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Speciation of mercury in liquid cosmetic samples by ionic liquid based
dispersive liquid–liquid microextraction combined with high-performance
liquid chromatography-inductively coupled plasma mass spectrometry
Xiaoyu Jia,ab Yi Han,ab Chao Wei,c Taicheng Duan*a and Hangting Chen*a
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Published on 17 March 2011 on http://pubs.rsc.org | doi:10.1039/C0JA00121J

Received 18th August 2010, Accepted 1st March 2011

DOI: 10.1039/c0ja00121j

Ionic liquid based dispersive liquid–liquid microextraction (IL-DLLME) combined with high
performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS)
for the determination of mercury species in liquid cosmetic samples is described. Firstly mercury (Hg2+),
methylmercury (MeHg+) and ethylmercury (EtHg+) were complexed with ammonium
pyrrolidinedithiocarbamate (APDC), and then the complexes were extracted into 1-hexyl-3-
methylimidazolium hexafluorophosphate ([C6MIM][PF6]) using DLLME. Under the optimized
conditions, the enrichment factors of 760, 115, 235 for Hg2+, MeHg+ and EtHg+ were obtained from
only 5.00 mL sample solution. The detection limits of the analytes (as Hg) were 1.3 ng L1 for Hg2+,
7.2 ng L1 for MeHg+ and 5.4 ng L1 for EtHg+, respectively. The relative standard deviation (n ¼ 10) of
0.5 ng mL1 Hg2+, MeHg+ and EtHg+ were 7.4%, 5.2% and 2.3%, respectively. Finally, the developed
method was successfully applied for the speciation of mercury in four liquid cosmetic samples, Hg2+ was
found as the main species in these samples, and a spike test was performed to verify the accuracy of the
method.

Introduction
Cosmetics are used worldwide by most of the population, and
these products are subjected to restrictive regulations, such as the
European Directive 76/768/EEC, in order to ensure their safety
and usefulness. Legislations include information requirements
about: (i) banned substances (these must not be found in the
finished product); (ii) restricted substances ; (iii) substances
subjected to positive listing (used in controlled concentration).1
Among these, all the compounds of Cd, Pb and Hg are pro-
hibited, because some of them might contain toxic substances to
achieve a certain specific efficiency. For example, mercury
compounds can spoil the leaven activity of the surface skin,
which consequently prevents the formation of melanin, so
mercury or mercury compounds were widely added in some
cosmetic products for whitening function. As we know, mercury
is considered one of the most toxic elements, and if cosmetics
containing mercury are used, it can enter the body through the
skin or directly through the mouth endangering one’s health
gradually. The toxicity of mercury is highly dependent on its
chemical forms, and alkylmercury compounds are deemed to be
a
National Analytical Research Center of Electrochemistry & Spectroscopy,
Changchun Institute of Applied Chemistry, Chinese Academy of Science,
Changchun, 130022, China. E-mail: tcduan@ciac.jl.cn; htchen@ciac.jl.
cn; Fax: +86-431-85262383; Tel: +86-431-85262017
b
Graduate School of Chinese Academy of Sciences, Beijing, 100039, China
c
National Institute of Metrology, Beijing, 100013, China
the most toxic among the various mercury species.2 Thereby,
monitoring the level of mercury, including each species in
cosmetics is of great importance to evaluate the risk of mercury
exposure to the user.
There are many analytical techniques reported for the speci-
ation of mercury, mostly are about chromatographic separation
combined with spectroscopic detection.3–6 For chromatographic
separation, high performance liquid chromatography (HPLC)
and gas chromatography (GC) have both been used for the
speciation of mercury in many previous studies, however,
because mercury species must be firstly derived to volatile
compounds with GC, HPLC was adopted in this work; while for
spectroscopic detection, inductively coupled plasma mass spec-
trometry (ICP-MS) was prefered, due to its high sensitivity and
rapidness. Hence, HPLC-ICP-MS has been used for mercury
speciation in various edible, biological and environmental
samples.7–14 As it is well known, the content of mercury species in
most samples is tiny, and maybe lower than the detection limits
of ICP-MS, thus, for samples containing very low levels of
mercury species, preconcentration of analytes is desirable prior
to HPLC-ICP-MS.
In many previous studies, various methods, including hollow
fiber liquid–liquid–liquid microextraction (HF-LLLME),15 solid
phase extraction (SPE),16 stir bar sorptive extraction (SBSE),17
single drop microextraction (SDME)18 and cloud point extrac-
tion (CPE)19 have been well used for the enrichment of mercury
species. Each of these methods had their unique advantages, but

