Trends in Analytical Chemistry 71 (2015) 282–292

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Trends in Analytical Chemistry

j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / t r a c

Theory and recent applications of coacervate-based extraction

techniques
A. Melnyk a,*, J. Namieśnik a, L. Wolska a,b
aDepartment of Analytical Chemistry, Chemical Faculty, Gdansk University of Technology (GUT), 11/12 G. Narutowicza Str., 80-233 Gdansk, Poland
bDepartment of Environmental Toxicology, Faculty of Health Sciences with Subfaculty of Nursing, Medical University of Gdansk, De˛bowa Str. 3, 80-204
Gdańsk, Poland

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

Keywords:
In recent years, development of sample-preparation techniques with significant advantages over con-
Cloud-point extraction
ventional methods for the extraction of organic compounds from different samples became increasingly
Coacervate
Coacervative extraction
important. Coacervates emerged as promising environment-friendly alternatives for sample prepara-
Green analytical chemistry tion. This review covers the basic theory of coacervates and summarizes recent applications (2010–15)
Metal ion of coacervates for preconcentration of organic compounds, metal ions and nanoparticles from various
Nanoparticle types of sample.
Organic © 2015 Elsevier B.V. All rights reserved.
Preconcentration
Sample preparation
Surfactant

Contents

1. Introduction ........................................................................................................................................................................................................................................................ 282

2. Basics of coacervate-based extraction process ....................................................................................................................................................................................... 284
2.1. Theory and concept of micelles ..................................................................................................................................................................................................... 284
2.2. Micellar solubilization ....................................................................................................................................................................................................................... 284
2.3. Coacervate-based extraction techniques .................................................................................................................................................................................... 285
3. Applications of coacervate-based techniques for sample preparation ........................................................................................................................................... 285
4. Limitations of coacervate-based techniques ........................................................................................................................................................................................... 290
5. Conclusions and future trends ..................................................................................................................................................................................................................... 290
References ............................................................................................................................................................................................................................................................ 291

1. Introduction samples with complex matrices, so it is necessary to introduce to

analytical procedures an additional step enriching analytes above
Available analytical techniques are not always sensitive enough the limit of detection (LOD) of the method.
to detect, to identify and to quantify compounds at trace levels in Sample preparation is a crucial step in the analytical process.
There are many techniques currently used for preconcentration, but
each, in addition to numerous advantages, also has some limita-
tions. Examples of limitations of commonly used sample-preparation
Abbreviations: ASTP, Aqueous surfactant two-phase; CAE, Coacervative extraction; techniques are presented in Table 1. These disadvantages result in
CMC, Critical micelle concentration; CPE, Cloud-point extraction; CPT, Cloud-point continual development of new, more effective extraction tech-
temperature; CTAB, Hexadecyltrimethylammonium bromide; DAD, Diode-array de-
niques and modifications of existing methods. These developments
tection; ET-AAS, Electrothermal atomic absorption spectrometry; GC, Gas
chromatography; HPLC, High-performance liquid chromatography; ICP-MS, Induc- are also driven by stricter environmental regulations and con-
tively coupled plasma – mass spectrometry; LC, Liquid chromatography; LLE, Liquid- cerns. Literature data provide the basis for the conclusion that so-
liquid extraction; LOD, Limit of detection; LOQ, Limit of quantification; NP, called green extraction techniques play a vital role in modern
Nanoparticle; PAH, Polycyclic aromatic hydrocarbon; SDS, Sodium dodecyl sulfate; techniques of sample preparation. Implementation of green ana-
SFVCDME, Solidified floating vesicular coacervative drop microextraction; SPE, Solid-
phase extraction; UV-vis, Ultraviolet-visible spectrophotometry.
lytical chemistry (GAC) principles is evidenced by the steadily
* Corresponding author. Tel.: +48 58 347 21 10; Fax: +48 58 347 26 94. growing number of publications on GAC [15]. The green character
E-mail address: Alma@Melnyk.pl (A. Melnyk). of the extraction process may be achieved by reducing consumption

