Back Accumulation of Diffusive Gradients in Thin-Films Devices with a Stack of Resin Discs To Assess Availability of Metal Cations to Biota in Natural Waters.

Determining species, concentrations, and physicochemical parameters in natural waters is key to improve our understanding of the functioning of these ecosystems. Diffusive Gradients in Thin-films (DGT) devices with different thicknesses of the resin or of the diffusive disc can be used to collect independent information on relevant parameters. In particular, DGT devices with a stack of two resin discs offer a simple way to determine dissociation rate constants of metal complexes from the accumulation of the target metal in the back resin disc. In this work, simple approximate expressions for the determination of the dissociation rate constant are reported and applied to a model Ni nitrilotriacetic complex as well as to Zn complexes in the Mediterranean Osor stream. Once the physicochemical parameters are known, one can plot the labile fraction of the metal complexes in terms of the thickness of the diffusion domain. These plots reveal a strong dependence on the nature of complexes as well as on the characteristics of the diffusion domain, and they are of high interest as predictors of availability to biota whose uptake is limited by diffusion.

Availability of metals to DGT devices with different configurations. The case of sequential Ni complexation.

The analytical technique DGT (Diffusive Gradients in Thin-films) is able to gain access to a wealth of information by carefully interpreting accumulation data from passive samplers with different configurations (i.e. different thicknesses of its constituent layers). A set of DGT devices were simultaneously deployed in solutions of Ni and nitrilotriacetic acid (NTA) of different concentrations to measure the availability of Ni in these solutions. Accumulations indicate that the availability of Ni depends on both the thickness of the resin and the thickness of the diffusive gel. In both cases, the lability degree increases as the thickness increases. As the formation of successive complexes (such as Ni(NTA)2) proceeds, the availability of the metal decreases, which is quantitatively explained by reducing the formulation to a case with only one complex, but with an effective dissociation rate constant that decreases as the concentration of NTA increases. Simple analytical expressions are reported to quantify the lability degree in the different DGT configurations. These results indicate that a set of different DGT devices can characterize the availability of a cation in a natural sample with uptake processes at different spatial or time scales. Alternatively, and from a more fundamental point of view, information on speciation, mobilities and labilities of the species present in natural samples can be obtained with a set of DGT configurations.

Extending the Use of Diffusive Gradients in Thin Films (DGT) to Solutions Where Competition, Saturation, and Kinetic Effects Are Not Negligible.

DGT (Diffusion Gradients in Thin films) was designed to sample trace metals in situ at their natural concentrations. The setup and the experimental deployment conditions were established to allow interpretation of a linear accumulation of metal with time, using a simple expression based on a steady-state flux under perfect sink conditions. However, the extension of DGT to a wide range of analytes and its use under varied conditions has shown that, in some situations, these conditions are not fulfilled, so that accumulations with time are nonlinear. Previously, when such curvature was observed, concentrations in solution could not be reliably calculated. Here, we present fundamentally derived equations that reproduce the time accumulation for three situations: (i) kinetic limitations in the binding to the resin, (ii) saturation or equilibrium effects, or (iii) non-negligible competitive effects. We show how the accumulations can be quantified, in terms of the required kinetic and thermodynamic constants, and provide practical guidance for their use to obtain reliable estimates of solution concentrations. Solutions containing Mg or Mn, where all three situations can prevail, are used as examples. Calculated concentrations show reasonable agreement with the experimentally known values and with the results of a numerical model of the system, significantly improving the estimations based on perfect sink conditions. Such an approach opens up the possibility of using DGT more widely in challenging systems and allows DGT data to be interpreted more fully.

Accumulation of Mg to Diffusive Gradients in Thin Films (DGT) Devices: Kinetic and Thermodynamic Effects of the Ionic Strength.

Availability of magnesium is a matter of concern due to its role in many environmental and biological processes. Diffusive Gradients in Thin Films (DGT) devices can measure Mg availability in situ. This work shows that Mg accumulation in water largely increases when ionic strength (I) decreases. This phenomenon can be explained from (i) the increase of both the association equilibrium (K) and rate (ka,R) constants for the reaction between Mg cations and resin sites, and (ii) the growing contribution of the partitioning of Mg cations at the resin-gel interface, as I decreases. Two theoretical models that take into account electrical interactions among Mg cations, background electrolyte, and resin sites can successfully be used to determine ka,R and K at each I. Both models yield similar ka,R values, which fulfill an expression for the kinetic salt effect. For freshwater (with a typical salinity of 10 mM and circumneutral pH), the binding of Mg is so fast and strong that the simplest perfect-sink DGT expression can be helpful to predict (overestimation lower than 5%) the accumulation in solutions with Mg concentrations up to 1 mM whenever the deployment time is below 9 h. Perfect sink conditions can still be applied for longer times, in systems with either a lower I or a lower Mg concentration.

