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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-
146
Scheme 1.
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
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
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
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)
148
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
Slope (107 )
Ord. origin
E0 (mV)
KAtr,j cj (105 )
3.75
Not significantly
different from 0
230
Not significantly
different from 0
pot
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
150
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
Slope (107 )
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)
151
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