Chinese Chemical Letters 25 (2014) 745748

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Chinese Chemical Letters

journal homepage: www.elsevier.com/locate/cclet

Original article

Combination of ionic liquid dispersive liquid-phase microextraction

and high performance liquid chromatography for the determination
of triazine herbicides in water samples
Qing-Xiang Zhou a,*, Yuan-Yuan Gao b
a
College of Geosciences, China University of Petroleum Beijing, Beijing 102249, China
b
College of Environmental Science and Engineering, Nankai University, Tianjin 300071, China

A R T I C L E I N F O A B S T R A C T

Article history: A temperature-controlled ionic liquid dispersive liquid-phase microextraction in combination with high
Received 16 December 2013 performance liquid chromatography was developed for the enrichment and determination of triazine
Received in revised form 31 December 2013 herbicides such as cyanazine, simazine, and atrazine in water samples. 1-Octyl-3-methylimidazolium
Accepted 13 January 2014
hexauorophosphate ([C8MIM][PF6]) was selected as the extraction solvent. Several experimental
Available online 24 January 2014
parameters were optimized. Under the optimal conditions, the linear range for cyanazine was in the
concentration range of 0.580 mg/L and the linear range for simazine and atrazine was in the range of
Keywords:
1.0100 mg/L. The limit of detection (LOD, S/N = 3) was in the ranges of 0.050.06 mg/L, and the intra day
Temperature-controlled ionic liquid
dispersive liquid-phase microextraction
and inter day precision (RSDs, n = 6) was in the ranges of 3.2%6.6% and 4.8%8.9%, respectively. Four real
Ionic liquid water samples were analyzed with the developed method, and the experimental results showed that the
Triazine herbicides spiked recoveries were satisfactory. All these exhibited that the developed method was a valuable tool
High performance liquid chromatography for monitoring such pollutants.
2014 Qing-Xiang Zhou. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights
reserved.

1. Introduction
Triazines have been widely used in agricultural domain due to
their high power for weed control for many years. The wide use of
these herbicides at large scale has resulted in serious pollution of
the environment [1]. Triazine herbicides and their degradation
products are highly toxic, mobile and soluble in water and can also
be strongly adsorbed onto soil, hence have been an environmental
safety concern [2,3]. In the United States, a joint project of the
Environmental Protection Agencys Ofce of Drinking Water
(ODW) and the Ofce of Pesticide Programs carried out a national
pesticide survey [4,5], and the results showed that atrazine and
simazine existed in the drinking water samples. In the EU, the
maximum allowed limit for each individual herbicide has been set
at 0.1 mg/L [6]. Hence it is necessary to develop simple, sensitive
and low cost monitoring techniques for the analysis of the residual
level of these pollutants.
Generally, a sample pretreatment step is used before an
effective method is developed, and up to now, many methods have
been established for the enrichment of triazines residues from
complex matrices. Liquidliquid extraction often needed a large
* Corresponding author.
E-mail address: zhouqx@cup.edu.cn (Q.-X. Zhou).
volume of the sample of interest and toxic organic solvents and
also was labor intensive, expensive, and time-consuming. Single-
drop microextraction is established based on the principle of
liquidliquid extraction and the enrichment is nished in a single
drop of solvent. However, the shortcomings of this method are the
instability of the drop when fast stirring or volatile organic solvents
are used [7]. This method is also time-consuming and the
equilibrium requires a long time to achieve in most cases.
Dispersive liquidliquid microextraction [8] is a recently con-
structed method and has many applications for the determination
of pesticides, pharmaceuticals, PCBs, PBDEs and PAHs, etc.
However, chlorobenzene, chloroform and carbon disulde were
often used as the extraction solvent and methanol, acetonitrile and
acetone etc. were used as the disperser solvents, which will result
in secondary pollution. In order to resolve such problems, ionic
liquid (IL) was an ideal selection because ionic liquids have many
interesting properties including wide liquid ranges, low volatility,
good stability, reusability, etc. Due to these useful properties, ILs
have been widely used in organic chemistry and analytical
chemistry. In dispersive liquidliquid microextraction processes,
ILs also have some applications for the analysis of hexabromocy-
clododecane diastereomers [9], polar herbicides [10], pesticides
and heavy metals, etc. [1113]. In this work, 1-octyl-3-methyli-
midazolium hexauorophosphate [C8MIM][PF6] was chosen as the
extraction solvent and triazine herbicides were selected as the