1380 | J. Anal. At. Spectrom., 2011, 26, 1380–1386 This journal is ª The Royal Society of Chemistry 2011
 View Online

commonly, all of them were mild, which meaned that no trans- Table 1 Operating conditions of the HPLC-ICP-MS system
formation of species occurs during the sample treatment. In
Parameters Value
addition, none of the objective species should be lost, and the
enrichment methods should be compatible with the subsequent ICP-MS system
analytical techniques. RF power/W 1400
Further, a novel enrichment method which was named Nebuliser gas flow/L min1 0.94
Auxiliary gas flow/L min1 0.80
dispersive liquid–liquid microextraction (DLLME) was intro- Cooling gas flow/L min1 13.0
duced by Assadi and his co-workers20 in 2006. At first, it used Sampling cone/mm 1.0, Pt cone
a binary mixture of a water miscible solvent, named the disperser, Skimmer cone/mm 0.7, Ni cone
202
and a high density one with very low water solubility, referred as Isotope monitored Hg
Dwell time/ms 200
the extractant, to extract the objective compounds from aqueous Resolution 0.7 amu
samples.21–23 In these studies, toxic chlorinated organic solvents Singal acquisition Time resolved analysis
such as carbon tetrachloride and chloroform were generally used HPLC system
Column C18 (5 mm,150 mm  4.6 mm)
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as the extractants, which were harmful to the operator. Thereby,
the ‘‘green’’ solvent named ionic liquid gradually took the place
of organic solvents, since it exhibited excellent properties,
including low volatility, good chemical and thermal stability, and
good solubility with most organic solvents. Up to the present
time, IL-DLLME has been successfully applied to the enrich-
ment of various organic compounds and inorganic metals in
environmental and biological samples.24–28 It exhibited high
performance features such as rapidness, cost effectiveness and
high concentrating ability, and most importantly, this pre-
concentration method was environmentally friendly and suitable
for batch sample analysis. However, there have been few reports
on the use of this method prior to HPLC-ICP-MS, especially for
element species analysis.
In this work, we attempt to use IL-DLLME combined with
HPLC-ICP-MS for the speciation of mercury at very low levels
in cosmetic samples. Hg2+, MeHg+ and EtHg+ were chosen as the
targets. Key factors such as the selection of extractant and
disperser and their volumes, the amount of chelating reagent,
pH, the extraction and centrifugation time, salt effect and
interferences from coexisting ions were investigated. To evaluate
the feasibility of the method, spike tests were carried out.
Experimental
Apparatus
An X series II ICP-MS (Thermo Fisher Corp., USA) was used in
this study. The instrument was operated in the time-resolved
analysis (TRA) mode. The chromatographic system consisted of
a Waters 626 pump and a rotary injection valve fitted with
a 20 mL sample loop. Separation was achieved using an
XBridge C18 column (150  4.6 mm id, 5 mm, Waters, MA,
USA). The outlet of the LC column was directly connected to the
sample introduction system of ICP-MS via 50 cm of 0.18 mm i.d.
Peek tubing. The optimized ICP-MS and HPLC operating
conditions are summarized in Table 1.
A centrifuge (model TDL-40B, China) was used to accelerate
the phase separation during DLLME. The pH values were
measured with a PHS-3C pH-meter (Shanghai Precision &
Scientific Instrument Co., Ltd, China).
Standard solutions and reagents
Ultrapure water with a resistivity of 18.2 MU cm, obtained from
a Milli-Q Plus water purification system (Millipore, Bedford,
MA, USA) was used throughout the experiment. A stock
Mobile phase 0.05 mol L1 ammonium acetate
4% v/v methanol
10 mM L-cysteine
Flow rate of the mobile phase 1.0 mL min1
Sample loop volume 20 mL
standard solution of 1000 mg L1 Hg2+ was purchased from
CRM Information Center (Beijing, China). Stock standard
solutions of methylmercury (1000 mg L1, as Hg) were prepared
by dissolving CH3HgCl, which was purchased from Dr Ehren-
storfer (Augsburg, Germany) in HPLC methanol (Fisher
Scientific, USA). A stock standard solution of C2H5HgCl was
supplied by National Institute of Metrology (Beijing, China).
Working standard solutions were prepared by successive dilution
of the stock solution with ultrapure water.
Three ionic liquids including 1-hexyl-3-methylimidazolium
hexafluorophosphate ([C6MIM][PF6]), 1-butyl-3-methyl-
imidazolium hexafluorophosphate ([C4MIM][PF6]) and 1-octyl-
3-methylimidazolium hexafluorophosphate ([C8MIM][PF6])
were purchased from Shanghai Cheng Jie Chemical Co. Ltd.
(Shanghai, China). Ammonium pyrrolidinedithiocarbamate
(APDC), diethyldithiocarbamate (DDTC) and L-cysteine as
chelating reagents were purchased form Sinopharm Chemical
Reagent Co., Ltd. (Shanghai, China). 2 mg mL1 APDC and
L-cysteine were prepared by dissolving the compound in ultra-
pure water, 0.05 mol L1 of DDTC was prepared by dissolving
the compound in HPLC grade methanol. All other reagents,
including ethanol, acetone and acetonitrile as disperser solvent
were at least of analytical grade.
All the glass and plastic (polypropylene) vessels used were
soaked in 25% nitric acid for 24 h and subsequently rinsed at least
five times with ultrapure water.
Sample preparation
Three skin refresheners and one hand moisturizing lotion
samples were diluted ten-fold with ultrapure water to reduce
their viscosity, then each of them was filtered through
a membrane of 0.22 mm pore size, after that they should be
treated with the DLLME procedure immediately, if not, kept in
a refrigerator at 4  C. For spike-recovery tests, the three mercury
species were added prior to any sample treatment including
dilution and filtration.