http://dx.doi.org/10.1016/j.trac.2015.03.013
0165-9936/© 2015 Elsevier B.V. All rights reserved.
 A. Melnyk et al./Trends in Analytical Chemistry 71 (2015) 282–292
Table 1
Examples of limitations of selected extraction techniques
Technique Principle of extraction process
Accelerated solvent A solid–liquid extraction process
extraction, performed at high temperatures
pressurized (50–200°C) and high pressures
liquid extraction (10–15 MPa).
(ASE, PLE)
Dispersive Analytes migrate to droplets of the
liquid–liquid extraction solvent which are dispersed
microextraction in the sample solution.
(DLLME)
Extraction with ionic Extraction occurs by contacting a
liquids sample with ionic liquid, which is low-
melting salt that form liquid composed
entirely of ions.
Headspace sampling Analytes are isolated from a solid or
(HS) liquid matrix to the gaseous or vapor
phase in equilibrium (or not) with the
matrix.
Microwave-assisted Microwaves penetrate into certain
extraction (MAE) materials and only selective and
targeted materials can be heated based
on their dielectric constant.
Microporous Extraction occurs through the
membrane membrane which is a barrier between
liquid–liquid two phases: 1) organic, filling the
extraction (MMLLE) membrane pores, 2) the aqueous
sample on the other side of the
membrane.
Stir-bar sorptive Analytes are extracted from the matrix
extraction (SBSE) to the polymer-coated stir-bar which is
put in contact with the solutes by
immersion or by headspace sampling.
Single drop The analyte is partitioning between
microextraction sample and the microdrop directly
(SDME) immersed in the sample, or in the
headspace above the sample.
Supercritical fluid Extraction media are fluids which,
extraction (SFE) when simultaneously heated and
compressed above their critical
temperature and pressure, gain
intermediate properties between a gas
and a liquid.
Solid-phase extraction Partitioning of solutes between two
(SPE) phases: a liquid (sample matrix) and a
solid (sorbent) phase.
Solid-phase Microfiber is immersed into the
microextraction sample for sorption and followed by
(SPME) direct desorption in the inlet of the GC.
Subcritical water Extraction solvent is water at
extraction (SWE) temperatures between 100 and 374°C
and at a pressure high enough to keep
it in the liquid state.
Ultrasound-assisted Extraction between two immiscible
emulsification phases (of which dispersed phase with
microextraction a good efficiency for emulsification is
(USAEME) in the aqueous sample in low volume)
supported with ultrasound.
Ultrasound assisted Mass transfer between two immiscible
liquid–liquid phases (i.e. liquid–liquid) supported by
extraction (USALLE) ultrasound.
283
Limitations Ref
- Expensive laboratory equipment is required. [1]
- Decrease of selectivity as a result of high temperature.
- Thermo-labile compounds may undergo disintegration and hydrolytic degradation in high
temperatures.
- Recovery depends on the pressure used.
- The number of extraction solvents available for use with the method is limited. [2]
- Low solubility and suitable density of the solvent is required.
- The solvent have to form a cloudy solution in the presence of the disperser solvent.
- The disperser solvent can complicate the process of phase separation.
- Sample preparation procedures that depend on pumping ionic liquids with common [3]
laboratory devices require viscosities <5 cP for normal operation.
- Ionic liquids with energetic groups are combustible.
- It is debatable whether all ionic liquids are characterized by a low toxicity.
- Room temperature ionic liquids are incompatible with GC.
- Limited to volatile compounds. [4]
- Dedicated instrumentation, cryoconcentration and tuning of several parameters are required.
- Poor recovery of very volatile components and polar analytes from complex or
multi-ingredient matrices.
- Vapor pressure of solvent in static HS-LPME must be low enough to avoid evaporation
during sampling but, it must be compatible with GC analysis.
- Requirement of additional clean up step to remove solvent from sample matrices. [5]
- Need to use only polar solvents in the system.
- The efficiency strongly relies on the selection of the conditions and the parameters affecting
the extraction mechanisms and yield.
- Memory effect problems. [6]
- Clogging of the membrane pores.
- Successful development of some procedures depends on the preparation of highly suitable
catalytic membranes for specific and selective reactions/extractions and the quantitative
control of the catalytic reaction rates.
- Preconditioning pre-treatment is needed in order to remove co-extractable interfering
compounds from the membrane material.
- The efficiency of SBSE can be strongly affected by the complexity of the sample matrix. [7]
- The possibility of physical damage of the coating due to the direct contact with the
bottom of the sample vial.
- The coated stir-bar cannot be directly desorbed in a simple GC injection port.
- Manual operations like removing the stir-bar from the sample, rinsing and drying are
time-consuming and can cause further errors, but automation of these steps increases the
cost and complexity of the hardware involved.
- Limited drop size. [8]
- Solvent solubility in the sample solvent and drop instability.
- The time required to reach equilibrium can be from seconds to hours, depending
on different terms, so it is often performed under non-equilibrium conditions.
- Only a limited number of solvents can be used for solvent microextraction.
- SFE with CO2 is not suitable for simultaneous extraction of a wide polarity range of analytes. [9]
- The need to maintain optimal parameters such as pressure and temperature
to achieve satisfying mass transfer.
- Some of the supercritical fluids are aggressive for the chromatograph.
- Frequently the chosen elution conditions will cause phase separation of supercritical fluid.
- High volumes of organic solvents are used for conditioning of the sorbent and [10]
elution of analytes.
- Strong adsorption of analytes to the sorbent can prevent the elution.
- Relatively high cost, due to a single use of the sorbent.
- The need to remove the suspension from the sample, which can cause clogging of
the channels between the grains of sorbent.
- The extraction fiber is expensive, fragile and has a limited lifetime. [11]
- Sample carry-over can be a problem.
- For aqueous matrices, fast sample flow, rapid fiber or vial movement, stirring or sonication
are required.
- An effect called “depletion zone” occurs close to the fiber as a result of fluid shielding and
slow diffusion of analytes in liquid media.
- The final product is a suspension of particles in water. [12]
- The drying of water from a suspension can be energy intensive.
- Slow evaporation of water may change the resulting morphology
of the precipitated compounds.
- Extraction may lead to the degradation of analytes.
- US probes (sonotrodes) are much more expensive and less robust than US cleaning baths but [13]
US cleaning bath are irreproducible and do not provide uniformity in the transmission of US.
- Low volumes of solvents used can be justified only for expensive reagents or when the
required preconcentration factor demands less extractant volume according to the
partitioning factor and sample volume.
- Different derivatization reactions are required.
- High demands on extractant (good efficiency for emulsification in the sample, high affinity
for the target analytes or their derivatization products, low solubility in water, compatibility
with determination techniques).
- US assistance raises the temperature. [14]
- Need to develop specific substances tailored to the particular liquid combination.
- Formation of emulsions.
284 A. Melnyk et al./Trends in Analytical Chemistry 71 (2015) 282–292
of chemicals, including organic solvents, which are toxic and harmful
to the environment. Procedures routinely used for sample prepa-
ration should therefore be environment friendly and characterized
by simplicity of implementation, low cost and short time to carry
out the whole process. These requirements are fulfilled by appli-
cation of coacervates in sample preparation, a coacervate being a
surfactant-rich liquid and an extraction medium.
The use of coacervates for extraction of analytes overcomes the
characteristics of some selected techniques shown in Table 1. No
expensive additional laboratory equipment is required [unlike in ac-
celerated solvent extraction (ASE), pressurized liquid extraction (PLE)
