Analytica Chimica Acta 503 (2004) 187194

Improving the lower detection limit of potentiometric sensors

by covalently binding the ionophore to a polymer backbone
Martin Pntener, Tams Vigassy, Ellen Baier, Alan Ceresa, Ern Pretsch
Laboratorium fr Organische Chemie, ETH Hnggerberg, CH-8093 Zrich, Switzerland

Received 2 July 2003; received in revised form 30 September 2003; accepted 14 October 2003

Abstract

The lower detection limit of polymeric membrane ion-selective electrodes (ISEs) is impaired by zero-current ion fluxes through the
organic phase. This adverse effect is largely eliminated by covalently attaching the ionophore to a polymer backbone. To this purpose, the
Pb2+ -selective ligand, 4-tert-butylcalix[4] arene-tetrakis(N,N -dimethylthioacetamide) is substituted on its upper rim by a diol derivative
which is subsequently copolymerized with poly(tetrahydrofuran)diol and 2,2,4-trimethylhexamethylene diisocyanate to the corresponding
polyurethane. By measurements on sandwich membranes, it is shown that through binding the ionophore to the polymer, the mobility of Pb2+
in the ISE membrane is strongly reduced. As a consequence, the response range of such an ISE is extended by several orders of magnitude.
This is the case even when using an internal electrolyte that with an ISE based on a mobile ionophore leads to strong deviations from the linear
response because of ion uptake from the sample into the membrane or ion release from the membrane into the sample. With a conventional
inner filling solution of 101 M Pb(NO3 )2 , a lower detection limit of 1.7 109 M Pb2+ has been achieved in the presence of 104 M Na+ .
2003 Elsevier B.V. All rights reserved.

Keywords: Potentiometric sensor; Polymeric ionophore; Detection limit; Ion fluxes

1. Introduction sample [810]. The lower detection limits and selectivities

During recent years, it has been well established that the
lower detection limit and selectivity behavior of traditional
polymer membrane ion-selective electrodes (ISEs) [1,2]
have been adversely affected by zero-current transmembrane
fluxes of the measuring (primary) ions [3]. Already very
small changes of <1% in the total ionic concentration of the
ISE membrane are sufficient to induce leaking of primary
ions into the sample and thus cause the concentration in the
contacting aqueous layer to be ca. 106 M, even if the bulk
sample virtually did not contain primary ions. Consequently,
the lower detection limit was found to be around 106 M
and the best selectivity coefficients were around 103 to
104 in practically all reported cases [4,5]. Usually, the ori-
gin of concentration gradients in the ISE membrane is the
coextraction of primary ions together with their counterions
from the internal solution [6,7] and/or the exchange of pri-
mary ions from the membrane by interfering ions from the
Corresponding author. Tel.: +41-1-632-29-26; fax: +41-1-632-11-64.
E-mail address: pretsch@org.chem.ethz.ch (E. Pretsch).
have been found to improve by many orders of magnitude
when compensating such gradients through careful adjust-
ment of the composition of the internal solution [9,1114]
or by applying a weak electric current [1517]. However, as
revealed by a detailed theoretical analysis [9,10,14,18], per-
fect compensation of transmembrane ion fluxes is only pos-
sible for individual sample compositions and any changes
in them inevitably leads to concentration gradients in one
or the other direction, and, consequently, to a biased ISE
response. For trace analysis of environmental samples [19],
ISEs have been optimized by a twofold strategy: (1) by
adjusting the composition of the inner solution so as to
induce an ion exchange similar to that caused by a typical
sample to be measured, and (2) by reducing transmembrane
ion fluxes through optimizing the membrane composition
[2022]. The topic of this contribution is to investigate
the influence on the response behavior of ISEs when the
ionophore is covalently bound to a polymer backbone.
Covalent bonding of electrically charged [2325] and
neutral [26] ionophores to polymer backbones has already
been reported many years ago. While the primary aim of
those contributions was to improve the lifetime of ISEs,

