], chlorophenols [16], phthalate esters [17,18], chlorophenoxy
acid herbicides [19] and inorganic species, such as palladium [20]
and cadmium [21], from water samples.
Due to the use of solvents which are lighter than water, SD-
DLLME overcomes some drawbacks of DLLME, mainly related to
the number of extraction solvents that are available to be used in
the method and to the ability to extract target analytes. In addi-
tion, when this technique is used, halogenated hydrocarbon sol-
vents are avoided. Another interesting advantage is the elimina-
tion of centrifugation, which is considered to be the most time-
consuming step of the method [8].
Because other solvents than the ones generally used in DLLME
extractions could be employed, this technique seems to be an in-
teresting option for the extraction of multiresidue compounds
from water samples, a fact that has been poorly explored when
this technique is used. With the aim of studying and expanding
the applicability of DLLME, a simple and fast method based on SD-
DLLME was developed by evaluating its performance when low-
density extraction solvents were applied, combined with liquid
chromatography-tandem mass spectrometry (LCMS/MS) for the
determination of 32 pesticides and 26 PPCPs in water samples.
To the best of our knowledge, there is no report on the use of
SD-DLLME for the multi-residue analysis of pesticides and PPCPs.
2. Experimental
2.1. Reagents
Salicylic acid, amitriptyline, avobenzone, sodium diclofenac, eu-
solex 6300, furosemide, mebendazole and sulfamethoxazole were
bought from United States Pharmacopeia (USP, USA). Albendazole,
carbamazepine, clarithromycin, diltiazem hydrochloride, urazepam
hydrochloride, chlorpropamide, gembrozil, glibenclamide, halo-
peridol, methylparaben, nimesulide, miconazole nitrate, propranolol
Please cite this article as: S.S. Caldas, et al., Talanta (2015), http://dx.doi.org/10.1016/j.talanta.2015.06.047i
and propylparaben were provided by Fiocruz (Fundao Oswaldo
Cruz, Brazil). Bisphenol A, 2,4-D, atrazine, atrazine-d5, azoxystrobin,
bentazone, carbofuran, carboxin, cyproconazole, clomazone, di-
chloran, diuron, difenoconazole, fenoxaprop-p-ethyl, pronil, ibu-
profen-d3, irgarol, malathion, metalaxyl-m, metsulfuron-methyl,
pyraclostrobin, pyrazosulfuron-ethyl, pirimiphos-methyl, propanil,
propiconazole, quinclorac, simazine, tau-uvalinate, tebuconazole
and trioxystrobin were bought from Sigma Aldrich (Brazil). Ibu-
profen, triclocarban, triclosan, bispyribac-sodium, cyhalofop-butyl,
chlorantraniliprole, epoxiconazole and penoxsulam were supplied by
Dr. Ehrenstofer GmbH (Germany).
Acetonitrile, methanol, toluene and acetone (HPLC grade) were
bought from J.T. Baker (USA). Hexyl acetate and octanol were
supplied by Sigma Aldrich (Brazil). Magnesium sulfate (anhydrous,
MgSO4) was purchased from J.T.Baker (Mexico) whereas sodium
chloride was bought from Merck (Germany). Calcium chloride,
sodium hydroxide and ammonium sulfate were provided by Synth
(Brazil). Glacial acetic acid and hydrochloridric acid were obtained
from Merck (Germany). The water was puried by an Ultrapure
Water System (USA).
2.2. Apparatus and software
LCMS/MS was performed by a Waters Alliance 2695 Separa-
tions Module (Waters, Milford, MA, USA) tted with an auto-
sampler, a membrane degasser and a quaternary pump. Mass
spectrometry was performed by a Micromass Quattro Micro API
(Waters, Milford, MA, USA) with an electrospray (ESI) interface.
The drying gas, as well as the nebulizing gas, was nitrogen, gen-
erated by pressurized air in an Genius NM32LA nitrogen generator
(Peak Scientic). The nebulizing gas ow was 50 L h
1 whereas
the desolvation gas ow was 550 L h
1.
To operate in the MS/MS mode, the collision gas was argon
99.99% (White Martins, Brazil) with a pressure of 3.5  10
3 mbar
in the collision cell. The optimized values were as follows: capil-
lary voltage, 4.0 kV; extractor voltage, 2 V; source temperature,
100 C; desolvation temperature, 400 C; and multiplier, 650 V.
For each compound, optimum collision energies, which aimed
at getting two characteristic multiple reaction monitoring (MRM)
transitions with the best signal intensity, were selected. After the
optimization of the collision cell energy of the triple quadrupole,
two different precursor ion-product ion transitions, for quanti-
cation and for conrmation, were selected for each analyte. Table 1
shows the optimized MRM transitions for the pesticides with their
respective retention times (tR). Analytical instrument control, data
acquisition and treatment were performed by the software Mas-
sLynx (Micromass, Manchester, UK), version 4.1.
The chromatographic separation was performed by a Kinetex
C8 (3.0 mm  50 mm i.d., 2.6 m lm thickness) column (Phe-
nomenex, USA). The mobile phase components were (A) ultra-
pure water with 0.1% acetic acid and (B) methanol, with elution in
the gradient mode. The initial composition was 20% B; it increased
linearly to 90% in 20 min, held until 23 min and, then, returned to
the initial composition (20% B) in 0.5 min and held for 6.5 min,
totaling a 30- min analysis. The ow rates were as follows: 0
20 min, 0.20.4 mL min
1; 2023 min, 0.4 mL min
1; 23
23.5 min, 0.4-0.2 mL min
1; 23.530 min, 0.2 mL min
1. The in-
jection volume was 10 L.
2.3. Analytes selection
Aiming at verifying the feasibility of the use of SD-DLLME for
the extraction of multiclass analytes from water samples, a large
number of analytes with different physicochemical properties, in
terms of polarity and water solubility, was chosen (Table 1)
[22,23]. Pesticides are widely used in agriculture. Pesticides and
 S.S. Caldas et al. / Talanta () 3

Table 1
Mass spectrometric parameters for the LCMS/MS determination of pesticides and PPCPs, water solubility and the partition coefcient octanol/water (Kow) [22,23].