1001-8417/$ see front matter 2014 Qing-Xiang Zhou. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.
http://dx.doi.org/10.1016/j.cclet.2014.01.026
746 [(Fig._1)TD$IG]
Q.-X. Zhou, Y.-Y. Gao / Chinese Chemical Letters 25 (2014) 745748

model analytes. A new temperature controlled ionic liquid 20

dispersive liquid phase microextraction was developed for the
sensitive determination of triazine herbicides. The possible factors 18
were optimized. 16

2. Experimental
Cyanazine, simazine, and atrazine were purchased from
AccuStandard, Inc. (New Haven, USA). The stock solutions with
a concentration of 1000 mg/L were prepared by dissolving the solid
bulk triazine herbicides in methanol and were stored at 4 8C in a
refrigerator. The working solutions were obtained by diluting the
stock solutions with methanol. 1-Octyl-3-methylimidazolium
hexauorophosphate ([C8MIM][PF6]) was synthesized in our
own laboratory [14]. HPLC grade methanol and acetonitrile were
purchased from Jiangsu Guoda Chemical Reagent Co., Ltd. (Huaian,
China). Ultrapure water used in the experiments was prepared on a
Milli-Q water purication system (Millipore, Bedford, MA, USA). All
glass used in the experiments was washed with pure water, then
soaked in 6 mol/L nitric acid for 24 h and cleaned with ultrapure
water before use.
The analysis and separation was carried out on a high
performance liquid chromatography system including two LC-10
ATvp pumps and an SPD-10 Avp ultraviolet detector (Shimadzu,
Kyoto, Japan). The analytical column was a reversed-phase SunFire
C18 column (150 mm  4.6 mm, particle size 5 mm) and the
software of Chromato Solution Light Chemstation for LC system
was used to acquire and process chromatographic data. The mobile
phase was a mixture of methanol and ultrapure water (40/60, v/v).
The mobile phase ow rate was set at 1.2 mL/min, the injection
volume was set at 20 mL, and the detection wavelength was set at
275 nm. An Anke 80-2 (Shanghai, China) centrifuge was utilized to
centrifuge the cloudy water solutions.
The enrichment procedure was as follows: 10 mL of ultra-pure
water was put in a 10 mL conical tube, the cyanazine, simazine and
atrazine were spiked with a concentration of 10 mg/L, 20 mg/L and
20 mg/L, respectively. Then sodium chloride was added at a
concentration of 20% (w/v). After 50 mL of [C8MIM][PF6] was
added, the tube was heated in a water bath with the temperature
controlled at 80 8C. When [C8MIM][PF6] was completely dissolved
in the aqueous solution and a nearly homogenous solution was
obtained, the tube was cooled with ice water. The tube was kept for
30 min to enhance the extraction of triazine herbicides from the
sample solution into the tiny droplets of [C8MIM][PF6]. Thereafter
the mixture was centrifuged for 20 min at 4000 rpm. The upper
aqueous phase was removed, and the residue was dissolved in
200 mL of methanol and 20 mL of the resulting solution was
injected into the HPLC system for analysis. Each experiment was
done in triplicate to ensure the accuracy of the experimental data.
Four water samples were collected for use. River water was
Concentration (g/L)
14
12
10
8
6
4
2
5 10 15 20 25
NaCl concentration (%, w/v)
Fig. 1. Effect of sodium chloride content. Conditions: [C8MIM][PF6] volume, 50 mL;
sample volume, 10 mL; spiked concentration, 10 mg/L for cyanazine, 20 mg/L for
simazine and atrazine; temperature, 70 8C; extraction time, 30 min; centrifuging
time, 20 min; (&) cyanazine, (*) simazine, (~) atrazine.
performance. In theory, addition of salt will decrease the solubility
of analytes in the aqueous sample and enhances their partitioning
into the adsorbent or organic phase. On the other hand, the
addition of salt increased the solubility of the ionic liquid in water,
which played a dominant role in the decrease of the volume of the
sediment phase. A series of experiments were designed to
investigate the salting-out effect (Fig. 1). The results demonstrated
that a signicant increase of the peak area occurred as the ionic
strength of the solution increased from 0% to 20% (w/v), but the
peak area decreased when the ionic strength of the solution was
further increased to the range of 20%25%. So 20% (w/v) NaCl was
added in the following experiments.
3.2. Effect of the volume of ionic liquid
The volume of extraction solvent was a key factor in the newly
developed procedure, which determines the enrichment perfor-
mance to a certain extent. In order to achieve the maximum
enrichment performance possible, the volume of extraction
solvent was optimized in the range of 4060 mL (Fig. 2). From
[(Fig._2)TD$IG] 2, we can see that the largest peak area of the targeted analytes
Fig.
20
18
16
Concentration (mg/L)

