933

Full Paper

Application of a Carbon Paste Electrode Modified with a Schiff

Base Ligand to Mercury Speciation in Water
Montserrat Colilla,a M. Antonia Mendiola,b Jesús R. Procopio,a M. Teresa Sevilla*a
a
Departamento de Qu
mica Anal
tica y Análisis Instrumental, Universidad Autónoma de Madrid, E-28049 Madrid, Spain
*e-mail: teresa.sevilla@uam.es
b
Departamento de Qu
mica Inorgánica, Universidad Autónoma de Madrid, E-28049 Madrid, Spain

Received: July 22, 2004

Accepted: October 21, 2004

Abstract
A carbon paste electrode, modified with benzylbisthiosemicarbazone is used for mercury speciation in water samples.
Mercury ion is selectively accumulated on the electrode surface at open circuit and its analysis was performed by
cyclic voltammetry or square-wave voltammetry (SWV). A detection limit of 8 mg L
1 (3s) was found for 15 min of
accumulation using SWV as measurement technique. The effect of several metallic ions and organic substances on
voltammetric signal is examined. For speciation purposes, a ligand competition methodology between ligands in
solution and electrode is used. Model mercury complexes are characterized as a function of their dissociation kinetics.
The method was applied to mercury speciation in water samples from the Jarama River in Madrid.

Keywords: Mercury, River water, CPE modified with Schiff base ligand, Cyclic voltammetry, Square-wave
voltammetry

1. Introduction
Mercury is a very toxic element due to its action as
enzymatic blocking agent and affinity for thiol groups. The
association of inorganic mercury with the natural compo-
nents in soils, sediments or waters is conducive to forming
complexes, specially with organic matter or sulfide groups,
that change the characteristics of toxicity respecting free ion
[1]. For these reasons analytical tools for determination of
this element and its chemical forms in environmental
samples at low concentration levels are necessary. Different
methods based principally on atomic techniques, with
adequate detection limits, can be used for determination
of total mercury [2, 3]. For determination of organic and
inorganic mercury species, high performance liquid chro-
matography or gas chromatography coupled with adequate
detection systems are used [1, 3, 4].
To distinguish the different complex species of the metals
in environmental samples, such as natural waters, are
necessary analytical methods that no exert alteration of
the samples, because the transformation of these species is
very easy. Electroanalytical methods are useful for discrim-
ination of metal species and a very interesting tool in
speciation studies in aquatic systems [5]. The lower detec-
tion limits of the stripping techniques as anodic stripping
voltammetry (ASV) allow the determination of different
metal species at very low concentrations in this kind of
samples. Using ASV techniques with chemical modified
electrodes (CMEs), sensitive and selective methods can be
developed [6, 7].The modified carbon paste electrodes
(MCPEs) are easy to prepare and renew the electrode
surface. For mercury determination have been used MCPEs
containing ion exchangers [8], humic acids [9] or ligands with
different heteroatoms [10, 11]. In our laboratory, we have
performed speciation schemes using MCPE voltammetric
measurements based on kinetic dissociation of natural
copper complexes [12, 14]. These schemes can be extended
to speciation studies of other transition metals. Also we have
developed a method for mercury speciation with a CME
based on the selective response of these species in the
voltammetric measurement step [15].
Schiff base macroligands derived from thiosemicarbazide
and their complexes are the great interest for their
pharmacological properties as antibacterial and anticancer
agents [16 – 20]. These ligands are suitable for complexing
metals because it has donor atoms (nitrogen and sulfur)
which bond to mercury, copper, cadmium and zinc. In
previous work, we have been interested in the synthesis and
characterization macroligands containing thiosemicarba-
zone moieties and their complexes with transition metals
[21 – 27] in order to their use as modifier in chemically
modified carbon paste electrodes. So copper complexes with
benzylbisthiosemicarbazone have been applied in the
development of potentiometric copper sensors [28]. Other
macrocyclic thiohydrazone ligand has been successfully
used in a modified carbon paste electrode for voltampero-
metric determination and speciation of copper in water
samples [14].
In this work, we explore the capability of a thiosemicar-
bazone, benzylbisthiosemicarbazone (BTC, Figure 1) as
modifier of a carbon paste electrode for mercury determi-
nation and the ability of such electrode to perform metal
determination and speciation studies. This ligand presents
reactivity with mercury(II) ions [22, 27] and other ions such

