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Biosensors and Bioelectronics 22 (2006) 145152

Potentiometric sensor for atrazine based on a


molecular imprinted membrane
G. DAgostino , G. Alberti, R. Biesuz, M. Pesavento
Dipartimento di Chimica Generale, Universit`a degli Studi di Pavia, Via Taramelli 12, 27100 Pavia, Italy
Received 24 January 2006; received in revised form 21 April 2006; accepted 4 May 2006
Available online 3 July 2006

Abstract
A molecular imprinted polymer (MIP) membrane for atrazine, not containing macropores, was synthesized and implemented in a potentiometric
sensor. It is expected to work like a solid ISE (where the specific carrier are the imprinted sites) the specific carrier being the imprinted site. The
active ion is the protonated atrazine, positively charged. To form this species the determination is carried out in acidic solution at pH lower than
1.8, in which atrazine is prevalently monoprotonated. At these conditions the membrane potential increases with atrazine concentration over a
wide concentration range (3 105 to 1 103 M). The slope of the function E versus log c is about 25 mV/decade, showing that the atrazine form
sorbed on MIP is the biprotonated one. The detection limit is determined by the relatively high concentration of atrazine released by the membrane
in the sample solution at the considered conditions. It seems to be independent of the atrazine concentration in the internal solution of the sensor,
but it depends on the acidity of the solution. The response time is less than 10 s and the sensor can be used for more than 2 months without any
divergence.
2006 Elsevier B.V. All rights reserved.
Keywords: Molecular imprinted polymer; Potentiometric sensor; Atrazine; Ion selective electrode

1. Introduction
The molecular imprinted polymers for triazinic herbicides,
particularly atrazine, were the subject of many investigations in
recent years with interesting application in chemical analysis.
MIPs are used for solid phase extraction (SPE) and chromatographic separation (Matsui et al., 1995, 1997; Muldoon and
Stanker, 1997; Olsen et al., 1998; Takeuchi et al., 1999; Lanza
and Sellergren, 1999; Bjarnason et al., 1999; Ferrer et al., 2000).
The use of these polymers for the recognition in chemical sensors has been also proposed. Some authors prepared polymeric
membranes for conductimetric and capacimetric (Piletsky et al.,
1995, 1998; Sergeyeva et al., 1999; Panasyuk-Delaney et al.,
2001), for piezoelectric detection (Pogorelova et al., 2002), and
other signal transduction methods, for example through the bulk
acoustic wave and fluorescence (Liang et al., 2000; Jenkins et al.,
2001) Despite the relatively simple transduction of the potentiometric signal, only a few sensing devices of this kind have

Corresponding author. Tel.: +39 0382 987 580; fax: +39 0382 528 544.
E-mail address: girolamo.dagostino@unipv.it (G. DAgostino).

0956-5663/$ see front matter 2006 Elsevier B.V. All rights reserved.
doi:10.1016/j.bios.2006.05.014

been developed. All of them were based on the use of very thin
membranes or films (Kitade et al., 2004; Zhou et al., 2005), with
problems due to difficulties in preparation, reproducibility and
possible interferences. Recently a sensor has been developed,
based on a MIP polymeric film deposited on the gate surface of
an ion-sensitive field-effect transistor, obtaining good results in
terms of specificity and low detection limit (Pogorelova et al.,
2002). Potentiometric sensors have been previously proposed
for charged analytes, for example for nitrate anion based on
imprinted conducting polymers (Hutchins and Bachas, 1995).
Atrazine is protonated, and as a consequence, positively
charged in aqueous solution at sufficiently low pH, its protonation constant being log Ka = 1.7 (Smolkova and Pacakova, 1978;
Skopalova and Kotoucek, 1995). The combination of the positively charged species with the imprinted membrane should
produce a variation of the membrane charge. This can be measured potentiometrically by a conventional ISE device, in which
the membrane is placed between an internal filling solution
at constant composition in contact with a reference electrode,
and the sample solution (Craggs et al., 1974). This configuration is very simple, and gives a Nernstian response to the
analyte concentration, which is very convenient for quantifi-

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G. DAgostino et al. / Biosensors and Bioelectronics 22 (2006) 145152

Scheme 1.