or ultrasound extraction (USE)]. Coacervates are incombustible as
are some ionic liquids (ILs) and are not limited to volatile com-
pounds, as headspace sampling is. Unlike microwave-assisted
extraction (MAE), an additional clean-up step is unnecessary. There
are no memory-effect problems as there are in microporous mem-
brane liquid-liquid extraction (MMLLE). Coacervate-based extraction
techniques do not involve so many manual operations as stir-bar
sorptive extraction (SBSE), so they are not so time-consuming and
do not cause further errors in the way SBSE does. Coacervates do
not require such specific parameters as pressure and temperature
as do supercritical fluid extraction (SFE) and subcritical water ex-
traction (SWE). Coacervate-based extraction is cheaper than solid-
phase microextraction (SPME), which involves expensive extraction
fiber, and does not use large volumes of organic solvents, as solid-
phase extraction (SPE) does.
Several review papers on the use of surfactants in sample prep-
aration have been published [16]. We focus on recent applications
(since 2010) of coacervate-based procedures for the isolation of
organic compounds, metal ions and nanoparticles (NPs) in samples
having various, often very complex, matrices. We also describe the
basics and the limitations associated with the use of coacervate-
based techniques.
2. Basics of coacervate-based extraction process
2.1. Theory and concept of micelles
Surfactants are conventionally composed of a polar head-
group and a long hydrophobic tail-group, and may be classified into
four different groups:
• anionic surfactants;
• cationic surfactants;
• zwitterionic surfactants; and,
• non-ionic surfactants [17].
Surfactant molecules are adsorbed at the interface: aqueous so-
lution – air or aqueous solution – organic solution, in such a way
that the hydrophilic part is always directed towards the aqueous
phase. When the concentration increases, amphiphilic molecules
dispersed in a solvent are under the influence of the amphiphilic
properties of hydrophobic chains, and self-assemble into colloidal-
sized clusters [16]. Association occurs after exceeding the
characteristic concentration for surface-active compounds, known
as the critical micelle concentration (CMC) [17]. Aggregates formed
above the CMC are called micelles.
Micelles are colloidal systems, transparent (aggregate size is
smaller than the length of visible light) in which the continuous
phase is water or solvent, and aggregated surfactant molecules are
the dispersed phase. Due to the size of the aggregates, micelles are
in a position between suspensions and proper solutions, where the
limit is not strictly defined. Micelles can have different forms from
roughly spherical to ellipsoidal, depending on the specific surfac-
tant and solution conditions. In polar solvents, micelles take a form
with hydrophilic heads outside of the structure so that tails are
protected from adverse effects of the solvent. In a non-polar solvent,
reverse micelles are formed, in which hydrophobic tails are outside
the structure [16].
The stability and the structure of the micelles are achieved due
to the presence of a load on the surface, in which the aggregate is
electrically neutral. The number of surfactant molecules present in
micelles has been called the degree or the aggregation number
(nagg). The numerical value of this parameter depends on certain
factors, such as:
• surfactant type;
• structure of the hydrophobic and hydrophilic groups;
• presence and concentration of electrolytes in the system;
• nature of the solvent;
• temperature; and,
• pH of the solution.
Micelles are in a dynamic equilibrium with monomers, where
each monomer can leave the micelle, and may be replaced by
another. The time of the monomer inside a micelle depends on the
type and the structure of the molecule, while the time of leaving
of the monomer is of the order of μs, and its return depends on the
diffusion rate. It is worth mentioning that the shape of micelles can
be altered by appropriate selection of solution parameters, such as
temperature, concentration of the surfactant, and the presence, the
type and the concentration of additional compounds in the solu-
tion [18]. Access to existing databases, after introducing information
regarding parameters of the solution and surfactant molecular struc-
ture, provides an opportunity to obtain information on:
• CMC parameter settings;
• surface tension; and,
• shape and size of the micelles obtained under certain conditions.
2.2. Micellar solubilization
For a long time, surface-active compounds are used in the ex-
traction process due to the observed increase in the solubility of
slightly-soluble compounds in the presence of amphiphilic com-
pounds [19]. The ability to absorb slightly-soluble compound particles
by micellar aggregates is given the term “solubilization”. Regions
of micelles, due to their diverse polarities, enhance the potential of
micelles to solubilize solutes in a wide range of polarities. In aqueous
solutions containing micelles, the solubility of many hydrophobic
compounds, or compounds only partially soluble in water, espe-
cially ionic and non-ionic organic compounds, is considerably higher
than in the pure aqueous solution. This process occurs through the
formation of a microemulsion in aqueous surfactant solutions (i.e.
a macroscopically homogeneous system, which is, however, mi-
croscopically heterogeneous). Hydrophobic solutes are solubilized
in the inner micellar core. Variants of solubilization of hydropho-
bic substances in the micellar aggregates are shown in Fig. 1.
Similarly, in organic solutions, the presence of reverse micelles
causes an increase in the solubility of hydrophilic substances, such
as amino acids or proteins. Solubilized chemical compounds of polar
character can be (in a dissociated form) located in the center of the
micelle or can undergo absorption near polar groups of the surfac-
tant (surface-active proteins) or near the micelle hydrophobic chains
(hydrophobic protein). Polar analytes are solubilized in the polar
region through a number of interactions (e.g. electrostatic, π-cation,
or hydrogen bond). Polar compounds that are incorporated into the
micelle structure may diffuse through the semi-permeable mem-
branes, which are used in drugs and cosmetics industries. Solutions
created in this way have an isotropic character and are thermody-
namically stable. Variants of solubilisate position in reverse micelles
are shown in Fig. 2. Amphiphilic solutes are incorporated into
 A. Melnyk et al./Trends in Analytical Chemistry 71 (2015) 282–292
Fig. 1. Possible methods for solubilizing particles in micelles; A – solubilisate mol-
ecules placed immediately adjacent to the micelle hydrophobic chain, B – solubilisate
molecules located inside the micelles to form a separate phase.
Fig. 2. Possible location of solubilized chemical compound molecules in reverse
micelles.
micelles through both hydrophobic and polar interactions, forming
mixed aggregates [16]. Solubilization of metal ions involves the prior
formation of a complex using suitable ligands [20].
As mentioned earlier, the structure of micellar aggregates gives
them the ability to form numerous bonds with dissolved sub-
stances, so that micelles are characterized by strong solvation properties
in relation to different compounds. The process of extracting begins
when the surfactant solution is introduced into the sample in a quan-
tity that allows formation of micelles (above the CMC value). After
solubilization of analytes into micellar aggregates, the creation of two
incompatible and immiscible isotropic phases is observed:
• a coacervative phase, which is a surfactant-rich phase with ex-
tracted compounds; and,
• a surfactant-poor phase, called equilibrium liquid or aqueous bulk
– a solution, which is in equilibrium with the coacervative phase.
The liquid-liquid phase separation is induced by environmen-
tal conditions, such as temperature, pH, and addition of an electrolyte
or a solvent that is miscible with water, in which macromolecules
are characterized by low solubility [16]. As a result, the analytes are
isolated from the sample matrix and concentrated in the small
volume of the coacervative phase, which is ready to be analyzed.
2.3. Coacervate-based extraction techniques
The first method to use surfactants for extraction was cloud-
point extraction (CPE), which was first studied in the 1970s [21] and
