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.

ISIDORE, a Probe for In Situ Trace Metal Speciation Based on the Donnan Membrane Technique and Electrochemical Detection Part 2: Cd and Pb Measurements during the Accumulation Time of the Donnan Membrane Technique.

The Donnan membrane technique (DMT), in which a synthetic or natural solution (the "donor") is separated from a ligand-free solution (the "acceptor") by a cation-exchange membrane, is a recognized technique for measuring the concentration of a free metal ion in situ, with coupling to electrochemical detection allowing for the quantification of the free metal ion directly on site. However, the use of the DMT requires waiting for the free metal ion equilibrium between the donor and the acceptor solution. In this paper, we investigated the possibility of using the kinetic information and showed that non-equilibrium experimental calibrations of Cd and Pb with the ISIDORE probe could be used to measure free metal concentrations under conditions of membrane-controlled diffusion transport. The application of this dynamic approach made it possible to successfully determine the concentration of free Cd in synthetic and natural river samples. Furthermore, it was found that the determination of free Cd from the slope was not affected by the Ca concentration ratio between the acceptor and donor solution, as opposed to the traditional approach based on Donnan equilibrium. This ISIDORE probe appears to be a promising tool for determining free metal ions in natural samples.

Absolute and Relative Positioning of Natural Organic Matter Acid-Base Potentiometric Titration Curves: Implications for the Evaluation of the Density of Charged Reactive Sites.

Potentiometric acid-base titration curves collected on humic (nano)particles as a function of pH and salt concentration reflect the electrostatics of the particles and the amount of chemical charges (Q) they carry. In turn, the interpretation of titration data helps quantify their reactivity toward metals provided that both intrinsic chemical and nonspecific electrostatic contributions to proton binding are correctly unraveled. Establishing a titration curve requires several steps, i.e., blank subtraction, relative curve positioning with respect to the electrolyte concentration, and absolute curve positioning achieved by the estimation of particle charge Q0 at low pH. Failure to properly establish each step may lead to the misevaluation of nanoparticle charging behavior. Here, we report (i) a simple procedure to measure and position titration curves for humic substances (HS) versus salt concentration and (ii) an original approach for absolute curve positioning upon the exploitation of proton affinity spectra. The latter do not depend on Q0 and they thus constrain the titration data analysis using the soft Poisson-Boltzmann-based titration (SPBT) formalism for nanoparticles in the thick electric double-layer regime. We illustrate the benefits of our approach by analyzing titration measurements for a large range of humic nanoparticles and by comparing the outcome with results from the literature.

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.

Chemodynamics of Soft Nanoparticulate Metal Complexes: From the Local Particle/Medium Interface to a Macroscopic Sensor Surface.

The lability of a complex species between a metal ion M and a binding site S, MS, is conventionally defined with respect to an ongoing process at a reactive interface, for example, the conversion or accumulation of the free metal ion M by a sensor. In the case of soft charged multisite nanoparticulate complexes, the chemodynamic features that are operative within the micro environment of the particle body generally differ substantially from those for dissolved similar single-site complexes in the same medium. Here we develop a conceptual framework for the chemodynamics and the ensuing lability of soft (3D) nanoparticulate metal complexes. The approach considers the dynamic features of MS at the intraparticulate level and their impact on the overall reactivity of free metal ions at the surface of a macroscopic sensing interface. Chemodynamics at the intraparticulate level is shown to involve a local reaction layer at the particle/medium interface, while at the macroscopic sensor level an operational reaction layer is invoked. Under a certain window of conditions, volume exclusion of the nanoparticle body near the medium/sensor interface is substantial and affects the properties of the reaction layer and the overall lability of the nanoparticulate MS complex toward the reactive surface.

Towards field trace metal speciation using electroanalytical techniques and tangential ultrafiltration.

In this work we propose a trace metal speciation methodology to determine the total, free and ultrafiltered (<1 KDa) metal fractions using electrochemical methods (SCP and AGNES) and tangential ultrafiltration (UF) experiments that can easily be carried out on-site. We tested our methodology spiking Cadmium ions into two natural waters samples from Itapanhau and Sorocabinha rivers in Sao Paulo State, Brazil. The limits of detection (LOD) was 1.6×10(-9) M for the total Cd(2+) determination performed by Stripping Chronopotentiometry (SCP) in the source and acidified ultrafiltered solution and 1.9×10(-9) M for the free Cd(2+) determination using Absence of gradients and Nernstian equilibrium stripping (AGNES), using a thin mercury film electrode. The total metal determination was performed by SCP in acidified samples and the results compared with graphite furnace atomic absorption spectroscopy (GF-AAS). The SCP results were adequate with a 96% of recovery from the known metal spike for the 12 samples tested. For the Itapanhau sample the free metal determined by AGNES and the ultrafiltered fraction are identical, while for the Sorocabinha the free metal in the source is significantly smaller than the ultrafiltered fraction, indicating that this sample must be rich in metal complexes with small inorganic ligands that are able to permeate the 1kDa membrane. The proposed metal speciation methodology validated in the laboratory combining UF and SCP/AGNES is able to be used in on-site experiments providing valid information regarding the total and free metal concentrations and additionally some insight on the role of small inorganic ligands to the metal complexation.