14
taken from Baihe, Nanyang City, Henan Province, China. Under-
ground water was obtained from Henan Normal University in 12
Xinxiang City, Henan Province, China. Drainage water was
collected from Gongchanzhuyi drainage, Xinxiang City, Henan 10
Province, China. Wastewater was obtained from the outlet of 8
Luotuowan sewage treatment plant, Xinxiang City, Henan
Province, China. Before use, all the environmental water samples 6
were ltered with 0.45 mm micro-pore membranes and then 4
stored in brown glass containers at a low temperature of 4 8C.
2
40 45 50 55 60
3. Results and discussion
Volume of ionic liquid (L)
3.1. Effect of ionic strength
Fig. 2. Effect of ionic liquid volume. Conditions: ionic liquid, [C8MIM][PF6]; sample
volume, 10 mL; spiked concentration, 10 mg/L for cyanazine, 20 mg/L for simazine
The salting-out effect is often present in many cases, and it is and atrazine; sodium chloride content, 20% (w/v); temperature, 70 8C; extraction
often utilized to enhance the enrichment or preconcentration time, 30 min; centrifuging time, 20 min. (&) cyanazine, (*) simazine, (~) atrazine.
 Q.-X. Zhou, Y.-Y. Gao / Chinese Chemical Letters 25 (2014) 745748
was obtained when the volume of extraction solvent was 50 mL.
The peak area increased with the increase of IL volume in the range
of 4050 mL. Even larger volume of IL led to the decrease of the
peak area when the volume of IL was increased in the range of 50
60 mL. The reason may be that the superuous IL could not be
dispersed into the solution as microdrops and absorb onto the wall
of the tube, and some analytes migrated into this part of IL. So this
part of the analytes would not be sedimented into the bottom of
the tube, leading to the decrease of the peak area. Hence, 50 mL of
[C8MIM][PF6] was chosen in the subsequent experiments.
3.3. Effect of dissolving temperature
In this newly established process, temperature is a crucial factor
because it is the driving force for the complete dispersion of
[C8MIM][PF6] into the aqueous solution. In order to achieve the best
temperature for extraction, temperature was investigated in the
range of 60100 8C (Fig. 3). From Fig. 3, we can see that 80 8C was the
reasonable temperature because the largest peak area occurred at
80 8C. The probable reasons are that the complete dispersion extent
of [C8MIM][PF6] into the aqueous solution increased along with the
rise of temperature and the migrating-in speed was equal to the
migrating-out speed into the ionic liquid phase. However the
recoveries of analytes obtained over 80 8C was lower than those
obtained at 80 8C. Maybe the migrating-in and migrating-out speeds
increased at the same time, but the migrating-out speed was larger
than the migrating-in speed to the ionic liquid phase. This resulted in
a decreased distribution of analytes in the ionic liquid phase. So the
dissolving temperature was set at 80 8C.
3.4. Sample pH value
The sample pH value is also a key factor in the sample
pretreatment techniques, which determines the present form of
the analytes and then affects extraction efciency. Therefore, the
effect of sample pH value has to be investigated. In this study, it
was carried out in the range of pH 311 (Fig. 4). The peak areas
increased with the increase of pH value in the range of pH 37, and
decreased in the range of pH 711. This phenomenon was closely
related to their chemical properties. Atrazine and simazine can be
hydrolyzed in strong acid or alkali environment at high tempera-
ture. Based on these results, sample pH value was set at 7 in all
[(Fig._3)TD$IG]subsequent experiments.
20
18
16
Concentraiton (g/L)
14
12
10
8
6
4
2 2
60 70 80 90 100
Temperature ( C)
Fig. 3. Effect of temperature. [C8MIM][PF6] volume, 50 mL; sample volume, 10 mL;
spiked concentration, 10 mg/L for cyanazine, 20 mg/L for simazine and atrazine;
sodium chloride content, 20% (w/v); extraction time, 30 min; centrifuging time,
20 min. (&) cyanazine, (*) simazine, (~) atrazine.
747
3.5. Effect of extraction time and centrifuging time
Mass transfer is a time dependent process and one of the salient
factors in most extraction procedures, especially in microextraction
methods such as SPME and LPME. In this procedure, the mass
transfer between the targeted compounds and ionic liquid phase
will be related to various factors and reach an equilibrium in the end.
The effect of extraction time was examined in the range of 10
50 min. The experimental results showed that the best extraction
performance was achieved at 30 min. Centrifugation is a critical step
in order to obtain two distinguishable phases, so it will affects the
size of the settled phase and the concentration of analytes in the
extraction phase. The centrifugation time was checked in the range
of 525 min at 4000 rpm after extraction. The results indicated that
the peak areas of the analytes increased as the centrifugation time
increased from 5 min to 15 min, and decreased after 15 min. The
sedimented volume increased along with the time, but longer
centrifuging time resulted in heat generation, which led to the re-
dissolution of some [C8MIM][PF6] into the water phase and a loss of
extraction performance. Finally, 15 min was used.
3.6. Analytical performance and real water sample analysis
To evaluate the proposed method, often-used parameters such
as linear range, precisions and detection limits were investigated
under the optimal experimental conditions (Table 1). The linear
range was in the range of 0.580 mg/L for cyanazine, and 1.0
100 mg/L for simazine and atrazine, respectively. Good linearity
with correlation coefcient (R2) in the range of 0.99390.9965 and
low detection limit in the range of 0.050.06 mg/L based on three
times of the ratio of signal to noise (S/N = 3) were achieved. The
intra day precision and inter day precision was in the ranges of
3.2%6.6% and 4.6%8.9% (RSD, n = 6), respectively.
To demonstrate the feasibility of the proposed method, it was
used to determine the triazine herbicides in four real water
samples (Table 2). The targeted analytes were not found in the
blank samples of the four samples. These samples were spiked with
two different concentrations, 1 mg/L for cyanazine, 2 mg/L for
simazine and atrazine, and 10 mg/L for cyanazine, 20 mg/L for
simazine and atrazine, respectively. As can be seen from Table 2,
good spiked recoveries in the range of 85.1%100% with the
precisions of 1.8%9.7% (RSD) were obtained. The typical
[(Fig._4)TD$IG]
chromatogram of water samples was showed in Fig. 5.
22
20
18
16
Concentration (g/L)
14
12
10
8
6
4
2 4 6 8 10 12
Sample pH
Fig. 4. Effect of sample pH. Conditions: ionic liquid, [C8MIM][PF6]; sample volume,
10 mL; spiked concentration, 10 mg/L for cyanazine, 20 mg/L for simazine and
atrazine; sodium chloride content, 20% (w/v); temperature, 80 8C; extraction time,
30 min; centrifuging time, 20 min. (&) cyanazine, (*) simazine, (~) atrazine.
748 Q.-X. Zhou, Y.-Y. Gao / Chinese Chemical Letters 25 (2014) 745748