Electroanalysis 2005, 17, No. 11  2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim DOI: 10.1002/elan.200403198
934
Fig. 1. Structure of benzylbisthiosemicarbazone (BTC).
as cobalt(II), zinc(II), cadmium(II) and copper(II) [22, 24,
25, 27]. The ligand is water insoluble, but it is light soluble in
alcohols and mineral oil and high soluble in N,N’ dimethyl-
formamide and dimethylsulfoxide.
2. Experimental
2.1. Reagents
Ultra pure water, obtained with a MilliRo-MilliQ system
(Millipore, USA) and analytical-grade reagents were used
throughout. The thiosemicarbazone of open chain benzyl-
bisthiosemicarbazone (BTC) was synthesized form benzyl
and thiosemicarbazide as described previously [21]. Humic
acids (HA, Aldrich, 600 – 1000 g mol
1) were dissolved in a
minimum amount of 0.001 mol dm
3 KOH until no pH
variation was observed and then filtered off a 0.45 mm
membrane filter (Durapore, Waters, USA). Stock standard
solutions, 0.01 mol dm
3 in concentration, of nitrilotriacetic
acid (NTA) (Merck, Germany), glycine (GLY), l-cysteine
(CYS), Urea (U), thiourea (TU) and salicylic acid (SAL)
(Fluka, Switzerland) were prepared in water. Iron colloids
(COL) were prepared as described previously [29]. The
100.0 mg L
1 mercury standard solution was prepared by
dissolution of the adequate amount of nitrate salt in water
acidulated with nitric acid at pH 2. All solutions were stored
in glass acid washed containers at 4 8C in the dark.
Solutions of model complexes containing mercury ion,
0.01 mol dm
3 KNO3 and the selected ligand concentration
were prepared. The pH of these solutions was adjusted
between 6.8 and 7.5 by using KOH or HNO3 solutions and
allowed to equilibrate for 24 h before their measurement.
2.2. Apparatus
A mAutolab (Ecochemie) voltammograph was used for
voltammetric studies. An Ag/AgCl reference electrode and
a platinum counter electrode were used. The total metal
concentrations in the water samples were measured by
flame and graphite furnace atomic absorption spectrometry
(AAS) using a Hitachi Zeeman Z-8200 Atomic Absorption
Spectrometer. Total mercury determination in samples was
performed by atomic fluorescence spectroscopy using a
PSA Merlin Fluorescence system. For decomposition of
organic matter from water samples, a Metrohm UV
Digester model 705 (Switzerland) was employed.
Electroanalysis 2005, 17, No. 11
M. Colilla et al.
2.3. Voltammetric Studies
The voltammetric studies of mercury and mercury-BTC
solutions on glassy carbon electrode were carried out in
0.01 mol dm
3 KNO3 supporting electrolyte containing
20% ethanol to keep ligand and complex in solution.
Before each measurement, the electrolyte was purged with
nitrogen for 5 min and a cyclic scan, between an initial
potential of
1.5 V and 1.0 V, was performed at a scan rate
of 100 mV s
1.
2.4. Carbon Paste Electrode Preparation
Chemical modified carbon paste was prepared by mixing
spectrographic graphite (> 40 mm diameter) and powdered
ligand in several w/w ratios in an agate mortar. Then paraffin
oil (Fluka, USA) was added and the mixture was mixed until
a uniform paste was obtained. The final graphite powered
and ligand: paraffin oil mass ratio was 5 : 2. The paste was
packed in the end (ca. 1 mm deep) of a PTFE tube (3 mm
i.d.) provided with an unmodified carbon paste-copper
contact. Appropriate packing of the carbon paste was
achieved by pressing the surface electrode against a filter
paper.
2.5. CMCPE Voltammetric Procedure
The mercury accumulation and measurement were ach-
ieved by the method of separate solutions. In the accumu-
lation step, the electrode was immersed in the accumulation
cell containing the mercury sample solution in 0.01 mol
dm
3 KNO3, stirred at 1000 min
1, at open circuit. After a
given accumulation time, the electrode was removed, rinsed
carefully with water and placed in the measurement cell
containing 0.1 mol dm
3 HCl as supporting electrolyte to
perform the measurement step. A constant potential of