PPCPs ESI MRM transition Cone voltage Collision energy Water solubility Log Kow

(m/z) (V) (eV) (mg L
1 at 2025 C)

Salicylic acid
136.9 492.8a 20 20 2240 2.26

136.9 464.8 20 25
Albendazole 266 4234 30 20 Practically insoluble 2.7
266 4191a 33 32
Amitriptyline 278.3 4233.3a 35 15 9.71 4.92
278.3 4116.9 35 15
Avobenzone 310.4 4135a 20 30 9.74
310.4 4161 20 30
Bisphenol A 227.17 4212.2a 43 19 120 3.183.7
2274133 43 25
Carbamazepine 236.9 4194.1a 26 12 17.7 2.45
237.4 4167.4 35 40
Clarithromycin 748.75482.8 35 59 0.33 3.16
748.754158.3a 35 29
Diltiazem hydrochloride 4154310 35 20 465 2.8
4154178a 35 20
Flurazepam hydrochloride 388.4 4315.2a 30 25 500,000 (HCl salt) 3.8
388.4 4288.2 30 25
Chlorpropamide 277 4175a 27 22 258 2.27
277 4110.9 27 29
Sodium diclofenac 293.6 4250.2a 20 10 50,000 (sodium salt) 4.51
294 4 214 20 25
Eusolex 6300 255.4 4157.2 25 20 4.95
255.4 4105a 25 30
Furosemide 328.8 4284.9 30 15 73.1 2.03
328.8 4205a 30 20
Gembrozil 249 4 121a 20 30 10,000 3.4
Glibenclamide 494 4369 30 18 4 4.7
494 4169a 30 38
Haloperidol 3764165a 30 21 14 4.30
376.4 4123 35 25
Ibuprofen 205.4 4161a 15 7 21 3.97
205.4 4125.8 15 7
Mebendazole 296.2 4264.2 35 30 71.3 2.83
296.2 4104.9a 35 30
Methylparaben 1514135.9 35 15 1.91
151491.6a 35 20
Nimesulide 3074229a 33 20 18.2 2.60
307.2 4198.1 30 25
Miconazole nitrate 417.14161 45 25 10,000 6.1
417.14159a 45 30
Propranolol 260 4 116a 30 18 61.7 3.48
260 4 183 30 20
Propylparaben 179.1 4137.1 30 15 2.94
179.1 491.8a 30 20
Sulfamethoxazole 254 4 92 30 25 610 0.89
254 4 156.1a 30 20
Triclocarban 3134160.1a 30 25 0.045 3.5
3154125.7 30 15
Triclosan 289 4 35 18 7 6.05 5.53, 4.98
287 4 35a 18 9
Pesticides
2,4-D 2194161a 15 20 23,180 (pH 7) 2.582.83
219489 15 30
Atrazine 2164174a 33 20 33 2.5
2164146 35 22
Azoxystrobin 404 4372a 20 20 6 2.5
404 4329 15 30
Bentazone 239 4 132a 35 25 570 0.77
239 4 197 35 20
Bispyribac-Sodium 453 4 297a 35 25 73,300 1.03 (
)
453 4 275 35 22
Carbofuran 222 4 165 20 25 351 1.52
222 4 123a 20 25
Carboxin 236 4 143a 35 15 147 2.3
236 4 87 14 33
Cyhalofop-Butyl 357.6 4256.1a 30 20 0.44 (pH 7.0) 3.31
357.9 4158 23 57
Cyproconazole 292 4 125 35 20 93 3.1
292 4 70a 35 20
Clomazone 2404125a 30 20 1100 2.5
2404219 26 20
Chlorantraniliprole 484.14453.1a 30 19 0.88 2.8626

Please cite this article as: S.S. Caldas, et al., Talanta (2015), http://dx.doi.org/10.1016/j.talanta.2015.06.047i
4 S.S. Caldas et al. / Talanta ()

Table 1 (continued )

PPCPs ESI MRM transition Cone voltage Collision energy Water solubility Log Kow

(m/z) (V) (eV) (mg L
1 at 2025 C)

484.14285.8 30 30
Dichloran [26] 205 4 175.2a 40 15 6.4 2.8
205 4 138.9 40 20
Difenoconazole 406 4251a 31 32 15 4.4
406 4337 32 20
Diuron 233472a 28 20 36.4 2.85
2334160 28 25
Epoxiconazole 3304123 27 30 0.663 3.44
3304121a 27 30
Fenoxaprop-p-ethyl 362.1 4288.1a 22 23 0.7 4.58
362.1 4121 22 37
Fipronil 4354330a 30 15 1.9 (pH 5), 2.4 (pH 9) 4.0
4354250 25 26
Irgarol 254 4 108 30 30 7 3.9
254 4 198a 30 19
Malathion 3314199 24 30 145 2.75
3314127a 24 10
Metalaxyl-M 280 4 220 16 17 26,000 1.71
280 4 192a 16 17
Metsulfuron-Methyl 380 4 139a 30 15 2790 (pH 7) 0.018
380 4 214 30 10
Penoxsulam 482 4109 35 40 408 (pH 7)
0.354
482 4179a 35 25
Pyraclostrobin 388.1 4194 20 19 1.9 3.99
388.1 4163a 20 19
Pyrazosulfuron-Ethyl 4134232a 35 15 9.76 3.16
4134154 35 26
Pirimiphos-Methyl 306 4136 40 33 10 (pH 7) 4.2
306 4108a 40 20
Propanil 2184127 25 28 130 3.3
2184162a 30 14
Propiconazole 342 4 159a 32 22 100 3.72
342 4 69 30 20
Quinclorac 2404196 15 6 0.065
1.15
Simazine 202 4 132a 35 18 6.2 2.1
202 4 124 35 18
Tau-Fluvalinate 503.4 4208.1a 20 11 0.00103 4.26
503.4 4180.9 20 11
Tebuconazole 308 4 70a 40 20 36 3.7
308 4 125 28 22
Trioxystrobin 409 4 206 35 15 0.610 4.5
409 4 145a 25 40

a
Quantication transitions.