This journal is ª The Royal Society of Chemistry 2011 J. Anal. At. Spectrom., 2011, 26, 1380–1386 | 1381

Extraction procedure
A sample solution (5 mL) containing Hg2+, MeHg+, EtHg+ and
30 mL of APDC (chelating reagent) was drawn into a 10 mL
screw-cap glass test tube with a conical bottom. 500 mL of
methanol (disperser solvent) containing 0.052 g of [C6MIM][PF6]
(extraction solvent) was then injected rapidly into the sample
solution. Quickly, a cloudy solution was observed, as fine
droplets were produced by the dispersion of the immiscible
extraction solvent in the aqueous sample, which greatly enlarged
the contact area between the extraction solvent and aqueous
phase. In this step, Hg2+, MeHg+ and EtHg+ reacted with APDC
and were extracted into the fine ionic liquid droplets. The mixture
was then centrifuged for 10 min at 4000 rpm, in this way, the
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dispersed fine ionic liquid droplets were sedimented at the
bottom of the test tube. The upper aqueous phase was removed
with a pipette, and the sedimented phase (about 8 mL) was dis-
solved in 60 mL methanol, of which 20 mL was injected into the
HPLC-ICP-MS system for separation and detection.
Results and discussion
Optimization of the mobile phase for HPLC
The optimization of the mobile phase for HPLC was according
to many previous studies, the aim is to shorten the separation
time and reduce the level of carbon and salt that enter the ICP
torch. Firstly, 0.1% 2-mercaptoethanol was used as the com-
plexing reagent in the mobile phase (containing 6% v/v methanol
and 0.06 mol L1 ammonium acetate), the results indicated that
the retention time for mercury species was too long (10 min for
MeHg+ and 12.5 min for Hg2+), which was undesirable for the
ICP-MS detection. Thus, 10 mM L-cysteine was tried as the
complexing reagent, the results showed that the retention time
for all the mercury species was greatly reduced (less than 6 min
for the three mercury species, when the concentration of meth-
anol and ammonium acetate were maintained constant).
Further, the concentrations of methanol and ammonium acetate
were also optimized, and the results showed that the retention
time was almost unchanged when the concentrations of meth-
anol and ammonium acetate were reduced to 4% v/v and
0.05 mol L1, respectively. Thereby, a solution containing 4% v/v
methanol and 10 mM L-cysteine with 0.06 mol L1 ammonium
acetate was chosen as the best mobile phase for this study. Fig. 1
illustrates the chromatographic separation of a mixed standard
solution containing Hg2+, MeHg+ and EtHg+.
Optimization of the DLLME procedure
Several key parameters, such as the types of extraction and
disperser solvent and their volumes, the amount of the chelating
reagent, pH value, the extraction and centrifugation time, salt
effect and interferences from coexisting ions were investigated
and optimized to obtain the best enrichment factor (EF), which is
calculated as the following equation.
Csed
EF ¼ (1)
Co
Where EF is the enrichment factor, Csed and Co are the
concentration of the analytes in the sedimented phase and the
1382 | J. Anal. At. Spectrom., 2011, 26, 1380–1386
View Online
Fig. 1 Chromatogram showing the separation of Hg2+, MeHg+ and
EtHg+. The concentration of each mercury species was 50 ng mL1 (as
Hg). The mobile phase contains 4% v/v methanol and 10 mM L-cysteine
with 0.05 mol L1 ammonium acetate.
initial concentration of the analytes in the aqueous solution,
respectively.
Effect of type and amount of ionic liquid. Generally, water
immiscible extraction solvents with density higher than water
were more favorable in DLLME. Three different ionic liquids,
including [C4MIM][PF6], [C6MIM][PF6] and [C8MIM][PF6]
were tested as extractants. The results showed that
[C6MIM][PF6] exhibited the highest extraction efficiency for all
the analytes among the three ionic liquids, as shown in Table 2.
Furthermore, the complex of inorganic mercury can only be
extracted by [C6MIM][PF6]. The reason might be that the
hydrophobic property of [C6MIM][PF6] is similar with the three
mercury complexes, thus, the analytes can be more soluble in
[C6MIM][PF6] when compared with the other ionic liquids.
Therefore, [C6MIM][PF6] was selected as the best extractant.
To investigate the effect of the extraction solvent amount on
the enrichment factor, solutions containing different amounts of
ionic liquid were subjected to the same DLLME procedure. The
experiments were performed by using 500 mL methanol con-
taining 30 mL APDC and different amounts of [C6MIM][PF6]
(0.026, 0.039, 0.048, 0.050, 0.052, 0.054, 0.056, 0.065, 0.078 and
0.091 g) in 5 mL aqueous sample spiked with 1.0 ng mL1 of each
analyte. The volume of the sedimented phase increased from 0 to
35 mL with the increase of the ionic liquid amount. The results
indicated that when the amount of the ionic liquid was less than
0.048 g, the cloudy solution could not be observed. And when the
amount of [C6MIM][PF6] increased from 0.048 g to 0.052 g, the
enrichment factor for the three mercury species firstly reached to
a plateau, then decreased with the increase of the ionic liquid
amount from 0.052 to 0.091 g. Thus, 0.052 g of [C6MIM][PF6]
was selected as the optimum amount.
Effect of type and volume of disperser solvent. Selecting an
appropriate disperser solvent which can be miscible in both the
extraction solvent and aqueous sample is of importance in
DLLME. For this purpose, acetone, acetonitrile and methanol
were studied as disperser solvents. A series of sample solutions
were studied by using 500 mL of each disperser solvent containing
This journal is ª The Royal Society of Chemistry 2011

