Journal of Chromatographic Science, 2020, Vol. 58, No.

2, 171–177
doi: 10.1093/chromsci/bmz080
Advance Access Publication Date: 4 November 2019
Article

Article

Combination of Counter Current Salting-Out

Homogenous Liquid–Liquid Extraction with

Downloaded from https://academic.oup.com/chromsci/article/58/2/171/5612250 by guest on 13 March 2022

Dispersive Liquid–Liquid Microextraction for the
High-Performance Liquid Chromatographic
Determination of Environmental Estrogens in
Water Samples
Zhihong Shi, Qingru Huai, Xinye Li, Hongyu Ma, Can Zhou,
Xiaoxue Chu and Hongyi Zhang*
College of Chemistry and Environmental Science, Hebei University, Key Laboratory of Analytical Science and
Technology of Hebei Province, 180 Wusi East Road, Lianchi District, Baoding, Hebei Province, 071002, China

Author to whom correspondence should be addressed. Email: hyzhang@hbu.edu.cn

Received 15 June 2018; Revised 21 February 2019; Accepted 27 August 2019

Abstract
In this paper, counter current salting-out homogenous liquid–liquid extraction was combined with
dispersive liquid–liquid microextraction for the determination of environmental estrogens in water
samples by high-performance liquid chromatography. In this method, initially, sodium chloride
was filled into a syringe and a mixture of water sample and acetonitrile was driven to pass
through the syringe. Due to salting-out effect, fine droplets of acetonitrile went up through the
remaining mixture and aggregated as a separated layer on the top. Then, the collected organic
phase (acetonitrile) was removed with a syringe and mixed with carbon tetrachloride (extraction
solvent). In the second step, the mixed organic phase was rapidly injected into 5 mL of distilled
water to further enrich the analytes. Good linearity was obtained in the concentration range of
2.0∼200 ng/mL for diethylstilbestrol (DES) and 8.0∼200 ng/mL for octylphenol (OP), respectively.
Limits of detection were 0.09 ng/mL for DES and 0.20 ng/mL for OP, respectively. Relative standard
deviations for intra- and inter-day precisions were less than 2.1 and 3.1%, respectively. Finally, the
established method was successfully applied to determine DES and OP in river water, well water,
bottled water and campus drinking water samples with recoveries in the range from 81.0 to 105.9%.

Introduction
As a group of endocrine-disrupting chemicals (1, 2), environmental
estrogens have attracted increasing concerns in recent years (3–5).
Environmental estrogens can be divided into two broad categories:
endogenous estrogens and synthetic estrogens (6). Synthetic estrogens
can mimic or hinder the behavior of natural hormones and thus affect
the biological growth, metabolism and reproduction and even cause
tumors such as breast cancer and ovarian cancer (7).
Diethylstilbestrol (DES) and octylphenol (OP) are two typical
synthetic estrogens. DES, a synthetic non-steroidal hormone, has been

© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com
171
172
Figure 1. The chemical structure of DES and OP.
used to treat gynecological disorders and to promote growth rate
of animals (8, 9). However, residues of DES and its metabolites in
animal tissues and organs may be transmitted to consumers through
the food chain, which can cause human cancer. DES and its metabo-
lites can also enter the environment through animal excreta (10).
OP is the main degradation product of alkylphenol polyethoxylate,
which is widely used in the production of emulsifiers for plasticizers,
detergents and pesticides (11). Finally, OP will enter the environment
through various paths. Therefore, monitoring of the levels of envi-
ronmental estrogens such as DES and OP in various water samples is
of great significance (12–17).
At present, gas chromatography (GC), high-performance liquid
chromatography (HPLC) and capillary electrophoresis (CE) are the
main analytical methods for the determination of environmental
estrogens (18, 19). GC is fast and sensitive; for most environmental
estrogens, which are nonvolatile compounds, the derivatization
process is often needed, which is cumbersome and susceptible to
sample loss (20). CE has good separation performance, but it lacks
stability and sensitivity for real samples with complex matrices (21).
Compared with GC and CE, HPLC does not require derivatization
and shows good reproducibility for the analysis of environmental
estrogens.
Due to the complexity of the sample matrix and the low con-
centration of environmental estrogens, sample preparation must
be performed prior to chromatographic analysis. Existing sample
preparation methods for the analysis of environmental estrogens
are liquid–liquid extraction (LLE) (22), solid phase extraction (SPE)
(23, 24), liquid phase microextraction (LPME) (25, 26), solid phase
microextraction (SPME) (27), stir bar sorptive extraction (SBSE)
(10), hollow fiber liquid-phase microextraction (HF-LPME) (28) and
so on. Among them, traditional sample preparation methods such
as LLE and SPE are time-consuming and require large amounts of
samples and organic solvents. For SBSE, the friction between the
stirring bar and the bottom of the vial during extraction can easily
cause coating losses. At the same time, the extraction time is relatively
long. SPME fibers are relatively fragile and their coatings tend to
swell or fall off when dissolved in organic solvents, which limits their
compatibility with HPLC (29).
In 2015, Farajzadeh et al. (30, 31) proposed counter current
salting-out homogeneous liquid–liquid extraction (CCSHLLE) com-
bined with dispersive liquid–liquid microextraction (DLLME) for the
extraction and preconcentration of pesticides from aqueous samples.
In the first step, analytes are extracted into the acetonitrile (ACN)
layer formed by counter current salting-out effect; in the second step,
ACN containing analytes was used as dispersant for the DLLME
procedure to enrich the analytes. Subsequently, CCSHLLE–DLLME
was applied to the extraction of drugs in urine samples (32).
The aim of this work is to combine CCSHLLE with DLLME
for the extraction and enrichment of environmental estrogens from
Shi et al.
various water samples. Several important factors influencing the
extraction efficiency have been studied in detail, including the type
Downloaded from https://academic.oup.com/chromsci/article/58/2/171/5612250 by guest on 13 March 2022
and volume of the extraction solvent, flow rate and pH of the sample
solution for CCSHLLE, the type and volume of preconcentration
solvent and ionic strength for DLLME. To the best of our knowledge,
this is the first application of CCSHLLE–DLLME for the determina-
tion of environmental estrogens in water samples, and the optimized
method has been successfully applied to detect DES and OP (Figure 1
shows the chemical structure of the analytes) in river water, well
water, bottled water and campus drinking water samples.
Materials and Methods
Chemicals and materials
DES (99.9%) was purchased from Sigma-Aldrich (USA), and
OP (97%) was purchased from J&K Technology Co., Ltd
(Beijing, China). Chloroform, dichloromethane, sodium chloride,
tetrachloroethane, carbon tetrachloride, dimethylformamide and
isopropanol were of analytical grade and obtained from Huaxin
Reagent & Apparatus Co., Ltd (Baoding, China). ACN was of
HPLC grade and was purchased from Concord Technology Co.,
Ltd (Tianjin, China). Doubly distilled deionized water was used
throughout the experiment.
For preparation of DES and OP stock solutions (1.0 mg/mL),
