Release of indium from In2O3 nanoparticles in model solutions and synthetic seawater.

Indium oxide (In2O3) nanoparticles (NPs) are used in electronic devices, from which indium (as its nanoparticulate form or as other generated chemical species) can be released to natural waters. To assess for the impacts of such releases (e.g. toxic effects), information on the kinetics and thermodynamics of the In2O3 dissolution processes is key. In this work, the evolution with time of the dissolution process was followed with the technique AGNES (Absence of Gradients and Nernstian Equilibrium Stripping) by measuring the free indium concentration ([In3+]). AGNES can determine the free ion concentration in the presence of nanoparticles without a prior separation step, as shown in the case of ZnO nanoparticles, a procedure that is more accurate than the typical sequence of centrifugation+filtration+elemental analysis. Excess of indium oxide NPs were dispersed in 0.1 mol L-1 KNO3 at various pH values ranging from 2 to 8. Additional dispersions with bulk In2O3 at pH 3 or NPs in synthetic seawater at pH 8 were also prepared. The temperature was carefully fixed at 25 °C. The dispersions were continuously stirred and samples were taken from time to time to measure free indium concentration with AGNES. 180-day contact of In2O3 to solutions at pH 2 and 3 was not enough to reach equilibrium. The dissolution of the NPs at pH 3 was faster than that of the bulk (i.e. non nanoparticulate) material. Equilibrium of the NPs with the solution was reached at pH 4 and 5 in KNO3 and at pH 8 in seawater, in shorter times for higher pH values, with free indium concentrations decreasing by a factor of 1000 for each increase in one pH unit. The solubility products of In(OH)3 and In2O3 were compared. Equilibration of NPs with synthetic seawater took <18 days, with an average free [In3+] (up to 196 days) of 1.03 amol L-1.

Direct determination of free Zn concentration in samples of biological interest.

The speciation of essential metal ions in biological fluids, such as blood plasma and serum, is of fundamental importance to understand the homeostasis of these elements. The activity of metal ions such as Zn2+ in extracellular media is thought to affect their interaction with membrane-bound transporters, and thus is critical for their cellular uptake. Previous approaches to determine "free" Zn2+ (i.e. the hexa-aquo ion) are based on separation by either chromatography or ultrafiltration, or on metallochromic dyes. However, both types of approach are prone to affect the relevant equilibria. These drawbacks can be circumvented with the electroanalytical technique AGNES (Absence of Gradients and Nernstian Equilibrium Stripping), since it can measure free zinc concentration without perturbing the sample speciation. Here, a Bovine Serum Albumin (BSA) + Zn synthetic mixture and Fetal Bovine Serum (FBS) are analyzed as proof of concept. Adsorption of BSA on the surface of the Hanging Mercury Drop Electrode (HMDE), despite the advantage of its renewal, is so intense that it blocks appropriate attainment of the required equilibrium, and only estimations of [Zn2+] can be derived. In contrast, a rotating disc electrode with a thin mercury film deposited on it (TMF-RDE) is advantageous because of its small volume and enhanced mass transfer. Protein adsorption can be prevented by covering the TMF-RDE with Nafion. A free Zn concentration [Zn2+] = 2.7 nmol L-1 was found at pH 7.0, total Zn 20 µµmol L-1 and BSA 600 µµmol L-1. A sample of FBS with fixed pH 7.2 (MOPS 0.08 mol L-1) yielded [Zn2+] = 0.25 nmol L-1. This methodology opens the way to free metal concentration determinations in biological fluids.

Effective concentration signature of Zn in a natural water derived from various speciation techniques.

The uptake of nutrients or toxicants by different organisms in aquatic systems is known to correlate with different fractions of the nutrient's or toxicant's total concentration. These fractions can be provided by different analytical techniques, from which the better correlation is expected to be found for those with a characteristic length comparable to that in the considered organism uptake. An effective concentration signature can be built up with the concentration values associated to the availability (i.e. fluxes in dynamic techniques) of the nutrient or toxicant measured by various analytical techniques with different characteristic lengths. Here, this new representation was obtained for the pool of Zn complexes in the Mediterranean stream Riera d'Osor (Girona, Catalonia, Spain) with a suite of four analytical techniques. Absence of Gradients and Nernstian Equilibrium Stripping (AGNES) and Polymer Inclusion Membrane (PIM) devices provided the free Zn concentration. Linear Anodic Stripping Voltammetry provided a labile fraction (defined here as cLASV, higher than the free concentration), related to the diffusion layer scale. Diffusion Gradients in Thin-films provided higher labile fractions (known as DGT concentrations, cDGT) connected to the different characteristic lengths of different configurations (e.g. one or two resin discs) longer, in any case, than that corresponding to LASV. The combination of the information retrieved by the techniques allowed to quantify lability degrees of the pool of Zn complexes and to build up the effective concentration signature for this water.

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.