pharmaceuticals and personal care products have been detected in
water samples [1].
2.4. SD-DLLME
Samples were divided into two subsamples: one of them had
the pH adjusted to 2 and the other one had the pH adjusted to 8.
Subsamples were extracted in triplicate and injected three times.
An aliquot of 10 mL water sample 1% MgSO4 (w/v) was placed in a
10 mL volumetric ask. The mixture of 120 mL organic solvent and
0.75 mL disperser solvent was quickly injected into the sample
solution with a 1.00 mL syringe. An emulsion (water, extraction
solvent, disperser solvent) was formed in the volumetric ask.
After, 0.75 mL demulsication solvent, serving as the demulsier,
was injected into the top surface of the aqueous bulk to break up
the emulsion. Then, the emulsion quickly cleared into two phases.
The upper layer was collected by a microsyringe and the volume of
the light phase was checked (120 75 mL). The lighter phase was
placed in an Eppendorf and the volume was increased to 250 mL
with methanol and placed in an insert. Then, 10 mL was injected
into the LCMS/MS.
2.5. Validation experiments and internal quality control criteria
The validation of the analytical method was performed by
using spiked control samples of drinking water and was assessed
in agreement with SANCO and INMETRO guidelines [24,25].
To ensure the quality of the results, some internal quality cri-
teria, which comprise the analyses of laboratory blanks (solvent
blank) and laboratory control samples, have been applied. When
the reagents were used, background levels of analytes were below
the detection limits. Furthermore, the daily set of samples under
analysis was processed together with a blank extract that elimi-
nated a false positive by contamination in the extraction process,
instrument or chemicals. Calibration curves were prepared daily in
blank matrix extracts to check both sensitivity and linearity in the
working range of concentrations. Thus, quantication mistakes
caused by possible matrix effects of instrumental uctuations
could be avoided.
Recovery was determined at least in three levels and in three
replicates. Calculations of recoveries were carried out by using the
peak areas according to the following equation: recovery (%)
[(C1
C2)/C3] x 100, where C1 concentration of the analyte in
the nal extract, C2 concentration of the analyte in the blank
sample and C3concentration of the analyte added to the sample.

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 S.S. Caldas et al. / Talanta ()
Fig. 1. Percentage of compounds within the recovery ranges ( o 20%, between 20%
and 50% and 450%) after extraction by SD-DLLME with different 1-octanol solvent
volumes (sample volume, 10 mL; sample pH, 6.6; disperser and demulsier (acet-
one) volume, 500 mL).
Precision was calculated as the relative standard deviation (RSD)
for each concentration level. Limits of detection (LODs) and limits
of quantication (LOQs) were determined as the lowest injected
compound concentration that yielded a signal-to-noise (S/N) ratio
of 3 and 10 (when the quantication ion was monitored), re-
spectively. Resulting values were also experimentally checked. The
linearity of the instrumental analytical curves was evaluated at a
concentration range from the LOQ to 1000 mg L
1 by at least ve
calibration solutions prepared in blank control sample extracts
and in the solvent. Overall SD-DLLME linearity was also in-
vestigated by using water samples fortied in a range from the
LOQ to 25 mg L
1.
The matrix effect (ME) was also investigated. The degree of ion
suppression/enhancement can vary from sample to sample, from
compound to compound, and may also depend on the analyte
concentration, as well as on the matrix-to-analyte concentration
ratio. Therefore, the evaluation of the ME was accomplished by
comparing the calibration curves in the solvent and the calibration
curves prepared in the matrix [26]. The effects of the matrix
components were rated according to the percentage of signal en-
hancement ( ) or suppression (
).
2.6. Statistical analysis
For the statistical analysis performed in the fractional factorial
design, at 95% level of condence, the Statistica 8.0 Portable U.S.
Headquarters: StatSoft, Inc., Tulsa, USA (2007) software was used.
3. Results and discussion
3.1. Disperser and extraction solvent optimization
Toluene, hexyl acetate and 1-octanol were tested as extraction
solvents, whereas methanol, acetonitrile and acetone were si-
multaneously tested as disperser solvents.
Toluene showed recoveries higher than 50% for 6 compounds
when combined with methanol and acetonitrile. Recoveries higher
than 50% were reached for 10 compounds when acetone was used
as disperser.
Although the recovery rates of many analytes were low in this
study, toluene had been successfully employed as an extraction
solvent in many microextraction techniques and several classes of
compounds had been extracted from different samples, such as
phthalate esters from bottled waters [18], organophosphorus from
farm-eld water and industrial wastewater [27], drugs of abuse
from urine [28] and chloropyrifos, lambda-cyhalothrin, cyuthrin,
Please cite this article as: S.S. Caldas, et al., Talanta (2015), http://dx.doi.org/10.1016/j.talanta.2015.06.047i
5
Fig. 2. Percentage of compounds within the recovery ranges (o 20%, between 20%
and 50% and 450%) after extraction by SD-DLLME with different acetone volumes
(sample volume, 10 mL; sample pH 6.6; 1-octanol volume, 120 mL).
cypermethrin, fenvalerate and deltamethrin [29], and carbamates
from water samples [10].
Among the solvents under study, hexyl acetate (as the extractor
with methanol as the disperser) had the worst results, i.e., re-
coveries higher than 50% were not obtained for any compound. In
other studies, hexyl acetate has been successfully employed for the
extraction of phenolic acids from fruits [30], acidic herbicides [31]
and phenols [32] from water samples, besides parabens from
water and cosmetics [33].
Octanol had the best results. Recoveries higher than 50% were
reached for 24 compounds when acetonitrile was used as the
disperser, 28 compounds when acetone was the disperser, and 30
compounds with methanol. The explanation could be related to
the dielectric constant of octanol (108). Its dielectric constant is
higher than those of toluene and hexyl acetate. The dielectric
constant is not only a key parameter for modeling solvent behavior
but also an important measure of the polarity of a solvent [34].
Octanol has been employed as an extraction solvent in other
studies for the extraction of chlorophenols from environmental
samples [35], endocrine disruptors from water samples [36], al-
kylphenols and bisphenol A from seawater samples [37], and for
the extraction of benzophenone from environmental water sam-
ples [38].
Octanol combined with acetone had the best results. Since si-
milar area values were obtained for some compounds by using the
solvent combination, a statistical test was also carried out to
evaluate whether there were signicant differences among the
area values and the solvent evaluation.
For some compounds (nimesulide, miconazole nitrate, triclo-
carban, azoxystrobin and tebuconazole), no signicant difference
(p 40.05) was found among the different extractor and disperser
solvents. When 1-octanol was used as extractor and acetone, as
disperser, a higher number of compounds with signicant higher
areas (p o0.05) was found. Considering the results, octanol and
acetone were chosen.
3.2. Extractor solvent volume optimization
In microextraction techniques that employ solvents that are
less dense than water, volumes of extractors between 8 and 145 mL
have been used [39,40].
In this study, volumes of 50, 80, 100, 120, 140 and 200 mL oc-
tanol were evaluated together with 500 mL acetone as the
disperser.
A high percentage of compounds with recoveries higher than
50% was obtained when extractor volumes were between 100 and
140 mL (Fig. 1). The Tukey test was carried out: from 100 to 120 mL,
for 7 compounds, a signicant difference was observed and high
6 S.S. Caldas et al. / Talanta ()

Fig. 3. Peak area after extraction by SD-DLLME with different demulsier solvents (0.75 mL) (sample volume, 10 mL; sample pH 2 and 8; 1-octanol volume, 120 mL; acetone
volume, 0.75 mL).

chosen.