10 mg of the standard was accurately weighed and dissolved in 10 mL
of methanol and stored at −20◦ C in the refrigerator. For preparation
of working standard solution, the stock solution was diluted with
ACN to the desired concentrations.
Apparatus
HPLC analysis was performed on an HPLC-20AD high-performance
liquid chromatographic system consisting of two LC-20AD pumps
and an SPD-20A multi-wavelength ultraviolet-visible detector (Shi-
madzu, Japan). Other apparatuses used were as follows: the D2012
Plus High Speed microcentrifuge, MX-S adjustable vortexer and
PT100-B magnetic stirrer were provided by Dragon Laboratory
Instrument Co. Ltd (Beijing, China). The HHKQ5200E ultrasonic
cleaner was purchased from Kunshan Ultrasonic Instrument Co. Ltd
(Kunshan, China). MS204S electronic analytical balance (Mettler,
Switzerland) and MTN-2800D pressure blowing instrument (Auto-
science Co. Ltd, Tianjin, China) as well as an HSE series solid-phase
extraction device with a vacuum pump (Tianjin Hengao Technology
Development Co., Ltd, Tianjin, China) were also used.
The HPLC separation was carried out on a Diamonsil C18 column
(150×4.6mm, 5 μm, Dikma) with column temperature kept at 30◦ C.
The mobile phase consisted of ACN and water (66:34, v/v), and the
flow rate was 1 mL/min. The detection wavelength was 230 nm. The
injection volume was 5 μL.
Analysis of Environmental Estrogens in Water
Figure 2. A schematic diagram of CCSHLLE-DLLME process for the determination of DES and OP in water sample.
Table I. Parameters of the Proposed Method for Quantitative Analysis
Compound Linear range (ng/mL) Regression equation
DES 2.0∼200.0 Y = 319.49X + 73.75
OP 8.0∼200.0 Y = 147.32X + 290.66
Extraction procedure
This method consists of two steps. In the first step, a membrane
syringe filter was placed at the bottom of a 10-mL syringe and
connected with an HSE series solid-phase extraction device. After-
wards, 5 g of sodium chloride was added into the syringe and slightly
compressed with the syringe plunger. Subsequently, 5 mL of water
sample was mixed with 2 mL of ACN and passed through the syringe
at a flow rate of 0.5 mL/min. By passing the above homogenous
solution (aqueous solution + ACN) through the syringe, fine droplets
of ACN formed at the interface of solid (NaCl) and solution due
to dissolvation of salt (salting-out effect). The produced droplets of
ACN moved through the remaining solution to the top of the syringe
and floated on the surface of the solution as a separated layer due to
the lower density of ACN (d = 0.786 g/mL) with respect to water.
During this step, the analytes were extracted into the fine droplets.
After all of the water sample passed through the syringe, the stopcock
was closed. The volume of the separated phase (ACN) on the top of
remained NaCl solid was 1 mL.
In the second step, the organic phase obtained in the first step
was removed using a syringe and mixed with carbon tetrachloride.
Then, the mixture was rapidly injected into 5 mL of deionized water
containing 5% of NaCl (w/v) placed in a 10-mL glass test tube with a
conical bottom. A cloudy solution, which resulted from dispersion of
the tiny droplets of carbon tetrachloride into the aqueous solution,
173
Downloaded from https://academic.oup.com/chromsci/article/58/2/171/5612250 by guest on 13 March 2022
R2 LOD (ng/mL) LOQ (ng/mL)
0.9995 0.09 0.31
0.9991 0.20 0.68
formed, and the analytes were extracted into carbon tetrachloride.
The mixture was then centrifuged at 5000 rpm for 5 min, which led
to sedimentation of the dispersed droplets of the extractant at the
bottom of the tube. The organic phase was obtained and dried under
the protection of nitrogen gas and finally reconstituted with 100 μL
of methanol. Five microliters was injected for HPLC analysis. The
extraction procedure is depicted in Figure 2.
Collection of water samples
Bottled water, river water, well water and drinking water were
collected as real samples to test the feasibility of the proposed method.
Bottled water was purchased from the local supermarket; river water
was collected from Yishui River; well water was collected from Boye
County, Baoding, Hebei Province; and drinking water was collected
from Hebei University. All water samples were filtered through a
0.45-μm membrane filter to remove suspended particles and stored
in a glass container for use.
Results
Analytical performance of the method
Linearity, limit of detection and limit of quantitation. Linearity was
studied via the external standard method in the concentration
range of 2.0∼200 ng/mL for DES and 8.0∼200 ng/mL for OP,
174 Shi et al.

