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We report here on the synthesis of poly(vinyl chloride) (PVC) covalently modified with ferrocene groups (FcPVC) and the electrochemical behavior of the resulting polymeric membranes in view of designing all solid state voltammetric ion sensors. The Huisgen cycloaddition ('click chemistry') was found to be a simple and efficient method for ferrocene attachment. A degree of PVC modification with ferrocene groups between 1.9 and 6.1 mol % was achieved. The chemical modification of the PVC backbone does not significantly affect the ion-selective properties (selectivity, mobility, and solvent casting ability) of potentiometric sensing membranes applying this polymer. Importantly, the presence of such ferrocene groups may eliminate the need for an additional redox-active layer between the membrane and the inner electric contact in all solid state sensor designs. Electrochemical doping of this system was studied in a symmetrical sandwich configuration: glassy carbon electrode [absolute value of FcPVC] glassy carbon electrode. Prior electrochemical doping from aqueous solution, resulting in a partial oxidation of the ferrocene groups, was confirmed to be necessary for the sandwich configuration to pass current effectively. The results suggest that only ~2.3 mol % of the ferrocene groups are electrochemically accessible, likely due to surface confined electrochemical behavior in the polymer. Indeed, cyclic voltammetry of aqueous hexacyanoferrate (III) remains featureless at cathodic potentials (down to -0.5 V). This indicates that the modified membrane is not responsive to redox-active species in the sample solution, malting it possible to apply this polymer as a traditional, single membrane. Yet, the redox capacity of the electrode modified with this type of membrane was more than 520 [micro]C considering a 20 [mm.sup.2] active electrode area, which appears to be sufficient for numerous practical ion voltammetric applications. The electrode was observed to operate reproducibly, with 1% standard deviation, when applying pulsed amperometric techniques. 10.1021/ac1010662

This paper reports on three-dimensional synchrotron radiation/Fourier transform-infrared microspectroscopy (SR/FT-IRM) imaging studies of water inclusions at the buried interface of solid-contact-ion-selective electrodes (SC-ISEs). It is our intention to describe a nondestructive method that may be used in surface studies of the buried interfaces of materials, especially multilayers of polymers. Herein, we demonstrate the power of SR/FT-IRM for studying water inclusions at the buried interfaces of SCISEs. A poly(methyl methacrylate)--poly(decyl methacyrlate) [PMMA--PDMA] copolymer revealed the presence of micrometer sized inclusions of water at the gold/ membrane interface, while a coupling of a hydrophobic solid contact of poly(3-octylthiopheue 2,5-diyl) (POT) prevented the accumulation of water at the buried interface. A similar study with a poly (3,4-ethylenedioxythiophene)/poly (styrenesulfonate) [PEDOT/PSS] solid contact also revealed an absence of distinct micrometer-sized pools of water; however, there were signs of absorption of water accompanied by swelling of the PEDOT/PSS underlayer, and these membrane zones are enriched with respect to water. 10.1021/ac101009n

We are demonstrating hero for the first time a thin layer coulometric detection mode for ionophore based liquid ion-selective membranes. Coulometry promises to achieve the design of robust, calibration free sensors that are especially attractive for applications where recalibration in situ is difficult or undesirable. This readout principle is here achieved with porous polypropylene tubing doped with the membrane material and which contains a chlorinated silver wire in the inner compartment, together with the fluidically delivered sample solution. The membrane material consists of the lipophilic plasticizer dodecyl 2-nitrophenyl ether, the lipophilic electrolyte ETH 500, and the calcium ionophore ETH 5234. Impertantly and in contrast to earlier work on voltammetric liquid membrane electrodes, the membrane also contains a cation-exchanger salt, KTFPB. This renders the membrane permselective and allows one to observe open circuit potentiometric responses for the device, which is confirmed to follow the expected Nernstian equation. Moreover, as the same cationic species is now potential determining at both interfaces of the membrane, it is possible to use rapidly diffusing and/or thin membrane systems where transport processes at the inner and outer interface of the membrane do not perturb each other or the overall composition of the membrane. The tubing is immersed in an electrolyte solution where the counter and working electrode are placed, and the potentials are applied relative to the measured open circuit potentials. Exhaustive current decays are observed in the range of 10 to 100 [micro]M calcium chloride. The observed charge, calculated as integrated currents, is linearly dependent on concentration and forms the basis for the coulometric readout of ion-selective membrane electrodes. 10.1021/ac100524z

The concept of locally heated polymeric membrane potentiometric sensors is introduced here for the first time. This is accomplished in an all solid state sensor configuration, utilizing poly(3-octylthiophene) as the intermediate layer between the ion-selective membrane and underlying substrate that integrates the heating circuitry. Temperature pulse potentiometry (TPP) gives convenient peak-shaped analytical signals and affords an additional dimension with these sensors. Numerous advances are envisioned that will benefit the field. The heating step is shown to give an increase in the slope of the copper-selective electrode from 31 to 43 mV per 10-fold activity change, with a reproducibility of the heated potential pulses of 1% at 10 [micro]M copper levels and a potential drift of 0.2 mV/h. Importantly, the magnitude of the potential pulse upon heating the electrode changes as a function of the copper activity, suggesting an attractive way for differential measurement of these devices. The heat pulse is also shown to decrease the detection limit by half an order of magnitude. 10.1021/ac902191h

Ion fluxes across polymeric ion-selective membranes are a decisive parameter dictating the lower detection limit of potentiometric ion sensors. An applied current was earlier proposed to counteract such fluxes and reduce the detection limit to ultratrace levels. So far, however, the method has not been used in practical situations since the correct current amplitude requires prior knowledge of the sample composition. This paper explores the use of the stir effect to evaluate the optimal current by theory and experiments. It is shown that the traditionally used steady-state model assuming a uniform distribution of ion exchanger in the membrane, fixed with time, violates the electroneutrality condition. A modified steady-state model is introduced that allows for a concentration tilt of the ion exchanger and predicts that a stir effect can indeed be utilized to find the optimal current. Ideally, by choosing the optimal current and very long measurement times, the thermodynamic detection limit might be obtained. However, in practice the stir effect declines at low concentrations and the conditions are far from steady state. Therefore, the improvement of the lower detection limit achievable by galvanostatic control is only about 1 order of magnitude. A numerical finite-difference approximation is shown to trace the experimental potential responses of silver-selective electrodes well and to reproduce the stir effect adequately, even for different conditioning protocols. The stir effect is successfully used to improve the detection limit of electrodes with ill-optimized inner solutions; however, significant improvements beyond what is commonly feasible by chemical optimization does not seem to be easily achievable. The results indicate that with conventional membranes the possibility of improving the detection limit by current polarization is much more limited than assumed so far.

A recent new direction in ion-selective electrode (ISE) research utilizes a stir effect to indicate the disappearance of an ion concentration gradient across a thin ion-selective membrane. This zeroing experiment allows one to evaluate the equilibrium relationship between front and backside solutions contacting the membrane by varying the backside solution composition. This method is attractive since the absolute potential during the measurement is not required, thus avoiding standard recalibrations from the sample solution and a careful control of the reference electrode potential. We report here on a new concept to alleviate the need to continuously vary the composition of the backside solution. Instead, transmembrane ion fluxes are counterbalanced at an imposed critical current. A theoretical model illustrates the relationship between the magnitude of this critical current and the concentration of analyte and countertransporting ions and is found to correspond well with experimental results. The approach is demonstrated with lead(II)-selective membranes and protons as dominating interference ions, and the concentration of [Pb.sup.2+] was successfully measured in tap water samples. The principle was further evaluated with calcium-selective membranes and magnesium as counterdiffusing species, with good results. Advantages and limitations arising from the kinetic nature of the perturbation technique are discussed.