3.3. Volume of disperser and demulsier solvent

To examine the effect of the disperser and demulsier solvent,

volumes of 250, 500, 750 and 1000 mL of acetone (disperser and
demulsier) were evaluated with 120 mL 1-octanol. The volume of
demulsier and disperser evaluated were the same (i.e., when
250 mL acetone was used as the disperser solvent, 250 mL acetone
was used as the demulsier). With 250 mL, lower recoveries were
obtained; the larger the volumes, the higher the recoveries. Thus,
higher recovery values were obtained with 750 and 1000 mL
(Fig. 2).
With 1000 mL, recoveries of some compounds, such as avo-
benzone, malathion, thiobencarb, pronil and glibenclamide,
showed signicant decrease. Thus, 750 mL was adopted for further
use.

3.4. pH

The inuence of pH on extraction recoveries was shown to be

Fig. 4. Steps and optimized extraction conditions of the proposed SD-DLLME.
area values were observed when 120 mL was employed. From 120
to 140 mL, only 3 compounds showed signicant differences when
140 mL was employed. Thus, based on these results, 120 mL was
very signicant. As most pesticides/PPCPs contain one or more
ionizable functional groups, the pH of the aqueous phase inu-
ences the extraction recovery because charged species are as-
sumed to avoid organic solvents [41].
Different behaviors of the analytes was observed. Some com-
pounds, such as quinclorac (pKa 4.34), metsulfuron-methyl (pKa

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 S.S. Caldas et al. / Talanta () 7

Table 2
Method limit of quantication (LOQ), slope (a) and intercept (b) for the analytical curves in the solvent and in the extract and matrix effect (ME).

PPCPs LOQ Solvent Matrix ME (%)

(lg L
1) a b a b

Albendazole 0.025 747,058 100.609 773.305 17.3543 4

Amitriptyline 0.125 322.008 47.1326 315.898 224.169
2
Avobenzone 0.25 44.407 334.097 40756.7 243.639
8
Bisphenol A 0.25 7689.5 43.357 6928.84 46.7468
10
Carbamazepine 0.025 274.312 83.7142 246,943 64.8114
10
Chlorpropamide 0.125 76940.2 157.967 67006.3 135.091
13
Clarithromycin 0.125 966.043 298.769 721.695 106.628
25
Diltiazem hydrochloride 0.0125 6.24E 06 1678.79 5.96E 06 1990.2
5
Eusolex 6300 1.25 30419.5 11.9771 28298.9 12.5305
7
Flurazepam hydrochloride 0.025 550.517 106.138 555.024 120.88 1
Furosemide 0.125 95029.5 22.1068 83852.8 53.8767
12
Gembrozil 0.25 46567.2 36.1276 41746.4 31.7285
10
Glibenclamide 0.25 154.988 329.358 119,081 119.333
23
Haloperidol 0.0125 1.34E 06 895.453 1.21E 06 727.162
9
Ibuprofen 0.125 85902.8 40.9775 68003.8 87.9626
21
Mebendazole 0.025 413.003 4.10616 433.436 136.741 5
Methylparaben 0.25 48962.9 41.1516 45.813 45.7173
6
Miconazole nitrate 0.125 498.723 222.322 449,900 405.97
10
Nimesulide 0.125 264.175 293.049 223.382 317.672
15
Propranolol 0.025 376.092 92.732 343,914 181.102
9
Propylparaben 0.125 48392.2 28.2522 44168.1 43.7894
9
Salicylic acid 0.25 43117.5 40.287 41825.5 70.2582
3
Sodium diclofenac 0.0125 308.471 11.7871 258.880 177.866
16
Sulfamethoxazole 1.25 17136.7 17.1644 15,893 37.8028
7
Triclocarban 0.025 407.710 403.656 376.532 365.503
8
Triclosan 1.25 2502.06 51.8209 1503.81 39.0416
40
Pesticides
2,4-D 0.25 34732.8 12.8854 31070.1 25.1036
11
Atrazine 0.125 238.123 68.384 223.412 82.8531
6
Azoxystrobin 0.0125 441.764 261.847 365.991 244.91
17
Bentazone 0.125 128.984 134.585 119.260 139.932
8
Bispyribac-sodium 0.125 328.437 241.925 239.207 268.642
27
Carbofuran 0.125 290.235 129.828 281.794 150.584
3
Carboxin 0.125 228.738 177.475 201.920 62.6734
12
Chlorantraniliprole 0.25 65114.3 43.2554 56.305 98.5393
14
Clomazone 0.125 419.062 50.6414 395.844 360.108
6
Cyhalofop-butyl 1.25 19902.5 136.779 15122.6 110.789
24
Cyproconazole 0.125 271.399 34.648 240.635 130.12
11
Dichloran 1.25 2011.53 12.934 1785.32 12.5396
11
Difenoconazole 0.125 363.418 89.8542 327.668 360.017
10
Diuron 0.25 53326.7 171.346 50703.9 183.738
5
Epoxiconazole 0.125 314.658 253.753 284.021 693.557
10
Fenoxaprop-p-ethyl 0.125 94622.2 58.1125 90913.9 106.641 4
Fipronil 0.025 847.267 100.531 757.628 80.941 11
Irgarol 0.0125 1.67E 06 281.37 1.61E 06 289.739 4
Malathion 0.25 4.92E 06 2977.79 3.77E 06 2895.49 23
Metalaxyl-M 0.125 228.358 91.7955 200.740 145.691 12
Metsulfuron-methyl 0.125 84251.9 18.6323 68595.7 30.2914 19
Penoxsulam 0.25 20.288 17.7606 17.932 18.4362 12
Pirimiphos-methyl 0.25 441296 1245.77 432.204 1435.89 2
Propanil 0.125 60,590 38.5425 57993.7 11.3197 4
Propiconazole 0.125 272.270 110.132 236467 318.317 13
Pyraclostrobin 0.125 140.355 190.218 119.155 202.441 15
Pyrazosulfuron-ethyl 0.025 286.414 4.61584 244.858 30.026 15
Quinclorac 0.25 7750.8 0.912524 5983.81 13.7731 23
Simazine 0.25 45953.6 31.3609 46522.1 1.01533 1
Tau-uvalinate 0.125 29021.4 16.5584 23819.4 15.5418 18
Tebuconazole 0.125 84895.2 1.14719 78189.7 58.1691 8
Trioxystrobin 0.125 263.615 310.779 240.404 391.59
9