Table II. The Results of Recovery Test (n = 3)

Compound Sample Added Recovery RSD (%)

(ng/mL) (%)

DES Bottled 8 97.2 1.7

water 25 90.7 1.8
100 95.7 1.0
River 8 88.7 3.2
water 25 88.2 3.9
100 94.7 2.5
Well 8 103.5 1.2
water 25 94.3 1.0
100 100.9 5.1
Drinking 8 105.5 2.0

Downloaded from https://academic.oup.com/chromsci/article/58/2/171/5612250 by guest on 13 March 2022

water 25 97.2 2.6
100 105.9 1.3
OP Bottled 8 84.8 4.1 Figure 3. The chromatograms of bottled water sample (a) and the bottled
water 25 85.2 2.7 water sample spiked with DES and OP after extraction (b). 1: DES, 2: OP.
100 81.0 7.6
River 8 82.0 6.1
water 25 94.4 5.8 Enrichment factor of the method. The enrichment factor (EF) of the
100 81.4 6.3 method was calculated according to Equation (1):
Well 8 84.9 4.2
water 25 99.0 2.7
100 88.0 5.3 C
EF = (1)
Drinking 8 85.4 3.5 Co
water 25 98.5 2.4
100 93.2 3.6
where C is the analyte concentration in the final organic solvent and
Co is the initial concentration of the analyte in the sample solution.
The EFs for DES and OP were 48 ± 1 and 44 ± 2, respectively.
respectively. Limit of detection (LOD) and limit of quantitation
(LOQ) were calculated based on the signal-to-noise ratio of 3 and
Analysis of water samples and recovery test
10, respectively. The linearity, LOD and LOQ values are listed in
Under the optimized conditions, the established method was used for
Table I.
the analysis of bottled water, drinking water, well water and river
water. All of the water samples were analyzed in triplicate, and no
Reproducibility of the method. Under the optimal conditions, the DES or OP was detected in these water samples.
intra-day and inter-day precision of the method was verified. The Recoveries of DES and OP from all these four kinds of water
intra-day precision of the method was evaluated by processing six samples were investigated at three concentration levels, and the
parallel samples in one day. The inter-day precision was determined experimental results are presented in Table II. The chromatograms
by treating parallel samples in six consecutive days. The relative of the bottled water sample and the spiked bottled water sample are
standard deviations (RSDs) for intra-day precision are 1.7% for DES shown in Figure 3. It could be seen from Figure 3 that good separation
and 2.2% for OP, respectively. The RSD values for inter-day precision between the analytes and coexisting compounds was obtained and
are 2.1% for DES and 3.1% for OP, respectively. the determination of DES and OP was not interfered.

Table III. Comparison of the Proposed Method with Other Methods

Sample Method Extraction time (min) LOD (ng/mL) Recovery (%) Ref.

DES OP

Environmental water and milk MMF-SPME-HPLC-DAD 40 0.10 0.11 75.6–118 (8)

Environmental water SBSE-HPLC-UV 30 0.26 0.24 72.2–122.8 (10)
Water SPME-HPLC-UV 45 0.3 0.80 (33)
Water μ-SPE-HPLC-UV 15 0.52 91–93 (34)
Environmental water TFME-HPLC-UV 30 0.40 86.0–96.6 (35)
Sea water SDME-HPLC-UV 60 9 101–104 (36)
Waste water FA-IT-DLLME HPLC-UV 4 0.50 97.2–101.2 (37)
Water CCSHLLE-DLLME-HPLC 40 0.09 0.20 81–105.9 This work
MMF-SPME: multiple monolithic fiber solid-phase microextraction; SBSE: stir bar sorptive extraction; SPME: solid-phase microextraction; μ-SPE: micro-solid phase extraction; TFME:
thin film microextraction; SDME: single-drop microextraction; FA-IT-DLLME: fatty-acid-based in-tube dispersive liquid–liquid microextraction; CCSHLLE- DLLME: counter current
salting-out homogenous liquid–liquid extraction-dispersive liquid–liquid microextraction.
Analysis of Environmental Estrogens in Water 175

Downloaded from https://academic.oup.com/chromsci/article/58/2/171/5612250 by guest on 13 March 2022

Fig. 5. Effect of flow rate in CCSHLLE. Extraction conditions: CCSHLLE: 5 mL
Figure 4. Effect of the volume of acetonitrile in CCSHLLE. Extraction con-
water sample; extraction solvent, 2 mL acetonitrile. DLLME: 5 mL deionized
ditions: CCSHLLE: 5 mL water sample; extraction solvent, acetonitrile; flow
water; extraction solvent, 50 μL carbon tetrachloride; ionic strength, 5% (w/v)
rate, 0.6 mL/min. DLLME: 5 mL deionized water; extraction solvent, carbon
NaCl; centrifugation rate and time, 3000 rpm and 5 min.
tetrachloride (50 μL); ionic strength, 5% (w/v) NaCl; centrifugation rate and
time, 3000 rpm and 5 min.

Comparison with other methods

The proposed method was compared with other reported methods
for the determination of DES and OP in the aspects of extraction
method, extraction time, detection method, LOD and recovery. As
shown in Table III, the established method demonstrates the merits
of comparable extraction time, low LOD and satisfactory recoveries.
Furthermore, the established CCSHLLE–DLLME method is easy to
operate, with no special equipment or material needed.