This study aimed to develop a novel approach for the production of analytically robust and miniaturized polymeric ion sensors that are vitally important in modern analytical chemistry (e.g., clinical chemistry using single blood droplets, modern biosensors measuring clouds of ions released from nanoparticle-tagged biomolecules, laboratory-on-a-chip applications, etc.). This research has shown that the use of a water-repellent poly(methyl methacrylate)/poly(decyl methacrylate) (PMMA/PDMA) copolymer as the ion-sensing membrane, along with a hydrophobic poly(3-octylthiophene 2,5-diyl) (POT) solid contact as the ion-to-electron transducer, is an excellent strategy for avoiding the detrimental water layer formed at the buried interface of solid-contact ion-selective electrodes (ISEs). Accordingly, it has been necessary to implement a rigorous surface analysis scheme employing electrochemical impedance spectroscopy (EIS), in situ neutron reflectometry/EIS (NR/EIS), secondary ion mass spectrometry (SIMS), and small-angle neutron scattering (SANS) to probe structurally the solid-contact/membrane interface, so as to identify the conditions that eliminate the undesirable water layer in all solid-state polymeric ion sensors. In this work, we provide the first experimental evidence that the PMMA/PDMA copolymer system is susceptible to water 'pooling' at the interface in areas surrounding physical imperfections in the solid contact, with the exposure time for such an event in a PMMA/ PDMA copolymer ISE taking nearly 20 times longer than that for a plasticized poly(vinyl chloride) (PVC) ISE, and the simultaneous use of a hydrophobic POT solid contact with a PMMA/PDMA membrane can eliminate totally this water layer problem.

Potentiometric sensors are today sufficiently well understood and optimized to reach ultratrace level (subnanomolar) detection limits for numerous ions. In many cases of practical relevance, however, a high electrolyte background hampers the attainable detection limits. A particularly difficult sample matrix for potentiometric detection is seawater, where the high saline concentration forms a major interfering background and reduces the activity of most trace metals by complexation. This paper describes for the first time a hyphenated system for the online electrochemically modulated preconcentration and matrix elimination of trace metals, combined with a downstream potentiometric detection with solid contact polymeric membrane ion-selective microelectrodes. Following the preconcentration at the bismuth-coated electrode, the deposited metals are oxidized and released to a medium favorable to potentiometric detection, in this case calcium nitrate. Matrix interferences arising from the saline sample medium are thus circumvented. This concept is successfully evaluated with cadmium as a model trace element and offers potentiometric detection down to low parts per billion levels in samples containing 0.5 M NaCl background electrolyte.

Polymer membrane ion-selective electrodes containing lipophilic ionophores are traditionally interrogated by zero current potentiometry, which, ideally, gives information on the sample activity of ionic species. It is shown here that a discrete cathodic current pulse across an [H.sup.+]-selective polymeric membrane doped with the ionophore ETH 5294 may be used for the chronopotentiometric detection of pH in well-buffered samples. However, a reduction in the buffer capacity leads to large deviations from the expected Nernstian response slope. This is explained by the local depletion of hydrogen ions at the sample--membrane interface as a result of the galvanostatically imposed ion flux in direction of the membrane. This depletion is found to be a function of the total acidity of the sample and can be directly monitored chronopotentiometrically in a flash titration experiment. The subsequent application of a baseline potential pulse reverses the extraction process of the current pulse, allowing one to interrogate the sample with minimal perturbation. In one protocol, total acidity is found to be proportional to the magnitude of applied current at the flash titration end point. More conveniently, the square root of the flash titration end point time observed at a fixed applied current is a linear function of the total acid concentration. This suggests that it is possible to perform rapid localized pH titrations at ion-selective electrodes without the need for volumetric titrimetry. The technique is explored here for acetic acid, MES and citric acid with promising results. Polymeric membrane electrodes based on poly(vinyl chloride) plasticized with o-nitrophenyl octyl ether in a 1:2 mass ratio may be used for the detection of acids of up to ca. 1 mM concentration, with flash titration times on the order of a few seconds. Possible limitations of the technique are discussed, including variations of the acid diffusion coefficients and influence of electrical migration.

We here report on the first example of an aptamer-based potentiometric sandwich assay of proteins. The measurements are based on CdS quantum dot labels of the secondary aptamer, which were determined with a novel solid-contact [Cd.sup.2+]-selective polymer membrane electrode after dissolution with hydrogen peroxide. The electrode exhibited cadmium ion detection limits of 100 pM in 100 mL samples and of 1 nM in 200 [micro]L microwells, using a calcium-selective electrode as a pseudoreference electrode. As a prototype example, thrombin was measured in 200 [micro]L samples with a lower detection limit of 0.14 nM corresponding to 28 fmol of analyte. The results show great promise for the potentiometric determination of proteins at very low concentrations in microliter samples.

Polymeric bulk optode microsphere ion sensors in combination with suspension array technologies such as analytical flow cytometry may become a power tool for measuring electrolytes in physiological samples. In this work, the methodology for the direct measurement of common blood electrolytes in physiological samples using bulk optode microsphere sensors was explored. The simultaneous determination of [Na.sup.+], [K.sup.+], and [Ca.sup.2+] in diluted sheep blood plasma was demonstrated for the first time, using a random suspension array containing three types of mixed microsphere bulk optodes of similar size, fabricated from the same chromoionophore without additional labeling. Sodium ionophore X, potassium ionophore III, and grafted AU-1 in poly(butyl acrylate) were the ionophores used in the bulk optode microsphere ion sensors for [Na.sup.+], [K.sup.+], and [Ca.sup.2+], respectively, in combination with the cation-exchanger NaTFPB (sodium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate) and the same concentration of the chromoionophore ETH 5294 (9-(diethylamino)-5-octadecanoylimino-5H-benzo[a]phen-oxazine) in plasticized poly(vinyl chloride). Excellent reproducibility was achieved for the sensing of potassium ions. The effect of sample pH was relatively small at nearphysiological pH and followed theoretical predictions, yet the sample temperature was found to influence the sensor response to a larger extent. Multiplexed ion sensing was achieved by taking advantage of the chemical tunability of the sensor response, adjusting the sensor compositions so that the three types of ion sensors responded with distinct levels of protonation of the chromoionophore. Consequently, three well-resolved peaks were simultaneously observed in the single-channel histogram during the multiplexed calibration as well as in the subsequent measurement of the three cations in 10-fold-diluted sheep plasma. The assigned peak positions corresponded very well to the physiological range of the measured ions.

A new direction in potentiometric sensing, termed backside calibration potentiometry, was recently introduced. It makes use of the fact that the stir effect disappears in the absence of an ion--ionophore complex concentration gradient across supported liquid ion-selective membranes. This method is especially suitable for measurements in which recalibration in the sample is not feasible, such as in remote monitoring applications. Here, a theoretical model is established to predict the working concentration range of the method. Lead(II)-selective Celgard membranes were used here with H+ as the dominant interfering ions. The emf difference for stirred and unstirred solutions was measured, and the magnitude of this emf change as a function of the sample [Pb.sup.2+] concentration was found to exhibit a bell shape that spans ~3 orders of magnitude. The concentration of interfering ions and the selectivity of the membrane were demonstrated to be important factors that affect the working range. Smaller ratios of primary ion concentrations at both aqueous sides of the membrane gave smaller emf difference values, and emf changes could still be observed with a logarithmic concentration ratio of 0.05. All experimental results correlated satisfactorily with the theoretical model.