is still the most popular coacervate-based extraction technique. In
the literature, it is sometimes also called micellar extraction (ME
285
or MEX), micelle-mediated extraction (MME) or liquid-coacervate
extraction (LCE). In the CPE technique, non-ionic and zwitterionic
surfactants are used as colloids, and the phase separation is induced
by temperature. There are three steps in the CPE process:
• solubilization of analytes for micellar aggregates – compounds
present in the original matrix bound to the micelles;
• clouding – cloudiness of the solution occurs and is described as
the cloud point (CP), which results from the scattering of visible
light that passes through the solution, and is the reason for the
presence of micellar aggregates; and,
• phase separation – after exceeding the temperature limit, known
as the cloud-point temperature (CPT), the solution separates into
two phases.
The CPT is a characteristic value of the surfactant. It can vary from
low (4.5°C) to quite high (100°C) values, depending on the surfac-
tant used (generally, for non-ionic surfactants, the temperature is
lower and, for zwitterionic surfactants, higher). CPT depends on the
concentration of the surfactant solution [22]. However, the pres-
ence of additives, such as salts, alkalis, acids, polymers, urea and
other surfactants, can modify (lower or raise) the CPT values [16].
Coacervates are also the extraction medium in another extrac-
tion technique, called coacervative extraction (CAE), which is often
treated in the literature as CPE, because both extraction techniques
are based on using the same physical and chemical processes. Fig. 3
shows the courses of these processes (CPE and CAE). The difference
between them lies in the conditions required to induce phase sep-
aration. In CAE, the coacervative phase forms as a result of the influence
of factors other than temperature (e.g. in the presence of salt or alcohol)
or it can be induced by pH [23]. Because of this, in the CAE tech-
nique, cationic and anionic surfactants can be used. In charged micelles,
electrostatic repulsion prevents phase separation. Only under the in-
fluence of the desolvation factor (e.g. by adding high concentrations
of salts or in the presence of hydrophobic counter ions), long-tailed
cationic surfactants self-assemble in aqueous solution into long, flex-
ible micelles. The formation of these long, worm-like micelles is more
favored, with an increase in the alkyl-chain length on the surfactant
head-group due to enhanced hydrophobic interactions [24]. For
example, cationic surfactants [e.g. hexadecyltrimethylammonium
bromide (CTAB)] undergo phase separation only at very low pH (1.0
optimal) [25] or high ionic strength (~40 kJ/mol) [24].
Apart from extraction using a single surfactant, mixtures of sur-
factants result in increased extraction efficiency. An extraction system,
called aqueous surfactant two-phase (ASTP), which uses mixtures
of cationic and anionic surfactants, is a separation technique offer-
ing properties superior to techniques using single surfactants because
of an increase in the effectiveness of extraction. In this technique,
after mixing dilute solutions of anionic and cationic surfactants, flat
bi-layer structures, such as vesicles, are formed and separation of
the solution occurs for two defined phases. As opposed to CPE and
CAE, phase separation occurs only at fairly limited compositions and
surfactant concentrations: concentrations of both surfactants must
be higher than their CMC, and the two surfactants must be in an
appropriate molar ratio to each other. However, ASTPs can be pre-
pared without any additives. The effectiveness of the extraction
procedure depends on only composition or concentration of the sur-
factants used. The ASTP extraction system is very promising due to
its potential application in wastewater treatment and separation of
charged substances, such as dyes or biomaterials [26].
3. Applications of coacervate-based techniques for sample
preparation
The wide range of surfactant properties resulted in surfactants
becoming an important area of science and technology. The
286 A. Melnyk et al./Trends in Analytical Chemistry 71 (2015) 282–292
Fig. 3. Extraction procedures using coacervates.
undeniable advantage of coacervate-based extraction techniques is
the low consumption of organic solvents used. Also, the economic
advantage, with reduced labor and time-consuming procedures, is
worth mentioning, because of the elimination of certain opera-
tions from sample preparation for analysis, which are necessary in
conventional extraction techniques, such as:
• an additional step of enrichment [e.g. using solid-phase extrac-
tion (SPE)];
• evaporation of the excessive amount of solvent [e.g. under a
stream of nitrogen]; and,
• an additional clean-up step for the extract.
The advantages of both preparation techniques include:
• the non-toxic character of surfactants;
• the significantly smaller amount of chemicals used, including
organic solvents for one analytical cycle, compared with con-
ventional extraction techniques, such as SPE and the liquid-
liquid extraction (LLE);
• the simultaneous extraction of several samples;
• the short time to reach extraction equilibrium, often only a few
minutes;
• the possibility of conducting extraction under mild conditions,
lowering energy consumption;
• the elimination of the problem of emulsion formation;
• the possibility of analyte extraction from solid and liquid samples,
characterized by the matrix being complex (plasma, beer, sludge,
and plants);
• the high extraction efficiency (high recovery values, high con-
centration factors, low values of LODs and LOQs);
• simple combination with analytical instruments used in the
sample analysis of extracts (e.g. atomic absorption, chromato-
graphic, and electrochemical analyses) [16].
As a result, coacervates carry out an extraction procedure that
is simple, cost-, energy- and time-saving, and environment-
friendly, and for analytes with wide ranges of polarity and charge.
At present, surfactants are used in molecular and atomic ab-
sorption and emission (luminescence) spectroscopy, potentiometry,
voltammetry, titrimetry, chromatography (thin-layer, high-
performance liquid, ion, affinity, supercritical fluid, and gel
permeation), extraction, flotation, ultracentrifugation, and capil-
lary and gel electrophoresis, and other methods for determination
and separation of organic and inorganic substances.
Coacervates are increasingly used in the determination of toxic
compounds and heavy elements at trace levels, including organic
compounds in different types of sample, often with complex ma-
trices. Obtaining good analysis parameters (high recoveries and low
LOD and LOQ values) is possible due to the high concentration of
the surfactant in some coacervates (up to 1 mg/μL). Moreover, as
there is no need to evaporate solvents after extraction, there is no
loss of analytes [16].
Table 2 lists works concerning preconcentration of organic com-
pounds by coacervative-based extraction techniques.
Coacervates were successfully used for determination of estro-
gens [estrone (E1), 17β-estradiol (E2β) and diethylstilbestrol (DES)]
in human urine. Determination methods of estrogens in urine are
characterized by poor interlaboratory reproducibility and limited
cross-reactivity. The procedure proved to be very simple and ef-
fective for their determination. Tergitol TMN-6 was used as the
surfactant. An aliquot of 0.5 mL of 10% (v/v) of the surfactant so-
lution was added to 10 mL of the urine sample and incubated for
45 min in a thermostatted ultrasonic cleaner at 45°C and an ultra-
sonic frequency of 35 kHz. To induce phase separation, 0.2 g NaCl
was added and the mixture was centrifuged at 4000 rpm for
3 min. The coacervative phase obtained was diluted to 1.0 mL
with acetonitrile and analyzed using high-performance liquid
chromatography/diode-array detection (HPLC-DAD). Dilution of co-
acervate with a solvent decreased the viscosity of the sample injected
into the feeder, prolonging the lifetime of the chromatographic
column. The relative standard deviations (RSD, n = 5) were below
5.3%. The LODs were 0.1 ng/mL, 0.2 ng/mL and 0.1 ng/mL for E2β,
E1 and DES, respectively, and are lower than LODs obtained with
other commonly used sample-preparation techniques for the de-
termination of estrogens in human urine samples – LODs obtained
 A. Melnyk et al./Trends in Analytical Chemistry 71 (2015) 282–292 287