Discussion
Optimization of extraction conditions
In order to achieve the best extraction efficiency, the type and volume
of extraction solvent, the flow rate and pH of the sample solution in
CCSHLLE and the type and volume of preconcentration solvent and
ionic strength in DLLME were investigated and optimized. In this
work, the extraction recovery (ER) was used to study the extraction
efficiency of the analytes. The calculation of the ER is based on
Equation (2):
CV
ER% = × 100
Co Vo
where C is the analyte concentration in the final organic solvent, Co
is the initial concentration of the analyte in the sample solution and
V and V o are the volume of the final organic solvent and the volume
of the sample solution, respectively.
Optimization of CCSHLLE
Selection of extraction solvent. In this experiment, the extraction
solvent played a double role: as an extractant in the CCSHLLE step
and as a dispersant in the next preconcentration step (DLLME).
The extraction solvent was selected on the basis of its extraction
efficiency for the analytes from the sample solution, its capability of
producing phase separation upon salt dissolvation and its miscibility
with both the extraction solvent of DLLME and the aqueous phase
(to form a homogenous solution). In this study, ACN, acetone,
Fig. 6. Effect of the volume of carbon tetrachloride in DLLME. Extraction condi-
tions: CCSHLLE: 5 mL water sample; extraction solvent, 2 mL acetonitrile; flow
rate, 0.4 mL/min. DLLME: 5 mL deionized water; extraction solvent, carbon
tetrachloride; ionic strength, 5% (w/v) NaCl; centrifugation rate and time,
(2) 3000 rpm and 5 min.
dimethylformamide and isopropanol were tested as extractants. The
results showed that only ACN could form phase separation upon
salting-out effect. This can be explained from the low donor number
of ACN (14.1) compared with the other tested solvents (isopropanol,
36; acetone, 17; and dimethylformamide, 26.6). Donor number is
a measure of the ability of a solvent to dissolve cations or Lewis
acids. In this study, NaCl was used as a salt to produce Na+ ions
in the aqueous solution, and while acetone, dimethylformamide and
isopropanol dissolved Na+ ions, phase separation could not happen
(24). ACN with the lowest donor number among the tested solvents
was separated from the aqueous phase and formed a separated
layer. Therefore, ACN was selected as extractant in the CCSHLLE
step.
176
Effect of the volume of ACN. The effect of the volume of ACN
on the extraction efficiency was investigated from 1 to 3 mL at
0.5-mL intervals. An insufficient amount of extractant may result
in incomplete extraction, and too much extractant may result in a
decrease in analyte concentration in the organic phase due to the
dilution effect. As shown in Figure 4, when the volume of extractant
was increased to 2 mL, the extraction efficiency was the highest, and
after 2 mL, the extraction efficiency was slightly lower. With 1 mL
of ACN, the emulsion did not form completely in the subsequent
DLLME procedure. However, with 1.5 to 3 mL of ACN, the emulsion
formed well. By using more than 2.0 mL of ACN, the solubility of
the analytes in the aqueous phase was increased and the extraction
efficiency was reduced. As the volume of ACN used was increased
from 1 to 3 mL, the collected ACN phase ranged from 0.2 to 2 mL.
Thus, according to the experimental results, 2 mL of ACN was used
for subsequent experiments. The collected ACN phase in CCSHLLE
was about 1 mL.
Effect of flow rate. Flow rate is one of the most important factors,
which can affect the whole analysis time and ER of the proposed
method. The flow rate of the sample solution must be low enough
to perform an effective salting-out process. On the other hand, it
must be high enough not to waste time (32). Therefore, the effect
of the flow rate was investigated in the range of 0.2 to 2 mL/min.
As shown in Figure 5, the extraction efficiency was increased when
the flow rate was raised up to 0.4 mL/min and then decreased. At
a higher flow rate, the extraction efficiency decreased rapidly. This
phenomenon can be explained as higher flow rate is not beneficial for
the sufficient interaction between the analytes and ACN. Therefore,
the flow rate of 0.4 mL/min was selected for further experiments, and
the extraction time was about 10 min.
Effect of pH. In this paper, the effect of pH on the extraction effi-