Table 2 Enrichment factors of different ionic liquids
Enrichment factors
Analytes [C4MIM][PF6] [C6MIM][PF6] [C8MIM][PF6]
Hg-APDC — 760 —
MeHg-APDC 52 115 96
EtHg-APDC 120 235 203
0.052 g of [C6MIM][PF6]. The results indicated that with aceto-
nitrile as the disperser, the cloudy solution could not be observed
and no sedimented phase could form at the bottom of the conical
tube, whereas with acetone as the disperser, the enrichment
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factors were lower than those with methanol, it might be because
that APDC is more soluble in methanol, thus the aqueous sample
can be dispersed well with methanol. More importantly, meth-
anol is the optimal solvent to dissolve the ionic liquid sedimented
phase, and the mobile phase for the subsequent HPLC also
contained methanol, thus, methanol was selected as the best
disperser.
The volume of methanol as the disperser solvent should also be
optimized. It was found that in order to obtain a constant sedi-
mented phase volume, the volumes of the disperser solvent and
the extraction solvent should alter simultaneously. The experi-
mental conditions were fixed and included the use of different
volumes of methanol (400, 500, 600, 700 and 800 mL containing
30 mL of APDC and 0.050, 0.052, 0.053, 0.055 and 0.057 g of
ionic liquid, respectively). It appeared that the disperser solvent
volume of 400 mL was not enough for a good dispersion of the
extraction solvent, for the cloudy solution could not form well;
while the extraction efficiency decreased when the volume of
methanol exceeded 500 mL, the reason was that the metal–chelate
complexes were more soluble in water with the increase of
methanol, as shown in Fig. 2. Thus, in order to achieve the best
extraction efficiency, as well as to obtain a stable cloudy solution,
500 mL methanol was chosen as the optimum volume.
Fig. 2 Effect of the volume of methanol on the enrichment factor of
Hg2+, MeHg+ and EtHg+. Extraction conditions: sample solution volume,
5.00 mL; [C6MIM][PF6], 0.052 g; APDC, 30 mL; pH 6. Analytes spiked,
1.0 ng mL1.
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Fig. 3 Effect of the APDC solution volume on the enrichment factor of
Hg2+, MeHg+ and EtHg+. APDC concentration, 2 mg mL1;
[C6MIM][PF6] amount, 0.052 g; methanol volume, 500 mL; pH 6. Ana-
lytes spiked, 1.0 ng mL1.
Effect of type and volume of chelating reagent. Selecting
a chelating reagent which can chelate well with the analytes, for
them to be efficiently extracted into the extractant is very
important in DLLME. For this purpose, L-cysteine, DDTC and
APDC were used as the chelating reagents. The experiments were
performed by using 500 mL of methanol containing 30 mL solu-
tion of each chelating reagent. The results indicated that when
+
L-cysteine was used as the chelating reagent, the peak of EtHg
could not be observed, while for DDTC, the enrichment factor
was lower than that with APDC. The reason might be that the
complexes with APDC are more soluble in ionic liquid than that
with DDTC. So APDC was chosen as the chelating reagent.
The effect of the amount of APDC on the enrichment factor
was also studied. Different volumes of the APDC solutions
(2 mg mL1 in ultrapure water) in the range of 26–38 mL (at 2 mL
intervals) were added into the sample solutions and the experi-
mental results are shown in Fig. 3. As it reveals, the enrichment
factor increases with the increase of the volume of APDC until
Fig. 4 Effect of pH on the enrichment factor of Hg2+, MeHg+ and
EtHg+. [C6MIM][PF6] amount, 0.052 g; methanol volume, 500 mL;
APDC volume, 30 mL. Analytes spiked, 1.0 ng mL1.
J. Anal. At. Spectrom., 2011, 26, 1380–1386 | 1383