0003-2670/$ see front matter 2003 Elsevier B.V. All rights reserved.
doi:10.1016/j.aca.2003.10.030
188 M. Pntener et al. / Analytica Chimica Acta 503 (2004) 187194
the extension of the linear response range by immobilizing
the ionophore was also noticed [24,26]. However, since the
lipophilicity of the parent mobile ionophores was rather
poor, the reason for this improvement was not necessarily
due to the reduction in transmembrane ion fluxes [27]. Sub-
sequently, different authors [2839] have covalently bound
ionophores but again in order to improve the lifetime of the
corresponding ISEs or to inhibit the leakage of components
into the sample during in vivo measurements [25,32], but not
to attain better lower detection limits. More recently, we have
shown that the mobility of ionophores and transmembrane
ion fluxes can be drastically reduced by covalent bonding of
a H+ -selective ionophore to polyurethane [40]. In the present
work, we describe the immobilization of the ionophore,
4 - tert - butylcalix[4]arene - tetrakis(N,N -dimethylthioacet-
amide), introduced in 1994 [41], which shows an extremely
high Pb2+ selectivity [42].
2. Experimental
2.1. Reagents
All solutions were prepared with freshly deionized water
(18 M cm) and acids and salts of puriss. p.a. or MicroSelect
quality (Fluka AG, CH-9471 Buchs, Switzerland). The plas-
ticizers, bis(2-ethylhexyl) sebacate (DOS) and o-nitrophenyl
octyl ether (o-NPOE), poly(vinyl chloride) (PVC), the
lipophilic salts, sodium tetrakis[3,5-bis(trifluoromethyl)phe-
nyl]borate (NaTFPB), potassium tetrakis[3,5-bis(trifluoro-
methyl)phenyl]borate (KTFPB), and tetradodecylammo-
nium tetrakis(4-chlorophenyl)borate (ETH 500), as well
as the ionophore, 4-tert-butylcalix[4]arene-tetrakis(N,N -di-
methylthioacetamide), were Selectophore products (Fluka).
The ion buffers, disodium ethylenediamine tetraacetic
acid (Na2 EDTA) and nitrilotriacetic acid (NTA), were
also from Fluka. Chemicals and solvents for the synthe-
ses were obtained from Aldrich, Fluka, Merck, Riedelde
Han, or Sigma in puriss. p.a. quality and were used
without further purification if not otherwise mentioned.
Poly(tetrahydrofuran) (PTHF) and 2,2,4-trimethylhexa-
methylene diisocyanate (TMDI) were from BASF AG
(D-67056 Ludwigshafen, Germany) and Hls Chemie
Schweiz AG (CH-8700 Ksnacht, Switzerland), respec-
tively. The diol derivative 2 was obtained in our laboratory
by a synthesis of seven steps, which will be described
elsewhere [43].
2.2. Synthesis of ionophore 3
A solution of dry 2 (0.25 g, 0.19 mmol, 1 equiv.) in dry
1,2-dichloroethane (80 ml) was stirred at RT under Ar for
10 min. The oligomeric diol PTHF (1.50 g, 0.75 mmol,
4 equiv.) was added and the yellowish solution was heated
to 70 C until PTHF was fully dissolved. The clear solu-
tion was then heated to 110 C and freed from water by
a 5 h Soxhlet extraction (4 molecular sieve), discarding
the first portion (30 ml) of the distillate. After the water
content was determined to be lower than 5 ppm, the reac-
tion mixture was cooled to 70 C. Freshly distilled TMDI
(0.20 g, 0.94 mmol, 5 equiv.) was added and the reaction
mixture was copolymerized at 70 C for 7 days. Once
GPC analysis showed no further increase in polymer chain
length after adding three drops of TMDI to the reaction
mixture, abs. MeOH (0.2 ml) was added and the mixture
was stirred for 1 h. The product was then slowly precipi-
tated with hexane cooled to 0 C (300 ml) under vigorous
stirring and stirring was continued for 2 h. The precipi-
tate was redissolved in CH2 Cl2 (50 ml) and precipitated
twice with cooled hexane (0 C, 300 ml) obtaining the
ionophore 3 (0.72 g, 37% yield) after drying for 1 week in
a drying oven at 40 C. Viscosimetric analysis of 3 gave a
weight-averaged molecular mass, Mw , of 28 850 g mol1
and Mw /Mn = 6.31, Mn being the number-averaged
molecular mass.
2.3. ISE membranes and electrodes
In general, the membrane components (typically 200 mg
of total mass) were dissolved in freshly distilled THF
(2.0 ml) during ca. 2 h and poured into a glass ring (24
or 26 mm i.d.) fixed on a Teflon support. After overnight
evaporation of the solvent at RT, 5 mm disks were punched
from the mother membrane (thickness, ca. 200 m) and
glued with THF to a plasticized PVC tubing, mechanically
fixed to a 1000 l pipette tip. On the basis of the mem-
branes containing DOS (ca. 75 mg), PVC (37 mg), KTFPB
(5 mmol kg1 ), and ionophore 3 (2.3422.21 mg), the ef-
fective concentration of 3 was calculated. The compositions
of all membranes are shown in Table 1. Inner filling so-
lutions were: for conventional ISEs, 101 M Pb(NO3 )2 ;
for ISE membranes showing strong ion uptake, 101 M
Na2 EDTA, 101 M NaOH, pH 7.5, 105 M Pb(NO3 )2 ; for
ISEs with optimized lower detection limits [19], 103 M
NTA, 2.1 101 M NaOH, pH 7.0, 104 M Pb(NO3 )2 . A
second inner filling solution, in contact with the first one
through a diaphragma, consisted of 101 M NaCl (conven-
tional ISEs and ISE membranes with strong ion uptake) or
2103 M NaCl (with optimized lower detection limit). The
ISEs were conditioned in 5 109 M Pb(NO3 )2 containing
103 M CaCl2 and 104 M HNO3 (Fig. 4) or 5 1010 M
Pb(NO3 )2 with 104 M NaCl and 104 M HNO3 (Fig. 5)
for 48 h.
2.4. EMF measurements
Measurements were performed with a 16-channel elec-
trode monitor (Lawson Labs Inc., Malvern, PA 19355)
in magnetically stirred solutions at ambient tempera-
ture (2022 C). Activity coefficients were obtained from
the DebyeHckel approximation and EMF values were
 M. Pntener et al. / Analytica Chimica Acta 503 (2004) 187194 189