ciency of the analytes was investigated in the range of 4 to 10. The
experimental results showed that pH of the aqueous solution had no
significant effect on the extraction efficiency of DES and OP. The pH
of the tested water sample was generally in the range of 5.5 and 6.5;
hence, there was no need to adjust pH in this study.
Optimization of DLLME
Selection of preconcentration solvent. The choice of an appropriate
preconcentration solvent plays a main role in this method in order
to achieve high EF. The preconcentration solvent should demonstrate
the following characteristics: higher density than water (preferably),
low solubility in water, high volatility, good extraction capability for
analytes and ability to form a cloudy solution in the presence of
ACN. Therefore, dichloromethane, chloroform, carbon tetrachloride
and tetrachloroethane were investigated. The experimental results
showed that carbon tetrachloride provided higher extraction effi-
ciency than did other extraction solvents. Therefore, carbon tetra-
chloride was selected for the following experiments.
Effect of the volume of carbon tetrachloride. Different volumes of
carbon tetrachloride (30, 40, 50, 60, 70, 80 μL) were tested to
optimize the extraction efficiency. Figure 6 shows that the extraction
efficiency of the analytes increased with the increase in the volume
from 30 to 50 μL and then kept almost constant with further increase
in the volume. Therefore, 50 μL of carbon tetrachloride was selected
for subsequent experiments.
Effect of ionic strength. The presence of salt in the aqueous solution
generally affects the solubility of the organic solute. When the soluble
Shi et al.
inorganic salt was added, the solubility of the target analytes in the
aqueous solution could be reduced, and the distribution ratio between
the organic phase and the aqueous phase could be increased; thereby,
the extraction efficiency was improved. In this experiment, different
concentrations of NaCl (0–20%) were investigated. When the con-
centration of NaCl was increased to 10%, the extraction efficiency
increased gradually and then kept almost constant. Therefore, 10%
NaCl was selected for further experiments.
Conclusion
In this work, CCSHLLE was combined with DLLME for the simul-
taneous determination of environmental estrogens in water samples
by HPLC-UV. Compared with traditional salting-out methods, phase
Downloaded from https://academic.oup.com/chromsci/article/58/2/171/5612250 by guest on 13 March 2022
separation of CCSHLLE was generated in the process of water flow
with the advantage of increased contact area, and no separation
equipment was needed. The target analytes were further concentrated
by using subsequent DLLME. This is the first application of CCSH-
LLE–DLLME in the determination of environmental estrogens, and
the established method has been successfully used for the analysis of
various water samples.
Acknowledgements
The authors acknowledge the financial support from the Department of Edu-
cation of Hebei Province (ZD2019089) and the Scientific Research Foundation
for the Returned Overseas Chinese Scholars, State Education Ministry of
China (No. 47). This work was also supported by the Laboratory Open
Project of Hebei University (sy201801) and the National College Students
Innovation and Entrepreneurship Training Program (201910075018) as well
as the College Students Innovation and Entrepreneurship Training Program of
Hebei University (2019191).
References
1. European Commission, European workshop on the impact of endocrine
disrupters on human health and wildlife. Report of the proceedings,
Weybridge, UK, 1996, Report EUR 17549.
2. Kabir, E.R., Rahman, M.S., Rahman, I.; A review on endocrine disruptors
and their possible impacts on human health; Environmental Toxicology
and Pharmacology, (2015); 40: 241–258.
3. Tapiero, H., Ba, G.N., Tew, K.D.; Estrogens and environmental estrogens;
Biomedicine & Pharmacotherapy, (2002); 56: 36–44.
4. Gong, S.X., Wang, X.L., Liu, W., Wang, M.L., Wang, X., Wang, Z.W. et al.;
Aminosilanized magnetic carbon microspheres for the magnetic solid-
phase extraction of bisphenol A, bisphenol AF, and tetrabromobisphenol A
from environmental water samples; Journal of Separation Science, (2017);
40: 1755–1764.
5. Yang, D.Z., Li, X.L., Meng, D.L., Wang, M., Yang, Y.L.; Supramolecu-
lar solvents combined with layered double hydroxide-coated magnetic
