Electroanalytical Trace Metal Cations Quantification and Speciation in Freshwaters: Historical Overview, Critical Review of the Last Five Years and Road Map for Developing Dynamic Speciation Field Measurements.

An historical overview covering the field of electroanalytical metal cations speciation in freshwaters is presented here, detailing both the notable experimental and theoretical developments. Then, a critical review of the progress in the last five years is given, underlining in particular the improvements in electrochemical setups and methodologies dedicated to field surveys. Given these recent achievements, a road map to carry out on-site dynamic metal speciation measurements is then proposed, and the key future developments are discussed. This review shows that electroanalytical stripping techniques provide a unique framework for quantitatively assessing metals at trace levels while offering access to both thermodynamic and dynamic features of metal complexation with natural colloidal and particulate ligands.

Developing On-Site Trace Level Speciation of Lead, Cadmium and Zinc by Stripping Chronopotentiometry (SCP): Fast Screening and Quantification of Total Metal Concentrations.

Electrochemical stripping techniques are interesting candidates for carrying out onsite speciation of environmentally relevant trace metals due to the existing low-cost portable instrumentation available and the low detection limits that can be achieved. In this work, we describe the initial analytical technique method development by quantifying the total metal concentrations using Stripping Chronopotentiometry (SCP). Carbon paste screen-printed electrodes were modified with thin films of mercury and used to quantify sub-nanomolar concentrations of lead and cadmium and sub-micromolar concentrations of zinc in river water. Low detection limits of 0.06 nM for Pb(II) and 0.04 nM for Cd(II) were obtained by the standard addition method using a SCP deposition time of 180 s. The SCP results obtained for Pb(II) and Cd(II) agreed with those of inductively coupled plasma mass spectrometry (ICP-MS). The coupling of SCP with screen-printed electrodes opens up excellent potential for the development of onsite speciation of trace metals. Due to the low analysis throughput obtained for the standard addition method, we also propose a new, more rapid screening Cd(II) internal standard methodology to significantly increase the number of samples that can be analyzed per day.

Electroanalytical metal sensor with built-in oxygen filter.

A rapid and reliable oxygen elimination system was evaluated here for the electroanalytical study of metals. Dissolved oxygen was removed locally in the vicinity of a sensor by the means of electrochemical oxygen filter constructed from platinum grids. Three metals (Cd, Pb, and Zn) were determined by stripping chronopotentiometry (SCP) at a mercury film screen-printed electrode. Limits of detection of metals were in the nanomolar range under optimized experimental conditions. The electrochemical device was also tested for metal quantification in simple electrolyte solutions and in a natural water matrix. The proposed combination of oxygen elimination system with the metal sensor completely removes the need to purge the sample before SCP measurement. This makes the determination of metals by SCP faster, portable and more suited for on-field applications.

Addressing the electrostatic component of protons binding to aquatic nanoparticles beyond the Non-Ideal Competitive Adsorption (NICA)-Donnan level: Theory and application to analysis of proton titration data for humic matter.

HYPOTHESIS: Charge descriptors of aquatic nanoparticles (NPs) are evaluated from proton titration curves measured at different salt concentrations and routinely analysed by the Non-Ideal Competitive Adsorption-Donnan (NICAD) model. This model, however, suffers from approximations regarding particle electrostatics, which may bias particle charge estimation. Implementation of Poisson-Boltzmann (PB) theory within consistent treatment of NPs protolytic data is expected to address NICAD shortcomings. EXPERIMENTS: An alternative to NICAD is elaborated on the basis of nonlinearized PB equation for soft particle electrostatics to properly unravel the electrostatic and chemical components of proton binding to NPs. A numerical package is developed for automated analysis of proton titration curves and proton affinity spectra at different salt concentrations. The performance of the method is illustrated for humic matter nanoparticles with different charge and size, and compared to that of NICAD. FINDINGS: Unlike NICAD, PB-based treatment successfully reproduces particle charge dependence on pH for practical salt concentrations from the thin to thick electric double layer limit. Donnan representation in NICAD leads to moderate to dramatic misestimations of proton affinity and binding heterogeneity depending on particle size to Debye layer thickness ratio. Interpretation of NPs protolytic properties with PB theory further avoids adjustment of the 'particle Donnan volume' empirically introduced in NICAD.