Table 2
Information on the use of coacervate-based extraction techniques for the preconcentration of organic compounds

Sample type Analytes Surfactants Conditions driving Determination LOD [ng/mL]a Recovery [%] Ref.
phase separation technique

biological aristolochic acids Genapol X-080 NaCl, T 50°C HPLC-UVb 10 94.5–105.4 [27]
curcumin Triton X-100 pH 4.5, T 70°C HPLC-UV 66 95.9–100.5 [28]
doxazosin, alfuzosin PONPE 7.5c, SDSd NaCl, pH 3.0 (alfuzosin) FLe 0.16 (alfuzosin) 96.73–104.24 [29]
or 4.0 (doxazosin) 0.21 (doxazosin)
drugs Triton X-114 pH 12.0, T 40°C HPLC-DADf 250–500 25.2–107.9 [30]
fluoroquinolones PONPE 7.5, SDS NaCl, pH 4.0, T 25°C FL 0.04–0.06 95.61–101.24 [31]
warfarin Tergitol 15-S-7 Na3PO4, pH 7.0, T 60°C FL 3.3 × 10−10 [mol/L] 94.8–105.0 [32]
food and drinks allura red CTAB, Triton X-114, KCl, pH 2.5, T 60°C UV-VISg 7.8 96.5–106.1 [33]
Triton X-100
carmoisine, brilliant Triton X-100 NaCl, pH 5.0, T 76°C UV-VIS 17 (carmoisine) 96–104 [34]
blue FCF 16 (brilliant blue FCF)
formaldehyde Triton X-114 NaCl, pH 4.0, T 60°C HPLC-UV 0.7 92.3–102.8 [35]
herbicides Triton X-114 pH 2.5, T 60°C CZEh 4–18 94.9–98.3 [36]
organophosphates Triton X-114 pH 2.0, T 85°C GC–MSi 0.03–0.47 [ng/g] 90–107 [37]
pesticides
phenolic fatty alcohol (NH4)2SO4, T 40°C HPLC-DAD 3.2–9.8 89.4–103.5 [38]
antioxidants polyoxyethylene ether-9
sulfonamides Triton X-100 Na2SO4, T 50°C HPLC–UV 2.23–9.79 67.0–105.7 [39]
xanthohumol Triton X-114 NaCl, pH 5.0, T 70°C HPLC–UV 3 90.7–101.9 [40]
pharmaceuticals ergotamine PONPE 7.5 pH 8.5 CEj 0.11 × 10−3 96.17–103.8 [41]
plants auxins Triton X-114 pH 5.0, T 40°C CE-ECLk 2.5 and 2.8 [nM] 89.7–111 [42]
bergenin Triton X-114 NaCl, T 70°C HPLC-UV 650 87.2 [43]
flavonoids Genapol X-080, CTAB NaCl, pH 8.0, T 55°C HPLC-UV 1.2–5.0 94.2–98.8 [44]
polycyclic aromatic Genapol X-080 NaCl, T 70°C HPLC-UV 0.01–0.05 67–107 [45]
hydrocarbons
saponins Triton X-100 Na2CO3, T 50°C UV-VIS n.m.l 98.4 [46]
water diazinon CTAB KI, T 35°C UV 0.02 85.0–93.6 [47]
naphthols Triton X-114 pH 3.0, T 30°C CZEg 0.20–0.24 92.43–103 [48]
phthalate esters decanoic acid, pH 2.0 HPLC-UV 0.22–0.30 87–94 [49]
tetrahydrofuran
triazole fungicides polyethylene glycol 600 Na2SO4, T 45°C HPLC–UV 0.0068–0.0345 82.0–96.0 [50]
monooleate
a Otherwise different unit is given.
b HPLC-UV, High-performance liquid chromatography with photometric detection.
c
PONPE 7.5, Polyoxyethylene(7.5)nonylphenylether.
d SDS, Sodium dodecyl sulfate.
e FL, Fluorimetry.
f HPLC-DAD, High-performance liquid chromatography with diode array detector.
g
UV-VIS, Ultraviolet-visible spectrophotometry.
h CZE, Capillary zone electrophoresis.
i GC-MS, Gas chromatography-mass spectrometry.
j
CE, Capillary electrophoresis.
k
CE-ECL, Capillary electrophoresis-electrochemiluminescence.
l n.m., not mentioned.