Table 3 Tolerable levels of coexisting ions
1
Matrix ions Tolerable levels/mg L
Nd2+ 10000
As3+, Al3+, Cr3+, Pt2+ 1000
F, Cl, Br 50000
SO42, NO3 50000
Fe3+, Zn2+, Pb2+, Bi3+, Au3+, Ti2+ 20
30 mL. The reason might be that the formed hydrophobic metal–
chelate complexes were partly dissolved in the excess of hydro-
philic APDC, which made them hard to be extracted into the
organic phase, thus, at larger amounts, the enrichment factor
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decreases; while at smaller amounts, the amount of APDC was
not enough to react with Hg2+, MeHg+ and EtHg+. Hereby,
30 mL of 2 mg mL1 APDC was optimum for this study.
Effect of pH. Hg2+, MeHg+ and EtHg+ in the aqueous phase
should firstly form hydrophobic complexes in order to be
extracted into [C6MIM][PF6]. Thus, to obtain the expected pre-
concentration effect, pH value plays an important role on the
metal–chelate formation and the subsequent extraction. To
investigate the pH effect, DLLME was performed at different pH
values (in the range of 4–8, which were adjusted by dilute HNO3,
NH3$H2O and CH3COONa). The results in Fig. 4 illuminate
that the enrichment factors for all the three species behaved
similarly, the EFs increased with the pH value to pH 6, then kept
almost constant, only a slight decrease can be observed. The
reason might be that the three mercury species can chelate well
with APDC at such acidity. Thus, pH 6 should be the best pH
value to obtain the highest enrichment factor.
Effect of the extraction time. The influence of the extraction
time was evaluated in the range of 0–20 min (at the intervals of 2,
5, 8, 10, 15, 20 min, respectively) with other experimental
conditions being constant. The results show that the extraction
time has no significant effect on the extraction efficiency. It is well
known that after the formation of the cloudy solution, the
interface area between the extraction solvent and the aqueous
phase is infinitely large. Thereby, the complexes of APDC-Hg,
APDC-MeHg and APDC-EtHg can transfer rapidly from the
aqueous phase into the extraction solvent. Thus, DLLME is
a very rapid preconcentration and separation technique. By
experiments, 2 min was found to be enough for a complete
extraction.
Effect of the centrifugation time. In this study, experiments
were performed by centrifuging for 3, 5, 8, 10, 12, and 15 min
respectively at 4000 rpm after extraction. With the centrifugation
time increased to 10 min, the volume of sedimented phase
increased from 3 mL to 8 mL, and then it kept almost constant
with longer centrifuging. Correspondingly, the enrichment
factors for the three mercury species also increased as the
centrifugation time increased until 10 min, and subsequently no
obvious change was observed until 15 min. Thus, 10 min was
selected as the final centrifugation time.
Effect of salt. The effect of salt concentration on the enrich-
ment factor was examined by adding different amounts of
1384 | J. Anal. At. Spectrom., 2011, 26, 1380–1386