3.8), pyrazosulfuron-ethyl (pKa 3.7), penoxsulam (pKa 5.1), bis-
pyribac-sodium (pKa 3.05), bentazone (pKa 3.3), salicylic acid (pKa
2.98), furosemide (pKa
1.05) and 2,4-D (pKa 2.73), were ex-
tracted only at pH equal to or lower than 4.
Amitriptyline (pKa 9.4), ditialzem chloridrate (pKa 8.18) and
urazepam chloridrate (pKa 8.71) showed signicantly increased
recoveries at pH higher than 6.
Results were in agreement with those found by other studies.
Some examples are given below.
Amitriptyline was recovered at pH between 6 and 12.
Amitriptyline was extracted by DLLME in another study where the
pH under study ranged from 10 to 13. The authors chose pH 12 as
the optimum for the extraction [42].
Carbofuran showed similar recoveries when the pH ranged
from 1 to 10. Carbamates were extracted by SD-DLLME in another
study in which the authors chose pH 7 [10].
Compounds that belong to the parabens class, methylparaben
and propylparaben, showed similar values for recovery in the pH
range from 1 to 8, with reduction at pH 10 and no extraction at pH
12. These results were in agreement with ndings reported in the

Please cite this article as: S.S. Caldas, et al., Talanta (2015), http://dx.doi.org/10.1016/j.talanta.2015.06.047i
 8
Table 3
Please cite this article as: S.S. Caldas, et al., Talanta (2015), http://dx.doi.org/10.1016/j.talanta.2015.06.047i

Recoveries (%) and relative standard deviations (RSD, %) in spiked drinking water samples.

Intra-day Inter-day

0.0125 0.025 0.125 0.25 1.25 2.5 12.5 0.125 0.25 1.25 2.5 12.5

PPCPs R RSD R RSD R RSD R RSD R RSD R RSD R RSD R RSD R RSD R RSD R RSD R RSD
(%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%) (%)
Albendazole 94 16 96 17 87 14 90 8 81 9 68 5 80 12 72 10 72 18 78 9 87 7
Amitriptyline 110 13 96 10 105 5 94 11 70 15 115 10 86 14 94 17 89 9 96 9
Avobenzone 121 9 117 13 108 19 74 13 117 12 102 16 105 19 93 13
Bisphenol A 79 18 115 13 117 14 90 16 92 21 97 20 88 13 81 12
Carbamazepine 117 22 104 20 96 22 118 7 116 21 84 13 75 19 100 19 99 11 99 10 94 12
Chlorpropamide 112 12 91 22 99 12 108 12 86 17 98 17 91 20 75 17 85 8
Clarithromycin 94 11 83 11 118 5 119 14 91 13 108 12 108 18 119 14 113 6 118 6
Diltiazem hydrochloride 88 17 68 7 99 9 91 9 118 3 110 15 75 14 96 11 97 13 103 15 99 8 99 10
Eusolex 6300 105 19 79 20 73 3 81 19 72 9 72 11
Flurazepam 77 8 89 17 86 11 113 7 107 13 76 5 93 17 93 16 94 16 94 8 99 9
hydrochloride
Furosemide 80 12 72 15 98 6 111 9 91 19 125 2 89 12 119 13 96 14 91 10
Gembrozil 84 12 89 10 89 18 70 9 72 10 68 14 69 6 64 4
Glibenclamide 99 13 88 12 86 13 73 19 106 15 80 18 75 14 90 9

S.S. Caldas et al. / Talanta ()

Haloperidol 108 21 73 8 101 18 88 9 109 7 104 15 71 15 97 14 92 15 105 16 95 7 95 12
Ibuprofen 106 14 83 13 118 5 116 2 99 19 106 19 93 12 120 13 111 20 95 8
Mebendazole 120 8 80 11 81 14 92 8 84 4 67 3 74 17 70 14 78 15 75 7 85 6
Methylparaben 84 15 100 7 112 7 100 19 119 10 106 15 107 8 117 7
Miconazole nitrate 94 11 75 18 97 11 104 11 84 21 84 20 69 16 87 19 74 16 84 12
Nimesulide 125 5 100 21 115 11 126 3 105 11 106 12 95 12 125 4 111 5 97 5
Propranolol 85 8 102 22 97 25 114 7 114 16 82 15 118 20 108 13 116 16 119 10 117 9
Propylparaben 73 23 77 25 88 13 82 8 65 9 86 19 71 15 76 18 75 8 81 8
Salicylic acid 110 14 112 8 116 11 103 18 147 12 120 4 107 13
Sodium diclofenac 116 19 80 11 118 10 93 19 116 3 119 10 92 20 99 9 82 16 101 12 99 19 89 11
Sulfamethoxazole 120 10 100 13 69 18 118 12 98 18 96 13
Triclocarban 120 12 113 12 111 13 118 13 98 12 74 6 99 11 100 12 104 6 106 5 79 9
Triclosan 85 26 106 14 87 3 108 16 117 18 88 9
Pesticides
2,4-D 73 19 97 10 104 10 102 17 118 12 118 19 99 19 96 11
Atrazine 92 17 84 18 97 10 103 8 98 18 99 19 109 20 87 15 90 7
Azoxystrobin 80 25 126 14 114 19 89 11 118 8 109 10 78 15 99 20 97 19 103 18 98 9 102 11
Bentazone 120 11 108 20 111 4 120 6 101 16 119 5 108 17 126 3 120 12 101 5
Bispyribac-sodium 108 19 99 16 120 15 107 7 99 19 118 9 79 12 77 7 72 22 70 17
Carbofuran 121 13 100 19 103 13 104 20 86 7 112 25 85 23 86 16 94 8 96 10
Carboxin 114 16 94 19 106 10 99 14 78 5 89 20 105 19 94 15 89 11 91 8
Chlorantraniliprole 115 7 115 15 99 15 73 8 107 18 88 14 91 9 100 7
Clomazone 89 15 73 25 94 7 102 10 90 19 98 19 79 20 100 15 97 20 95 7
Cyhalofop-butyl 92 25 106 22 77 3 75 21 85 13 84 13
Cyproconazole 86 9 70 11 70 7 81 9 71 16 119 3 83 18 74 17 70 11
Dichloran 106 21 109 19 72 13 107 17 97 16 84 14
Difenoconazole 93 14 74 19 96 3 104 11 89 19 116 21 84 11 119 10 104 16 102 8
Diuron 78 29 97 16 110 18 85 19 103 20 79 9 91 20 72 11
Epoxiconazole 124 11 106 10 94 16 90 22 72 17 93 15 71 16 72 18 96 9
Fenoxaprop-p-ethyl 100 26 97 18 108 8 93 20 70 2 83 17 85 17 95 9 92 7 81 10
Fipronil 90 17 118 7 87 16 105 5 112 10 87 18 82 13 70 12 90 17 73 16 70 10
Irgarol 114 13 102 18 87 15 77 7 90 8 80 7 71 1 77 14 72 5 76 15 78 10 90 5
Malathion 107 11 105 11 92 7 123 4 119 5 87 6 94 18 107 11 116 15 111 9 112 14
Metalaxyl-M 61 14 73 20 89 12 91 10 83 19 116 13 120 9 104 14 97 9
Metsulfuron-methyl 111 10 88 20 105 5 113 10 93 18 108 15 82 17 117 14 99 16 88 10
Penoxsulam 95 20 108 9 130 7 104 20 121 6 105 17 108 13 102 9
Pirimiphos-methyl 117 10 84 11 75 16 74 4 84 10 56 7 72 7 79 11
 S.S. Caldas et al. / Talanta () 9