with SBSE-liquid desorption (LD)-HPLC/DAD, LC-tandem mass spec-
trometry (MS/MS) and molecularly-imprinted solid-phase extraction
(MISPE)-HPLC/DAD were 0.3–10 ng/mL, 0.1–0.5 ng/mL and 0.47–
1.26 ng/mL, respectively [51].
For many years, coacervate-based extraction techniques have been
used for extraction of polycyclic aromatic hydrocarbons (PAHs),
which are ubiquitous environmental contaminants. For many of these
compounds, their genotoxicity and mutagenicity were confirmed,
so PAHs are classified as priority pollutants by the US Environmen-
tal Protection Agency (US EPA). Preconcentration of PAHs in
environmental samples using conventional methods involves
large amounts of potentially hazardous organic solvents, is
time-consuming and requires an additional clean-up step for the
extracts. Coacervate-based techniques have been used for the
preconcentration of PAHs in different types of sample and achieved
low LODs – ng/L per to even sub-ng/L, in some cases. Recently, for
determination of four PAHs {benzo[a]anthracene, chrysene,
benzo[b]fluorantene and benzo[a]pyrene}, López-Jiméne et al. pro-
posed vesicles made of octanoic acid/tetrabutylammonium octanoate.
5 g of octanoic acid was mixed with 11.25 mL of 40%Bu4NOH and
40 mL of distilled water and the mixture was centrifuged at 3000 rpm
for 5 min. An aliquot of 200 μL of coacervative phase was mixed with
200 mg of food sample. The mixture was vortex-shaken at 2500 rpm
for 10 min, centrifuged at 7000 rpm for 10 min at room tempera-
ture, and the extract was analyzed by LC with fluorescence detection.
Recoveries of PAHs were over 92% and the LODs were 0.1–0.2 μg/kg
[52].
Liu et al. [53] recently described CPE/HPLC analysis of triazine
herbicides in milk, which had not been published before. The pro-
cedure included the use of Triton X-100 solution and was applied
for determination of four triazines (atrazine, cyanazine, simazine,
simetryn). Within the procedure, 10 mL of milk were mixed with
the surfactant, 40 μL of glacial acetic acid and 0.90 g of anhydrous
sodium sulfate, and diluted to 15 mL with pure water. The solu-
tion obtained was centrifuged at 9917 g for 10 min at 5°C. The
supernatant was filtered through a 0.45-μm membrane and ad-
justed to pH 5 with NaOH. An aliquot of 9 mL of supernatant was
incubated at 60°C for 30 min to induce phase separation. The
coacervative phase was diluted to 2 mL with methanol, centri-
fuged at 6953 g for 5 min, filtered through a 0.22-μm membrane
and analyzed by HPLC with a UV-vis detector. The procedure ap-
peared to be useful and efficient. The calibration curves of the four
triazines showed satisfactory linearity in the range 50–2000 μg/L.
LODs were 6.79–11.19 μg/L and average recoveries were 70.5–96.9%.
In the literature, more research relates to coacervate-based
microextractions, in which the amount of surfactant-rich phase is
288 A. Melnyk et al./Trends in Analytical Chemistry 71 (2015) 282–292
less than 100 μL. One such example was proposed in 2011 – a new
procedure for determination of parabens using solidified floating
vesicular coacervative drop microextraction (SFVCDME). The
coacervative phase was decanoic acid aqueous vesicles in the pres-
ence of tetrabutylammonium (Bu4N+). HPLC-UV was used for
determination. The SFVCDME method was used for the extraction
of four esters of p-hydroxybenzoic acid (methyl paraben, ethyl
paraben, propyl paraben and butyl paraben) from water samples
and commercial cosmetic products. The extraction process was
carried out for 30 min, and the analytes were extracted into the
30-μL coacervate droplet. The LODs for the analytes were
0.2–0.5 μg/L and the relative recoveries were 91.2–104.4% for water
samples and 92.6–110.7% for cosmetic products. The proposed
SFVCDME technique, due to its simplicity, sensitivity, analytical pre-
cision, low consumption of organic solvents, low cost and short
sample-preparation time, proved to be a very valuable tool for
microextraction of parabens from water samples and cosmetic
products [23].
Coacervates also proved to be very useful in determination of
metal ions from environmental samples. Conventional LLE tech-
nique used for determination of trace metals require large amounts
of sample and involve high volumes of solvents, so coacervate-
based extraction techniques are the leading techniques for metal-
ion detection in the samples. The CPE technique is characterized by
the lowest values of LODs for preconcentration of Co, Fe, Mn, Mo,
Pd and Zn. In comparison with organic compounds, metals require
for determination the use of a chelating agent to form a complex
with sufficient hydrophobicity to be extracted to the small volume
of the surfactant-rich phase.
Amjabi et al. [54] published on the use of coacervates made of
decanoic acid reverse micelles in a water/tetrahydrofuran mixture
for the determination of arsenic in water samples. Electrothermal
atomic absorption spectrometry (ETAAS) was used as the detec-
tion technique. Maximal absorbance signal was obtained in the
following conditions:
• 80 mg of decanoic acid in 10% (v/v) tetrahydrofuran to obtain
reverse micelles;
• 0.05 mol/L of O,O-diethyl dithiophosphate (DDTP) as a chelat-
ing agent for the extraction of As(III); and,
• 1.25% (v/v) HCl solution, as a medium to stimulate As(III)-
DDTP complex formation.
The volume of the coacervate obtained was 120 μL, and, in the
optimum conditions, equilibrium was reached after about 10 min.
The LOD was 0.04 μg/L and the enhancement factor was 64. Re-
coveries of As(III) were 98–102% in real water samples (subterranean
canal water, river water, and underground water).
In 2012, Meeravali and Kumar used mixed micelles for
preconcentration of Cd, Pb, Cu, Ni and Mn in effluents and natural
waters. In the procedure, 5 mL of sample solutions (with pH ad-
justed to 5.5) were mixed at room temperature with 0.2 mL of 10%
ammonium pyrrolidine dithiocarbamate (APDC), 0.4 mL of 10% Triton
X-114, 0.3 mL of 10% sodium dodecyl sulfate (SDS) and 2 mL of 30%
NaCl, and diluted to 10 mL. The surfactant-rich phase obtained was
made up to 0.5 mL by adding 0.3 mL of methanol and nitric-acid
mixture. With this method, LODs of 5 pg/mL, 10 pg/mL, 30 pg/mL,
50 pg/mL and 80 pg/mL were achieved for Cd, Mn, Cu, Ni and Pb,
respectively. Recoveries were 95–102% with 2–5% RSD [55].
Two years later, the same authors, together with two other re-
searchers, successfully used coacervates for preconcentration of
platinum, cisplatin and carboplatin. Water samples (2–5 mL) were
mixed with 1 mL of 10% tri-octyl methyl ammonium chloride
(TOMAC) and 0.5 mL of 10% Triton X-114, and filled up to 20 mL with
Milli-Q water at room temperature. The surfactant-rich phase was
diluted with 1 mL of methanol and analyzed using continuum-source
ETAAS (CS-ETAAS). Recoveries were 97–99%, 77–80% and 96–
110% for carboplatin, cisplatin and hexachloroplatinate, respectively.
The LOD was 0.02 ng/mL [56].
Table 3 summarizes other analytical characteristics of metal-
ion preconcentration using coacervates.
Very recently, coacervates were successfully used in
preconcentration and quantification of NPs. Nanotechnology in-
cludes the use of materials with dimensions of the order of 100 nm
or less. Use and production of NPs has been rapidly growing, since
they show a number of properties that make them interesting in
various fields, such as medicine, nutrition and energy. Applications
of metallic NPs in biomedical field are numerous due to their an-
tibacterial, antifungal, antiviral and anti-parasite properties. As a result,
nanotechnology is a rapidly developing area and increasing release
of NPs to the environment may cause a serious problem. The risk of
exposure and the toxicity of NPs are not exactly known [84].
Studies on preconcentration of NPs usually involve relatively large
amounts of organic solvent, complex and costly instrumentation,
and analyte loss, or do not allow for the quantification of particles
with diameter less than 20 nm. Hartmann and Schuster described
a simple, cost-effective separation technique for the determina-
tion of gold NPs (AuNPs) in aquatic systems [85]. AuNPs have a
unique ability to conjugate ligands, such as oligonucleotides, pro-
teins and antibodies, containing functional groups, such as thiols,
mercaptans, phosphines, and amines, and appear to be useful in rec-
ognition and detection of human cancer cells. Within the procedure,
to 40 mL of sample containing citrate-capped AuNPs, 400 μL of 0.1 M
HCl (achieved pH 3.1), 400 μL 1% citric acid and 800 μL of 10% Triton
X-114 were added. If species separation was needed, 400 μL of 1 M
sodium thiosulfate (TS) solution was also introduced to the sample.
The mixture obtained was vortexed and heated to 40°C and cen-
trifuged at 3000 rpm for 15 min. The coacervate formed a viscous
droplet attached to the bottom of the tube. The droplet was dis-
solved in 200 μL of ethanol and analyzed with ETAAS. With this
procedure, a low LOD (5 ng/L) was achieved. The enrichment factor
was 80 and the SD of 12 replicates at an AuNP concentration of
100 ng/L was 9.5%. Recoveries for spiked environmental samples
(river water and wastewater-treatment-plant effluent) were
91.1 ± 2.7% to 103.2 ± 6.9% [84].
In recent decades, there was rapid growth of silver NPs (AgNPs)
in commercial use. About 30% of consumer products with NPs
contain AgNPs. The production of these NPs is about 320 tons per
year. As a result, increased amounts of AgNPs can get into the
environment. The procedure of determination of AgNPs in envi-
ronmental waters spiked with antibacterial products was as follows:
0.02–1 mL of the sample, 0.1 mL of 1 M Na2S2O3 (or 0.1 mL of 3.5 M
NaNO3 to avoid the interference of S2O32-) and 0.2 mL of 10% Triton
X-114 were added to water with pH 3.0. The mixture was diluted
to 10 mL with water and heated for 30 min in 40°C. After that, the
solution was centrifuged at 2000 rpm for 5 min at room tempera-
ture. The coacervative phase formed (100 μL) was analyzed. For
identification, a UV-Vis spectrometer was used and, for quantifi-
cation, an ICP-MS technique was used. The LOQ value for AgNPs was
20 μg/L for aqueous suspensions and 10 μg/L for AgNO3 solutions
with an RSD less than 5%. Recoveries were 74.5–108%. This method
was an efficient separation method for speciation of AgNPs in en-
vironmental samples [86]. The extraction procedure proposed by
López-García et al. [87] involved 20 mL of sample solution, which
was heated to 80°C with 0.1 mL of a 0.1 mol/L nitric acid solution.
Aliquot of 0.5 mL of the Triton X-114 solution (30% w/v) was in-
corporated into the mixture, shaken for a few seconds and incubated
at 40°C, and, after 10 min, centrifuged at 4000 rpm for 5 min. 30 μL
of surfactant-rich phase were injected into ETAAS. The LOD 2 ng/L,
preconcentration factor 242 and recovery 96–105% indicated that
proposed methodology was a reliable analytical tool for discrimi-
nation AgNPs at very low concentrations in water samples.
 A. Melnyk et al./Trends in Analytical Chemistry 71 (2015) 282–292 289