Comparison of different speciation techniques to measure Zn availability in hydroponic media.

Four analytical techniques are compared: AGNES (Absence of Gradients and Nernstian Equilibrium Stripping), LASV (Anodic Stripping Voltammetry with Linear stripping), DGT (Diffusive Gradients in Thin films) and PIM (Polymer Inclusion Membranes). These techniques have been designed to provide the free ion concentration or a labile fraction, complementarily contributing to an integrated description of speciation and availability. Their simultaneous application to the determination of free Zn concentrations or labile fluxes in seven solutions of a hydroponic medium reveals characteristics of each technique and correlations between their results. All dynamic results can be interpreted in terms of a general theoretical framework on fluxes. Indeed, in techniques under diffusion-limited conditions in the sample, the flux can be split into the free contribution (linearly proportional to the free fraction), plus the contribution of the complexes (where mobility, lability and abundance of complexation are intertwined). A methodology to compute lability degrees is developed. Measurements with PIM devices confirm that diffusion in the sample solution is not rate limiting, so its flux is proportional to the free metal in the donor solution. A proportionality between the responses of any given two techniques is observed, which suggests that, for the low ligand-to-metal concentration ratios used in the present work, any of these techniques would correlate similarly with uptake, toxic or nutritional measurements.

Free indium concentration determined with AGNES.

Indium is increasingly used in electronic devices, from which it can be mobilized towards environmental compartments. Speciation of In in waters is important for its direct ecotoxicological effects, as well as for the fate of this element in the environment (e.g. fluxes from or towards sediments). Free indium concentrations in the environment can be extremely low due to hydrolysis, especially important in trivalent cations, to precipitation and to complexation with different ligands. In this work, the free indium concentration (which is a toxicologically and geochemically relevant fraction) in aqueous solutions at pH3 has been measured with an adapted version of the electroanalytical technique AGNES (Absence of Gradients and Nernstian Equilibrium Stripping). Speciation measurements in mixtures of indium with the ligands NTA (nitrilotriacetic acid) and oxalate indicate that the values of their stability constants in the NIST46.6 database are less adequate than those published in some more recent literature. The extraordinary lability and mobility of In-oxalate complexes allow the measuring of free indium concentrations below nmol/L in just 25s of deposition time.

Absence of Gradients and Nernstian Equilibrium Stripping (AGNES) for the determination of [Zn(2+)] in estuarine waters.

Zinc (Zn) has been classified as a "Specific Pollutant" under Annex VIII of the EU Water Framework Directive by two thirds of the EU member states. As a result, the UK Environmental Quality Standard (EQS) for Transitional and Coastal (TrAC) Waters has been reduced from 612 nM to 121 nM total dissolved Zn. It is widely accepted that the free metal ion ([Zn(2+)]) is the most bioavailable fraction, but there are few techniques available to determine its concentration in these waters. In this work, Absence of Gradients and Nernstian Equilibrium Stripping (AGNES) has been applied, for the first time, to determine [Zn(2+)] in estuarine waters. The AGNES method had a mean RSD of ±18%, a (deposition time dependent) limit of detection of 0.73 nM and a [Zn(2+)] recovery of 112 ± 19% from a certified reference material (BCR-505; Estuarine Water). AGNES results for 13 estuarine samples (salinity 0.1-31.9) compared well (P = 0.02) with Competitive Ligand Exchange Cathodic Stripping Voltammetry (CLE-AdCSV) except for one sample. AGNES requires minimal sample manipulation, is unaffected by adsorption of interfering species at the electrode surface and allows direct determination of free zinc ion concentrations. Therefore AGNES results can be used in conjunction with ecotoxicological studies and speciation modelling to set and test compliance with water quality standards.

The chronic toxicity of ZnO nanoparticles and ZnCl2 to Daphnia magna and the use of different methods to assess nanoparticle aggregation and dissolution.

In this study, the effect of ZnO nanoparticles and ZnCl2 on growth, reproduction and accumulation of zinc in Daphnia magna was determined in a 21-day chronic toxicity test. A variety of techniques were used to distinguish the free zinc ion, dissolved, nanoparticle and aggregated zinc fraction in the Daphnia test medium. The results showed similar chronic effects on growth, reproduction and accumulation for the ZnO nanoparticles (EC10, 20, 50 reproduction: 0.030, 0.049, 0.112 mg Zn/l) and the ZnCl2 (EC10, 20, 50 reproduction: 0.014, 0.027, 0.082 mg Zn/l). A large fraction of the nanoparticles rapidly dissolved after introduction in the exposure medium. Aggregation of nanoparticles was also observed but within 48 h of exposure most of these ZnO aggregates were dissolved. Based on the combined dissolution kinetics and toxicity results, it can be concluded that the toxicological effects of ZnO nanoparticles at the chronic level can be largely attributed to the dissolved fraction rather than the nanoparticles or initially formed aggregates.