cation. Moreover it reduces potential interferences, for example


those from redox active substances (Hutchins and Bachas, 1995),
or generally from substances active at the transductor.
In the present work, a molecular imprinted polymeric membrane for atrazine was prepared, The membrane was formed
directly at the end of a small Teflon tube which can be filled
with a solution at constant composition, in contact with an internal reference electrode, as in a classical potentiometric cell
for ion selective electrode (ISE). (Craggs et al., 1974), like
that depicted in the Scheme 1 reported below. This preparation
method for potentiometric sensors for charged species is here
applied because it is simple and straightforward. The affinity
of the protonated form of atrazine for the imprinted sites in the
MIP membrane can be different from that of the neutral atrazine,
which is the template. For this reason the effect of the solution
acidity is particularly investigated in the present work.

1 h. The membrane was conditioned in 3.7 104 mol l1 aqueous atrazine solution (pH 1.5, HCl solution, KCl 0.02 mol l1 )
for 16 h. In some cases the membrane was treated in 0.1 mol l1
HCl before conditioning.
2.3. Sensor construction and potential measurements
The Teflon tube bearing the membrane at the end was filled
with a solution containing 3.7 104 mol l1 atrazine, HCl
0.03 M (pH 1.5) and saturated with KCl. A Ag/AgCl reference
electrode was contacted with the internal filling solution to transduce the potentiometric signal.
All the potential measurements were carried out at 25.0 C
(0.1 C) in the cell as shown in Scheme 1.
Test solution: HCl pH 1.5, KCl 2 102 mol l1 , atrazine;
Membrane: MIP membrane.

2. Experimental
2.1. Materials
Methacrylic acid [79-41-4] and ethylene glycol dimethacrylate [97-90-5] (SigmaAldrich)) were distilled in vacuum prior
to use in order to remove stabilizers. Atrazine [1912-249] (SigmaAldrich)) and 2,2 -azobisisobutyronitrile [78-67-1]
(AIBN) (SigmaAldrich), were of reagent grade and were used
without any further purification. All other chemicals were of
analytical reagent grade and the solutions were prepared with
ultrapure water (Milli-Q).

The cell potential was measured at different atrazine concentration in the test solution in the range 1.7 107 to
1.7 104 mol l1 . All pH adjustments were made with HCl
37% or NaOH 50%.
The liquid junction potential Ej at the interface between
the filling solution of the double junction reference electrode,
0.34 mol l1 KNO3 , and the test solution at the usual composition depends on the proton activity according to the experimental
relation: Ej = 24aH = 24 H cH . The correction for the liquid
junction potential was done when appropriate.
3. Results and discussion

2.2. Preparation of the MIP membrane


3.1. Potentiometric response of the MIP membrane
The molecular imprinted polymer membranes for atrazine
was prepared from a reagent mixture obtained by mixing 48 l
of methacrylic acid, 95.2 l of ethylene glycol dimethacrylate,
25 mg of atrazine and 28 mg of AIBN. The mixture was uniformly dispersed by sonication (sonic bath model Transsonic
T420Elma) and then deaerated with nitrogen for 10 min. Ethylene glycol dimethacrylate acts not only as cross-linker, but also
as solvent. A 25 l aliquot of reagent mixture was placed in
a home made Teflon device that controls the shape of the liquid during the polymerization. This device was introduced in
an oven (model M710) at 70 C for 17 h. A glassy membrane is
obtained with 5 mm diameter, tightly fixed at the end of a small
Teflon tube 10 mm long. This is directly used for assembling the
sensor without any further manipulation of the membrane. The
thickness of the membrane can be varied by placing different
amounts of reagent mixture into the Teflon device for the polymerization. The template was removed by washing the membrane successively in 10 ml of a methanol/acetic acid solution
(4:1, v/v, of 98% methanol and pure acetic acid) for three times,
each time for 1 h, then in 10 ml of water/acetic acid (5:1) for three
times for 1 h and finally in 10 ml of pure water for three times for