Polymeric membrane ion-selective electrodes are normally interrogated by zero current potentiometry, and their selectivity is understood to be primarily dependent on an extraction/ion-exchange equilibrium between the aqueous sample and polymeric membrane. If concentration gradients in the contacting diffusion layers are insubstantial, the membrane response is thought to be rather independent of kinetic processes such as surface blocking effects. In this work, the surface of calcium-selective polymeric ion-selective electrodes is coated with polyelectrolyte multilayers as evidenced by [zeta] potential measurements, atomic force microscopy, and electrochemical impedance spectroscopy. Indeed, such multilayers have no effect on their potentiometric response if the membranes are formulated in a traditional manner, containing a lipophilic ion exchanger and a calcium-selective ionophore. However, drastic changes in the potential response are observed if the membranes are operated in a recently introduced kinetic mode using pulsed chronopotentiometry. The results suggest that the assembled nanostructured multilayers drastically alter the kinetics of ion transport to the sensing membrane, making use of the effect that polyelectrolyte multilayers have different permeabilities toward ions with different valences. The results have implications to the design of chemically selective ion sensors since surface-localized kinetic limitations can now be used as an additional dimension to tune the operational ion selectivity.

Potentiometric sensors based on polymer membrane electrodes, if properly optimized, are useful for measurements at trace levels. The expected independence of the electrochemical signal of the sample size makes them extremely attractive for measurements in small volumes. Here, we report on electrodes for the potentiometric detection of cadmium ions that reach a detection limit of 6 nM and utilize a [Na.sup.+]-selective electrode as pseudoreference in order to facilitate measurements in 150-[micro]L samples. A potentiometric immunoassay of mouse IgG is performed via CdSe quantum dot labels on a secondary antibody according to a sandwich immunoassay protocol in a microtiter plate format. The CdSe quantum dots are found to be easily dissolved/oxidized in a matter of minutes with hydrogen peroxide, allowing us to maintain the pH at a near-neutral value. The potentiometric protein immunoassay exhibits a log--linear response ranging from 0.15 to 4.0 pmol of IgG, with a detection limit of

Multicolor quantum dot-encoded polymeric microspheres are prepared with controllable and uniform doping levels that function as chemical sensors on the basis of bulk optode theory. TOP/TOPO-capped CdSe quantum dots and CdTe quantum dots capped with CdS ([[lambda].sub.em] = 610 and 700 nm, [[lambda].sub.ex] = 510 nm) are blended with a THF solution of poly(methyl methacrylate-co-decyl methacrylate), poly-(n-butylacrylate), or poly(vinyl chloride) plasticized with bis(2-ethylhexyl) sebacate without a need for ligand exchange. Polymeric microspheres are generated under mild, nonreactive conditions with a particle caster that breaks down a polymer stream containing the quantum dots into fine droplets by the vibration of a piezocrystal. The resulting microspheres exhibit uniform size and fluorescence emission intensities. Fluorescent bar codes are obtained by subsequent doping of two quantum dots with different colors and mass ratios into the microspheres. A linear relationship is found between the readout fluorescence ratio of the two types of nanocrystals and the mixing ratio. Quantum dot-encoded ion sensing optode microspheres are prepared by simultaneous doping of sodium ionophore X, chromoionophore II, a lipophilic tetraphenylborate cation exchanger, and TOPO-capped CdSe/CdS quantum dot as the fluorescent label. A net positive charge of the quantum dots is found to induce an anion-exchange effect on the sensor function, and therefore, an increased concentration of the lipophilic cation exchanger is required to achieve proper ion sensing properties. The modified quantum dot-labeled sodium sensing microspheres show satisfactory sodium response between [10.sup.-4] and 0.1 M at pH 4.8, with excellent selectivity toward common interferences. The amount of the carried positive charges of the CdSe quantum dots is estimated as 2.8 [micro]mol/g of quantum dots used in this study.

In direct potentiometry, the magnitude of the measured potentials is used to determine the composition of the sample. While this places rather formidable demands on the required reproducibility of the associated potential measurements, typically on the order of microvolts, in vitro clinical analyses of blood samples are today successfully performed with direct potentiometry using ion-selective electrodes (ISEs). Unfortunately, most other analytical situations do not permit the sensor to be recalibrated every few minutes, as in environmental monitoring or in vivo measurements, and direct potentiometry is often bound to fail as an accurate method in these circumstances. This paper introduces a novel direction for potentiometric sensing, termed backside calibration potentiometry. Chemical asymmetries across thin supported liquid ISE membranes are assessed by determining the direction of potential drift upon changing the stirring rate on either side of the membrane. Disappearance of this drift indicates the disappearance of concentration gradients across the membrane and is used to determine the sample composition if the solution composition at the backside of the membrane and the interfering ion concentration in the sample are known. For practical determinations, the concentration of either the primary or the interfering ion is varied in the reference solution until the stirring effect disappears. The procedure is demonstrated with a [Ca.sup.2+]-selective membrane using [Ba.sup.2+] as the dominant interfering ion. Another example includes the determination of [Pb.sup.2+] in environmental samples where the pH is adjusted to a known level. At pH 4.0, [H.sup.+] turns out to be the dominant interfering ion. The practical applicability of the method is shown with different environmental water samples, for which the results obtained with the novel method are compared with those obtained by traditional calibration using standard additions. The limitations of the novel method in terms of accuracy and applicable concentration ranges are discussed.

Ion-selective electrodes ideally operate on the basis of the Nernst equation, which predicts less than 60- and 30-mV potential change for a 10-fold activity change of monovalent and divalent ions measured at room temperature, respectively. Typical concentration ranges in extracellular fluids are quite narrow for the electrolytes of key importance. A range of 2.2-2.6 mM for calcium ions, for instance, translates into just a 2.2-mV potential change. The direct potentiometric measurement of physiological electrolytes is certainly possible with direct potentiometry and is done routinely in clinical analyzers and handheld measuring devices. It places, however, strong demands on the precision of the reference electrode and requires careful temperature control and frequent calibration runs. In this paper, a robust 10-20-fold sensitivity enhancement for calcium measurements is attained by departing from the classical response mechanism and operating in a non-Nernstian response mode. Stable and reproducible super-Nernstian responses of these so-called pulstrodes in a narrow calcium activity range can be controlled by instrumental means in good agreement with theory. The potentials may be measured during a galvano-static excitation pulse (mode I) or immediately after it (mode II), under open-circuit conditions. Subtraction of the potentials, sampled at different times during a single pulse, allows one to obtain a sensitive differential peak-shaped signal at a critical and fully adjustable analyte activity range. Calcium pulstrodes based on the diamide ionophore AU-1 were characterized and applied to the measurement in model physiological liquids. Super-Nernstian responses exceeding 700 mV/decade were observed in a physiological range of calcium concentration. Such remarkable sensitivity of the pulstrodes, complemented with the well-documented high selectivity of these potentiometric sensors, may provide a significant increase in the accuracy and precision of electrolyte measurements in clinical analysis.

A simple procedure for the development of a range of polymeric ion-selective electrodes (ISEs) with low detection limits is presented. The electrodes were prepared by using a plasticizer-free methyl methacrylate-decyl methacrylate copolymer as membrane matrix and poly(3-octylthiophene) as intermediate layer deposited by solvent casting on gold sputtered copper electrodes as a solid inner contact. Five different electrodes were developed for [Ag.sup.+], [Pb.sup.2+], [Ca.sup.2+], [K.sup.+], and [I.sup.-] , with detection limits mostly in the nanomolar range. In this work, the lowest detection limits reported thus far with solid contact ISEs for the detection of silver (2.0 x [10.sup.-9] M), potassium ([10.sup.-7] M), and iodide ([10.sup.-8] M) are presented. The developed electrodes exhibited a good response time and excellent reproducibility.