literature [43]. The authors suggested that, due to the pKa values
(methylparaben, pKa 8.2 and propylparaben, pKa 8.4), at high
11
4
4
15
15
11
5
12
9
pH values, the compounds are in the ionic form and cannot be
extracted by the organic solvent. In addition, in alkaline solutions
99
94
94
73
87
114
93
101
86
(high values of pH), parabens can hydrolyze.
Triclosan and triclocarban were extracted over a wide range of
pH values. In agreement with other studies, these compounds
12
10
10
16
13
18
5
16
5

were found to be very stable in acidic and medium conditions;

their recoveries decreased when pH values higher than 11 were
97
118
91
76
116
83
118
71
97

used [11,44].
Because the compounds showed different recoveries in the pH
range under study, two values of pH were chosen for the
20
8
12
8
11
20
4
9
8

extraction.
95
89
109
87
117
98
115
86
96

Salicylic acid, bisphenol A, chlorpropamide, sodium diclofenac,

furosemide, ibuprofen, methylparaben, nimesulide, miconazole
nitrate, 2,4-D, atrazine, bentazone, bispyribac-sodium, cyproco-
nazole, clomazone, difenoconazole, diuron, pronil, metalaxyl-M,
3
13
12
14

16
8
2
10

metsulfuron-methyl, penoxsulam, pyraclostrobin, pyrazosulfuron-

135
93
73
71

119
118
155
97

ethyl, propanil, propiconazole, quinclorac, simazine and tebuco-

nazole were extracted at pH 2, whereas albendazole, amitriptyline,
avobenzone, carbamazepine, clarithromycin, diltiazem hydro-
18
19
12

23

18

chloride, urazepam hydrochloride, eusolex 6300, gembrozil,

glibenclamide, haloperidol, mebendazole, propranolol, propylpar-
120
108
94

78

100

aben, sulfamethoxazole, triclocarban triclosan, azoxystrobin, car-

bofuran, carboxin, cyhalofop-butyl, chlorantraniliprole, dichloran,
epoxiconazole, fenoxaprop-p-ethyl, irgarol, malathion, pirimiphos-
16
19
15
14
16
19
18
20
3

methyl, tau-uvalinate and trioxystrobin were extracted at pH 8.

90
100
74
88
84
89
86
93
73

3.5. Effect of addition of salt

The addition of salt to aqueous solutions generally causes de-

7
8
15
9
10
19
6
14
17

crease in the solubility of the organic compounds in water;

113
118
90
119
101
102
114
107
103

therefore, addition of salt has been widely used to enhance the

extraction recovery of analytes. When salt is added to the solution,
the water molecules can form hydration spheres around the ionic
15
12
6
5
7
16
4
11
11

salt molecules. These hydration spheres reduce the amount of

water available to dissolve analyte molecules, driving additional
109
112
88
109
99
90
119
85
112

analytes into the organic phase. This effect is observed mainly for
highly polar compounds [21]. Usually, most studies employ NaCl to
investigate the inuence of ionic strength on SD-DLLME perfor-
13
13
12
17
11

16
14
17
25

mance. To verify the inuence of other salts on the recoveries, 1%

(w/v) magnesium sulfate (33.7 g/100 mL), ammonium sulfate
82

70
92

76
112
87
107
66

106

(75.4 g/100 mL), calcium chloride (74.5 g/100 mL) and sodium
chloride (35.9 g/ 100 mL) were evaluated.
In general, the use of salt showed better, or similar, recoveries
21
9
2
10

13
14
10

than the experiment without salt. Calcium chloride and magne-

sium sulfate showed similar results (p 40.05), but when calcium
89

114
119

105
85
108
104

chloride was used, the sample got cloudy, thus, making the droplet
withdrawal difcult. When NaCl was used, results were statisti-
cally similar to other salts for 69% of the analytes, but the area
19

values were lower for all analytes.

106

Improvement in the areas was observed when magnesium sul-

fate was used. It can be explained by the lyotropic series concept, i.e.,
an empirical order of ions based on their ability to precipitate water-
loving substances. DobryDuclaux reports an order for the cations
Mg2 4Sr2 4Ca2 4Ba2 4K 4Na 4NH4 4Li and for the
anions SO42
4C2H3O2
4Cl
4NO3
4Br
4I
4CNS. The ions
that comprise the salts under study are among the ones with the
highest ability to precipitate water-loving substances [45]. Thus,
Pyrazosulfuron-ethyl

magnesium sulfate 1% (w/v) was used in the extractions.

Tau-uvalinate

Trioxystrobin
Pyraclostrobin
Propiconazole

Tebuconazole

3.6. Demulsier volume and type

Quinclorac
Simazine
Propanil

In this study, the 1:1 volume ratio of disperser solvent and

demulsier was used (i.e., when 500 mL acetone was used as the
disperser solvent, 500 mL acetone was used as the demulsier).

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10 S.S. Caldas et al. / Talanta ()

Table 4
Concentrations of compounds in collected waters (g L
1) and Relative Standard Deviation (RSD, %).