0.60 V was applied for 20 s for metal reduction. For cyclic
voltammetric measurement, a potential sweep towards
positive values at a scan rate of 500 mV s
1 was then
performed causing the metal reoxidation leading to the
voltammetric signal. For square wave voltammetry, a scan
rate of 750 mV s
1, a potential increase of 8 mV and a
frequency of 94 Hz were used. The peak height, for all
mercury solution assayed, was plotted as a function of the
accumulation time. Applying a potential of 1.00 V during
1 min in the stirred supporting electrolyte did regeneration
of the electrode. The electrode was stored at the air when not
in use and in this conditions can be used during several
weeks without further surface renewal.
Prior to the mercury measurements, the new electrode
surface was activated by successive potential sweeps at
500 mV s
1 in the measurement electrolyte, followed by a
few accumulation-measurement cycles in the same condi-
tions and techniques that the mercury analysis procedure
described above, in 100 mg L
1 mercury solutions, until a
reproducible response was obtained.
 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Carbon Paste Electrode Modified with a Schiff Base Ligand
Fig. 2. Cyclic voltammograms of 1  10
3 mol dm
3 Hg(II) con-
taining BTC in concentrations 0 (—) and 1  10
3 mol dm
3 (– – –).
For conditions see text.
3. Results and Discussion
3.1. Voltammetry of Mercury in the Presence of BTC
The cyclic voltammetric behavior of mercury on glassy
carbon electrode, in the absence and presence of BTC in
solution, is shown in Figure 2. In absence of ligand, a
cathodic wave at
0.16 V, due to electrorreduction of
Hg(II) to Hg8, and an anodic peak at 0.25 V corresponding
to electrooxidation of Hg8, are observed. After adding 1 
10
3 mol dm
3 BTC into the cell containing 1  10
3 mol
dm
3 Hg(II), the cathodic peak is shift to
0.32 V whereas
the anodic peak increase in intensity. The peak at
0.32 V is
explained as the reduction of mercury complex to metal
mercury and free ligand.
The composition and overall stability constant of Hg(II)-
BTC was determined using the Lingane method [30]
measuring the potential shift observed in the mercury
reduction wave with the increase of ligand concentration.
The relation between potential shift (DEp) and the complex
stoichiometry, p, and stability constant, b, is expressed in
excess of ligand for Equation 1
DEp ¼ (RT/nF)(ln b þ p ln [L]t) (1)
A plot of DEp versus ln [L]t would give a straight line from
which p and b can be estimated. From results, shown in
Figure 3, a stoichiometry 1 : 1 L : M and a logb of 9.47 were
obtained.
3.2. Voltammetric Measurements on CMCPE
The parameters that influence the accumulation of metal on
electrode surface were studied to characterize how these
factors affect to the voltammetric signal of the metal. For pH
values lower than 5 (Figure 4) a decrease of mercury signals
is observed, indicating a weak acid behavior of the ligand on
the electrode surface. For pH greater of the indicated, a
Electroanalysis 2005, 17, No. 11
935
Fig. 3. Variation of DEp versus ln[L]t for evaluation of equili-
brium constant for the reaction of mercury with BTC in solution.
Fig. 4. Effect of accumulation solution pH on the voltammetric
peak height using cyclic voltammetry. Conditions: modified
percentage 40%, mercury concentration 100 mg L
1; accumulation
time: 2 min; reduction potential:
0.6 V; reduction time: 30 s; scan
rate: 500 mV s
1.
maximum voltammetric response of metal is observed, up to
pH values of 9. It was studied the effect of the ionic strength
of the medium on the signal, in potassium nitrate solutions in
concentrations between 10
4 and 0.2 mol dm
3, and no
significant variations in the signal and in the signal-to-noise
ratio were observed in the tested concentration range.
The response of modified carbon paste electrode can be
varied by changing the percentage of modifier incorporated
to carbon paste. Then the percentage of modifier was
studied using different electrodes containing percentages of
modifier between 10 and 50%. The electrodes were
activated of similar way until a reproducible response was
obtaining and were evaluated their better responses in
signal-to-noise ratio basis. As it was expected, the signal
increase when modifier percentage increase, up to a value of
the 40%. For higher percentage values, the signal-to-noise
ratio decrease and the electrode became less conductive and
unstable [14, 31]. These results can indicate that whereas the
metal is accumulated by complexation with the superficial
ligand, with the subsequent increase of the signal when
modifier percentage increase, the redissolution occurs from
 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
936 M. Colilla et al.