Supported liquid membranes are used here to establish steady-state concentration profiles across ion-selective membranes rapidly and reproducibly. This opens up new avenues in the area of nonequilibrium potentiometry, where reproducible accumulation and depletion processes at ion-selective membranes may be used to gain valuable analytical information about the sample. Until today, drifting signals originating from a slowly developing concentration profile across the ion-selective membrane made such approaches impractical in zero current potentiometry. Here, calcium- and silver-selective membranes were placed between two identical aqueous electrolyte solutions, and the open circuit potential was monitored upon changing the composition of one solution. Steady state was reached in ~1 min with 25-[micro]m porous polypropylene membranes filled with bis(2-ethylhexyl) sebacate doped with ionophore and lipophilic ion exchanger. Ion transport across the membrane resulted on the basis of nonsymmetric ion-exchange processes at both membrane sides. The steady-state potential was calculated as the sum of the two membrane phase boundary potentials, and good correspondence to experiment was observed. Concentration polarizations in the contacting aqueous phases were confirmed with stirring experiments. It was found that interferences (barium in the case of calcium electrodes and potassium with silver electrodes) induce a larger potential change than expected with the Nicolsky equation because they influence the level of polarization of the primary ion (calcium or silver) that remains potential determining.

We describe here in detail the first reversible electrochemical sensors for the polyion protamine. Potentiometric sensors were proposed in recent years, mainly for the determination of the polyions heparin and protamine. Such potentiometric polyion sensors functioned on the nonequilibrium extraction of polyions into a hydrophobic membrane phase via ion pairing with lipophilic ion exchangers. This made it difficult to design sensors that operate in a truly reversible fashion. The reversible sensors described here utilize the same basic response mechanism as their potentiometric counterparts, but the processes of extraction and ion stripping are now fully controlled electrochemically. Spontaneous polyion extraction is avoided by using membranes containing highly lipophilic electrolytes that possess no ion-exchange properties. Reversible extraction of polyions is induced if a constant current pulse of fixed duration is applied across the membrane, followed by a baseline potential pulse. The key theoretical response principles of this new class of polyion sensors are discussed here and compared to those of its classical potentiometric counterpart. The electrochemical sensing system is characterized in terms of optimal working conditions, membrane composition, selectivity, and influence of sample stirring and organic-phase diffusion coefficient on the response characteristics. Excellent potential stability and reversibility of the sensors are observed, and measurements of heparin concentration in whole blood samples via protamine titration are demonstrated.

An optical microsensor array is described for the rapid analysis of silver ions at low parts per trillion levels. Because the ionophore o-xylylenebis(N,N-diisobutyldithio-carbamate) (Cu--I) was reevaluated and shown to exhibit excellent selectivity for silver ions, ion-selective electrode (ISE) membranes were optimized and found to exhibit the lowest reported detection limit so far (3 x [10.sup.10] M). A corresponding [Ag.sup.+]-selective fluorescent optical microsensor array for the rapid sensing of trace level [Ag.sup.+] was then developed. It was fabricated using plasticized PVC-based micrometer-scale fluorescent microspheres that were produced via a sonic particle casting device. They contained 156 mmol/kg Cu--I, 10 mmol/kg 9-(diethylamino)--5-[4-(15-buty1-1,13-dioxo-2,14-dioxanodecyl) phenylimino]-benzo[a]phenoxazine (chromoionophore VII, ETH 5418), 2.3 mmol/kg 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate (internal reference dye), and 14 mmol/kg sodium tetrakis[3,5-bis(trifluoromethyl)phenyl]borate and were deposited onto the etched distal end of a 3200-[micro]m-diameter optical fiber bundle. The microarray was characterized by fluorescence spectroscopy in samples containing [10.sup.12]--[10.sup.8] M AgN[O.sub.3] at pH 7.4, with selectivity characteristics comparable to the corresponding ISEs. The response time of the microsensor array was found to be less than 15 min for [10.sup.9] M AgN[O.sub.3], which is drastically shorter than earlier data on optode films (8 h) and corresponding ISEs (30 min). A detection limit of 4 x [10.sup.11] M for [Ag.sup.+] was observed, lower than any previously reported optode or silver-selective ISE. The microsensor array was applied for measurement of free silver levels in buffered pond water samples.

A numerical solution for the prediction of the time-dependent potential response of a polymeric-based ion-selective electrode (ISE) is presented. The model addresses short- and middle-term potential drifts that are dependent on changes in concentration gradients in the aqueous sample and organic membrane phase. This work has important implications for the understanding of the real-time response behavior of potentiometric sensors with low detection limits and with nonclassical super-Nernstian response slopes. As a model system, the initial exposure of membranes containing the well-examined silver ionophore O,O'-bis[2-(methylthio)ethyl]-tert-butylealix[4]arene was monitored, and the large observed potential drifts were compared to theoretical predictions. The model is based on an approximate solution of the diffusion equation for both aqueous and organic diffusion layers using a numerical scheme (finite difference in time and finite elements in space). The model may be evaluated on the basis of experimentally available parameters and gives time-dependent information previously inaccessible with a simpler steady-state diffusion model. For the cases studied, the model gave a very good correlation with experimental data, albeit with lower than expected diffusion coefficients for the organic phase. This model may address numerous open questions regarding the response time and memory effects of low-detection-limit ion-selective electrodes and for other membrane electrodes where ion fluxes are relevant.

The spontaneous hydroxy-bridged dimer formation of metalloporphyrins in ion-selective membranes gives rise to a short sensor lifetime (typically days), triggered by solubility problems, the occurrence of a super-Nernstian response slope, and a pH cross response. This dimer formation is eliminated here by covalent attachment of the ionophore to the polymer matrix. Specifically, two different [indium.sup.III]porphyrins containing polymerizable groups, the chloride-selective chloro(3-[18-(3-acryloyloxypropyl)7,12-bis (1-methoxyethyl)-3,8,13,17-tetramethylporphy-rin-2-yl]propyl ester)indium(III) and the nitrite-selective Chloro(5-(4-acryloyloxyphenyl)-10,15,20-triphenylporphyrinato)indium(III), were synthesized and copolymerized with methyl methacrylate and decyl methacrylate. The covalent attachment of the ionophore to the polymer matrix indeed prevents the metalloporphyrin from forming dimeric species, as confirmed by UV/visible spectroscopy. The ion-selective membranes with grafted indium porphyrin showed Nernstian response slopes to chloride, nitrite, perchlorate, and thiocyanate anions, with a selectivity comparable to membranes with freely dissolved or underivatized metalloporphyrin. The membranes containing grafted ionophores showed a lifetime of at least two months, apparently since crystallization of the poorly soluble dimeric species may no longer occur. This is one of the first examples where the covalent attachment of an ionophore drastically improves on a number of important sensor characteristics.

The extent of optimization of the lower detection limit of ion-selective electrodes (ISEs) can be assessed with an elegant new method. At the detection limit (i.e., in the absence of primary ions in the sample), one can observe a reproducible change in the membrane potential upon alteration of the aqueous diffusion layer thickness. This stir effect is predicted to depend on the composition of the inner solution, which is known to influence the lower detection limit of the potentiometric sensor dramatically. For an optimized electrode, the stir effect is calculated to be exactly one-half the value of the case when substantial coextraction occurs at the inner membrane side. In contrast, there is no stir effect when substantial ion exchange occurs at the inner membrane side. Consequently, this experimental method can be used to determine how well the inner filling solution has been optimized. A rotating disk electrode was used in this study because it provides adequate control of the aqueous diffusion layer thickness. Various ion-selective membranes with a variety of inner solutions that gave different calculated concentrations of the complex at the inner membrane side were studied to evaluate this principle. They contained the well-examined silver ionophore O,O'-bis[2-(methylthio)ethyl]-tert-butylcalix[4]arene, the potassium ionophore valinomycin, or the iodide carrier [9]mercuracarborand-3. Stir effects were determined in different background solutions and compared to theoretical expectations. Correlations were good, and the results encourage the use of such stir-effect measurements to optimize ISE compositions for real-world applications. The technique was also found to be useful in estimating the level of primary ion impurities in the sample. For an iodide-selective electrode measured in phosphoric acid, for example, apparent iodide impurity levels were calculated as 5 x [10.sup.-10] M.

