Electrochemical sensing of blood proteins for mild traumatic brain injury (mTBI) diagnostics and prognostics: towards a point-of-care application.

Traumatic Brain Injury (TBI) being one of the principal causes of death and acquired disability in the world imposes a large burden on the global economy. Mild TBI (mTBI) is particularly challenging to assess due to the frequent lack of well-pronounced post-injury symptoms. However, if left untreated mTBI (especially when repetitive) can lead to serious long-term implications such as cognitive and neuropathological disorders. Computer tomography and magnetic resonance imaging commonly used for TBI diagnostics require well-trained personnel, are costly, difficult to adapt for on-site measurements and are not always reliable in identifying small brain lesions. Thus, there is an increasing demand for sensitive point-of-care (POC) testing tools in order to aid mTBI diagnostics and prediction of long-term effects. Biomarker quantification in body fluids is a promising basis for POC measurements, even though establishing a clinically relevant mTBI biomarker panel remains a challenge. Actually, a minimally invasive, rapid and reliable multianalyte detection device would allow the efficient determination of injury biomarker release kinetics and thus support the preclinical evaluation and clinical validation of a proposed biomarker panel for future decentralized in vitro diagnostics. In this respect electrochemical biosensors have recently attracted great attention and the present article provides a critical study on the electrochemical protocols suggested in the literature for detection of mTBI-relevant protein biomarkers. The authors give an overview of the analytical approaches for transduction element functionalization, review recent technological advances and highlight the key challenges remaining in view of an eventual integration of the proposed concepts into POC diagnostic solutions.

In-Line Seawater Phosphate Detection with Ion-Exchange Membrane Reagent Delivery.

There is an urgent need for reliable seawater phosphate measuring tools to better assess eutrophication. Today, most accepted sensing approaches are based on the established colorimetric molybdenum blue assay. It requires one to modify the sample to strongly acidic conditions and to add various reagents, principally molybdate and reducing agent (e.g., ascorbic acid), to form a blue colored phosphate complex that is subsequently detected spectrophotometrically. The associated need for large sample and mobile phase reservoirs and mixing coils are, unfortunately, not ideally adapted for the development of operationally simple in situ sensing instruments. It is here demonstrated for the first time that the key reagents needed to achieve phosphate detection by the molybdate method may be delivered by passive counter transport across ion-exchange membranes. A cation-exchange Donnan exclusion membrane placed in contact with a sample flow (450 µm thick) is shown to provide the strongly acidic conditions (pH ∼ 1) necessary for phosphate determination. Proton transport is driven, via cation-exchange, by the high sodium content of the seawater sample. Molybdate was similarly released through an anion-exchange membrane by chloride counter transport. Consequently, an in-line flow system containing the two membrane modules in series was used for delivering both hydrogen and molybdate ions into the sample to form the desired phosphomolybdate complex for subsequent spectrophotometric detection. A linear calibration  in the range of 0.1-10 µM phosphate (3-300 ppb inorganic P) was achieved, which is sufficiently attractive for environmental work. A range of seawater samples was tested and the results from this membrane delivery device showed no significant differences compared to the classical molybdate assay chosen as the reference method.

Fast Potentiometric CO2 Sensor for High-Resolution in Situ Measurements in Fresh Water Systems.

We present a new potentiometric sensor principle and a calibration protocol for in situ profiling of dissolved CO2 with high temporal and spatial resolution in fresh water lakes. The sensor system is based on the measurement of EMF between two solid-contact ion selective electrodes (SC-ISEs), a hydrogen ion selective and a carbonate selective sensor. Since it relies on SC-ISEs, it is insensitive to changes in pressure, thus suitable for in situ studies. Also, as it offers a response time ( t95%) of <10 s, it allows for profiling applications at high spatial resolution. The proposed optimum in situ protocol accounts for the continuous drift and change in offset that remains a challenge during profiling in natural waters. The fast response resolves features that are usually missed by standard methods like the classical Severinghaus CO2 probe. In addition, the insensitivity of the presented setup to dissolved sulfide allows also for measurements in anoxic zones of eutrophic systems. Highly resolved CO2 concentration profiles obtained by the novel and robust SC-ISE setup along with the developed optimum in situ protocol allow investigating hotspots of biogeochemical processes, such as mineralization and primary production in the water column and help improving estimates for CO2 turnover in freshwater systems.

In Situ Detection of Macronutrients and Chloride in Seawater by Submersible Electrochemical Sensors.