The molecular imprinted polymer membrane was prepared


from a reagent mixture containing only methacrylic acid as the
functional monomer. Not any porogen solvent was used, in order
to avoid the formation of macropores through which a rapid
diffusion of atrazine across the membrane would take place. A
large fraction of cross-linker, EDMA, was employed, 88%, and
in this way a rigid membrane, resistant to stress, and well sealed
to the Teflon tube, was obtained.
Some examples of the potentiometric response of MIP membranes are displayed in Fig. 1.
Increasing concentrations of atrazine (cadd ) were added to
the test solution. The potential increases immediately after
the atrazine addition at concentrations higher than around
3 105 M, in the case of the membrane conditioned as
described in the experimental part. For comparison the potentiometric response obtained with a similar membrane, but not
previously conditioned in atrazine solution, is reported in Fig. 1.
It shows that in the first point, at cadd = 1 107 mol l1 , a long
time is required to reach the equilibrium potential. Besides, the
slope of the function E versus log cadd increases with the con-

G. DAgostino et al. / Biosensors and Bioelectronics 22 (2006) 145152

Fig. 1. Cell potential at different atrazine concentrations in function of


time. Conditions reported in Scheme 1. Atrazine concentration 1.7 107 to
1.7 104 mol l1 . Membrane 0.5 mm thick. () MIP membrane conditioned;
(*) MIP membrane non conditioned; () NIP membrane conditioned.

centration, being in any case much lower than that obtained with
the conditioned membrane. Probably when the concentration of
atrazine in the membrane is low, as it is in the not conditioned
membrane, some atrazine is sorbed from the test solution into
the membrane, producing a potential decrease. It has been previously reported, for example in the case of a potentiometric
sensor for nitrate (Hutchins and Bachas, 1995), that the sensor
conditioning, reduces the potential drift due to the diffusion of
the template in the bulk of the membrane and shortens the equilibration time, even in the case of relatively thick membranes,
as seen in Fig. 1.
A membrane synthesized, washed and conditioned like the
MIP membrane but in the absence of template (NIP membrane)

147

gave a much lower potential variation in function of the atrazine


concentration, as seen in Fig. 1. This indicates that atrazine interacts only weakly with the NIP membrane because of the absence
of specific molecular interaction sites. Besides, the potential of
the not imprinted membrane is considerably lower than that of
the MIP membrane, for example 24 mV instead of 99 mV.
The experiments reported in Fig. 1 were carried out in
acidic solution, HCl 0.031 mol l1 (pH 1.5) containing KCl
0.02 mol l1 . Atrazine is at least partially protonated at pH 1.5,
according to the protonation constant reported in the literature (Smolkova and Pacakova, 1978; Skopalova and Kotoucek,
1995), while it is not protonated at neutral pH. Indeed not any
potentiometric response was obtained in neutral 0.02 mol l1
KCl. This shows that the membrane potential depends on the
concentration of charged species, similarly to the usual ion selective electrodes (Bard and Faulkner, 1980). The effect of the
proton concentration will be further discussed below.
Other membranes were treated with 0.1 mol l1 HCl, after the
usual washing with acetic acid/methanol, and before the conditioning procedure. The E versus log cadd straight line is shown
in Table 1. While the slope was similar to that obtained for the
membranes not treated with HCl 0.1 M, E0 and E(p) were about
40 mV lower. In this case, 2 days were required for conditioning in 3.6 104 mol l1 atrazine solution. The lower measured
potential could be related to the higher protonation of the carboxylic groups in the membranes treated in more acidic solution.
Anyhow the potentiometric response of the membranes treated
with 0.1 mol l1 HCl to increasing atrazine concentration was
similar to that of the not treated ones both in terms of slope and
detection limit. After the HCl treatment, the selectivity of the
membrane was improved, as it will be shown below.
A lower potential was measured using a thicker membrane,
1 mm instead of 0.5 mm, as seen in Fig. 2. The equilibration time
was near to that of the thinner membranes, only a few seconds,
suggesting that the potentiometric equilibrium is established at
the interface membraneaqueous solution in the case of the conditioned membranes.
The membrane potential was constant at 0.1 mV after equilibration.
The sensors can be used for at least 1 month with no modification of the potentiometric response. The sensors were stored

Table 1
Standardization curve of different atrazine MIP membranes, at pH 1.5, in KCl 0.02, and results from Eq. (3)

Membrane 0.5 mm
Membrane 1 mm
Membrane 0.5 mm,
treated with 0.1 M HCl
Membrane 0.5 mm,
atrazine concentration
internal solution
c = 5 105 M
Average of four different
membranes 0.5 mm