Determination of the Free Metal Ion Concentration Using AGNES Implemented with Environmentally Friendly Bismuth Film Electrodes.

Ex situ plated Bi film electrodes (Bi-FE) have been employed, for the first time, to measure the free concentration of Pb(II) in aqueous solutions using absence of gradients and Nernstian equilibrium stripping (AGNES) with stripping chronopotentiometry (SCP) quantification. The amount of deposited Pb°, below a certain threshold, follows a Nernstian relationship with the applied potential. This threshold can be interpreted as the frontier of transition from surface deposition to solid (bulk) formation of Pb°. AGNES with Bi-FE yielded a very good detection limit (3σ) for Pb(II) of 6.0 × 10(-9) M with an applied gain of 398 and a deposition time of 400 s. The ability of the Bi film electrode to perform speciation measurements was demonstrated for Pb(II)-PSS and Pb(II)-IDA systems. The measured values with the Bi-FE were in good agreement with the values obtained using the Hg film electrode and/or the values reported in the literature.

The effect of phosphate on the nuclease activity of vanadium compounds.

The nuclease activity of VO(acac)2 (1, acac = acetylacetone) and its derivatives VO(hd)2 (2, hd = 3,5-heptanedione), VO(Cl-acac)2 (3, Cl-acac = 3-chloro-2,4-pentanedione), VO(Et-acac)2 (4, Et-acac = 3-ethyl-2,4-pentanedione) and VO(Me-acac)2 (5, Me-acac = 3-methyl-2,4-pentanedione), is studied by agarose gel electrophoresis, UV-visible spectroscopy, cyclic and square wave voltammetry and (51)V NMR. The mechanism is shown to be oxidative and associated with the formation of reactive oxygen species (ROS). Hydrolytic cleavage of the phosphodiester bond is also promoted by 1, but at much slower rate which cannot compete with the oxidative mechanism. The generation of ROS is much higher in the presence of phosphate buffer when compared with organic buffers and this was attributed to the formation of a mixed-ligand complex containing phosphate, (V(IV)O)(V(V)O)(acac)2(HnPO4(n-3)), presenting a quasi-reversible voltammetric behavior. The formation of this species was further observed by Electrospray Ionization Mass Spectrometry (ESI-MS). Phosphate being an essential species in most biological media, the importance of the formation of mixed-ligand species in other vanadium systems is emphasized.

Toxicokinetics and tissue distribution of cadmium-based Quantum Dots in the marine mussel Mytilus galloprovincialis.

Environmental health hazards of Quantum Dots (QDs) are of emergent concern, but limited data is available about their toxicokinetics (TK) and tissue distribution in marine bivalves. This study investigated the QDs behavior in seawater, their TK and tissue distribution in Mytilus galloprovincialis, in comparison with soluble Cd. Mussels were exposed to CdTe QDs and soluble Cd for 21 days at 10 µgCd L(-1) followed by a 50 days depuration. TK of QDs in mussels is related to the homo-aggregate uptake, surface charge, aggregation and precipitation as key factors. There were tissue- and time-dependent differences in the TK of both Cd forms, and soluble Cd is the most bioavailable form. Digestive gland is a preferential site for QDs storage and both Cd forms are not eliminated by mussels (t1/2>50 days). Results indicate that the TK model of CdTe QDs in marine mussels is distinct from their soluble counterparts.

Partitioning of humic acids between aqueous solution and hydrogel. 3. Microelectrodic dynamic speciation analysis of free and bound humic metal complexes in the gel phase.