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NaCl (0–6%, w/v) into 5.00 mL of the solutions spiked with
1.0 ng mL1 each of Hg2+, MeHg+ and EtHg+, while other
experimental conditions were kept constant. The experimental
results showed that no matter how much NaCl was added, the
solution mixture presented was uniformly transparent, and no
sedimented phase could form after centrifugation. The reason
might be that the solubility of the ionic liquid in the aqueous
phase increases with the increase of the ionic strength of the
aqueous solution, so it was more difficult to be dispersed by
methanol in the aqueous phase. Hence, in this study, no salt was
needed.
Effect of coexisting ions. The effects of common coexisting ions
on the recoveries of Hg2+, MeHg+ and EtHg+ were studied. For
this experiment, 5.00 mL of the solution containing 1.0 ng mL1
each of the three mercury species and various amounts of
interfering ions (as shown in Table 3) were treated with the whole
analytical procedure, and the recoveries of Hg2+, MeHg+ and
EtHg+ were in the range of 89.7%–108.3%, which meant that the
interference from the matrices and co-existing ions can be
ignored.
Figures of merit and comparison with other methods
The analytical characteristics of the optimized method are
summarized in Table 4, including the linear range, the limit of
detection, reproducibility and EFs. Three replicate measure-
ments were performed for each concentration level. The cali-
bration graph was linear in the range of 0.005–2 ng mL1 for
Hg2+, 0.01–2 ng mL1 for MeHg+ and EtHg+. The limit of
detection (LOD), defined as CL ¼ 3 SB/m (where CL is the limit of
detection, SB is standard deviation of the blank values and m is
the slope of the calibration graph), were 1.3 ng L1 for Hg2+,
7.2 ng L1 for MeHg+ and 5.4 for EtHg+, respectively. While the
EFs for the three species obtained from seven replicate
measurements of samples spiked with 1 ng mL1 of analytes
ranged from 115 to 760.
A comparison of the present method with other reported
methods is made and the results are listed in Table 5. Obviously,
this method has the distinct advantages of low detection limits,
high enrichment factors, short extraction time and low sample
consumption, and most importantly, it is environmental friendly,
since volatile toxic extractants are avoided. Therefore, IL-
DLLME combined with HPLC-ICP-MS is a simple, fast,
Table 4 Analytical characteristics
Analytical features
Parameters Hg2+ MeHg+ EtHg+
Linear range/ng mL1 0.005–2 0.01–2 0.01–2
Correlation coefficient (R2) 0.9986 0.9987 0.9999
Limit of detection/ng mL1 (3s, n ¼ 7) 0.0013 0.0072 0.0054
Repeatability (R.S.D.a, %) (n ¼ 10) 7.4 5.2 2.3
Enrichment factor (EF) 760 115 235
Sample volume/mL 5 5 5
Extraction time/min 2 2 2
a
The solution contained 1 ng mL1 each of Hg2+, MeHg+ and EtHg+.
This journal is ª The Royal Society of Chemistry 2011
 View Online

sensitive and environmentally friendly analytical strategy for the

This work
Reference
speciation of mercury in liquid cosmetic samples.