Standardization line
0
E = Eadd
+ Nlog cadd

E(p)
(mV)

DL
(105 M)

E = 211(3) + 24.2(7)log c
E = 135(2) + 32.2(5)log c
E = 171(4) + 22(1)log c

99
3.5
62.7

E = 177(5) + 26(1)log c

E = 209(12) + 23(3)log c

Linearized Eq. (1)




KAtr,j cj (105 )
pot

Slope

Ord. origin

E0 (mV)

2.3
5.0
1.2

3.4(1) 107
1.3(1) 104
5.8(2) 106

1.9(1) 103
0.5(1)
85(7)

228.2
121.3
205.8

3.5(2)
4.1(9)
1.5(4)

61.0

3.2

2.8(2) 106

31(1)

196.4

1.1(0)

106(1)

3(1)

8(2) 107

1.2(4) 103

239(3)

1.9(8)

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G. DAgostino et al. / Biosensors and Bioelectronics 22 (2006) 145152

3.2. Selectivity of the atrazine MIP membrane

Fig. 2. Standardization curves of atrazine at different conditions. () Membrane


0.5 mm thick, pH 1.5; () membrane 0.5 mm thick, pH 7; () membrane 1 mm
thick, pH 1.5.

in aqueous solution containing 3.6 104 mol l1 atrazine,


0.02 mol l1 KCl and 0.031 mol l1 HCl.
Atrazine diffuses through the MIP membrane because of the
concentration difference at the two interfaces of the membrane.
The diffusion was checked in the following way: the Teflon
tube carrying the membrane, previously conditioned in a concentrated atrazine solution, was filled with 0.1 ml of atrazine
solution 3.6 104 mol l1 , containing KCl 0.02 mol l1 and
HCl 0.031 mol l1 , and the membrane was contacted with 1 ml
of solution at the same composition, but not containing atrazine.
The contact was maintained for 48 h at room temperature. After
this time, the concentration of atrazine in the outer solution
was 5 106 mol l1 . The determination was made spectrophotometrically at = 225 nm, at which the molar absorptivity
of atrazine is 2.07 104 cm1 l mol1 . This demonstrates that
actually some atrazine diffuses from the inner to the outer solution. The diffusion rate is relatively low, since the concentration
in the inner and outer solution is still different after 48 h. This is
favorable for the potentiometric determination, and is due in part
to the absence of macropores in the membrane. The diffusion
can affect the detection limit.

The effect of increasing concentrations of simazine (6chloro-N2,N4-diethyl-1,3,5-triazine-2,4-diamine), a chlorinated


triazine with a structure very similar to atrazine, was evaluated
for a MIP membrane washed only with methanol/acetic acid
and for a membrane further treated with 0.1 mol l1 HCl solution before conditioning in concentrated atrazine solution. The
standardization curve obtained for the membrane only treated
in methanol/acetic acid is reported in Table 2, it is similar to
that obtained for atrazine (see Table 1). On the contrary, there
was not any potential variation for simazine concentration in the
case of the membrane treated with HCl 0.1 M. The results are
also reported in Table 2 for comparison. The same holds for two
other triazines: ametryne (N2-ethyl-N4-isopropyl-6-methylthio1,3,5-triazine-2,4-diamine), which contains a SCH3 group
instead of a chlorine atom, and has protonation properties different from atrazine (log Ka = 4.1), and 2-hydroxy-atrazine bearing
an OH instead of SCH3 or chlorine, which is the principal
degradation product of atrazine. Evidently the membrane is more
specific for atrazine after treatment in HCl 0.1 mol l1 .
3.3. Response of the MIP membrane based sensor to
atrazine concentration
Some typical Nernstian plots, E versus log cadd are reported
in Fig. 2.
At pH 7, the slope is always near to zero, while at pH 1.5 the
curve is composed of two straight lines, one of which, at lower
atrazine concentration, has slope near to zero and the other has a
positive slope near to 25 mV/decade. The slope and the ordinate
at the origin determined by fitting the linear part of the calibration plot to the Nerst equation are reported in Table 1, together
with the potential corresponding to the part with slope equal to
zero (E(p) ). The detection limit (DL), i.e. the intercept of the two
straight lines (Buck and Lindner, 1994) is also reported. Four
different membranes were synthesized and used at similar con-