The hydrogel/water partitioning of the various species in the cadmium(II)/soil humic acid (HA) system is studied for two types of gel, using in situ microelectrodic voltammetry. Under the conditions of this work, with HA particles of ca. 25 and 125 nm radius, the CdHA complex is shown to be close to nonlabile toward a 12.5 µm radius microelectrode. This implies that its kinetic contribution to Cd(2+) reduction at the medium/microelectrode interface is practically negligible. The polyacrylamide (PAAm) gels equilibrate with the aqueous medium under significant sorption of HA at the gel backbone/gel medium interface, which in turn leads to induced sorption of Cd(II) in the form of immobilized gel-bound CdHA. The rather high total Cd content of the PAAm gel suggests that the binding of Cd(2+) by the hydrophobically gel-bound HA is stronger than that for dispersed HA particles. Still, the intraparticulate speciation of Cd(II) over Cd(2+) and CdHA corresponds to an intrinsic stability constant similar to that for simple monocarboxylate ligands such as acetate. Alginate gels are negatively charged, and their free [Cd(aq)(2+)] is higher than that in the medium by the corresponding Donnan coefficient. On top of that, Cd(2+) is specifically sorbed by the gel backbone/gel medium interface to reach accumulation factors as high as a few tens. HA and CdHA accumulate in the outer 20 µm film of gel at the gel/water interface of both gels, but they do not penetrate into the bulk of the alginate gel. Overall, the gel/water interface dictates drastic changes in the speciation of Cd/HA as compared to the aqueous medium, with distinct features for each individual type of gel. The results have broad significance, for example, for predictions of reactivity and bioavailability of metal species which inherently involve partitioning and diffusion into diverse gel layers such as biointerfacial cell walls, biofilm matrices, and mucous membranes.

Immunocytotoxicity, cytogenotoxicity and genotoxicity of cadmium-based quantum dots in the marine mussel Mytilus galloprovincialis.

There is an increased use of Quantum Dot (QDs) in biological and biomedical applications, but little is known about their marine ecotoxicology. So, the aim of this study was to investigate the possible immunocytotoxic, cytogenotoxic and genotoxic effects of cadmium telluride QDs (CdTe QDs) on the marine mussel Mytilus galloprovincialis. Mussels were exposed to 10 µg L(-1) of CdTe QDs or to soluble Cd [Cd(NO3)2] for 14 days and Cd accumulation, immunocytotoxicity [hemocyte density, cell viability, lysosomal membrane stability (LMS), differential cell counts (DCC)], cytogenotoxicity (micronucleus test and nuclear abnormalities assay) and genotoxicity (comet assay) were analyzed. Results show that in vivo exposure to QDs, Cd is accumulated in mussel soft tissues and hemolymph and induce immunotoxic effects mediated by a decrease in LMS, changes in DCC, as well as genotoxicity (DNA damage). However, QDs do not induce significant changes in hemocytes density, cell viability and cytogenetic parameters in opposition to Cd(2+). Soluble Cd is the most cytotoxic and cytogenotoxic form on Mytilus hemocytes due to a higher accumulation of Cd in tissues. Results indicate that immunotoxicity and genotoxicity of CdTe QDs and Cd(2+) are mediated by different modes of action and show that Mytilus hemocytes are important targets for in vivo QDs toxicity.

Polyhydroxyfullerene binds cadmium ions and alleviates metal-induced oxidative stress in Saccharomyces cerevisiae.

The water-soluble polyhydroxyfullerene (PHF) is a functionalized carbon nanomaterial with several industrial and commercial applications. There have been controversial reports on the toxicity and/or antioxidant properties of fullerenes and their derivatives. Conversely, metals have been recognized as toxic mainly due to their ability to induce oxidative stress in living organisms. We investigated the interactive effects of PHF and cadmium ions (Cd) on the model yeast Saccharomyces cerevisiae by exposing cells to Cd (≤5 mg liter(-1)) in the absence or presence of PHF (≤500 mg liter(-1)) at different pHs (5.8 to 6.8). In the absence of Cd, PHF stimulated yeast growth up to 10.4%. Cd inhibited growth up to 79.7%, induced intracellular accumulation of reactive oxygen species (ROS), and promoted plasma membrane disruption in a dose- and pH-dependent manner. The negative effects of Cd on growth were attenuated by the presence of PHF, and maximum growth recovery (53.8%) was obtained at the highest PHF concentration and pH. The coexposure to Cd and PHF decreased ROS accumulation up to 36.7% and membrane disruption up to 30.7% in a dose- and pH-dependent manner. Two mechanisms helped to explain the role of PHF in alleviating Cd toxicity to yeasts: PHF decreased Cd-induced oxidative stress and bound significant amounts of Cd in the extracellular medium, reducing its bioavailability to the cells.

Interactions of core-shell quantum dots with metal resistant bacterium Cupriavidus metallidurans: consequences for Cu and Pb removal.