15
16
18
19
Analysis of real cosmetic samples

Procedure time/min
To test the applicability of the established method to real sample
analysis, four liquid cosmetic samples (named hand moisturizing
lotion, skin refreshener 1, skin refreshener 2 and skin refreshener
3, respectively) purchased form local markets were analysed. For
Hg speciation, 5 mL of each sample was firstly treated with the
>20
>7
25

20
proposed IL-DLLME technique, then was analysed by the
2 HPLC-ICP-MS method (the typical chromatograms of the three
species in skin refreshener 1 are shown in Fig. 5); and then the
Sample consumption/mL
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total Hg content in each sample was directly determined by ICP-

MS. For each sample, the extraction was repeated three times. As
Table 6 illustrates, Hg2+ was found as the main species in these
four samples, the content of MeHg+ was small and was only
found in hand moisturizing lotion and skin refreshener 1, EtHg+
100–500

was not detected in any sample, and the sum of the three mercury
3.8

species were almost equal to the content of total Hg. The accu-
12
25
5

racy of the method was verified by a spike-recovery test, 0.20 ng

mL1 of each mercury species were spiked into various cosmetic
1000 (Hg2+) 1000 (MeHg+) 1000 (EtHg+)

samples prior to any sample preparation. The results in Table 6

760 (Hg2+) 115 (MeHg+) 235 (EtHg+)

show that the recoveries for spiked analytes were between 86.7%
3 (Hg2+) 27 (MeHg+) 31 (EtHg+)

and 101.2%, demonstrating satisfactory accuracy of the estab-

lished method for determination of mercury species in cosmetic
120 (MeHg ) 215 (EtHg )

samples.
+

42 (Hg2+) 21 (MeHg+)
Enrichment factor

Conclusion
+

IL-DLLME was for the first time used for the enrichment of
mercury species prior to HPLC-ICP-MS, and the combination of
the two techniques results in a robust analytical method for
speciation of mercury at very trace levels in liquid cosmetic
samples. Distinct advantages over previous studies are observed:
high sensitivity, fast, environmentally friendly and low sample
consumption. Potentially, with suitable adjustments, this method
22.8 (Hg2+) 1.0 (MeHg+) 1.6 (EtHg+)
0.8 (Hg2+) 4.3 (MeHg+) 1.4 (EtHg+)

0.0013 (Hg2+) 0.0072(MeHg+)

0.004 (Hg2+) 0.01 (MeHg+)
3.8 (MeHg ) 0.7 (EtHg )
+

0.0054(EtHg+)
LOD/ng mL1
+
Table 5 Comparison with other methods

IL-DLLME-HPLC-ICP-MS
HF-LLLME-HPLC-UV

IL-SDME-HPLC-DAD
CPE-HPLC-ICP-MS
SPE-HPLC-AFS

Fig. 5 Chromatograms of mercury species in skin refreshener 1 (a)

Method

spiked with 0.20 ng mL1 of each analytes, (b) with no spiking, and (c) the
reagent blank. The separation conditions for HPLC were the same as in
Fig. 1.

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Table 6 Analytical results of mercury in cosmetic samples and spike-recovery tests

Concentration/ng mL1, mean  S.D.a Recovery (%)

Sample Hg2+ MeHg+ EtHg+ Total Hg b/ng mL1 Spiked c/ng mL1 Hg2+ MeHg+ EtHg+

Hand moisturizing lotion 0.093  0.008 0.032  0.006 ND 0.128  0.017 0.20 98.6 89.7 92.3
Skin refreshener 1 0.089  0.007 0.010  0.002 ND 0.106  0.011 0.20 101.2 87.1 90.4
Skin refreshener 2 0.115  0.010 ND ND 0.121  0.013 0.20 96.6 88.3 93.5
Skin refreshener 3 0.104  0.008 ND ND 0.108  0.009 0.20 97.1 86.7 88.2
a
Standard deviation (n ¼ 3). b Total content of Hg in cosmetic samples. c Spiked 0.20 ng mL1 of mercury species in cosmetic samples.

also can be applied to high sensitive speciation of other metals in 10 D. S. Vidler, R. O. Jenkins, J. F. Hall and C. F. Harrington, Appl.
cosmetic samples with similar matrices. Organomet. Chem., 2007, 21, 303–310.
Downloaded by Universidade Federal de Sao Joao Del Rei on 24 August 2011
Published on 17 March 2011 on http://pubs.rsc.org | doi:10.1039/C0JA00121J