Table 2
Standardization curve of atrazine MIP membranes 0.5 mm thick, in KCl 0.02 M and pH 1.5, for different triazines

Simazine membrane
washed in methanol/
acetic acid
Simazine Membrane
washed in methanol/
acetic acid and treated
in HCl 0.1 M
2-OH atrazine membrane
washed in methanol/
acetic acid and treated
in HCl 0.1 M
Ametrine membrane
washed in methanol/
acetic acid and treated
in HCl 0.1 M

Standardization line
0
E = Eadd
+ Nlog cadd

E(p)
(mV)

DL
(105 M)

E = 223(4) + 29(1)log c

95.6

4.65

E = 81(1) + 4.0(2)log c

62.70

E = 68.5(2) + 0.64(5)log

66.6

E = 68.2(1) + 0log c

68.1

Linearized Eq. (1)




Slope (107 )

Ord. origin

E0 (mV)

KAtr,j cj (105 )

3.75

Not significantly
different from 0

230

Not significantly
different from 0

pot

G. DAgostino et al. / Biosensors and Bioelectronics 22 (2006) 145152

ditions in a short time (2 days) to investigate the reproducibility


of the membranes. The average values obtained for the parameters are reported in Table 1, with the corresponding standard
deviation, which is relatively low.
The slope (N) of the Nernstian part of the standardization curve E versus log cadd is similar for all the considered
membranes, and is near to 25 mV/decade. The same slope
was obtained with an ISFET/MIP based sensor, with a different imprinted polymer (Pogorelova et al., 2002). According to
the classical model for ISE this should indicate that a doubly charged ion is responsible for the potentiometric signal.
However atrazine is not doubly protonated at the considered conditions, as calculated from the protonation constant of atrazine
reported in the literature, log Ka = 1.7 (Smolkova and Pacakova,
1978; Skopalova and Kotoucek, 1995). Only 63% of atrazine in
solution is monoprotonated at pH around 1.5.
E0 is noticeably different in some of the experiments reported
in Table 1. For example, when an internal solution at lower
atrazine concentration is used, a lower potential is obtained. This
variation corresponds to the concentration difference in the internal solution. The detection limit is only slightly affected by the
atrazine concentration in the internal solution, probably because
of the very slow diffusion rate.
Considering that interfering cations may be present, and that
the potentiometrically active form of atrazine is ionic, say zcharged or z-protonated, the following relationship can be used
for modeling the potentiometric response of the electrode:
 pot
E = E0 + N ln([Hz Az+ ] + KAtr,j cjz/zI )


 pot
cadd
z/zI
0
+
KAtr,j cj
E = E + N ln
(1)

Hz A

RT
N=
nF
where z is the charge of the species which actually distributes
pot
between the two phases, Hz Az+ . KAtr,j is the atrazine/jion (with
charge zj ) potentiometric selectivity coefficient, and Hz A is the
ratio of the total atrazine concentration in the solution phase to
the concentration of the z-protonated form (side reaction coefficient of protonated atrazine):

za [H]z
Hz A =
za [H]z


za [H]z
[Hz Az+ ]
cAtr = [Hz Az+ ] = [Hz Az+ ] 
za =
za [H]
[H]z [A]
(2)

za is the z-protonation constant of atrazine, and the summation is extended from z = 0 to n (n is the maximum protonation number), and 0a = 1. Charges are omitted for simplicity.
The concentration of the active species depends on the total
atrazine concentration, and on the acidity of the solution. Possible interfering species are protons and atrazine released by the
membrane, the concentration of which, indicated by the symbol cr , is unknown. Its potentiometric selectivity coefficient is
pot
KAtr,Atr = 1.
Eq. (1) may be written in the following linearized way, to
differentiate the two terms in the logarithm, one depending on