In the present study we address the interactions of carboxyl-CdSe/ZnS core/shell quantum dots (QDs), as a model of water dispersible engineered nanoparticles, and metal resistant bacteria Cupriavidus metallidurans, largely used in metal decontamination. The results demonstrate that QDs with average hydrodynamic size of 12.9 nm adhere to C. metallidurans. The percentage of bacterial cells displaying QD-fluorescence increased proportionally with contact time and QD concentration in bacterial medium demonstrating the association of QDs with the metal resistant bacteria. No evidence of QD internalization into bacterial cytoplasm was found by transmission electron microscopy with energy dispersive X-ray spectrometry, however QD clusters of sizes between 20 and 50 nm were observed on the bacterial surface and in the bacterial periplasmic compartment; observations consistent with the losses of membrane integrity induced by QDs. The presence of 20 nM QDs induced about 2-fold increase in Cu and Pb uptake fluxes by C. metallidurans exposed to 500 nM Pb or Cu, respectively. Overall, the results of this work suggest that when present in mixture with Cu and Pb, low levels of QDs originating from possible incidental release or QD disposal could increase metal accumulation in metal resistant bacterium.

Dynamics and heterogeneity of Pb(II) binding by SiO2 nanoparticles in an aqueous dispersion.

Pb(II) binding by SiO(2) nanoparticles in an aqueous dispersion was investigated under conditions where the concentrations of Pb(2+) ions and nanoparticles are of similar magnitude. Conditional stability constants (log K) obtained at different values of pH and ionic strength varied from 4.4 at pH 5.5 and I = 0.1 M to 6.4 at pH 6.5 and I = 0.0015 M. In the range of metal to nanoparticle ratios from 1.6 to 0.3, log K strongly increases, which is shown to be due to heterogeneity in Pb(II) binding. For an ionic strength of 0.1 M the Pb(2+)/SiO(2) nanoparticle system is labile, whereas for lower ionic strengths there is loss of lability with increasing pH and decreasing ionic strength. Theoretical calculations on the basis of Eigen-type complex formation kinetics seem to support the loss of lability. This is related to the nanoparticulate nature of the system, where complexation rate constants become increasingly diffusion controlled. The ion binding heterogeneity and chemodynamics of oxidic nanoparticles clearly need further detailed research.

Modeling of Cd uptake and efflux kinetics in metal-resistant bacterium Cupriavidus metallidurans.

The Model of Uptake with Instantaneous Adsorption and Efflux, MUIAE, describing and predicting the overall Cd uptake by the metal-resistant bacterium Cupriavidus metallidurans CH34, is presented. MUIAE takes into account different processes at the bacteria-medium interface with specific emphasis on the uptake and efflux kinetics and the decrease in bulk metal concentration. A single set of eight parameters provides a reasonable description of experimentally determined adsorbed and internalized Cd, as well as the evolution of dissolved Cd concentrations with time, for an initial Cd concentration between 10(-8) and 10(-4) M, covering the situation of contaminated environments and heavily polluted effluents. The same set of parameters allowed successful prediction of the internalized and adsorbed Cd as a function of the measured free Cd ion concentration in the presence of natural and anthropogenic ligands. The findings of the present study reveal the key role of Cd efflux and bulk depletion on the overall Cd uptake by C. metallidurans, and the need to account for these processes to understand and improve the efficiency of the metal removal from the contaminated environment.

Evaluation of thin mercury film rotating disk electrode to perform absence of gradients and Nernstian equilibrium stripping (AGNES) measurements.

In the present work, the applicability of thin mercury film on a rotating disk electrode (TMF-RDE), to assess the free metal ion concentration by the absence of gradients and Nernstian equilibrium stripping (AGNES), is evaluated. The thickness of the mercury film and several AGNES parameters has been optimized. A nominal 16 nm film is chosen due to the higher signal (faradaic current) relative to the value of the noise (capacitive current). Due to the smaller volume to area ratio, the deposition time needed to reach a certain preconcentration factor (Y) is much shorter than in larger electrodes, like the HMDE. The limit of detection (3 sigma) for lead(II) is 7.4 x 10(-9)M and 7.2 x 10(-8)M for a Y of 5000 (deposition time of 150 s) and 1000 (deposition time of 100 s), respectively. A specific mathematical treatment is developed in order to subtract a corrected blank taking into account the degradation of the thin film (presumably, falling down of drops). The couple TMF-RDE/AGNES is successfully applied for speciation purposes in the systems Pb(II)-latex nanospheres and Pb(II)-IDA (iminodiacetic acid), where the stability constants calculated for both systems agree with values reported in the literature.