Table 1
Composition of ISE membranes
Membrane Ionophore, mg (mmol kg1 , wt.%) Anionic sites, mg (mmol kg1 ) Plasticizer, mg Polymer, mg (wt.%) 102 M Pb
no. (NO3 )2 , l
1 3 NaTFPB KTFPB o-NPOE DOS PVC PU

Iaa 2.11 (9.85) 0.36 (1.96) 134.8 66.4 (32.5)

Iba 0.37 (2.05) 133.6 66.4 (33.1)
IIaa 21.53 (7.87, 9.61) 0.63 (3.12) 135.7 66.2 (29.6)
IIba 0.57 (3.10) 135.8 66.4 (32.8)
IIIb 2.13 (9.36) 0.94 (4.86) 144.2 68.2 (31.7)
IVb 1.95 (8.03) 0.97 (4.68) 139.2 67.0 (29.1) 21.4 (9.27)
Vb 22.80 (8.18, 9.98) 0.99 (4.81) 137.5 67.2 (29.4)
VIb,c 1.96 (8.02) 0.98 (4.69) 140.0 67.4 (29.2) 20.9 (9.05)
VIIb,c 22.21 (8.17, 9.96) 1.04 (5.17) 133.1 66.6 (29.9)
VIIId 5.01 (10.38) 2.01 (4.95) 299.1 151.2 (33.1) 113.0
IXd 29.67 (10.42, 12.71) 1.03 (5.00) 134.6 66.1 (28.6) 57.5
a For double membranes.
b For selectivity measurements.
c For Fig. 4.
d For Fig. 5.