11 I. Cattani, S. Spalla, G. M. Beone, A. A. M. Del Re, R. Boccelli and

Acknowledgements
This work was financially supported by National Natural
Science Foundation of China (20605021, 20975099), Key
projects in the National Science & Technology Pillar Program
during the Eleventh Five-Year Plan Period (2009ZX09308-006-
5) and Specific researches in the National Science & Innovation
Method (2010IM030400).
References
1 I. Lavilla, N. Cabaleiro, M. Costas, I. de la Calle and C. Bendicho,
Talanta, 2009, 80, 109–116.
2 J. G. Chen, H. W. Chen, X. Z. Jin and H. T. Chen, Talanta, 2009, 77,
1381–1387.
3 R. Ito, M. Kawaguchi, N. Sakui, H. Honda, N. Okanouchi, K. Saito
and H. Nakazawa, J. Chromatogr., A, 2008, 1209, 267–270.
4 B. Jackson, V. Taylor, R. A. Baker and E. Miller, Environ. Sci.
Technol., 2009, 43, 2463–2469.
5 K. J. Chen, I. H. Hsu and Y. C. Sun, J. Chromatogr., A, 2009, 1216,
8933–8938.
6 Y. G. Yin, Z. H. Wang, J. F. Peng, J. F. Liu, B. He and G. B. Jiang,
J. Anal. At. Spectrom., 2009, 24, 1575–1578.
7 S. C. Hight and J. Cheng, Anal. Chim. Acta, 2006, 567, 160–172.
8 B. Vallant, R. Kadnar and W. Goessler, J. Anal. At. Spectrom., 2007,
22, 322–325.
9 M. Wang, W. Y. Feng, J. W. Shi, F. Zhang, B. Wang, M. T. Zhu,
B. Li, Y. L. Zhao and Z. F. Chai, Talanta, 2007, 71, 2034–2039.
M. Trevisan, Talanta, 2008, 74, 1520–1526.
12 M. Lemes and F. Y. Wang, J. Anal. At. Spectrom., 2009, 24, 663–668.
13 J. S. dos Santos, M. de la Guardia, A. Pastor and M. L. P. dos Santos,
Talanta, 2009, 80, 207–211.
14 S. S. de Souza, J. L. Rodrigues, V. C. de Oliveira Souza and
F. Barbosa Jr., J. Anal. At. Spectrom., 2010, 25, 79–83.
15 L. B. Xia, B. Hu and Y. L. Wu, J. Chromatogr., A, 2007, 1173, 44–51.
16 J. Margetınova, P. Houserova -Pelcova and V. Kubaň, Anal. Chim.
Acta, 2008, 615, 115–123.
17 R. Ito, M. Kawaguchi, N. Sakui, N. Okanouchi, K. Saito, Y. Seto and
H. Nakazawa, Talanta, 2009, 77, 1295–1298.
18 F. Pena-Pereira, I. Lavilla, C. Bendicho, L. Vidal and A. Canals,
Talanta, 2009, 78, 537–541.
19 H. T. Chen, J. G. Chen, X. Z. Jin and D. Y. Wei, J. Hazard. Mater.,
2009, 172, 1282–1287.
20 M. Rezaee, Y. Assadi, M. R. M. Hosseini, E. Aghaee, F. Ahmadi and
S. Berijani, J. Chromatogr., A, 2006, 1116, 1–9.
21 R. Montes, I. Rodrıguez, E. Rubı and R. Cela, J. Chromatogr., A,
2009, 1216, 205–210.
22 M. Rezaee, Y. Yamini, S. Shariati, A. Esrafili and M. Shamsipur,
J. Chromatogr., A, 2009, 1216, 1511–1514.
23 T. A. Kokya and K. Farhadi, J. Hazard. Mater., 2009, 169, 726–733.
24 Y. Liu, E. C. Zhao, W. T. Zhu, H. X. Gao and Z. Q. Zhou,
J. Chromatogr., A, 2009, 1216, 885–891.
25 Q. X. Zhou, X. G. Zhang and J. P. Xiao, J. Chromatogr., A, 2009,
1216, 4361–4365.
26 M. H. Mallah, F. Shhmirani and M. G. Maragheh, Environ. Sci.
Technol., 2009, 43, 1947–1951.
27 P. Berton, E. M. Martinis, L. D. Martinez and R. G. Wuilloud, Anal.
Chim. Acta, 2009, 640, 40–46.
28 P. Berton and R. G. Wuilloud, Anal. Chim. Acta, 2010, 662, 155–
162.

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