Table 1
Linear ranges, precisions, detection limits for the enrichment of triazine herbicides by temperature controlled ionic liquid dispersive liquid phase micro-extraction.

Compound Linear range (mg/L) R2 Intra day precision (RSD%, n = 6) Inter day precision (RSD%, n = 6) Detection limit (mg/L)

Cyanazine 0.580 0.9965 3.2 4.8 0.05

Simazine 1.0100 0.9939 6.6 8.2 0.05
Atrazine 1.0100 0.9965 5.4 8.9 0.06

Table 2
Spiked recoveries in real water samples by the proposed method.

Compounds Spiked level (mg/L) Underground water Luotuowan sewage treatment plant outlet Drainage water Baihe River

Cyanazine 1 88.9  6.2 89.7  4.7 94.5  5.0 88.0  3.9

10 90.3  2.1 93.2  2.8 88.7  2.9 88.7  2.9
Simazine 2 87.8  3.9 87.7  5.4 97.5  3.0 87.7  2.1
20 94.1  2.3 91.0  1.8 95.7  2.4 95.7  2.4
Atrazine 2 85.2  5.2 85.5  9.3 96.8  3.2 85.1  9.7
20 100  2.2 93.7  4.9 99.9  2.2 99.9  3.2

[(Fig._5)TD$IG]

Fig. 5. Typical chromatogram of Baihe water sample. (A) Blank; (B) spiked concentration, 1 mg/L for cyanazine, 2 mg/L for simazine and atrazine; (C) spiked concentration,
10 mg/L for cyanazine, 20 mg/L for simazine and atrazine. Peak identication, (1) cyanazine, (2) simazine, (3) atrazine.

4. Conclusion
This paper described a simple and sensitive temperature-
controlled ionic liquid dispersive liquid phase microextraction in
combination with HPLC for the analysis of triazine herbicides in
environmental water samples. This developed extraction system is
a cheap, convenient sample pretreatment technique, and reduces
the exposure danger to the toxic solvents used in the conventional
extraction procedure. Under the best extraction conditions, low
limit of detection, good linearity and repeatability were achieved.
The analysis of real water samples indicated that the method was
excellent and would be very competitive in the routine analysis of
environment pollutants.
Acknowledgments
This work was nancially supported by the National Natural
Science Foundation of China (No. 21377167), Program for New
Century Excellent Talents in University (No. NCET-10-0813).
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