Investigating the Binding Heterogeneity of Trace Metal Cations With SiO2 Nanoparticles Using Full Wave Analysis of Stripping Chronopotentiometry at Scanned Deposition Potential.

Silica oxides nano- and microparticles, as well as silica-based materials, are very abundant in nature and industrial processes. Trace metal cation binding with these bulk materials is generally not considered significant in speciation studies in environmental systems. Nonetheless, this might change for nanoparticulate systems as observed in a previous study of Pb(II) with a very small SiO2 particle (7.5 nm diameter). Besides, metal binding by those nanoparticles is surprisingly characterized by a heterogeneity that increases with the decrease of metal-to-particle ratio. Therefore, it is interesting to extend this study to investigate different trace metals and the influence of the nanoparticle size on the cation binding heterogeneity. Consequently, the Cd(II), Pb(II), and Zn(II) binding by two different sized SiO2 nanoparticles (Ludox LS30 and TM40) in aqueous dispersion was studied for a range of pH and ionic strength conditions, using the combination of the electroanalytical techniques Scanned Stripping ChronoPotentiometry and Absence of Gradients and Nernstian Equilibrium Stripping. The coupling of these techniques provides the free metal concentration in the bulk (AGNES) and information of the free and complex concentration at the electrode surface for each Stripping Chronopotentiometry at Scanned deposition Potential (SSCP). A recent mathematical treatment allows the reconstruction of a portion of the metal to ligand binding isotherm with the included heterogeneity information using the full SSCP wave analysis. In this work, we observed that the Zn(II) binding is homogeneous, Cd(II) is slightly heterogeneous, and Pb(II) is moderately heterogeneous, whereas the results obtained with the 7.5 nm diameter nanoparticle are slightly more heterogeneous than those obtained with the one of 17 nm. These findings suggest that the Zn(II) binding is electrostatic in nature, and for both Cd(II) and Pb(II), there should be a significant chemical binding contribution.

Towards improving the electroanalytical speciation analysis of indium.

The geochemical fate of indium in natural waters is still poorly understood, while recent studies have pointed out a growing input of this trivalent element in the environment as a result of its utilisation in the manufacturing of high-technology products. Reliable and easy-handling analytical tools for indium speciation analysis are, then, required. In this work, we report the possibility of measuring the total and free indium concentrations in solution using two complementary electroanalytical techniques, SCP (Stripping chronopotentiometry) and AGNES (Absence of Gradients and Nernstian Equilibrium Stripping) implemented with the TMF/RDE (Thin Mercury Film/Rotating Disk Electrode). Nanomolar limits of detection, i.e. 0.5 nM for SCP and 0.1 nM for AGNES, were obtained for both techniques in the experimental conditions used in this work and can be further improved enduring longer experiment times. We also verified that AGNES was able (i) to provide robust speciation data with the known In-oxalate systems and (ii) to elaborate indium binding isotherms in presence of humic acids extending over 4 decades of free indium concentrations. The development of electroanalytical techniques for indium speciation opens up new routes for using indium as a potential tracer for biogeochemical processes of trivalent elements in aquifers, e.g. metal binding to colloidal phases, adsorption onto (bio)surfaces, etc.

Impact of intracellular metallothionein on metal biouptake and partitioning dynamics at bacterial interfaces.