149

the atrazine concentration added to the solution and the other


independent of it, but possibly depending on the atrazine concentration released by the membrane:


 pot
cadd
E/N
E0 /N
z/zI
(3)
= 10
+
KAtr,j cj
10
Hz A
N is assumed to be 29.5 mV/decade at 25 C, near to the
experimental value.
The results obtained by the linearization procedure are also
reported in Table 1. The logarithm of the slope of the linearized
function is E0 N log Hz A , equal to that obtained directly from
the function E versus log cadd (E0 ). It is also reported in Table 1
for comparison. The differences are due to the slope of the function, which is slightly different from 29.5, as seen in the second
column of Table 1.
The term accounting for the interferences, expressed as the
 pot
summation KAtr,j cj , obtained from the parameters of Eq. (3),
is reported in Table 1. It is similar in the different experiments,
because the composition of the solutions was the same. It is
slightly lower in the case of lower atrazine concentration in the
internal solution.
The detection limit, defined as the intercept of the two straight
lines in the E versus log cAtr plot, is relatively high. On the basis
of the experiments described in this paragraph the interfering
substances responsible for this high detection limit cannot be
identified.
3.4. Effect of proton concentration on the sensor response
Some curves E versus log cadd obtained with the same membrane, but at different acidity of the test solution are reported in
Fig. 3.
It can be seen that the potential is higher at higher acidity.
At pH 4, the potential does not increase at increasing atrazine
concentration, while at lower pH the curve E versus log cadd
is composed of two straight lines. The parameters of the part
of the curve E versus log cadd with slope different from zero are
reported in Table 3, together with those obtained from Eq. (3). At
the pH of the experiments, the liquid junction potential variation
is low, so that not any correction was made.

Fig. 3. Standardization curves of atrazine at different acidities. Membrane


0.5 mm thick, treated in HCl 0.1 M. () pH 1.0; () pH 1.45; () pH 1.8;
(*) pH 4.0.

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G. DAgostino et al. / Biosensors and Bioelectronics 22 (2006) 145152

Table 3
Standardization curve of atrazine MIP membrane 0.5 mm thick, treated in 0.1 M HCl, in KCl 0.02 at different pH, and results from Eq. (3)

pH 1.00
pH 1.40
pH 1.85

Standardization line
0
E = Eadd
+ Nlog cadd

E(p) (mV)

E = 218(10) + 23(3)log c
E = 214(6) + 25(1)log c
E = 198(9) + 24.2(2)log c

115
104
96

DL (105 M)

3.30
4.90
6.60

The slope of the curve E versus log cadd is almost independent of the pH, while the ordinate at the origin (E0 ) decreases
with increasing pH. This can be ascribed to the protonation
of atrazine, if the z-protonated form, is that potentiometrically
active. Its concentration becomes higher at higher proton activity, so that the corresponding potential should also increase, as
experimentally observed. If the completely deprotonated form
prevails, E0 depends on the solution pH as here reported
E0 = E0 + N log za N z pH
The plot of E0 versus pH is actually a straight line, with these
parameters obtained by least square method:
E0 = 269.2(6) 27.3(4)pH
The expected slope is 59.16 mV/decade, since z corresponds
to the charge of the ionic form of atrazine. This is the same effect
observed in the case of the E versus log catr plot.
The difference between the constant potential E(p) and E0
is constant at different pH, around 105 mV, a value welll corresponding to Nlog(DL). It can be deduced that protons do
not influence significantly the detection limit. The other possible interfering ion is the atrazine released by the membrane,
or diffusing from the internal solution to the sample solution,
pot
the potentiometric selectivity coefficient of which, KAtr,Atr , is
equal to 1, as atrazine is the primary ion. So the interference
term represents the concentration released by the membrane at
the interface membrane/aqueous solution. This is a reasonable
value compared to the detection limit, and is near to that reported
in Table 1, obtained at fixed pH.
Since the detection limit increases slightly at decreasing H+
concentration, it is possible that more atrazine is released in
solution at higher pH. This could explain why at sufficiently
high pH, for instance at pH 4, the membrane potential does not
increase at increasing atrazine concentration.
The effect of the acidity on the potentiometric response of the
atrazine MIP membrane is seen also in Fig. 4, in which the potential in function of the solution pH is reported for different membranes, conditioned in the usual way but not containing atrazine
in the test solution, so that the measured potential corresponds
to E(p) . The cell potential reported in ordinates is corrected for
the liquid junction potential variation at pH lower than around
1.2, as reported in the experimental part. At pH lower than 2.0
the potential increases at decreasing pH, while at higher pH it
is constant. The potential versus pH function for pH lower than
around 2 is a straight line with slope near to 23 mV/decade, and
ordinate at the origin approximately 135 mV. One of these lines,
obtained from one of the experiments in Fig. 4, is here given