Pesticides and Riacho Do Gelo So Gonalo Canalete

PPCPs
LOQm May August May August May August

(mg L
1) Conc. RSD (%) Conc. RSD (%) Conc. RSD (%) Conc. RSD (%) Conc. RSD (%) Conc. RSD (%)
(mg L
1) (mg L
1) (mg L
1) (mg L
1) (mg L
1) (mg L
1)

Salicylic acid 0.25 1.67 9 n.d. n.d. n.d. n.d. n.d.

Avobenzone 0.25 5.93 18 n.d. n.d. n.d. n.d. n.d.
Bisphenol A 0.25 n.d. 0.418 22 n.d. n.d. n.d. 4.42 34
Clarithromycin 0.125 n.d. n.d. n.d. n.d. 0.065 9 n.d.
Ditialzem 0.0125 n.d. n.d. 0.07 29 n.d. n.d. n.d.
chloridrate
Sodium diclofenac 0.0125 0.939 9 0.036 17 n.d. n.d. n.d. 0.02 11
Haloperidol 0.0125 n.d. n.d. n.d. n.d. n.d. 0.04 12
Methylparaben 0.0250 n.d. n.d. n.d. 0.805 17 n.d. 0.32 24
Nimesulide 0.0125 n.d. n.d. n.d. 0.102 18 n.d. n.d.
Miconazole nitrate 0.1250 0.265 21 n.d. n.d. n.d. 0.207 12 n.d.
Triclocarban 0.0250 0.575 21 oLOQ n.d. n.d. o LOQ n.d.
Azoxystrobin 0.0125 n.d. n.d. 0.07 20 n.d. n.d. n.d.
Bentazone 0.1250 n.d. oLOQ n.d. oLOQ 6,7 9 oLOQ
Cyproconazole 0.0250 n.d. n.d. n.d. 0.06 9 n.d. n.d.
Clomazone 0.0125 n.d. n.d. n.d. oLOQ n.d. n.d.
Difenoconazole 0.0125 n.d. n.d. n.d. oLOQ n.d. n.d.
Fipronil 0.0250 0.030 35 n.d. n.d. n.d. n.d. n.d.
Irgarol 0.0125 n.d. n.d. n.d. 0.25 20 n.d. n.d.
Malathion 0.0125 n.d. n.d. n.d. oLOQ n.d. n.d.
Propiconazole 0.0250 n.d. n.d. n.d. oLOQ n.d. n.d.
Tebuconazole 0.0250 n.d. n.d. n.d. oLOQ n.d. n.d.

This ratio had also been used in previous reports [10,11,14,20].
In the SD-DLLME technique, a volume of the disperser solvent
is injected to separate the emulsion into its constituent phases at a
given time. When the solvent is injected, the emulsion quickly
clears into two phases. In this way, the separation of the organic
phase from the aqueous bulk is achieved without centrifugation.
In this study, for the rst time, the use of water, rather than, an
organic solvent was proposed to break the emulsion.
No signicant difference was found for many compounds and
for other high recoveries when water, rather than acetone, was
used (Fig. 3).
The high recoveries could be attributed to the different solu-
bility of 1-octanol in water and in acetone. When acetone is in-
jected, the emulsion is broken but a little solubilization of the
extractor solvent can occur, leading to lower extraction recoveries.
Since one of the characteristics of the extractor solvent is to be
insoluble in water, when water is added to break the emulsion,
losses by the redissolution of the analytes decrease, thus, achiev-
ing higher recoveries.
Therefore, in this step, water was chosen as the demulsication
solvent. The addition of water, rather than an organic solvent
makes the method environmentally friendlier and less expensive
than the methods that were previously published.
3.7. Optimal conditions
After the optimization, the optimal conditions included the
extraction of 10 mL of an acidic (pH 2) and basic aliquots (pH 8),
both with 1% MgSO4 (w/v). The extraction was carried out using a
mixture of 120 mL octanol and 750 mL acetone which were rapidly
injected into the sample solution. A cloudy solution (water, octanol
and acetone) was formed in the volumetric ask. After, 750 mL
water was injected into the aqueous bulk to break up the emul-
sion. Then, the cloudy solution cleared into two phases quickly and
the upper layer was collected. The light phase was placed in an
Eppendorf and the volume was increased to 250 mL with methanol
(Fig. 4).
3.8. Method validation
Calibration curves were prepared in the extract and in the
solvents; results are shown in Table 2. Correlation coefcients (r)
ranging from 0.9849 to 0.9962 indicated excellent linearity for all
compounds. The SD-DLLME linearity was also evaluated with a
series of fortied samples at concentration levels ranging from the
LOQ of each compound to 25 mg L
1, to consider the extraction
efciency depending on the concentration (working curve). In all
cases, correlation coefcients ranging from 0.9846 to 0.9979 were
reached.
LOQ was experimentally estimated from the analysis of real
samples as the concentration of analyte giving a signal-to noise
ratio of 10. The LOQs for all target analytes ranged from 0.0125 to
1.25 mg L
1. Regarding pesticides, the LOQs were lower than the
maximum contaminant level (MCL) allowed by the UE for in-
dividual pesticide (0.1 mg L
1) and for total pesticides (0.5 mg L
1)
in water [46] except for 2,4-D, chlorantraniliprole, diuron, pe-
noxsulam, pirimiphos-methyl, quinclorac and simazine, which is
0.25 mg L
1, and for cyhalofop-p-butyl and dichloran, which was
1.25 mg L
1. Concerning pharmaceuticals and PPCPs, no MCLs were
found in the literature. Furthermore, values reported in Table 2
were similar to or even better than the values obtained for other
applications of DLLME [4749].
The evaluation of the SD-DLLME efciency was accomplished
by means of extraction recovery (R%). Recoveries ranged from 61%
to 130% for the intra-day and from 64% to 147%, for the inter-day
(Table 3).
The precision of the method was expressed as RSD by treating
drinking water samples at least in three levels. The method
showed good intra-day and inter-day precision with RSD values
between 129% and 223%, respectively, at all levels of
fortication.
Results of the matrix effect evaluation showed that clari-
thromycin, glibenclamide, triclosan, malathion and quinclorac are
negatively inuenced by the matrix (suppression of the signal
during ionization). Results from the evaluation of ME by DLLME in

Please cite this article as: S.S. Caldas, et al., Talanta (2015), http://dx.doi.org/10.1016/j.talanta.2015.06.047i
 S.S. Caldas et al. / Talanta () 11

other studies have also found a suppression effect for the analytes
[37] and the matrix-matched calibration approach has been used

Reference

Proposed
method
[52]

[53]
to compensate for the ME [50].