The traditional cation exchangers used in ion-selective electrodes and optodes are tetraphenylborate derivatives, which are generally adequate for most analytical applications but may in some cases suffer from decomposition by acid hydrolysis, oxidants, and light. Recently, halogenated dodecacarboranes were found to be improved cation exchangers in terms of lipophilicity and chemical stability. This forms the basis for the convenient covalent attachment of the cation exchanger to the polymeric backbone of the sensing material. This is a challenge that has not satisfactorily been solved and which is especially important in view of developing ultraminiaturized sensing arrays. Here, a C-derivative of the closo-dodecacarborane anion (C[B.sub.11][H.sub.12.sup.-]) with a polymerizable group was synthesized as a chemically stable cation exchanger. This new derivative was copolymerized with methyl methacrylate and decyl methacrylate (MMA-DMA) to fabricate a plasticizer-free polymer with cation-exchange properties. This polymer could be conveniently blended with traditional plasticized poly(vinyl chloride) or with noncrosslinked methacrylic polymers to give solvent cast films that appear to be clear and homogeneous and that could be doped with ionophores. Optode leaching experiments supported the covalent grafting of the carborane anions. Ion-selective membranes and optode thin films were evaluated in terms of response function, response time, and selectivity. In all cases, the new material exhibited behavior similar to free tetraphenylborate derivative-based membranes. As a result of these studies, an all-polymeric plasticizer-free calcium-selective membrane was fabricated on the basis of the covalently attached carborane, a recently introduced grafted calcium ionophore, and an MMA-DMA polymer matrix. The resulting ion-selective electrodes showed Nernstian response slopes and rapid response times, demonstrating that covalent grafting of all sensing components is a feasible approach to the development of ion sensors.

This paper describes a pulsed galvanostatic technique to interrogate ion-selective electrodes (ISEs) with no intrinsic ion-exchange properties. Each applied current pulse is followed by a longer baseline potential pulse to regenerate the phase boundary region of the ion-selective membrane. The applied current fully controls the magnitude and sign of the ion flux into the membrane, thus offering instrumental control over an effect that has become very important in ion-selective electrode research in recent years. The resulting chronopotentiometric response curves essentially mimic traditional ISE behavior, with apparently Nernstian response slopes and selectivities that can be described with the Nicolsky equation. Additionally, the magnitude and sign of the current pulse may be used to tune sensor selectivity. Perhaps most important, however, appears to be the finding that the extent of concentration polarization near the membrane surface can be accurately controlled by this technique. A growing number of potentiometric techniques are starting to make use of nonequilibrium principles, and the method introduced here may prove to be very useful to advance these areas of research. The basic characteristics of this pulsed galvanostatic technique are here evaluated with plasticized poly(vinyl chloride) membranes containing the sodium-selective ionophore tert-butyl calix[4]arene tetramethyl ester and a lipophilic inert salt.

The lower detection limit and the selectivity behavior of anion-selective electrodes (ISEs) are improved by using optimized inner solutions and membrane compositions. With a membrane based on the recently described ionophore [9]mercuracarborand-3, a detection limit of 2 x [10.sup.-9] M has been achieved for iodide. Nevertheless, the improvements are less pronounced than in the case of cation ISEs. This is mainly due to the fact that so far no anion ISE is known with the extremely high selectivities of cation ISEs. If the membrane does not contain an ionophore, leaching of the ion exchanger from the membrane into the sample is also a relevant limiting factor except for ion exchangers of very high lipophilicity.

A derivative of a known [Ca.sup.2+]-selective ionophore, ETH 129, was synthesized to contain a polymerizable acrylic moiety (AU-1) and covalently grafted into a methyl methacrylate-co-decyl methacrylate polymer matrix. The polymer containing AU-1 was prepared via a simple one-step homogeneous polymerization method. It exhibited mechanical properties suitable for the fabrication of plasticizer-free ion-selective membrane electrodes and bulk optode films by solvent-casting and spin-coating techniques, respectively. The segmented sandwich membrane technique was utilized to assess the binding constant of free and covalently bound ionophores to calcium and to study their diffusion coefficients in the membrane phase. Diffusion was greatly diminished for the bound ionophore. This was confirmed in ion-selective electrode membranes containing no calcium ions in the inner solution, which should normally show apparent super-Nernstian response slopes in dilute calcium solutions. The response slope was Nernstian down to submicromolar concentration levels, indicating slow mass transport of calcium in the membrane. Optical-sensing films with the new copolymer matrix, unblended and blended with PVC-DOS, also confirmed that covalently bound ionophores are fully functional for maintaining selective ion extraction and binding properties of the sensing membrane.

Recently, it has been discovered that carba-closo-dodecaborates can be used as cation exchangers in neutral carrier-based ion-selective chemical sensors. Because of their inherent chemical stability and versatile functionalization chemistries, they offer many advantages that may potentially be exploited for ion analyses that require nontraditional sample conditions, including strongly acidic media. In this work, trimethylammonium salts of undecachlorinated (UCC), undecabrominated (UBC), hexabrominated (HBC), and undecaiodinated (UIC) carborane anions were prepared and evaluated for their potential use in solvent polymeric membrane-based sensors. Computational methods including Natural population analysis and electrostatic mapping were used to predict the ion-exchanging ability of each lipophilic anion. In addition, the sandwich membrane technique was used to evaluate the ion-pairing ability of each carborane anion in situ (i.e., within bis(2-ethylhexyl) sebacate (DOS)- and 2-nitrophenyl octyl ether (o-NPOE)-plasticized ISE membranes). The results of the computational and potentiometric studies found that binding affinity of the anions followed the generalized trend HBC > UCC > UBC > UIC. PVC-DOS bulk optode thin films containing the chromoionophore ETH 5315 and a respective anion were used to determine the chemical stability/lipophilicity of the carboranes and tetrakis[3,5-bis(trifluoromethyl)phenyl] borate (TFPB) in acidic media (0.2 M HOAc) under flowing conditions. The studies found that in terms of stability/lipophilicity UIC > UBC > TFPB [approximately equal to] UCC [much greater than] HBC. Electrodes containing a [Pb.sup.2+]-selective ionophore, tert-butylcalix[4]arene-tetrakis-(N,N-dimethylthioacetamide)(lead IV), were used to evaluate the functionality of each cation exchanger. An evaluation of response characteristics such as slope and selectivity found that UIC and UBC were quite comparable to the behavior of TFPB. Interestingly, both UIC and UBC showed a marked selectivity improvement over cadmium, with log [K.sup.pot.sub.Pb,Cd] values of -7.19 and -7.29, respectively, with TFPB giving a value of -5.89. Demonstrating excellent stability and suitable electrostatic properties, the carboranes, UIC in particular, are a very promising

Fluorescent sensing microspheres based on perhaps the most selective and practically useful chloride ionophore known, the recently reported [9]mercuracarborand-3 (MC-3), have been prepared and optimized for physiological measurements. In initial work, this ionophore was shown to yield functional optical sensing films in combination with an electrically neutral chromoionophore, ETH 5418. Unfortunately, however, these optodes suffered from unacceptably high levels of sodium interference under physiological conditions. To better understand the sensing mechanism, optical and potentiometric binding experiments were used to characterize the stoichiometry and the complex formation constants for this ionophore. It was found that the preferred stoichiometry is 1:2, rather than 1:1 as assumed earlier. The 1:2 complex is extremely stable (log [[beta].sub.2] = 13.4), but a relatively strong 1:1 complex also exists (log [K.sub.1] = 9.9). These characteristics were used to fabricate chloride optodes that make use of the stepwise ion-ionophore decomplexation equilibrium, by adding a calculated amount of lipophilic anion exchanger to the polymer film. Such optodes showed dramatically reduced sodium interference while maintaining the excellent selectivity of the traditional formulation. The optimized composition also shifted the measuring range to physiological conditions, making them useful for the assessment of chloride in undiluted and 10-fold-diluted blood at pH 7.4. After necessary alterations of the particle preparation procedure and sensor formulation, the new insights were used to fabricate mass-produced optical sensing microspheres with characteristics essentially identical to those of the optode sensing films.