corrected for liquid-junction potentials with the Hender-
son equation. Sample pH values were determined with a
Metrohm glass electrode (No. 6.0133.100, Metrohm AG,
CH-9010 Herisau, Switzerland). The reference electrode
was a Metrohm double junction Ag/AgCl reference elec-
trode (No. 6.0729.100) with 3 M KCl as reference elec-
trolyte and 1 M KCl as bridge electrolyte.
2.5. Selectivity measurements
Prior to selectivity measurements, the ISEs were condi-
tioned in 102 M NaCl for 2 days. The sequence of the
investigated ions was: Na+ , H+ , K+ , Mg2+ , Ca2+ , Cd2+ ,
Pb2+ , Cu2+ , and Ag+ . For each of these ions, the mem-
brane was conditioned in the corresponding 102 M ni-
trate solution (during 0.5 h), after which calibration curves
were taken using a Liquino 711 (Metrohm) for automatic
dilution. Even with this procedure, sub-Nernstian slopes
were obtained for some highly discriminated cations. In
these cases, the selectivity values represent upper limits
only.
2.6. Sandwich membrane measurements
Membranes were individually conditioned in 102 M
Pb(NO3 )2 for 12 h. The two membranes (without and with
ionophore 1 or 3) for the sandwich membranes were se-
lected if their EMF values agreed within 1 mV. Then,
one membrane disk ( 6 mm) without and another with
ionophore (1 or 3) were stacked together and mounted in a
Philips IS 561 electrode body (Mller Glasblserei, Zrich,
Switzerland), with the ionophore-loaded membrane on the
outer side. It took <1 min between assembling the sand-
wich membrane and starting the potential measurements.
Again, 102 M Pb(NO3 )2 served as internal electrolyte and
sample solution.
3. Results and discussion
In the presence of zero-current transmembrane ion fluxes,
the primary ion activity near the membrane surface, aI ,
differs from that in the bulk of the sample, aI (bulk). At
steady state, this difference given by the second term on the
right-hand side of Eq. (1) depends on the difference between
the concentration of the primary ion complex on the inner
([IL] ) and outer ([IL]) side of the membrane, the diffusion
coefficients, D, of ionic species in the membrane (m) and
the aqueous sample (aq), and the thicknesses, d, of the dif-
fusion layers of both phases:
Dm daq
aI = aI (bulk) + ([IL] [IL]) (1)
Daq dm
In this approximate equation, derived for the steady state
[8,14,18], activities are set equal to concentrations. At
submicromolar sample activities of Iz i, already very small
changes (below 1%) in the concentration of primary ion
complexes, ILz i, on either side of the membrane induce
relevant deviations between aI and aI (bulk). If the concen-
tration of ILz i in the membrane decreases toward the sample
phase, the lower detection limit, aI (DL), is determined by
the primary ions leaking from the membrane [14]:


1 z I + zJ
log aI (DL) = +
2 6

 
Dm daq RT zI /zJ  pot zI /zJ
log KIJ aJ (2)
Daq dm zI
Here, zI and zJ are the charges of primary (I) and interfering
pot
(J) ions, KIJ is the potentiometric selectivity coefficient,
and RT is the concentration of ion exchanger sites in the
membrane. It is assumed that [IL] = RT /zI . On the other
hand, if [IL] = 0 so that [IL] strongly decreases toward the
190 M. Pntener et al. / Analytica Chimica Acta 503 (2004) 187194