[51]
3.9. Applicability

40 and 5

11.6 and
The proposed SD-DLLME method was applied to verify the

46 and
ME (%) occurrence of 58 PPCPs and pesticides in three water samples

r 13
collected in Rio Grande, RS, Brazil. The quantication was carried

85

95
out by the standard addition method to compensate for any matrix
effect. Atrazine-d5 and ibuprofen-d3 were added as surrogates to
Accuracy (%) Precision (%) LOQ range

12.51250
check the extraction efciency in the sample analysis. Samples

505000
201000
(ng L
1)

0.150
were collected in May and August 2014. Samples of surface water
were collected directly in their water sources. One liter of the
sample was collected with the aid of a stainless steel mug and
stored in an amber glass bottle which had been rinsed with
acetone and baked at 100 C. Before sampling, bottles were rinsed
o 8, 9

with the sample. Samples were stored at 4 C until analysis, which

129
112
o 17

was performed on the same day. Results are shown in Table 4.

The rst sampling spot (So Gonalo Channel) is near an agri-
15 min ultrasound 10 min 74.6107.5

cultural area where water is captured for the water supply after
94.5116
93101

61147

treatment. Results showed that the region is mostly inuenced by

agriculture and little inuenced by domestic sewage because most
detected contaminants are pesticides. Azoxystrobin, bentazone,
clomazone, cyproconazole, difenoconazole, irgarol, malathion,
propiconazole and tebuconazole were detected in concentrations
3 h 800 rpm 15 min

from oLOQ to 0.25 mg L
1. These pesticides are recommended for

the rice culture, the main agricultural activity in that region.
Extactor solvent (volume) Extraction time

The second sampling spot is located downtown (Canalete). The

centrifugation

surface water was collected from a drainage channel used for

ultrasound
5580 min

storm water runoff; some domestic sewage is illegally discharged

10 min

there. In this place, bisphenol A, chlaritomicin, sodium diclofenac,

haloperidol, methylparaben, miconazole nitrate and triclocarban
were detected, showing the inuence of domestic discharge on
that water.
acetonitrile:methanol

The third sampling spot is in another drainage channel (Riacho

(50:50, v/v) 200 mL

Acetonitrile 10 mL
Comparison of SD-DLLME with other methods for the determination of compounds in water samples.

1-octanol 120 mL
Methanol 10 mL

do Gelo) constructed for storm water runoff but it also collects

some domestic efuents. In addition, this channel ows onto a
beach. Salicylic acid, avobenzone, bisphenol A, sodium diclofenac,
miconazole nitrate and triclocarban were detected in this sam-
pling spot.
In these drainage channels, the inuence of domestic sewage is
Wastewater (250 mL, pH 2) groundwater

Surface water (10 mL, pH 2 and pH 8, 1%

Tap, leaching and sewage water (10 mL,

clearly visible because some PPCPs were detected. PPCPs and

pesticides were identied at concentration levels of mg L
1.
and surface water (500 mL, pH 2)
Filtered water (20 mL, 10 % NaCl)

Data in Table 4 show that the new multiresidue method is

Sample (volume, pH and salt)

suitable for the environmental monitoring of PPCPs and pesticides

in surface waters.
To verify the accuracy, precision and matrix effect in different
surface water samples and guarantee the analytical quality, these
samples were fortied in one level and extracted in triplicate.
Table S1 describes the recoveries, RSD and ME of the samples.
2 g NaCl)

Results show that recoveries and RSD are within the range re-
MgSO4)

commended for residue analysis (70 and 120% with RSDs values
lower than 20%). For the matrix effect, different behaviors were
observed for each analyte and in each sample, highlighting the
58 pesticides and PPCPs
PPCPs, EDCs, and arti-

importance of carrying out the quantication with the standard

addition method to compensate the ME.
cial sweeteners
Ultrasound 48 pesticides
15 pesticides

3.10. Comparison of SD-DLLME with other methods

analytes

The extraction and determination of pesticides and PPCPs in

water samples by this method was compared with those of other
Technique

SD-DLLME

methods in Table 5 [5153]. The advantages of the method de-

scribed in this paper over the others include shorter extraction
Table 5

SBSE

SPE

times and less solvent consumption. Besides, the method showed

accuracy, precision and ME similar to what has been proposed by

Please cite this article as: S.S. Caldas, et al., Talanta (2015), http://dx.doi.org/10.1016/j.talanta.2015.06.047i
12 S.S. Caldas et al. / Talanta ()
other studies. In addition, most of the other proposed studies have,
at the end, an evaporation step, which increases not only the
number of steps during the extraction, but also time, cost and
errors during the extraction. In the method proposed in this paper,
there is no evaporation step; the nal volume only needs to be
completed to 250 mL with methanol.
In sum, the SD-DLLME technique is simple, fast and en-
vironmentally friendly because it uses less toxic extractor solvent
(alcohol) and de-emulsication solvent (water).
4. Conclusions
The use of 120 mL 1-octanol and 750 mL acetone for the ex-
traction of 58 PPCPs and pesticides from 10 mL samples, with
750 mL water as the demulsication solvent, led to suitable re-
coveries. Some interesting features of this method could be en-
hanced. An organic solvent with density lower than water was
used as the extraction solvent; solvent demulsication, rather than
centrifugation, was used to break up the emulsion after extraction,
and the emulsion was broken with water rather than with an or-
ganic solvent.
By comparison with the usual DLLME methods usually em-
ployed, the SD-DLLME has shown some advantages, such as easi-
ness, quickness and low cost. Overall, this study demonstrated, for
the rst time, that SD-DLLME and LCMS/MS can be considered an
important, innovative and efcient method for multiresidue PPCPs
and pesticide extraction and determination in water samples. SD-
DLLME was successfully applied to multiresidue PPCPs and pesti-
cide extraction and determination in real water samples.
Acknowledgments
The authors acknowledge the nancial support and fellowships
granted by the Brazilian agencies CAPES, FINEP, CNPq and FURG.
Part of this study was supported by a grant from the Brazilian
Agency CNPq/CAPES (Process number 552318/2011-6), FAPERGS
(Process number 810-25.51/13-3) and FAPERGS (Process number
831-25.51/13-0). E.G. Primel received a productivity research fel-
lowship from the Brazilian Agency CNPq (DT 310517/2012-5).
Appendix A. Supplementary material
Supplementary data associated with this article can be found in
the online version at http://dx.doi.org/10.1016/j.talanta.2015.06.
047.
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