The use of flow cytometry as a rapid, high-throughput diagnostic tool for the analysis of ions is described. Monodisperse, uniform microspheres, which obey bulk optode theory and are governed by bulk extraction processes rather than surface phenomena, were prepared under mild, nonreactive conditions using a sonic stream particle casting apparatus. As an initial example demonstrating the utility of this approach, microspheres that contained a [H.sup.+]-selective fluorescent chromoionophore (ETH 5294), a cation-exchanger (NaTFPB), and either a highly sodium-selective (sodium ionophore X) or a potassium-selective ionophore (BME-44) were prepared. Separate solution analysis of sodium- and potassium-selective microspheres resulted in the generation of functional response curves using peak channel fluorescence intensities. The selectivity observed for both types of particles is sufficient for the clinical determination of [Na.sup.+] and [K.sup.+]. Furthermore, sodium- and potassium-selective microspheres were analyzed in parallel using sodium sample solutions, resulting in the successful determination of sodium ion concentrations and providing important information about the selectivity of the potassium-selective sensors over sodium. This work demonstrates the potential applicability of flow cytometry as a means for developing multiplexed, rapid, high-throughput analyses for clinically relevant ions.

The development and characterization of small, uniform, and mass-produced plasticized PVC-based sensing microspheres in view of rapid trace level analysis of lead ions is reported. Micrometer-sized particles obtained via an automated casting process were rendered selective for lead ions by doping them with highly selective components in a manner analogous to traditional optode sensing films. Single particles that contained the lipophilic ionophore N,N,N',N'-tetradodecyl-3-6-dioxaoctane-1-thio-8-oxodiamide (ETH 5493), the chromoionophore ETH 5418 together with a lipophilized indocarbocyanine derivative as internal reference dye (DiI[C.sub.18]), and lipophilic ion-exchanger sites sodium tetrakis[3,5-bistrifiuoromethylphenyl]borate, yielded measurable lead responses at the low nanomolar level in pH buffered solutions. The detection limit for single particles was 3 x [10.sup.-9] M at pH 5.7. The microspheres were fabricated via a reproducible formation of polymer droplets within a flowing aqueous phase followed by collection of spherical particles of ~13 [micro]m in size. The particles were immobilized and assayed individually in a microflow cell via fluorescence microscopy. Selectivity patterns found were in agreement with those reported earlier for the lead-selective ligand ETH 5493, and all response functions were fully described by theory. In contrast to optode films that necessitated very long equilibration times and large sample volumes in diluted samples of analyte, particles exhibited extremely enhanced equilibrium response times. Thus, for lead sample concentrations at and above 5 x [10.sup.-8] M, response times were ~3 min, whereas at the detection limit, complete equilibrium was recorded after just 15 min, with required sample volumes on the order of 1 mL. This new class of microspheres appears to be suitable for rapid and sensitive ion detection at trace levels in environmental and biological applications.

Submicromolar to picomolar lower detection limits have recently been obtained with various polymer membrane ion-selective electrodes by minimizing biases due to ion fluxes through the membrane. For the best performance, the compositions of the membrane and inner solution should be optimized for each application. Given the number of parameters to be adjusted, it has been difficult to find the best parameters for a target sample. In this paper, a much simplified and more practical steady-state model of zero-current ion fluxes is derived, which is based on measurable parameters. The model allows one to predict achievable lower detection limits for a membrane with given selectivities. It can also be used to predict the optimal composition of the inner filling solution for the measurement of samples with a known, typical ionic background. Selectivity coefficients of monovalent and divalent analyte ions required for desired detection limits in drinking water are calculated. As an application of the proposed general recipe, a silver-selective electrode is developed on the basis of the ionophore O,O'-bis[2-(methylthio)ethyl]-tert-butylcalix[4]arene. With the predicted optimal composition of the inner electrolyte, its lower detection limit is found to be [10.sup.-9] M or 100 ppt [Ag.sup.+] with an ionic background of [10.sup.5] M LiN[O.sub.3], which is very close to the expected value.

The upper detection limit of polar ionophore-based ion-selective electrode membranes is predicted by utilizing the coextraction constant of dissociated electrolyte, the stability constant of the ionophore, and the membrane composition. The coextraction constant of dissociated electrolytes into the polar poly(vinyl chloride) membrane plasticized with o-nitrophenyl octyl ether (PVC-NPOE) is here measured by a novel approach. The sandwich membrane technique is utilized, with one membrane segment containing a lipophilic cation exchanger and the other containing an anion exchanger. This yields information about the coextraction constant and the free ion concentrations of the electrolyte in the two segments. Predictions correlate quantitatively with the upper detection limit observed for ion-selective electrodes based on the ionophores valinomycin, tert-butylcalix[4]arene tetraethyl ester, and calcimycin. The difficulties of the prediction of the upper detection limit for nonpolar poly(vinyl chloride) membranes plasticized with bis(2-ethylhexyl sebacate) (PVC-DOS) due to ion association are discussed in detail. A thermodynamic cycle experiment with a series of sandwich membranes shows that the principal processes governing the upper detection limit of PVC-DOS membranes are identical to those for the PVC-NPOE membranes. However, the stability of the ion pairs between the ionophore-metal ion complexes and the extracted anion are different from that of ion pairs formed between the same anion and the lipophilic anion exchanger. This makes it difficult to quantitatively predict the upper detection limit on the basis of simple apparent coextraction and complexation data alone. The approach reported herein is useful not only for mechanistic purposes but also to shed light onto the many cases where coextraction effects need to be understood but are not directly experimentally accessible.

The 2,3,4,5,6,7,8,9,10,11,12-undecabromocarborane anion, 1-HC[B.sub.11][Br.sub.11.sup.-] (UB[C.sup.-]) has been evaluated for its suitability as an ion exchanger in solvent polymeric membrane electrodes and bulk optodes. Experiments comparing the chemical stability of the perhalogenated carborane anion to that of the best lipophilic tetraphenylborate, 3,5-[bis(trifluoromethyl)phenyl]borate (TFP[B.sup.-]), demonstrated that in the presence of 0.2 M acetic acid TFP[B.sup.-] was completely lost within 6 h, while the concentration of UB[C.sup.-] decreased by less than 10% in the same time period. Thin-film bulk optodes containing BME-44 as potassium-selective ionophore, ETH 5294 as chromoionophore, and UB[C.sup.-] as ionic sites exhibited a [K.sup.+] response similar to analogous optodes containing TFP[B.sup.-], with comparable selectivities over [Na.sup.+] and [Ca.sup.2+]. Potentiometric measurements evaluating the selectivity behavior of UB[C.sup.-] in both ionophore-free and ionophore-containing electrodes were performed. Ionophore-free PVC membranes containing UB[C.sup.-] as ion exchanger and either DOS or NPOE as plasticizer also demonstrated selectivity similar to TFP[B.sup.-]-containing membranes. Sodium-selective membranes containing the ionophore 4-tertbutylcalix[4]arenetetraacetic acid tetraethyl ester (sodium ionophore X) and UB[C.sup.-] as ionic sites showed a Nernstian response for sodium and selectivity comparable to that found in analogous electrodes containing TFP[B.sup.-].