A new submersible probe for the in situ detection of nitrate, nitrite, and chloride in seawater is presented. Inline coupling of a desalination unit, an acidification unit, and a sensing flow cell containing all-solid-state membrane electrodes allows for the potentiometric detection of nitrate and nitrite after removal of the key interfering ions in seawater, chloride and hydroxide. Thus, the electrodes exhibited attractive analytical performances for the potentiometric detection of nitrate and nitrite in desalinated and acidified seawater: fast response time ( t95 < 12 s), excellent stability (long-term drifts of <0.5 mV h-1), good reproducibility (calibration parameter deviation of <3%), and satisfactory accuracy (uncertainties <8%Diff compared to reference technique). The desalination cell, which can be repetitively used for about 30 times, may additionally be used as an exhaustive, and therefore calibration-free, electrochemical sensor for chloride and indirect salinity detection. The detection of these two parameters together with nitrate and nitrite may be useful for the correlation of relative changes in macronutrient levels with salinity cycles, which is of special interest in recessed coastal water bodies. The system is capable of autonomous operation during deployment, with routines for repetitive measurements (every 2 h), data storage and management, and computer visualization of the data in real time. In situ temporal profiles observed in the Arcachon Bay (France) showed valuable environmental information concerning tide-dependent cycles of nitrate and chloride levels in the lagoon, which are here observed for the first time using direct in situ measurements. The submersible probe based on membrane electrodes presented herein may facilitate the study of biogeochemical processes occurring in marine ecosystems by the direct monitoring of nitrate and nitrite levels, which are key chemical targets in coastal waters.

Fluorinated tripodal receptors for potentiometric chloride detection in biological fluids.

Fluorinated tripodal compounds were recently reported to be efficient transmembrane transporters for a series of inorganic anions. In particular, this class of receptors has been shown to be suitable for the effective complexation of chloride, nitrate, bicarbonate and sulfate anions via hydrogen bonding. The potentiometric properties of urea and thiourea-based fluorinated tripodal receptors are explored here for the first time, in light of the need for reliable sensors for chloride monitoring in undiluted biological fluids. The ion selective electrode (ISE) membranes with tren-based tris-urea bis(CF3) tripodal compound (ionophore I) were found to exhibit the best selectivity for chloride over major lipophilic anions such as salicylate ( [Formula: see text] ) and thiocyanate ( [Formula: see text] ). Ionophore I-based ISEs were successfully applied for chloride determination in undiluted human serum as well as artificial serum sample, the slope of the linear calibration at the relevant background of interfering ions being close to Nernstian (49.8±1.7mV). The results of potentiometric measurements were confirmed by argentometric titration. Moreover, the ionophore I-based ISE membrane was shown to exhibit a very good long-term stability of potentiometric performance over the period of 10 weeks. Nuclear magnetic resonance (NMR) titrations, potentiometric sandwich membrane experiments and density functional theory (DFT) computational studies were performed to determine the binding constants and suggest 1:1 complexation stoichiometry for the ionophore I with chloride as well as salicylate.

In-Line Acidification for Potentiometric Sensing of Nitrite in Natural Waters.

We report on a novel approach for in-line sample acidification that results in a significant improvement in the limit of detection of potentiometric anion-selective electrodes aiming at determining nutrients in natural waters. The working principle of the developed acidification module relies on the cation-exchange process between the sample and an ion-exchange Donnan exclusion membrane in its protonated form. The resulting in-line acidification of natural waters with millimolar sodium chloride level (freshwater, drinking water, and aquarium water, as well as dechloridized seawater) decreases the pH down to ∼5. By using the acidification module, the limit of detection of nitrite-selective electrodes significantly improves by more than 2 orders of magnitude with respect to that observed at environmental pH. The originality of the proposed flow cell lies in the possibility to adjust the pH of the sample by modifying its exposure time with the membrane by varying the volumetric flow rate. Facile coupling with a detection technique of choice, miniaturized configuration and simple implementation for long-term monitoring with submersible probes for environmental analysis are possible analytical configurations. This approach was here successfully applied for the potentiometric detection of nitrite in aquarium and dechloridized seawater samples.

SCHeMA EU Project Summer School Report (Bilbao June 16 - 17, 2016).

This conference report describes the training activities that took place in the frame of the Integrated in Situ Chemical MApping probe (SCHeMA) summer school organized from the 14th to the 16th of June 2016 in Bilbao (Spain).

Local Acidification of Membrane Surfaces for Potentiometric Sensing of Anions in Environmental Samples.