the carbon interface, showing a lower signal at lower carbon

percentages [14, 31].
As for the accumulation step, the parameters that affect to
the measure step were also studied. The voltammetric
measures were performed changing the working electrode
to a cell containing a supporting electrolyte. Various types
and concentrations of electrolyte solutions were tested in
terms of analytical signal and background, reproducibility
and electrode stability. The mercury voltammetric response
is optimum in mineral acid media, with sharp redissolution
waves and lower background currents. As measurement
medium a 0.1 mol dm
3 HCl solution was chosen because in
this electrolyte the highest analytical signal and reproduci-
bility, the lowest residual current and the easier electrode
regeneration are obtained.
The instrumental parameters in the measurement step Fig. 5. Typical SWV (– – – ) and CV (—) voltammograms
recorded using optimized experimental conditions for an accu-
were also tested. First the potential and the reduction time mulation time of 2 min. Mercury concentrations in both cases
were optimized. Reduction potentials between
0.1 and were 10 (lower line) and 50 (upper line) mg L
1.

0.8 V and reduction times between 2 and 60 s were tested.
The signal increases quickly when a more negative reduc-
tion potential is applied to the electrode. A stable and estimation of the constant of interaction between mercury
maximum value for peak height is reached at a potential and BTC on the electrode surface. This estimation was
higher than
0.6 V. Upon increasing the reduction time the carried out by the Kalcher method [14, 32] which considers
voltammetric signal increases. The highest peak current is that the equilibrium metal concentration on electrode
obtained at reduction times higher than 20 s. This last value surface is proportional to the measured voltammetric
was chosen in subsequent measurements. These negative current intensity, Ieq. Otherwise the total ligand concentra-
potential and reduction time are necessary for complete tion on the surface can be estimated through voltammetric
reduction of mercury from complex formed on the electrode measure saturating the surface of the electrode with the
surface. metal, Imax, by accumulation of the analyte during a
The following redissolution step was performed both by sufficiently long time. The difference between Imax and Ieq
cyclic voltammetry and square-wave voltammetry. For permits to determine the free BTC in the equilibrium on the
cyclic voltammetry (CV) the potential sweep was accom- surface electrode. According with these conditions, the
plished to scan rates varying between 100 and 600 mV s
1, stability constant for the complex formed on the surface
obtaining a linear increase in the peak intensity when the electrode can be expressed as
scan rate increase. This result corresponds to the oxidation
of species deposited on the electrode surface. The signal-to-
noise ratio keeps maxima and constant for scan rates lower ½HgBTCe Ieq
Keq ¼ ¼ ð2Þ
or equal to 500 mV s
1. We choose such scan rate to perform ½Hgs ½BTCe ½Hgs ðImax
Ieq Þ
the analytical measure. In square-wave voltammetry (SWV)
the studied parameters were scan rate, pulse height, scan where [Hg]s is the mercury concentration in the bulk of
increment and frequency. The optimized parameters, based solution. Reordering Equation 2 we have
in the best signal-to noise, were: pulse height, 40 mV;
frequency, 94 Hz; scan increment, 8 mV and scan rate, 1/[Hg]s ¼ (Keq Imax/Ieq)
Keq (3)
750 mV s
1. Figure 5 shows voltammograms obtained for
both measurement techniques at two concentration levels. So a plot of 1/[Hg]s versus 1/Ieq would give a straight line,
whose ordinate intercept is
Keq and its intercept with the
abscissa axis is 1/Imax. The Figure 6 reflects this situation (r ¼
0.997), and allowing from the value of the ordinate intercept
3.3. Characteristics of the Complex Formed on the
to estimate a value for log Keq of 6.94. This value is lower
Electrode Surface
than log b obtained in solution. The difference between both
The stoichiometry of the complex formed on the electrode values may result mainly from the different characteristics
surface can be suggested by the possibilities of ligand and conditions of the experiments.
coordination. These possibilities can be restricted by the
conditions in which the step of accumulation is accomplish-
ed: excess of ligand and aqueous solution with a moderately
3.4. Calibration and Interferences
basic pH value. As it is shown in the voltammetric studies on
glassy carbon electrode, the stoichiometry of the complex in Mercury calibration was performed in the optimum ana-
solution is 1 : 1. Based in this result, it was carried out an lytical conditions and the linearity of peak current versus