A convenient method for the preparation of monodisperse, plasticized poly(vinyl chloride) particles based on an automated particle casting technique is described. The particles are made highly selective for a number of ions by doping them with ionophores and other active components, in complete analogy to thin-film or fiber-optic chemical sensors. The approach used here produces spheres of high monodispersity at a rate of ~20 000 particles/s. The casting process is based on a reproducible polymer drop formation and precipitation process, and the particles are formed under very mild, nonreactive conditions. This allows one to conveniently incorporate known amounts of different active components into the polymers. As an initial example, the particles are doped with three optical sensing components, the sodium ionophore tert-butylcalix[4]arene tetraethyl ester, the [H.sup.+]-chromoionophore ETH 5294, and the anionic additive sodium tetrakis[3,5-bis(trifluoromethyl)phenyl] borate. The particles are found to be of spherical shape with a diameter of ~10 [micro]m. They respond individually and selectively to sodium according to classical optode theory, as determined by fluorescence microscopy. With a RSD of 1.6%, sensing reproducibility from particle to particle is excellent. This technique may allow the development of mass-produced chemically selective microspheres on the basis of bulk extraction processes.

The equilibrium partitioning of lipophilic ion-exchanger salts from ion-selective polymeric membrane electrodes (ISEs) and its possible effect on the lower detection limit of these sensors is described. Predictions are made on the basis of various parameters, including the knowledge of tetraphenylborate potassium salt partitioning constants, the selectivity of ionophore-free ion-exchanger membranes, and ionophore stability constants in the membrane. Ion-exchanger lipophilicities are significantly increased if the membrane contains an ionophore that strongly binds the primary ion. Predicted detection limits are on the order of [10.sup.-5]-[10.sup.-8] M for ionophore-free membranes, and may reach levels as low as [10.sup.-18] M with adequate ionophores in the membrane. Experiments are performed for well-described lead-selective membranes containing different tetraphenylborate derivatives, and detection limits appear to be independent of the ion-exchanger used. However, they are much higher if a more hydrophilic carborane cation-exchanger is incorporated in the membrane. The first finding confirms recent theory, which states that transmembrane ion fluxes, given by a small level of ion-exchange at the sample side by interfering ions, normally dictate the detection limit of these sensing systems. Predicted detection limits on the basis of ion-exchanger leaching alone are here listed for a number of analytically relevant cases. For potassium-selective electrodes containing BME-44 and tetraphenylborate as ion-exchanger, the experimental detection limits are in agreement with predicted values. These results suggest that the detection limit of many current ISEs for ultratrace level analysis are, in optimal cases, dictated by transmembrane ion fluxes; however, because improved chemical solutions are being developed to reduce such effects, simple ion-exchanger partitioning may indeed become an important mechanism that can give higher detection limits than practically desired, and should not be ruled out.

Polymeric membrane ion-selective electrodes exhibit an apparently super-Nernstian response at low sample activities if inner solutions are used that induce strong zero-current fluxes of primary ions toward the inner compartment This is due to the limited ion fluxes in the aqueous boundary layer near the membrane. In the presence of labile complexes, the effective flux rate is increased and the emf depends on the total concentration of the ions. The concept is illustrated experimentally with calcium-selective electrodes based on the ionophore N,N-dicyclohexyl-N',N'-dioctadecyl-3-oxapentanediamide (ETH 5234) that either respond to total or free ion concentrations. Samples can be distinguished that contain varying levels of total calcium but are all buffered with EDTA to the same free calcium concentration of 5 x 10(super -8) M.

Selectivities of solvent polymeric membrane ion-selective electrodes (ISEs) are quantitatively related to equilibria at the interface between the sample and the electrode membrane. However, only correctly determined selectivity coefficients allow accurate predictions of ISE responses to real-world samples. Moreover, they are also required for the optimization of ionophore structures and membrane compositions. Best suited for such purposes are potentiometric selectivity coefficients as defined already in the 1960s. This paper briefly reviews the basic relationships and focuses on possible biases in the determination of selectivity coefficients. The traditional methods to determine selectivity coefficients (separate solution method, fixed interference method) are still the same as those originally proposed by IUPAC in 1976. However, several precautions are needed to obtain meaningful data. For example, errors arise when the response to a weakly interfering ion is also influenced by the primary ion leaching from the membrane. Wrong selectivity coefficients may be also obtained when the interfering agent is highly preferred and the electrode shows counterion interference. Recent advances show how such pitfalls can be avoided. A detailed recipe to determine correct potentiometric selectivity coefficients unaffected by such biases is presented.

A potentiometric method to determine ionophore complex formation constants in solvent polymeric membrane phases, proposed originally by Russian researchers, is critically evaluated and compared to other established methods. It requires membrane potential measurements on two-layer sandwich membranes, where only one side contains the ionophore. The resulting initial membrane potential reflects the ion activity ratio at both aqueous phase - membrane interfaces and can be conveniently used to calculate complex formation constants in situ. This method is potentially useful, since it does not require the use of a reference ion or second ionophore in the measurement. In this paper, the five ionophores valinomycin, BME-44, ETH 2120, tert-butylcalix[4]arene tetraethyl ester, and S,S'-methylenebis(diisobutyldithiocarbamate) are characterized in poly(vinyl chloride) (PVC) plasticized with dioctyl sebacate (DOS) and compared with other established methods. The resulting formation constants correspond well to literature values. The influence of varying membrane concentrations and different anionic site additives is studied and found to be relatively small. Experiments are also performed with and without lipophilic inert electrolytes and with ionophore-free sandwich membranes to illustrate the effect of ion pairing and the membrane internal diffusion potential on the response of such sandwich membranes. These experiments suggest that ions are completely associated in PVC - DOS membranes, but that such ion pairs are rather nonspecific. Diffusion potentials seem to play a minor role with these systems. The results are explained with theory. This work indicates that the characterization of electrically charged ionophores, anion-selective ionophores, and ionophores in membrane matrixes other than PVC plasticized with DOS may now be experimentally accessible.

Polymer membrane-based potentiometric sensors have been developed earlier to provide a rapid and direct method of analysis for polyions such as heparin, a natural anticoagulant administered to prevent thrombus formation during cardiovascular surgery. These heparin sensors are irreversible, requiring a membrane renewal procedure between measurements which currently prevents the sensors from being used for continuous monitoring of blood heparin. A newly developed heparin sensor is shown here to allow an alternate and more practical method of membrane renewal. The electrically charged H(super +) ionophore 5-(octadecanoyloxy)-2-(4-nitrophenylazo)phenol (ETH 2412) is incorporated as an additional ionophore into a heparin-sensing membrane. This membrane will respond to pH only at low H(super +) concentrations, while sample anions are coextracted with H(super +) ions into the membrane at physiological pH. In buffered samples at physiological pH, the sensors will therefore respond to heparin via an ion-exchange mechanism with chloride anions. The pH cross-sensitive heparin-sensing membranes are shown to give an excellent potentiometric response toward heparin in aqueous samples at physiological pH and Cl(super -) levels as well as in undiluted whole blood with no loss of heparin response. The membrane renewal is accomplished by moderately increasing the pH of the sample, causing heparin to diffuse out of the membrane with H(super +) ions. Reproducibilities are, with less than 1 mV standard deviation, improved over the classical system. Unlike the high NaCl concentration used to strip heparin from the previously established heparin sensor, the pH change used here could ultimately be performed locally at the sample-membrane interface, allowing the sensor to be used for automated long-term monitoring of heparin in blood. A theoretical model is presented to explain the experimental results.