Linearized Eq. (1)




Slope (107 )

Ord. origin (102 )

E0 (mV)

15(1)
5.9(4)
2.4(2)

23(6)
5(2)
4(1)

241
229
218

KAtr,j cj (105 )
pot

1.5(4)
0.9(4)
1.5(4)

as an example: E = 136(1)24(1)pH (R2 = 0.991). These results


are in good agreement with those obtained in the experiments
reported above, and the explanation is that the concentration of
protonated atrazine, which is the potentiometrically active form,
is higher at higher acidity.
The effect of the pH variation on a NIP membrane is completely different, as shown in Fig. 4 for comparison. Here the
potential increases at increasing pH, contrary to what observed in
the case of the MIP membrane. This could indicate that protons
easily diffuse through the membrane not containing atrazine,
while their mobility is reduced in presence of atrazine. This
increases the conductivity of the membrane. It will be seen below
that actually the resistance of the membrane is lower at higher
atrazine concentration in the solution phase.
3.5. Electrical characterization of the membrane
The MIP membrane was characterized by the impedance
spectroscopy technique (De Marco and Pejcic, 2000) at different
atrazine concentration in the external solution. The instrument
utilized was a Solartron SI 1260 impedance/gain-phase analyzer, in the 0.1 Hz10 MHz frequency range. The usual cell,
that reported in Scheme 1, was investigated. The impedance
spectrum shows that only one resistance and capacitance in
parallel configuration constitute the equivalent circuit of the
electrochemical system considered, to be attributed to the polymeric material. The electrical capacitance of the membrane was
13.4 pF, and does not significantly vary in function of the atrazine
concentration.
The membrane electrical resistance at different atrazine concentration in the external solution is reported in Fig. 5. The
membrane resistance is relatively low, and decreases at concentration of atrazine higher than 3 105 mol l1 , which is near

Fig. 4. Dependence of the potential of cell 1 on the pH of the test solution. No


added atrazine. (,,) MIP membrane 0.5 thick; (*) NIP membrane 0.5 thick.

G. DAgostino et al. / Biosensors and Bioelectronics 22 (2006) 145152

151

References

Fig. 5. Resistance of the conditioned MIP membrane at different atrazine concentration obtained by impedence spectroscopy.

to the detection limit of the potentiometric determination. The


variation of the resistance is almost linear at the considered concentrations. The decrease of the membrane resistance with the
atrazine concentration is probably sufficiently high to reduce the
slope of the Nernst function with respect to 59.16 mV/decade.
4. Conclusions
A potentiometric sensor for atrazine was developed, based
on a molecular imprinted polymer membrane. It works as an
ion selective electrode highly specific for atrazine. The membrane was rigid enough to bear the filling solution in contact
with the internal reference electrode. To obtain a good potentiometric response, the membrane must be previously conditioned.
An interesting characteristics of the potentiometric sensor is that
a short time, only a few seconds, is required to reach the equilibrium potential.
Since the potential variation is produced by the charged
species, the solution pH must be lower than pH 1.7 to ensure
the protonation of atrazine,. Protons do not interfere according
to an ion exchange mechanism, but they have an influence on
the measured potential, since the concentration of the protonated species of atrazine increases with increasing acidity. The
slope of the potentiometric curve E versus log cadd is lower than
expected for a monocharged ion. This is probably due to the
variation of the electrical resistance of the membrane with the
atrazine concentration. Atrazine is sorbed more strongly at low
pH, at which the carboxylic groups in the polymeric membrane
are completely protonated, as demonstrated by the fact that the
detection limit slightly decreases with pH. This is probably the
reason for which at pH higher than approximately 1.7 the membrane potential does not change with the atrazine concentration.
The detection limit is around 2 105 mol l1 , and it appears to
be determined by the distribution coefficient of atrazine between
the acidic aqueous solution and MIP. Thus the best way to lower
the detection limit is using membranes with stronger sorbing
properties at the considered conditions.
Acknowledgment
This work was financially supported by the project FIRB01
(MURST, Italy).

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