Genetically engineered microorganisms are alternatives to physicochemical methods for remediation of metal-contaminated aquifers due to their remarkable bioaccumulation capacities. The design of such biosystems would benefit from the elaboration of a sound quantitative connection between performance in terms of metal removal from aqueous solution and dynamics of the multiscale processes leading to metal biouptake. In this work, this elaboration is reported for Escherichia coli cells modified to overexpress intracellular metallothionein (MTc), a strong proteinaceous metal chelator. Depletion kinetics of Cd(ii) from bulk solution following biouptake and intracellular accumulation is addressed as a function of cell volume fraction using electroanalytical probes and ligand exchange-based analyses. It is shown that metal biouptake in the absence and presence of MTc is successfully interpreted on the basis of a formalism recently developed for metal partitioning dynamics at biointerfaces with integration of intracellular metal speciation. The analysis demonstrates how fast sequestration of metals by intracellular MTc bypasses metal excretion (efflux) and enhances the rate of metal depletion to an extent such that complete removal is achieved at sufficiently large cell volume fractions. The magnitude of the stability constant of nanoparticulate metal-MTc complexes, as derived from refined analysis of macroscopic bulk metal depletion data, is further confirmed by independent electrochemical measurement of metal binding by purified MTc extracts.

Structural effects of soft nanoparticulate ligands on trace metal complexation thermodynamics.

Metal binding to natural soft colloids is difficult to address due to the inherent heterogeneity of their reactive polyelectrolytic volume and the modifications of their shell structure following changes in e.g. solution pH, salinity or temperature. In this work, we investigate the impacts of temperature- and salinity-mediated modifications of the shell structure of polymeric ligand nanoparticles on the thermodynamics of divalent metal ions Cd(ii)-complexation. The adopted particles consist of a glassy core decorated by a fine-tunable poly(N-isopropylacrylamide) anionic corona. According to synthesis, the charges originating from the metal binding carboxylic moieties supported by the corona chains are located preferentially either in the vicinity of the core or at the outer shell periphery (p(MA-N) and p(N-AA) particles, respectively). Stability constants (KML) of cadmium-nanoparticle complexes are measured under different temperature and salinity conditions using electroanalytical techniques. The obtained KML is clearly impacted by the location of the carboxylic functional groups within the shell as p(MA-N) leads to stronger nanoparticulate Cd complexes than p(N-AA). The dependence of KML on solution salinity for p(N-AA) is shown to be consistent with a binding of Cd to peripheral carboxylic groups driven by Coulombic interactions (Eigen-Fuoss mechanism for ions-pairing) or with particle electrostatic features operating at the edge of the shell Donnan volume. For p(MA-N) particulate ligands, a scenario where metal binding occurs within the intraparticulate Donnan phase correctly reproduces the experimental findings. Careful analysis of electroanalytical data further evidences that complexation of metal ions by core-shell particles significantly differ according to the location and distribution of the metal-binding sites throughout the reactive shell. This complexation heterogeneity is basically enhanced with increasing temperature i.e. upon significant increase of particle shell shrinking, which suggests that the contraction of the reactive phase volume of the particulate ligands promotes cooperative metal binding effects.

Kinetic and thermodynamic determinants of trace metal partitioning at biointerphases: the role of intracellular speciation dynamics.

There is a large body of work evidencing the necessity to evaluate chemical speciation dynamics of trace metals in solution for an accurate definition of their bioavailability to microorganisms. In contrast, the integration of intracellular metal speciation dynamics in biouptake formalisms is still in its early stages. Accordingly, we elaborate here a rationale for the interplay between chemodynamics of intracellular metal complexes and dynamics of processes governing metal biouptake under non-complexing outer medium conditions. These processes include the conductive diffusion of metal ions to the charged soft biointerphase, metal internalisation, excretion of intracellular free metal species and metal depletion from bulk solution. The theory is formulated from Nernst-Planck equations corrected for electrostatic and reaction kinetic terms applied at the biosurface and in the intracellular volume. Computational illustrations demonstrate how biointerfacial metal distribution dynamics inherently reflects the chemodynamic properties of intracellular complexes. In the practical limits of high and weak metal affinity to biosurface internalisation sites, the metal concentration profile is explicitly solved under conditions of strong intracellular complexing agents. Exact analytical expression is further developed for metal partitioning at equilibrium. This provides a way to evaluate the metal biopartition coefficient from refined analysis of bulk metal depletion measured at various cell concentrations. Depending on here-defined dimensionless parameters involving rates of metal internalisation-excretion and complex formation, the formalism defines the nature of the different kinetic regimes governing bulk metal depletion and biouptake. In particular, the conditions leading to an internalisation flux limited by diffusion as a result of demanding intracellular metal complexation are identified.