This paper describes basic response features of solvent polymeric membrane ion sensors with voltammetric and amperometric transduction. The model systems used here contain no ionophore for simplicity reasons. Reasonable simplifications of the theory are introduced that allow one to understand the response mechanism in view of a practical application of these sensors. It is shown that ion-sensing membranes preferentially contain no ion-exchanger properties in order to function optimally in a voltammetric mode. As with the systems studied by Kihara, both liquid-polymer interfaces of the membrane are preferably polarizable. Specifically, they contain the highly lipophilic electrolyte tetradodecylammonium tetrakis(4-chlorophenyl)borate (ETH 500) in the membrane to improve lifetime, increase the magnitude of the potential window, and prohibit exchange reactions with sample ions. An ohmic behavior that is associated with an assisted electrolyte-transfer process is observed only above a threshold potential which can be quantitatively predicted by theory. The threshold potential depends on the nature and activity of sample anions and cations in the sample and inner filling solution of the membrane electrode. Within the experimental conditions discussed in this paper, these sensors seem to measure sample ion activities, not concentrations, since the rate-limiting step is the diffusion of extracted ions away from the interface into the membrane bulk. Similarly, no effect of sample stirring on the measured current is observed. This contrasts to work done on liquid-liquid electrolyte-transfer reactions, where large diffusion coefficients in the organic phase often lead to substantial sample depletion effects. The detection of anions and cations with the same membrane is demonstrated in a cyclic voltammetric mode. Direct continuous detection of one type of anion is accomplished by pulsed amperometry to ensure a rapid, repetitive renewal of the membrane composition between measurements.

An examination of the influence of the composition of the internal electrolyte solution on the response of Pb(super 2+)- and Ca(super 2+)-selective membrane electrodes has demonstrated the improvement of the lower detection limit via the generation of ionic gradients in the membrane. These ionic gradients result in a flux of primary ions toward the inner reference electrolyte solution. Results verify the contribution of both ion exchange and coextraction to the induced fluxes and show the suitability of different internal electrolyte solutions for determining true selectivity coefficients and measuring submicromolar activities.

A quantitative steady-state ion flux model that describes the underlying processes in ion-selective electrodes (ISEs) has been developed. It has been proven to be useful in understanding the basic factors influencing the lower detection limit of carrier-based ISEs. Concentration gradients within the organic phase are induced by ion-exchange and coextraction equilibria on both sides of the membrane, which also affect the lower detection limit via the resulting ion fluxes. These findings show that, although trace level measurements with ISEs are feasible, they frequently require greater membrane selectivities than expected from the Nicolskii equation.

Solvent polymeric membranes containing electrically charged [H.sup.+] cariers show dramatic variations in their pH response behavior depending on the charge type of lipophilic ionic sites present in the membrane. These shifts are predicted with a simplified phase boundary potential model. Membranes without ion-exchanger sites are expected to show half-Nernstian electrode slopes with extended measuring ranges if the ionophore has acidic as well as basic properties. Relevant experiments with membrane electrodes on the basis of the electrically charged carriers 4[prime],5[prime]-dibromofluorescein octadecyl ester (ETH 7075) and 5-(octadecanoyloxy)-2-(4-nitrophenylazo)phenol (ETH 2412), lipophilic indicators formerly used in optode films, confirm these predictions. It is shown that the incorporation of tridodecylmethylammonium chloride into NPOE-PVC membranes with ETH 7075 induces a dynamic range from 4 to 10.4 in 0.1 M KCl while membranes with ETH 2412 show a dynamic range that is shifted by [approximately]4 orders of magnitude toward higher pH values. Experimental EMF responses correlate satisfactorily to analogous absorbance experiments on thin polymeric films, although deviations in the ion-exchange region at high sample pH are larger than ordinarily observed with neutral carrier-based systems. Electrode membranes based on ETH 7075 are characterized and optimized. Anion interferences quantitatively follow the Hofmeister selectivity pattern of ionophore-free membranes. These electrodes show no discrimination among the tested alkali metal ions, indicating that the lipophilicity of cations is not a major factor governing the lower detection limit. The electrodes are stable and exhibit lifetimes of over half a year without appreciable loss in analytical performance after continuous storage in electrolyte solutions and may in some cases be a viable alternative to electrode membranes containing neutral carriers.

Solvent polymeric membranes containing electrically charged [H.sup.+] cariers show dramatic variations in their pH response behavior depending on the charge type of lipophilic ionic sites present in the membrane. These shifts are predicted with a simplified phase boundary potential model. Membranes without ion-exchanger sites are expected to show half-Nernstian electrode slopes with extended measuring ranges if the ionophore has acidic as well as basic properties. Relevant experiments with membrane electrodes on the basis of the electrically charged carriers 4[prime],5[prime]-dibromofluorescein octadecyl ester (ETH 7075) and 5-(octadecanoyloxy)-2-(4-nitrophenylazo)phenol (ETH 2412), lipophilic indicators formerly used in optode films, confirm these predictions. It is shown that the incorporation of tridodecylmethylammonium chloride into NPOE-PVC membranes with ETH 7075 induces a dynamic range from 4 to 10.4 in 0.1 M KCl while membranes with ETH 2412 show a dynamic range that is shifted by [approximately]4 orders of magnitude toward higher pH values. Experimental EMF responses correlate satisfactorily to analogous absorbance experiments on thin polymeric films, although deviations in the ion-exchange region at high sample pH are larger than ordinarily observed with neutral carrier-based systems. Electrode membranes based on ETH 7075 are characterized and optimized. Anion interferences quantitatively follow the Hofmeister selectivity pattern of ionophore-free membranes. These electrodes show no discrimination among the tested alkali metal ions, indicating that the lipophilicity of cations is not a major factor governing the lower detection limit. The electrodes are stable and exhibit lifetimes of over half a year without appreciable loss in analytical performance after continuous storage in electrolyte solutions and may in some cases be a viable alternative to electrode membranes containing neutral carriers.

Individual carrier-based ion-selective electrodes and bulk optides are characterized. Selectivity coefficients were determined by a number of methods, including fixed interference, matched potential, mixed solution and separate solution. The characterization of the response mechanism of optodes were based on Valinomycin and the alkyl-acridine orange derivative called Ind-2. A number of sensors were also determined for NH4+ as an alternative response mechanism.

Lipophilic inert electrolytes, i.e., salts without ion-exchange properties, may influence the selectivity of ionophore-based liquid membrane electrodes by affecting the activity coefficients in the organic phase. It is expected by a theoretical model that the addition of a lipophilic salt renders the ion-selective electrode more selective for divalent over monovalent ions. These predictions are confirmed with [Ca.sup.2+]-responsive membranes containing the ionophores ETH 2120, ETH 1001, and ETH 129. The effect is especially pronounced with nonpolar membrane phases containing a low concentration of charged species, where up to 2 orders of magnitude selectivity improvement is observed.

A spectropotentiometric approach is used to examine time-dependent processes created by acidic solutions in solvent polymeric membranes characterized by an H+-selective chromoionophore. Imaging tests show nonlinear steady-state concentration profiles and rapid diffusion processes that are attributed to the parallel extraction of undissociated acid into the membrane. A description of the reaction of H+ chromoionophore-based anion optodes is provided.

The detection limit of carrier-based ion-selective electrodes is explained by the presence of a locally elevated concentration of measuring ions at the sample-membrane phase boundary. Since ion-selective electrodes are responsive to phase boundary activities, such elevated concentrations render the potentiometric sensor insensitive to dilute bulk concentration changes. Different mechanisms for the continuous release of measuring ions from the membrane are conceivable. The extraction of inner electrolyte into the backside of the ion-selective membrane is predicted to lead to a concentration gradient of electrolyte across the membrane and therefore to a net flux of measuring ions from the inner filling solution to the sample. This effect is described by an extended model that respects the relevant extraction and diffusion processes. The extent of coextraction at the backside is predicted on the basis of potentiometric measurements on the range of anion interference. These predictions are found to relate well to experimental results with valinomycin electrodes. The presence of lipophilic anions in the inner electrolyte is found to increase detection limits owing to the increased extraction into the membrane. An upper limit apparently exists beyond which the detection limit is no longer increased upon increasing the inner filling solution concentration. Stirring the sample decreases the detection limit owing to increased mass transport from the membrane surface to the bulk sample.