inner solution, a sudden step-like potential drop is observed
at aI (step) [18]:
Dm daq RT
aI (step) =
Daq dm zI
To construct ISEs for submicromolar activity measurements,
the composition of the internal solution has to be designed
so as to minimize the transmembrane ion gradients [44].
Since, however, any change in the sample composition in-
evitably induces some concentration gradients, as a second
measure, the value of the right-hand side of Eq. (3) (the same
term also occurs in Eq. (2)) must be minimized. The benefi-
cial influence of various measures including the increase in
the polymer content or the thickness of the membrane [20],
the decrease in RT [19,20], strong stirring, application of a
flow-through cell or rotating electrodes [4547] as well as
the introduction of lipophilic particles into the membrane
matrix [22] have been documented. As a further possibil-
ity, the present work investigates the influence of covalently
binding the ionophore to a polymer backbone on the re-
sponse behavior of the corresponding ISE membranes. Co-
valent attachment of an ionophore is also a requirement for
developing rugged miniaturized ISEs [33,34].
Recently, we have shown that covalent attachment of a
H+ -selective ionophore to a polyurethane backbone can
eliminate the ion-flux-induced sudden potential drop caused
by a strong transmembrane pH gradient [40]. As an ex-
tension of that study, we have modified the Pb2+ -selective
ionophore, 4-tert-butylcalix[4]-tetrakis(N,N -dimethylthio-
acetamide) [41,42] by introducing on its upper rim a diol
(3) derivative (2) and subsequently copolymerizing it with
poly(tetrahydrofuran)diol and 2,2,4-trimethylhexamethylene
diisocyanate to the corresponding polyurethane (see Fig. 1).
Although synthetically more demanding, the modification
of the upper rim was chosen in order not to interfere with
the coordination sphere. A similar strategy was applied re-
cently for modifying a related Na+ -selective calix[4]arene
[39]. The synthesis of the diol precursor 2 required seven
steps and will be described elsewhere.
The copolymerization reaction yielded a mixture of
polymers with a weight mean molecular mass of Mw =
28.850 g mol1 (see Section 2). As shown by the ratio
Mw /Mn = 6.31 (with Mn as the numerically weighted
molecular mass), the polymer mixture has a rather high dis-
persity. The effective molar concentration of the complexing
calixarene units has been determined potentiometrically by
pot
evaluating the selectivity coefficient, KPbNa , as a function of
the concentration of ionophore 3 relative to that of the cation
exchanger sites (TFPB ), kept constant at 5 mmol kg1
(Fig. 2). As long as the molar ratio of binding sites to ion
exchanger, [L]/[TFPB ] is <0.5, about the same potentio-
metric selectivity is expected as for a ligand-free membrane.
Since the concentration of the uncomplexed Pb2+ ions in
the membrane sharply decreases at [L]/[TFPB ] = 0.5, a

O
O O + HO O
O OH OH
9 27
OH PTHF
CH2 CH2
O O O +

N S N S N S
4 3 O C N
N C O

TMDI
2
1
1,2-dichloroethane,
o
70 C, 7 d, 37%

O
O O polymer
O O
O
CH2 polymer
O O

N S N S
3

3
Fig. 1. Structure of the mobile ionophore 1 and scheme of the synthesis of the covalently immobilized ionophore 3. The number of poly(tetrahydrofuran)
(PTHF) linear-chain polymer units is an average.
 M. Pntener et al. / Analytica Chimica Acta 503 (2004) 187194 191

ionophore per 100 mg of total membrane is only marginal,

4 further experiments were made with an excess of ionophore
3 (10 mg/100 mg of total membrane) in order to avoid selec-
2 tivity changes due to small variations of the ionophore con-
centration.
log KPpbotNa