Electroanalysis 2005, 17, No. 11  2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Carbon Paste Electrode Modified with a Schiff Base Ligand 937

sweep is always good and allows mercury determination

without this interference mode.

3.5. Mercury Speciation Studies

Fig. 6. Variation of 1/[Hg]s versus 1/Ieq for evaluation of equili-
brium constant for the reaction of mercury with BTC on electrode
surface.
mercury concentration was checked. The linear working
ranges and the statistical data obtained for both measure-
ment techniques are reflected in Table 1. It can be observed
that the square-wave voltammetry presents a lower detec-
tion limit and greater linear range that to cyclic voltamme-
try. The limits of detection obtained with this sensor are
lower [11] or similar than the LoD observed for other
chemically modified electrodes described in the literature
[8, 9, 15]. The precision, expressed as relative standard
deviation, was also better for SWV (2.4%) than CV (4.8%)
for 12.5 mg L
1 Hg(II) and preconcentration time of 15 min.
Inorganic salts commonly present in natural water
systems were examined. No interference was observed in
aqueous solution of calcium and magnesium when present
in concentration lower than 100 mg mL
1. Carbonate does
not interfere for concentration lower than 0.01 mol dm
3.
Other electroactive cations, such as copper, zinc, lead and
cadmium were investigated. These metallic ions can also be
simultaneously incorporated at the electrode surface and
compete with mercury ions for the active groups, because
these metals can form complexes with BTC [22, 27]. The
variation of peak current for a 100 mg L
1 mercury solution
was investigated with the variation of interfering metal/
mercury concentration ratio. No significant signal variations
are observed in presence of metals for metal/mercury
concentration ratios up to 50. For interfering metal concen-
trations higher than indicated, a lower mercury linear range
was observed, because interfering ion compete with mer-
cury for the ligand. Although simultaneous reduction-
oxidation at the electrode surface occurs for the mentioned
electroactive cations, peak resolution during the reoxidation
In previous articles [12, 14] we presented copper speciation
studies in natural water based on voltammetric measure-
ments using chemically modified carbon paste electrodes.
The methodology employed in these procedures consists of
the comparison of metal uptake rate by the electrode in the
absence and in the presence of a competing natural ligand in
solution. Therefore, the speciation study is based on the
chemical competition between the ligand (the discrimina-
tor) attached to the electrode surface and the complexing
agents in the sample solution. The dissociation of a 1 : 1
complex formed in solution between a metal ion M and a
natural ligand Y, and the subsequent adsorption of M by
reaction with the discriminator L at the electrode surface,
Le, can be described by the general reactions
kf
MY $ M þ Y (4)
kd
ka
Le þ Ms
! MLe (5)
Where kd, kf and ka are the rate constant for dissociation and
formation of MY in solution and for formation of the adduct
on electrode surface, MLe, respectively. Assuming that,
under the experimental conditions, the complex MY is not
directly adsorbed and ka is fast compared with kd, the
accumulation of M on the electrode surface is only affected
by the speed of the dissociation of MY.
During the competing ligand reaction, the modifier
concentration at the electrode surface can be considered
constant and we can define the pseudo-first order reaction
rate at the initial step of the reaction for both cases by the
following equations
d½Me
v0 ¼ ¼ k0 ½Ms ð6Þ
dt
d½Me
v Y0 ¼ ¼ k Y0 ½MYs ð7Þ
dt
where v0 and v Y0 are the reaction rates, and k0 and k Y0 are the
pseudo-first order reaction rate constants, in absence and in
presence of the ligand Y in solution, respectively. The values

Table 1. Statistical treatment of calibration plot and detection limits ( LoD ).