Structure of Multiresponsive Brush-Decorated Nanoparticles: A Combined Electrokinetic, DLS, and SANS Study.

Particles consisting of a glassy poly(methyl methacrylate) core (ca. 40 nm in radius) decorated with a poly(N-isopropylacrylamide) anionic corona are synthesized using either methacrylic acid (MA) or acrylic acid (AA) as reactive comonomers in the shell. The different reactivity ratios of MA and AA toward N-isopropylacrylamide originates p(MA-N) and p(N-AA) particles with carboxylate charges supposedly located, preferentially, in the close vicinity of the core and at the shell periphery, respectively. The corresponding swelling features of these nanoparticles are addressed over a broad range of pH values (4 to 7.5), NaNO3 concentrations (3 to 200 mM), and temperatures (15 to 45 °C) by dynamic light scattering (DLS) and small angle neutron scattering (SANS). DLS shows that the swelling of the particle shells increases their thickness from ∼10 to 90 nm with decreasing temperature, ionic strength, or increasing pH, with the effect being more pronounced for p(N-AA) whose lower critical solution temperature is shifted to higher values compared to that of p(MA-N). Potentiometric titration and electrokinetic results further reflect the easier dissociation of carboxyl groups in p(N-AA) and a marked heterogeneous interfacial swelling of the latter with decreasing solution salt content. The DLS response of both particles is attributed to the multiresponsive nature of a peripheral dilute shell, while SANS only probes the presence of a quasi-solvent-free dense polymer layer, condensed on the core surface. The thickness of that layer slightly increases from ∼6 to 9.5 nm with increasing temperature from 15 to 45 °C (at 15 mM NaNO3 and pH 5) due to the collapse of the outer dilute shell layer. Overall, results evidence a nonideal brush behavior of p(MA-N) and p(N-AA) and their microphase segregated shell structure, which supports some of the conclusions recently formulated from approximate self-consistent mean-field computations.

Evaluation of metal biouptake from the analysis of bulk metal depletion kinetics at various cell concentrations: theory and application.

Bioavailability of trace metals is a key parameter for assessment of toxicity on living organisms. Proper evaluation of metal bioavailability requires monitoring the various interfacial processes that control metal partitioning dynamics at the biointerface, which includes metal transport from solution to cell membrane, adsorption at the biosurface, internalization, and possible excretion. In this work, a methodology is proposed to quantitatively describe the dynamics of Cd(II) uptake by Pseudomonas putida. The analysis is based on the kinetic measurement of Cd(II) depletion from bulk solution at various initial cell concentrations using electroanalytical probes. On the basis of a recent formalism on the dynamics of metal uptake by complex biointerphases, the cell concentration-dependent depletion time scales and plateau values reached by metal concentrations at long exposure times (>3 h) are successfully rationalized in terms of limiting metal uptake flux, rate of excretion, and metal affinity to internalization sites. The analysis shows the limits of approximate depletion models valid in the extremes of high and weak metal affinities. The contribution of conductive diffusion transfer of metals from the solution to the cell membrane in governing the rate of Cd(II) uptake is further discussed on the basis of estimated resistances for metal membrane transfer and extracellular mass transport.

Dynamics of metal uptake by charged soft biointerphases: impacts of depletion, internalisation, adsorption and excretion.

A comprehensive theory is elaborated for the dynamics of metal ion uptake by charged spherical microorganisms. The formalism integrates the interplay over time between bulk metal depletion, metal adsorption, metal excretion (efflux) and transport of metals by conductive diffusion toward the metal-consuming biomembrane. The model further involves the basic physicochemical features of the microbial interphase in terms of size, distribution of electrostatic charges and thickness of peripheral soft surface appendage. A generalization of the Best equation is proposed and leads to the expression of the time-dependent concentration of metal ions at the active membrane surface as a function of bulk metal concentration. Combination of this equation with the metal conservation condition over the sample volume allows a full evaluation of bulk metal depletion kinetics and the accompanying time-dependent uptake and excretion fluxes as a function of metal-microorganism electrostatic interaction, microbe concentration and relevant biophysicochemical features of the interphase. Practically tractable expressions are derived in the limit where the Biotic Ligand Model (BLM) is obeyed and in situations where conductive diffusion transport of metals significantly determines the rate of biouptake. In particular, the plateau value reached at sufficiently long times by bulk metal concentration is rigorously expressed in terms of the key parameters pertaining to the adsorption process and to the kinetics of metal uptake and excretion. The theory extends and unifies previous approximate models where the impacts of extracellular metal transport and/or metal efflux on the overall rate of uptake were ignored.