1
0
-1
-2
-3
0 2 4 6 8 10
Ionophore 3 [mg/100 mg membrane]
pot
Fig. 2. Potentiometric selectivity coefficient, log KPbNa , as a function of
the concentration of ionophore 3 (cf. Fig. 1 and Table 1) relative to a
constant concentration of TFPB (5 mmol kg1 ).
strong selectivity gain is expected at this concentration. In
the range of 0.5 < [L]/[TFPB ] < 1, the ligand concentra-
tion is not sufficient to form a 1:1 complex with the monova-
lent interfering ion. Therefore, the Pb2+ selectivity is better
in this range and decreases somewhat because of the forma-
tion of Na+ complexes at higher ionophore concentrations.
The experimental trend (Fig. 2) nicely fits these expecta-
tions except that the change in selectivity at [L]/[TFPB ] =
0.5 is less sharp than presumed for a strongly complexing
ionophore (see below). This might be due to the fact that
the ionophore is a mixture of polymers, with possibly some-
what varying complexation behavior. Nevertheless, from this
study, the effective ligand concentration can be determined
as 8.21 105 mmol mg1 , which corresponds to the in-
corporation of 84% of the initially used diol into the poly-
mer backbone. Since the selectivity gain around 6 mg of
Next, we determined the effective transmembrane ion
mobilities of Pb2+ in o-NPOE membranes by using the
sandwich membrane method [4850]. For this experiment,
two membranes, one with the ionophore (1 or 3) and the
other without, were prepared and conditioned separately.
The EMF of the cell was measured using the same solu-
tion (102 M Pb(NO3 )2 ), also used for conditioning, on both
sides of the membrane. Since in this symmetrical setup, the
transmembrane potential ideally is zero, the measured value
of ca. 300 mV (cf. Fig. 3) for the individual membranes is
mainly due to different internal filling solutions. Then, the
two kinds of membranes were joined together and mounted
in an electrode body, with the ionophore-loaded membrane
on the sample side. The initial potential reflecting the com-
plex formation constant [48,50] (see below) was stable for
10 and 30 min with 1 and 3, respectively, and then started to
drift toward the value obtained with the single membranes
(Fig. 3). The apparent large difference between the time re-
sponses found with ionophores 1 and 3 reflects the differ-
ence in their mean mobility. With the mobile ionophore 1,
the equilibrium value is approached after 4 h, while with
the polymeric ionophore 3, it is still not reached even af-
ter 11 h. Since the latter corresponds to a mixture of rather
high dispersity, no attempt has been made to fit a diffusion
coefficient to the observed curves.
The initial EMF difference across double membranes
is directly related to the effective ion complex formation
constants of the ionophore in the membrane [48,50]. The

800

700

3
EMF / mV

600

500
1

400

300
0 1 2 3 4 5 6 7 8 9 10 11 12
Time / h

Fig. 3. Time response of ISEs with sandwich membranes (see Section 2). At t = 0, an ionophore-free ion exchanger membrane (inner side) was joined
to an analogous membrane additionally containing ionophore 1 or 3 (cf. Fig. 1 and Table 1).
192 M. Pntener et al. / Analytica Chimica Acta 503 (2004) 187194

Table 2
pot
Potentiometric selectivity coefficients, log KPbJ , obtained with the separate solution method for ionophores 1 and 3 (cf. Fig. 1) on polymer membranes
plasticized with o-NPOE or DOSa
Ion J 1, o-NPOE 1, o-NPOE with PU 1, DOS 1, DOS with PU 3, o-NPOE 3, DOS (membrane VII)
(membrane III) (membrane IV) [19] (membrane VI) (membrane V)
H+ 5.3 0.3 (60.2) 2.9 0.3 (53.4) 7.5 0.2 2.3 0.1 (56.1) 1.3 0.2 (49.3) 1.7 0.6 (50.7)
Na+ 7.2 0.2 (56.6) 4.3 0.6 (53.6) 7.5 0.2 3.6 0.1 (56.4) 3.5 0.2 (41.0) <4.4 0.5 (42.6)
K+ 7.8 0.1 (54.8) 5.3 0.2 (49.7) 6.9 0.2 4.4 0.1 (55.1) 4.1 0.2 (38.0) <4.3 0.4 (44.8)
Ag+ >13.9 2.9 (25.4)b >12.9 0.7 (28.3)b 14.7 0.2 (55.1)b >5.9 2.1 (11.7) >6.5 0.9 (12.9)
Mg2+ <14.3 0.2 (11.4) 9.4 0.2 (22.7) 13.9 0.2 8.4 0.1 (25.8) <6.6 0.3 (17.2) <6.6 0.6 (19.5)
Ca2+ 12.9 0.1 (23.4) 8.0 0.1 (26.2) 13.1 0.2 7.5 0.1 (26.9) 6.1 0.2 (23.7) 6.4 0.6 (25.0)
Cu2+ 6.1 0.1 (30.7) 5.2 0.04 (30.8) 4.0 0.2 5.0 0.1 (30.1) <2.3 0.2 (14.7) <2.3 0.6 (15.6)
Cd2+ 4.1 0.1 (30.9) 3.9 0.1 (27.3) 6.3 0.1 3.6 0.1 (23.8) 4.5 0.2 (24.3) 5.5 0.6 (24.3)
a Standard deviations obtained with three ISEs. In parentheses: response slopes (mV) for the measuring range of 102 to 104 M.
b Slopes for 105.2 to 106 M Ag+ .