The work dramatically improves the lower detection limit of anion selective membranes at environmental pH by using local acidification to suppress hydroxide interference at the membrane surface. Three separate localized acidification strategies are explored to achieve this, with ionophore-based membrane electrodes selective for nitrite and dihydrogen phosphate as guiding examples. In a first approach, a concentrated acetic acid solution (ca. 1 M) is placed in the inner filling solution of the PVC-based membrane electrode, forcing a significant acid gradient across the membrane. A second strategy achieves the same type of passive acidification by using an external proton source (fast diffusive doped polypropylene membrane) placed in front of a potentiometric solid contact anion selective electrode where the thin layer gap allows one to observe spontaneous acidification at the opposing detection electrode. The third approach shares the same configuration, but protons are here released by electrochemical control from the selective proton source into the thin layer sample. All three protocols improve the limit of detection by more than 2 orders of magnitude at environmental pH. Nitrite and dihydrogen phosphate determinations in artificial and natural samples are demonstrated.

All-solid-state potentiometric sensors with a multiwalled carbon nanotube inner transducing layer for anion detection in environmental samples.

While ion to electron transducing layers for the fabrication of potentiometric membrane electrodes for the detection of cations have been well established, similar progress for the sensing of anions has not yet been realized. We report for this reason on a novel approach for the development of all-solid-state anion selective electrodes using lipophilic multiwalled carbon nanotubes (f-MWCNTs) as the inner ion to electron transducing layer. This material can be solvent cast, as it conveniently dissolves in tetrahydrofuran (THF), an important advantage to develop uniform films without the need for using surfactants that might deteriorate the performance of the electrode. Solid contact sensors for carbonate, nitrate, nitrite, and dihydrogen phosphate are fabricated and characterized, and all exhibit comparable analytical characteristics to the inner liquid electrodes. For example, the carbonate sensor exhibits a Nernstian slope of 27.2 ± 0.8 mV·dec(-1), a LOD = 2.3 µM, a response time of 1 s, a linear range of four logarithmic units, and a medium-term stability of 0.04 mV·h(-1) is obtained in a pH 8.6 buffered solution. Water layer test, reversibility, and selectivity for chloride, nitrate, and hydroxide are also reported. The excellent properties of f-MWCNTs as a transducer are contrasted to the deficient performance of poly(3-octyl-thiophene) (POT) for carbonate detection. This is evidenced both with a significant drift in the potentiometric measures as well as a pronounced sensitivity to light (either sunlight or artificial light). This latter aspect may compromise its potential for environmental in situ measurements (night/day cycles). The concentration of carbonate is determined in a river sample (Arve river, Geneva) and compared to a reference method (automatic titrator with potentiometric pH detection). The results suggest that nanostructured materials such as f-MWCNTs are an attractive platform as a general ion-to-electron transducer for anion-selective electrodes.

Potentiometric sensing array for monitoring aquatic systems.

Since aquatic environments are highly heterogeneous and dynamic, there is the need in aquatic ecosystem monitoring to replace traditional approaches based on periodical sampling followed by laboratory analysis with new automated techniques that allow one to obtain monitoring data with high spatial and temporal resolution. We report here on a potentiometric sensing array based on polymeric membrane materials for the continuous monitoring of nutrients and chemical species relevant for the carbon cycle in freshwater ecosystems. The proposed setup operates autonomously, with measurement, calibration, fluidic control and acquisition triggers all integrated into a self-contained instrument. Experimental validation was performed on an automated monitoring platform on lake Greifensee (Switzerland) using potentiometric sensors selective for hydrogen ions, carbonate, calcium, nitrate and ammonium. Results from the field tests were compared with those obtained by traditional laboratory analysis. A linear correlation between calcium and nitrate activities measured with ISEs and relevant concentrations measured in the laboratory was found, with the slopes corresponding to apparent single ion activity coefficients γ(*)(Ca(2+)) = 0.55(SD = 0.1mM) and γ(*)(NO(3)(-)) = 0.75(SD = 4.7µm). Good correlation between pH values measured with ISE and CTD probes (SD = 0.2 pH) suggests adequate reliability of the methodology.

Environmental Sensing of Aquatic Systems at the University of Geneva.

Aquatic environments are complex living systems where biological and chemical constituents change rapidly with time and space and may exhibit synergistic interactions. To understand these processes, the traditional approach based on a typically monthly collection of samples followed by laboratory analysis is not adequate. It must be replaced by high-resolution autonomous in situ detection approaches. In our group at the University of Geneva, we aim to develop and deploy chemical sensor probes to understand complex aquatic systems. Most research centers around electrochemical sensing approaches, which involves: stripping voltammetry at gel-coated microelectrode arrays for direct measurements of bioavailable essential or toxic trace metals; direct potentiometry for the measurement of nutrients and other species involved in the nitrogen and carbon cycles; online desalination for oceanic measurements; the development of robust measurement principles such as thin layer coulometry, and speciation analysis by tandem electrochemical detection with potentiometry and dynamic electrochemistry. These fundamental developments are combined with instrument design, both in-house and with external partners, and result in field deployments in partnership with environmental researchers in Switzerland and the European Union.