nanoparticles for extraction of bisphenols and 4-tert-octylphenol from
fruit juices; Food Chemistry, (2017); 237: 870–876.
6. Chen, B.B., Huang, Y.L., He, M., Hu, B.; Hollow fiber liquid-
liquid-liquid microextraction combined with high performance liquid
chromatography-ultraviolet detection for the determination of various
environmental estrogens in environmental and biological samples; Journal
of Chromatography A, (2013); 1305: 17–26.
7. Xu, Z.G., Yang, Z.L., Liu, Z.M.; Development of dual-templates molec-
ularly imprinted stir bar sorptive extraction and its application for the
analysis of environmental estrogens in water and plastic samples; Journal
of Chromatography A, (2014); 1358: 52–59.
8. Mei, M., Yu, J., Huang, X.J., Li, H.N., Lin, L., Yuan, D.X.; Monitoring
of selected estrogen mimics in complicated samples using polymeric
ionic liquid-based multiple monolithic fiber solid-phase microextraction
Analysis of Environmental Estrogens in Water
combined with high-performance liquid chromatography; Journal of
Chromatography A, (2015); 1385: 12–19.
9. Socas-Rodríguez, B., Herrera-Herrera, A.V., Hernández-Borges, J.,
Rodríguez-Delgado, M.Á.; Multiresidue determination of estrogens
in different dairy products by ultra-high-performance liquid
chromatography triple quadrupole mass spectrometry; Journal of
Chromatography A, (2017); 1496: 58–67.
10. Hu, C., He, M., Chen, B.B., Zhong, C., Hu, B.;
Polydimethylsiloxane/metal-organic frameworks coated stir bar sorptive
extraction coupled to high performance liquid chromatography-
ultraviolet detector for the determination of estrogens in environmental
water samples; Journal of Chromatography A, (2013); 1310: 21–30.
11. Lin, W.C., Wang, S.L., Cheng, C.Y., Ding, W.H.; Determination of
alkylphenol residues in breast and commercial milk by solid-phase
extraction and gas chromatography–mass spectrometry; Food Chemistry,
(2009); 114: 753–757.
12. Zhou, Q.X., Lei, M., Li, J., Zhao, K.F., Liu, Y.L.; Sensitive determination
of bisphenol A, 4-nonylphenol and 4-octylphenol by magnetic solid phase
extraction with Fe@MgAl-LDH magnetic nanoparticles from environ-
mental water samples; Separation and Purification Technology, (2017);
182: 78–86.
13. Acir, I.H., Guenther, K.; Endocrine-disrupting metabolites of alkylphenol
ethoxylates-A critical review of analytical methods, environmental occur-
rences, toxicity, and regulation; Science of the Total Environment, (2018);
635: 1530–1546.
14. Liu, Y.Y., Lin, Y.S., Yen, C.H., Miaw, C.L., Chen, T.C., Wu, M.C. et al.;
Identification, contribution, and estrogenic activity of potential EDCs in
a river receiving concentrated livestock effluent in Southern Taiwan; The
Science of the Total Environment, (2018); 636: 464–476.
15. Kapelewska, J., Kotowska, U., Karpinska, J., Kowalczuk, D., Arciszewska,
A., Swirydo, A.; Occurrence, removal, mass loading and environmental
risk assessment of emerging organic contaminants in leachates, ground-
waters and wastewaters; Microchemical Journal, (2018); 137: 292–301.
16. Skrbic, B.D., Kadokami, K., Antic, I.; Survey on the micro-pollutants
presence in surface water system of northern Serbia and environmen-
tal and health risk assessment; Environmental Research, (2018); 166:
130–140.
17. Valcarcel, Y., Valdehita, A., Becerra, E., Lopez de Alda, M., Gil, A., Gorga,
M. et al.; Determining the presence of chemicals with suspected endocrine
activity in drinking water from the Madrid region (Spain) and assessment
of their estrogenic, androgenic and thyroidal activities; Chemosphere,
(2018); 201: 388–398.
18. Ternes, T.A., Andersen, H., Gilberg, D., Bonerz, M.; Determination of
estrogens in sludge and sediments by liquid extraction and GC/MS/MS;
Analytical Chemistry, (2002); 74: 3498–3504.
19. Grześkowiak, T., Czarczyńska-Goślińska, B., Zgoła-Grześkowiak, A.; Cur-
rent approaches in sample preparation for trace analysis of selected
endocrine-disrupting compounds: Focus on polychlorinated biphenyls,
alkylphenols, and parabens; TRAC-Trends in Analytical Chemistry,
(2016); 75: 209–226.
20. Grover, D.P., Zhang, Z.L., Readman, J.W., Zhou, J.L.; A comparison of
three analytical techniques for the measurement of steroidal estrogens in
environmental water samples; Talanta, (2009); 78: 1204–1210.
21. Kuehnbaum, N.L., Britz-McKibbin, P.; Comprehensive profiling of free