Table 3
Information on the use of coacervate-based extraction techniques for preconcentration of metal ions

Sample type Metal Chelating agent Surfactants pH of buffer Temperature Determination LOD Recovery [%] Ref.
solution driving phase technique [ng/mL]a
separation [°C]

biological Al(III) PANb Triton X-114 8.0 40 GFAASc 0.06 92.3–94.7 [57]
Bi(III) 8-HQd Triton X-114 7.0 50 ICP-OESe 0.12 92.3–94.7 [58]
Cr(III) PAN Triton X-114 11.0 50 GFAAS 0.02 92.0–94.7 [59]
Cr(III) Magneson If Triton X-114 6.5 70 FAASg 2.3 103 [60]
Cu(II) 2-amino-4-(m- Triton X-114 4.5 50 ICP-AESh 1.2 99.73–100.51 [61]
tolylazo)pyridine-3-ol
(ATAP)
Sb(III), Sb(V) VPB(+)i Triton X-114 10.0 45 FAAS 0.25 Sb(V) 96.3–103.9 [62]
5.15 Sb(III)
food and drinks Cd(II) methyl green Triton X-114 5.0 50 FAAS 0.9 90.0–110 [63]
Co(II), Cu(II), 1-Phenylthiosemicarbazide Triton X-114 9.0 50 FAAS 0.10 [μg/g] (Co) n.d. [64]
Pb(II) (1-PTSC) 0.15 [μg/g] (Cu)
0.50 [μg/g] (Pb)
Cu (II), Fe(III) Eriochrome Cyanine R Triton X-114 4.5 (Cu) 70 FAAS 0.57 (Cu) 95.6–104.3 [65]
6.0 (Fe) 0.33 (Fe)
Fe(III), Pb(II) Magneson I Triton X-114 6.5 70 FAAS 2.5 (Pb) 95.2–101 [59]
1.9 (Fe)
Mg(II) 1,2,5,8- Triton X-114 8.5 50 UV-VISj 0.80 96.0–102.5 [66]
tetrahydroxyanthracene-
9,10-dione (quinalizarin)
Mo(VI) VPB(+) Triton X-114, 2.0 45 FAAS 2.18 97.5–102.3 [67]
CPCk
Se(IV) Pyronine B Ponpe 7.5, SDS 4.0 40 FAAS 3.81 94.0–101.8 [68]
Sn(II), Sn(IV) calcon carboxylic acid Ponpe 7.5, CPC 8.0 50 FAAS 2.86 97.4–102.3 [69]
(CCA)
geological Ag(I), Au(III), 4-(p-chlorophenyl)-1- Triton X-114 6.0 25 ETAAS 0.08 (Ag) n.m.l [70]
Pd(II) (pyridin-2- 0.30 (Au)
yl)thiosemicarbazide 0.12 (Pd)
(HCPTS)
As(III) APDC Triton X-114 4.5 35 ETAAS 0.025 97.3–98.8 [71]
Cr(III), Cr(VI) 1-(2-thiazolylazo)-2- Triton X-114 5.5 40 HPLC-UV 7.5 Cr(III) 96.4–104 [72]
naphthol (TAN) 3.5 Cr(VI)
Tl(I) - Triton X-114 - 90 CS-ETAAS 0.2 [ng/g] 95–102 [73]
pharmaceutical V(V) 8-HQ Triton X-114 4.0 45 ETAAS 0.042 98.5–99.0 [74]
formulations
tobacco As, Cd(II), APDC Triton X-114 6.0 (As) 50 ETAAS 0.021 (As) 98.1–99.2 [75]
products Ni(II), Pb(II) 6.0–7.0 (Cd and Pb) 0.22 (Cd)
5.5–6.5 (Ni) 0.47 (Ni)
1.04 (Pb)
water Au(III) - Triton X-114 - room ICP-MSm 0.0011 98.1 [76]
temperature
Cd(II), Co(II), 8-HQ Triton X-114 7.0 55 ICP-OES 0.01–0.34 90.0–110.0 [77]
Cu(II), Ni(II),
Pb(II), Zn(II)
Cd (II), Ni (II), PAN Triton X-114 8.0 60 FAAS 0.37 (Cd) 95–102 [78]
Zn(II) 2.3 (Zn)
2.6 (Ni)
Co(II), Cu(II), 2-(5-bromo-2-pyridylazo)- TritonX-114 9.0 50 FAAS 1.5 (Cu) 96–103 [79]
Ni(II) 5- (diethylamino) phenol 1.7 (Ni)
(5-Br-PADAP) 2.4 (Co)
Hg(II) 3-nitro benzaldehyde Triton X-114 8.0 55 ICP-OES 1.1 96.5–103.5 [80]
thiosemicarbazone (3-
NBT)
Rh(III) 5-(4′-nitro-2′,6′- Triton X-114 4.75 50 UV-VIS 0.15 99.00–100.75 [81]
dichlorophenylazo)-6-
hydroxypyrimidine-2,
4-dione (NDPHPD)
Se dithizone Triton X-114 1.0 (RS-CPEn) 50 UV–VIS 0.2 (RS-CPE) 93.1–105 [82]
0.8 (UA-CPEo) 0.3 (UA-CPE)
U(VI) P,P-di(2-ethylhexyl) CTAB, 3.5 room ICP-MS 0.003 95–107 [83]
methanediphosphonic acid Triton X-114 temperature
(H2DEH[MDP])
a Otherwise different unit is given.
b PAN, 1-(2-pyridylazo)-2-naphthol.
c
GFAAS, Graphite furnace atomic absorption spectrometry.
d 8-HQ, 8-hydroxyquinoline.
e
ICP-OES, Inductively coupled plasma - optical emission spectrometry.
f Magneson I, p-nitrophenylazoresorcinol.
g
FAAS, Flame atomic absorption spectroscopy.
h
ICP-AES, Inductively-coupled plasma - atomic emission spectroscopy.
i VPB(+), Victoria Pure Blue BO.
j UV-VIS, Ultraviolet-visible spectrophotometry.
k
CPC, Cetylpyridinium chloride.
l n.m., not mentioned.
m
ICP-MS, Inductively coupled plasma - mass spectrometry.
n
RS-CPE, Rapidly synergistic CPE.
o
UA-CPE, Ultrasound-assisted CPE.
290 A. Melnyk et al./Trends in Analytical Chemistry 71 (2015) 282–292
Copper(II)-oxide NPs (CuO-NPs) are widely used in sensors,
catalysts, surfactants, and antimicrobials. Translocation and bio-
transformation of CuO-NPs in plants can cause metal contamination
in the food chain. Under optimal conditions [i.e. 5% (w/v) Triton-
114, pH 9.0, temp. 40°C, ionic strength adjusted to 10 mM with 0.2 M
NaCl] and, with ICP-MS as a determination technique, the LOD was
0.02 μg/L. The enrichment factor was 82 and recoveries for water
samples were 59.2–108.2% [88].
An interesting approach is preconcentration of metal ions using
coacervates and NPs. For example AgNPs can be used for
preconcentration of chromium. During the extraction procedure
using coacervates, both chromium ions and AgNPs are transferred
from the surfactant-rich phase. In this case, AgNPs go beyond the
role of the complexing agent. In the absence of AgNPs, the extrac-
tion will not occur due to there being no transfer of metal ions into
the coacervate. The required concentration of AgNPs for complete
transfer of Cr(III) to the coacervate is at the μg/L level. This AgNP-
CPE–ETAAS technique detected low amounts of chromium in
samples, such as beer, wine and water, with an LOD of 2 ng/L and
recoveries of 95–105% [89].
Pourreza and Naghdi described a combined technique – cloud-
point-SPE by dispersion of TiO2-NPs in micellar media. The procedure
were as follows: 5 mg of TiO2-NPs was stirred for 1 min with 8 mL
of 5% (v/v) of Triton X-100. To the mixture obtained with dis-
persed and suspended TiO2-NPs were added a zinc solution, 1 mL
of phthalate buffer (pH 7) and 1 mL of 0.5 mol/L of NaCl. The so-
lution was diluted to 50 mL with water and kept in water bath at
80°C for 40 min. Phase separation occurred and, in the coacervate
phase, zinc ions adsorbed on the TiO2-NPs were present. Next, the
solution was cooled in an ice-bath for 5 min and the aqueous phase
was decanted. The surfactant-rich phase obtained was separated
from TiO2-NPs by centrifugation for 3 min. An aliquot of 500 μL of
5.85 × 10−4 mol/L of dithizone (chelating agent) was added to TiO2-
NPs phase and shaken for 2 min to desorb the zinc ions and to create
a complex. After centrifugation for about 3 min, Zn dithizonate was