values obtained with Pb2+ according to the formalism used
in Ref. [48] are log = 18.4 0.5 and 17.3 0.3 for 1
and 3, respectively, in a PVC/o-NPOE membrane. Earlier,
this complex formation constant of 1 in a PVC/DOS and a
PVC/o-NPOE matrix was determined as log = 15.9 0.2
[42] and 21.3 0.3 [27], respectively. Although the attach-
ment of the coordinating calix[4]arene unit to the polymer
backbone leads to some loss of complex stability, this re-
duction is not substantial and the polymeric ionophore 3 still
belongs to the compounds forming the strongest 1:1 com-
plexes in membranes found so far [49].
pot
The potentiometric selectivity coefficients, log KPbJ , were
determined for ionophores 1 and 3 using o-NPOE or DOS as
plasticizer (Table 2). With the mobile ionophore (1), the se-
lectivities obtained with o-NPOE were somewhat better than
those reported earlier with DOS. Such differences have been
observed repeatedly, and were ascribed to the coordinating
ability of DOS [27,51]. Interestingly, the same trend was
not found with the polymeric ionophore (3). This ionophore
gave a significant loss of selectivity with respect to all inter-
fering ions. As shown by the similar value of the complex
formation constant with Pb2+ (see above), the selectivity
loss is a consequence of stronger interactions with the inter-
(A) consisted of 101 M Pb(NO3 )2 , which induces a rather
strong leaking of Pb2+ to the sample due to its coextraction
on the inner side of the membrane. Indeed, with this internal
solution, the lower detection limit of the ISE membrane with
1 was only ca. 106.5 M Pb2+ . In contrast, with the poly-
meric ionophore 3, the lower detection limit was improved
by about one logarithmic unit (Fig. 4, two top curves). The
second internal solution (B) consisted of 105 M Pb(NO3 )2
and 101 M Na2 EDTA, its pH being adjusted to 7.5 with
NaOH. The ISE with the mobile ionophore 1 showed the ex-
pected apparently super-Nernstian response between 106
and 107 Pb2+ (Fig. 4, bottom curve). Also this bias was
eliminated when using the polymeric ionophore 3. It must
be noted that neither of these internal solutions of extreme
compositions would be applied to practical issues because
EMF / mV

fering ions and not due to weaker interactions with Pb2+ .