Kinetic features of metal complexes with polysaccharide colloids: impact of ionic strength.

The dynamic features of metal binding by a gel-like polysaccharide, carboxymethyldextran (CMD), are investigated by stripping chronopotentiometry (SCP). This technique measures the diffusive flux properties of the metallic species in the ligand dispersion as defined by their concentration, mobility, and lability. Cadmium(II) forms only 1:1 complexes with CMD, the lability of which is well described by Eigen mechanism principles, that is, the removal of a water molecule from the inner hydration sphere of the metal ion is limiting the complex formation rate. Lead(II) and copper(II), however, also form intramolecular bidentate complexes with CMD, which requires a conformational reorganization of the polymeric chain. The reorganization process appears to be the rate-limiting step of the overall complexation reaction, which takes place on a time scale of hours. The influence of ionic strength on the rate of bidentate complex formation is insignificant. In contrast, its impact on the stability of the monodentate complex follows the corresponding Donnan potential of the soft CMD particle.

Kinetics of metal ion binding by polysaccharide colloids.

The dynamics of metal sorption by a gel-like polysaccharide is investigated by means of the electrochemical technique of stripping chronopotentiometry (SCP). The measured response reflects the diffusive flux properties of the metallic species in the dispersion. The colloidal ligand studied here is a functionalized carboxymethyldextran. Its complexation with Pb(II) reveals a time dependence that identifies strong differences in the dynamic nature of the successive metal complexes formed. Apparently, the formation of intramolecular bidentate complexes requires a slow conformational reorganization of the macromolecule that becomes the rate-limiting step in the complexation reaction. The relevant parameters for metal binding and release kinetics are computed and thus provide knowledge of the time-dependent stability and lability of metal polysaccharide complexes.

Relationship between swelling and the electrohydrodynamic properties of functionalized carboxymethyldextran macromolecules.

The electrostatic, hydrodynamic, and swelling properties of a well-defined, functionalized carboxymethyldextran (CMD) polysaccharide are investigated in aqueous NaNO3 solution over a broad ionic strength range. The impact of the polycarboxylate charge and molar mass of the CMD macromolecules on their electrohydrodynamic features is thoroughly examined by combined protolytic titration, dynamic light scattering, and electrokinetic analyses. Electrophoretic mobility data obtained for sufficiently high electrolyte concentrations reveal a typical soft particle behavior. Upon decrease of the ionic strength, mobilities strongly increase in magnitude while significant electrostatic swelling takes place, as reflected in a decrease in the diffusion coefficients. CMD entities undergo conformational transitions from compact random coil at large ionic strengths to swollen coil and possibly a wormlike structure at lower NaNO3 concentrations. The magnitude of the variations in size and mobility with electrolyte concentration strongly depends on the overall charge of the CMD entity as well as on its molar mass. These factors control the stiffness of the constituent polymer chains and thus the degree of macromolecular permeability ("softness"). Using the soft-diffuse interface formalism previously developed for the electrohydrodynamics of charged permeable macromolecules, a quantitative analysis of the electrophoretic mobility data is presented. The measured values of the diffusion coefficient and space charge density Gamma degrees, as evaluated independently from the modeling of potentiometric titration curves, are taken into account in a self-consistent manner. It is found that large CMD entities of low charge densities are the most permeable to flow penetration with a limited heterogeneous electrostatic stiffening of the chains, whereas small CMD entities of larger Gamma degrees significantly expand upon lowering the ionic strength, giving rise to a strong anisotropy for the spatial distribution of polymer chain density.