collected and analyzed with a UV-vis spectrophotometer. Under
optimal conditions, the LOD was 0.33 μg/L, the RSD was 1.47% and
the enrichment factor was 80. This procedure was successfully
applied for determination of Zn2+ in tap water, powder milk and zinc-
sulfate tablets. Recoveries were 96–105% [90].
4. Limitations of coacervate-based techniques
Extraction techniques involving the use of surface-active com-
pounds have a number of advantages in comparison to other, much
more common analyte-extraction techniques. However, as other
techniques, they have their limitations that affect their usability:
• detection problems due to the extreme complexity of commer-
cial mixtures available for non-ionic surfactants;
• low extraction efficiency of non-ionic surfactants for polar
compounds;
• low numerical values for analyte enrichment;
• relatively low extraction efficiency for thermally-labile com-
pounds; and,
• the need for further treatment of the extract before final deter-
mination of the analyte.
It is necessary to remove completely the aqueous surfactant phase
from the micellar phase containing isolated analytes, which is
achieved by cooling the test tube and then decanting supernatant
or by evaporation under a stream of neutral gas (e.g. nitrogen). The
viscous, micelle-rich phase obtained has then to be diluted with
aqueous or organic solvent to enable injection into the analytical
instrument, significantly increasing use of reagents [91].
In coacervate-based techniques, optimization of the procedure
plays a significant role. A suitable pH determines coacervate for-
mation and extraction efficiency. Also, a suitable temperature is
crucial for phase separation to occur. The next important factor is
selecting a suitable concentration of surfactant. If the concentra-
tion is too high, the analytical signal deteriorates, because the final
volume of surfactant is too large; furthermore, when using lower
surfactant concentrations, conditions of analysis, such as accuracy
and reproducibility, deteriorate.
Disadvantages associated with the use of the coacervates also
apply to GC as a technique used for final determination of results.
However, if it is necessary to derive the analyte present in the
coacervative phase, there is still the possibility of contaminating the
dispenser and shortening the life of the column [92]. As a result,
the most commonly-used determination techniques are LC and UV-
vis and, in case of metal ions, AAS and ICP-MS.
A further major disadvantage of coacervate-based techniques is
that they are difficult to automate.
5. Conclusions and future trends
For the above analysis, we conclude that there are several pos-
sibilities of extending the range of applications for coacervate-
based extraction techniques. Modification of the extraction medium
is one possibility.
One example is the use of surface-active ILs (SAILs) as surfac-
tants. They have properties similar to traditional surfactants and can
assemble aggregates, such as micelles or vesicles [93]. In the litera-
ture, research on new SAIL systems is described {e.g. made of 1-butyl-
3-methylimidazolium tetrafluoroborate ([Bmim][BF4]) and an anionic
surfactant (sodium dodecylbenzenesulfonate, SDBS) [94] or 1-hexyl-
3-methylimidazolium tetrafluoroborate ([Hmim][BF4]) and cationic
surfactant (3-p-nonylphenoxy-2-hydroxypropyltrimethylammonium
bromide, NPTAB) [95]}.
In 2015, Dai et.al. used synthesized IL N-dodecyl-N-
methylpiperidinium bromide (C12MDB) as a cationic surfactant and
SDS to form the ASTP system [96]. Some of SAILs present even better
surface-active properties than traditional surfactants {e.g. N-butyl-
N-methylpyrrolidinium dodecylsulfate ([C4MP][C12SO4]}, a halogen-
free, low-cost, alkyl sulfate-based SAIL, has properties superior to
SDS [97]. Recently, so-called “catanionic” solutions, which are salt-
free mixtures of anionic and cationic surfactants in water, were
investigated. They show surface activity superior to the correspond-
ing single-chained SAILs and conventional anionic surfactant
analogues [93].
In recent years also, polymer/surfactant ATPS systems were ex-
plored. Due to the steric hindrance effect in the presence of polymer
molecules, two phases (surfactant-rich and polymer-rich) occur and
the form and the amount of the micelles in these phases depends
on the system temperature. Polyethylene glycol (PEG) as a hydro-
philic polymer and non-ionic surfactant is usually used to form the
two-phase system [98]. An interesting direction in this area is the
use of other polymer/surfactant systems.
Modifications of extraction techniques also involve reducing the
amounts of chemical reagents used. Although the amounts of sol-
vents used are relatively small compared to those of many other
techniques for sample preparation, there is a constant desire to
reduce them by developing new procedures, in which the volume
of the extraction medium is less than 100 μL. These techniques
include microextraction, in which the surfactant-rich phase has the
form of a droplet (the SFVCDME technique described above) or a
thin film.
There are developments in hand relating to modification of
coacervate-based techniques based on implementing derivatization
prior to chromatographic analysis, ultrasound-assisted or microwave-
assisted back-extraction or diverse chromatographic columns types
 A. Melnyk et al./Trends in Analytical Chemistry 71 (2015) 282–292
that would enable GC analysis of sample extracts and obtain even
better results.
Coacervate-based extraction techniques create a great alterna-
tive to routine preconcentration techniques, which usually involve
large amounts of chemicals. High recoveries and low LODs provide
excellent opportunities applying coacervate-based extraction tech-
niques to preconcentrate organic compounds, metal ions and NPs.
By achieving such results, coacervates can play an important role
in analytical chemistry.
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