Technique Accumulation time (min) Linear range (mg L
1) Sensitivity (mA/mg
1) LoD (mg L
1) r
CV 2 40 – 160 0.121 30 0.998
15 10 – 35 0.420 9 0.993
SWV 2 20 – 170 0.282 16 0.996
15 10 – 40 1.77 8 0.996

Electroanalysis 2005, 17, No. 11  2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
938 M. Colilla et al.

Table 2. Comparison of experimental kinetic data for various

complex and concentration ratios of mercury , Xexp, with the
values of calculated free metal fraction, Xcal, at experimental
conditions. Mercury concentration 100 mg L
1.
Ligand [ L] (mol dm
3) Xcal Xexp Kd (s
1)
NTA 10
5 0.584 0.654 0.930
10
3 0.013 0.043 0.390
TU 10
5 0 0.046 0.604
10
3 0 0 Inert
CYS 10
5 0 0 Inert
10
3 0 0 Inert
GLY 10
3 0.635 0.733 4.21
U 10
3 – [a] 1.0 Labile
HA [b] 2  10
6 – [a] 0 Inert
COL 10
7 [c] – [a] 0 Inert

[a] Without equilibrium constant data.

Fig. 7. Influence of natural ligands in solution in the variation of
peak height of voltammetric signal with the accumulation time.
Solutions contain 100 mg L
1 mercury, All solutions were 1 
10
5 mol dm
3 in ligand, except 1.0 mg L
1 HA and 1.8 
10
7 mol dm
3 in iron for COL: mercury alone (^), GLY (&), U
(*); SAL (~); NTA (&); HA (^), COL (*), and TU (~).
for v0 and v Y0 can be evaluated by means of the voltammetric
signal variation versus accumulation time. At low accumu-
lation times, the time-signal relationship will result in a
straight line which slope is related to the reaction rate. The
relative variation of v Y0 vs. v0 (variation of slopes) will allow
us to evaluate the importance of influence of a ligand in
kinetic terms, that is to say the metal availability.
The availability of metal at the discriminator at the time t,
X, can be defined as equal to [M]/[M]total ratio, where [M] is
the metal concentration bound to the discriminator at the
time t and [M]total is the total metal concentration in solution;
that is to say, X is the molar fraction of dissociated MY at
time t. The availability can be related to the dissociation rate
constant, kd, by means of the equation, if a first order
reaction is assumed
1/(1
x) ¼ exp(kdt) (8)
where t is the time scale of the measurement [5]. In
voltammetric measurements made in stirred solutions, as
is the case, the parameter controlling the measurement time
is the effective diffusion layer thickness d [33].
t ¼ d 2/(p DHg) (9)
Under these conditions, the measurement time can be
considered as the time allowed for MY to diffuse to the
electrode, to dissociate and to be adsorbed on the electrode
surface. The value of d can be determined by striping
voltammetry of mercury means the equation
DHg te ½Hg
d ¼ 0:29 n2 F 2 Av ð10Þ
RTIp
[b] Mr 1000.
[c] mol Fe L
1.
were A is the effective area of electrode, v the scan rate, te the
accumulation time, Ip the peak intensity and the others
parameter have their habitual meaning. The effective area
of electrode was estimated by cyclic voltammetry of
potassium ferrocianide, Fer, in 0.1 mol L
1 KCl, on the
modified electrode. The effective area is related with the
peak current by the equation
A ¼ Ip/2.7  105 n2/3(DFerv)1/2 [Fer] (11)
A d of 14 mm and a measurement time of 0.078 s, for used
stirring conditions, can be estimated. The kd values of
studied complexes can be estimated substituting the calcu-
lated labile fraction and measurement time in Equation 8.
Solutions of 100 mg L
1 mercury in the complexing media
described, containing 0.01 mol dm
3 potassium nitrate, at
neutral pH, were checked by the speciation method
proposed. Figure 7 shows a typical Ip vs. time plots for a
mercury solution in absence and in presence of selected
ligands. The results obtained for available mercury by
CMCPE are compared in Table 2 with the calculated
noncomplexed metal fraction (Xcal) in the solution con-
ditions by means MINEQLþ program (USEPA).
Equilibrium calculation shows that more than 98% of
mercury is in the complexed form in solutions of TU in all
ligand concentrations, however a measurable availability is
observed at 10
5 mol dm
3 TU, as a consequence of
dissociation of the complexes. Introducing these values of
X in Equation 8, a dissociation rate constant of 0.604 s
1 for
this complex can be calculated (Table 2). The availability
observed for Hg-GLY and Hg-NTA solutions is higher than
Xcal obtained by equilibrium calculations. These results
indicate that Hg-CYS and Hg-NTA complexes are moder-
ately labile to the discriminator employed in this work. The
dissociation rate constants are reflected in Table 2. A
decrease of availability and kinetic constant is observed
with an increase of ligand/metal ratio for NTA solutions.
No mercury availability is observed in mercury solutions
containing CYS, HA and COL when the ligand is at higher