Obviously, the other functional groups of the polyurethane
1
are rather good coordinating sites for the interfering alkali A
and alkaline earth metal cations. As a further evidence, the 3
selectivities were also determined with membranes based
on the mobile ionophore 1, which contained about the same
amount of the polyurethane (PU) backbone as membranes
based on 3 (Table 2, columns 3 and 5). The strong loss of 50 mV
selectivity due to the presence of this PU further confirms 3
the above interpretation. It must be emphasized that the se- B
lectivities of the ISE membranes with 3 (Table 2, columns 1
6 and 7) are still very good and sufficient for obtaining a
-9 -8 -7 -6 -5 -4 -3
Pb2+ response down to submicromolar concentrations.
The influence of the reduced mobility of 3 on the response log aPb
to dilute Pb2+ samples was tested with two internal solu-
Fig. 4. Potentiometric response curves of ISEs based on ionophore 1
tions of extreme compositions. In order to ensure the same or 3 (cf. Fig. 1 and Table 1) and having as internal electrolyte either
viscosity, the membrane with 1 contained the same amount A (101 M Pb(NO3 )2 , conventional) or B (105 M Pb(NO3 )2 , 101 M
of PU as that with 3 (see Table 1). The first internal solution Na2 EDTA, 101 M NaOH, pH 7.5).
 M. Pntener et al. / Analytica Chimica Acta 503 (2004) 187194
100 mV
EMF / mV
bg I
bg II
-12 -11 -10 -9 -8 -7 -6 -5 -4
log aPb
Fig. 5. Potentiometric response curves of ISEs based on the ionophore 1
and with an internal solution for optimized lower detection limit (105 M
Pb(NO3 )2 , 103 M NTA, 2.1 101 M NaOH, pH 7.0) in comparison
with those of ISEs based on ionophore 3 and having a conventional inner
electrolyte (101 M Pb(NO3 )2 ). Sample background: bg I, 103 M CaCl2 ,
104 M HNO3 ; bg II, 104 M NaCl, 104 M HNO3 .
much smaller ionic gradients are sufficient to avoid biases
caused by transmembrane ion fluxes.
As a further test, the response of an ISE based on
ionophore 1 and an optimized inner solution [19] was
compared with that of an ISE with ionophore 3 and a
conventional internal electrolyte solution (Fig. 5). With a
103 M CaCl2 solution as sample background, this latter
internal solution with the ISE membrane based on the mo-
bile ionophore 1 gave a lower detection limit of 106.2 M
Pb2+ (Fig. 4). On the other hand, when it was combined
with the immobilized ionophore 3 using the detection limit
was significantly improved but still less good by about one
order of magnitude than that obtained with ionophore 1
and the optimized inner solution (Fig. 5, top two curves).
As indicated by the selectivity coefficients (Table 1), this
is due to the combined interference of H+ (KPbH (aH )2
pot
= 5 1010 ) and Ca2+ (kPbCa aCa = 5.9 1010 ). If, how-
pot
ever, a 104 M NaCl solution is used as sample background
(KPbNa (aNa )2 = 2.6 1012 ), the response of the ISE with
pot
3 and the conventional internal electrolyte is as good as that
of 1 with the optimized inner solution (Fig. 5, two bottom
curves). This result indicates that covalent attachment of
ionophores to a polymer has a high potential in constructing
rugged electrodes since they can efficiently help to reduce
transmembrane ion fluxes. From the various methods for
improving the lower detection limits [3,11,14,20], cova-
lent binding to a polymer is especially relevant in view of
constructing miniaturized sensors having long lifetimes.
Although the polyurethane applied is not optimal regard-
193
ing the selectivity behavior, it must be emphasized that the
selectivity coefficients of <106 for Ca2+ and Mg2+ are
1
still much better than those achieved prior to recent devel-
3 opments of the past 5 years. Also, with the lower detection
limit of 109 M Pb2+ in the presence of 104 M NaCl at
pH 4.0, the new sensor currently belongs to the best ones.
1 In spite of this, there is room for improvements by using
other polymers instead of polyurethanes.
3
4. Conclusions
It is shown that the covalent binding of an ionophore to
a polymer backbone significantly reduces the adverse influ-
ence of zero-current ion fluxes in an ISE membrane. With the
new immobilized Pb2+ -selective ionophore, the lower detec-
-3
tion limit for ISEs having a conventional inner electrolyte has
became comparable to that obtained with the corresponding
mobile ionophore and an optimized inner solution. Although
the use of a polyurethane backbone strongly reduced the se-
lectivities, the general strategy of reducing transmembrane
ion fluxes by covalent attachment of ionophores to polymers
looks very promising on the basis of the present results.
Acknowledgements
This work was financially supported by the Swiss National
Science Foundation, an internal research grant of the ETH
Zrich, Orion Research Inc. (Beverly, MA), and the National
Institutes of Health (grant R01-GM59716). We thank Dr. D.
Wegmann for careful reading of the manuscript.
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