and conjugated estrogens by capillary electrophoresis-time of flight/mass
spectrometry; Analytical Chemistry, (2011); 83: 8063–8068.
22. Xu, X., Roman, J.M., Issaq, H.J., Keefer, L.K., Veenstra, T.D., Ziegler,
R.G.; Quantitative measurement of endogenous estrogens and estrogen
metabolites in human serum by liquid chromatography-tandem mass
spectrometry; Analytical Chemistry, (2007); 79: 7813–7821.
177
23. Qian, L.L., Li, X.Q., Qi, F.F., Li, J., Lu, L.G., Xu, Q.; An amino-
functionalized grooved nanofiber mat for solid-phase extraction of phe-
nolic pollutants; Microchimica Acta, (2017); 184: 2861–2870.
24. Omar, T.F.T., Aris, A.Z., Yusoff, F.M., Mustafa, S.; An improved SPE-LC-
MS/MS method for multiclass endocrine disrupting compound determi-
nation in tropical estuarine sediments; Talanta, (2017); 173: 51–59.
25. Chang, C.C., Huang, S.D.; Determination of the steroid hormone levels
in water samples by dispersive liquid-liquid microextraction with solidi-
fication of a floating organic drop followed by high-performance liquid
chromatography; Analytica Chimica Acta, (2010); 662: 39–43.
26. Yang, D.Z., Li, G.X., Wu, L., Yang, Y.L.; Ferrofluid-based liquid-phase
microextraction: Analysis of four phenolic compounds in milks and fruit
juices; Food Chemistry, (2018); 261: 96–102.
27. Lan, H.Z., Gan, N., Pan, D.D., Hu, F.T., Li, T.H., Long, N.B. et al.;
An automated solid-phase microextraction method based on magnetic
Downloaded from https://academic.oup.com/chromsci/article/58/2/171/5612250 by guest on 13 March 2022
molecularly imprinted polymer as fiber coating for detection of trace
estrogens in milk powder; Journal of Chromatography A, (2014); 1331:
10–18.
28. Socas-Rodriguez, B., Asensio-Ramos, M., Hernandez-Borges, J.,
Rodriguez-Delgado, M.A.; Analysis of estrogenic compounds in dairy
products by hollow-fibre liquid-phase microextraction coupled to liquid
chromatography; Food Chemistry, (2014); 149: 319–325.
29. Nerin, C., Salafranca, J., Aznar, M., Batlle, R.; Critical review on recent
developments in solventless techniques for extraction of analytes; Analyt-
ical and Bioanalytical Chemistry, (2009); 393: 809–833.
30. Farajzadeh, M.A., Feriduni, B., Afshar Mogaddam, M.R.; Development
of counter current salting-out homogenous liquid-liquid extraction for
isolation and preconcentration of some pesticides from aqueous samples;
Analytica Chimica Acta, (2015); 885: 122–131.
31. Farajzadeh, M.A., Feriduni, B., Afshar Mogaddam, M.R.; Develop-
ment of a new extraction method based on counter current salting-out
homogenous liquid–liquid extraction followed by dispersive liquid–liquid
microextraction: Application for the extraction and preconcentration of
widely used pesticides from fruit juices; Talanta, (2016); 146: 772–779.
32. Akramipour, R., Fattahi, N., Pirsaheb, M., Gheini, S.; Combination of
counter current salting-out homogenous liquid–liquid extraction and
dispersive liquid-liquid microextraction as a novel microextraction of
drugs in urine samples; Journal of Chromatography B, (2016); 1012-1013:
162–168.
33. Peñalver, A., Pocurull, E., Borrull, F., Marcé, R.M.; Method based on solid-
phase microextraction high-performance liquid chromatography with UV
and electrochemical detection to determine estrogenic compounds in water
samples; Journal of Chromatography A, (2002); 964: 153–160.
34. Naing, N.N., Li, S.F.Y., Lee, H.K.; Evaluation of graphene-based sorbent in
the determination of polar environmental contaminants in water by micro-
solid phase extraction-high performance liquid chromatography; Journal
of Chromatography A, (2016); 1427: 29–36.
35. Sun, M., Wu, Q.H., Wang, C., Wang, Z.; Thin-film microextraction for
the preconcentration of some endocrine disrupting chemicals in aqueous
samples before chromatographic analysis; Analytical Methods, (2014); 6:
6316–6321.
36. López-Darias, J., Germán-Hernández, M., Pino, V., Afonso, A.M.; Disper-
sive liquid-liquid microextraction versus single-drop microextraction for
the determination of several endocrine-disrupting phenols from seawaters;
Talanta, (2010); 80: 1611–1618.
37. Shih, H.K., Shu, T.Y., Ponnusamy, V.K., Jen, J.F.; A novel fatty-acid-based
in-tube dispersive liquid-liquid microextraction technique for the rapid
determination of nonylphenol and 4-tert-octylphenol in aqueous sam-
ples using high-performance liquid chromatography-ultraviolet detection;
Analytica Chimica Acta, (2015); 854: 70–77.