Electroanalysis 2005, 17, No. 11  2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Carbon Paste Electrode Modified with a Schiff Base Ligand
Fig. 8. Variation of peak height of voltammetric signal with the
accumulation time for a 100 mg L
1 mercury standard solution
(^), and spiked natural (~) and spiked irradiated (&) Jarama river
water sample.
Table 3. Physicochemical parameters of studied water sample.
Parameter
pH
Conductivity (mS cm
1)
Dissolved oxygen (ppm)
Alkalinity (mg L
1)
Chloride (mg L
1)
DOC (mg L
1)
Sulfate (mg L
1)
Nitrate (mg L
1)
Calcium (mg L
1)
Magnesium (mg L
1)
Iron (mg L
1)
Copper (mg L
1)
Lead (mg L
1)
Mercury (mg L
1)
concentration than the metal. This result indicates that
mercury complexes formed in these solutions are inert to the
discriminator. Urea complex is labile at 10
3 mol dm
3
concentration. As expected, the mercury complexes formed
with sulfur containing ligands (CYS, TU) are inert or less
labile than complexes formed with oxygen containing
ligands (U, GLY).
3.6. Mercury Availability in Natural Water Sample
The modified electrode was tested for the speciation of
mercury in natural water samples from Jarama River
(Spain). The sample was taken and immediately filtered
over a 0.45 mm cellulose acetate filter and stored in a
polyethylene bottle without pH modification for speciation
analysis and was analyzed within 48 h. The characteristics of
the sample are shown in Table 3. In the sample, the total
mercury concentration was measured by cold-vapor atomic
fluorescence spectrometry. As mercury concentration in the
sample is lower than detection limit of the method, the
speciation model was applied to the filtered nontreated
Electroanalysis 2005, 17, No. 11
939
sample spiked with 100 mg L
1 mercury and allowed to
equilibrate for 48 h. Figure 8 shows the variation of peak
height with accumulation time for the spiked sample and a
100 mg L
1 standard mercury solution. Calculating their
slopes we obtained a value for mercury availability of
0.0185. The average rate constant, estimated using Equa-
tion 8, was 0.078 s
1. Taking into account the presence of a
high concentration of organic mater in the samples, we
suppose that mercury is organically bound in the samples. In
order to confirm this assumption, the sample was irradiated
in the UV digestor for 4 hours. After the irradiation stage, an
availability of 0.815 is obtaining. These results confirm that
the metal is mainly in an organic form in the samples, and
they are in agreement with those found in the literature for
mercury speciation in waters where the percentage of
organic mater is high [1, 34].
4. Acknowledgements
The authors acknowledge the financial support from
6.7 Ministerio de Educación, Ciencia y Tecnolog
a (Plan
82 Nacional I þ D þ I, Proyect n8 BQU 2001-0151).
12.8
183
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