Influence of the settling of the resin beads on diffusion gradients in thin films measurements.

Binding resin beads used in DGT (diffusion gradients in thin films) tend to settle to one side of the resin during casting. This phenomenon might be relevant for metal accumulation when partially labile complexes dominate the metal speciation, especially after recognizing the important role played by complex dissociation in the resin domain. The influence of the inhomogeneity of the binding agent distribution on metal accumulation is here assessed by numerical simulation of DGT devices with binding beads in only one half of the resin disc, as a reasonable model of the standard resin discs. Results indicate that a decrease in mass accumulation of less than 13% can arise in these inhomogeneous devices (as compared with an ideal disc with homogeneous dispersion of the resin beads) when complexes with stability constant K<10(2)m(3)mol(-1) (K<10(5)Lmol(-1)) dominate the metal speciation. The loss increases as K increases, but the percentage of mass loss always remains lower than the volume fraction of resin disc without beads. For very labile or inert complexes, the impact of the inhomogeneous distribution of binding resin beads is negligible. As kinetic dissociation constants of complexes can be estimated from the distribution of the metal accumulation in a DGT device with a stack of two resin discs, the influence of the inhomogeneity on the recovered kinetic constant is also assessed. For the cases studied, the recovered kinetic dissociation constant, kd,recovered, retains the correct order of magnitude, being related to the true kd by kd≈f(-1)kd,recovered, quite independently of K and kd values, being f the fraction of volume of the resin disc where resin beads are dispersed.

Limits of the linear accumulation regime of DGT sensors.

A key question for the practical application of DGT (Diffusive Gradients in Thin films) as dynamic sensors in the environmental monitoring of trace metals is the influence of pH and dissolved ligands over the linear accumulation regime. Protons compete with metal ions for the binding to the DGT resin sites at relatively low pH, whereas high affinity dissolved ligands compete with resin sites for the binding of metals. Any of the two phenomena can lead to a departure from the linear accumulation regime and an underestimation of the actual species concentration in solution. These effects are studied here through numerical simulation of the diffusion-reaction processes in both gel and resin domains using a detailed chemical model of metal ions and protons interacting with resin sites. Results were tested successfully against experimental data of the Cd-NTA representative system. Charts to delimitate the range of experimental conditions (pH, ligand concentration and strength) where the linear accumulation regime prevails, can be helpful for designing sampling strategies in field conditions. For example, it is foreseen that perturbations of linear regime within 10 h of deployment are negligible above pH 5 and weak complexation (log K' < 0) or above pH 7 and strong complexation (log K' < 3), where K' is the effective stability constant. These plots can also be approximately used for partially labile systems whenever the time is replaced with the product lability degree times t.

Kinetic mixture effects in diffusion gradients in thin films (DGT).

The penetration of complexes into the resin domain of the DGT devices has a large influence on the lability degree of these complexes, since the reaction layer (the layer where there is net dissociation) extends from the diffusive gel into the resin domain. Numerical simulation shows that, typically, the contribution to the metal accumulation from dissociation of complexes inside the resin domain is dominant. As a consequence, in excess of ligand, the influence of the ligand concentration on the lability degree is much reduced, in comparison with this effect in the voltammetric sensors. The presence of a mixture of ligands leads to parallel complexes that mutually influence their lability degrees. In general, the interaction between the complexes has an impact on the lability degree of each one, but the total metal accumulation is less sensitive due to cancellation (mutually opposite effects of a couple of complexes). This result paves the way to predict the metal accumulation from the lability degree available for each complex in a single ligand system. Maximum discrepancies of 10% have been found in these predictions which can still be reduced if thicker resin gels are used.

Lability criteria in diffusive gradients in thin films.

The penetration of metal complexes into the resin layer of DGT (diffusive gradients in thin films) devices greatly influences the measured metal accumulation, unless the complexes are either totally inert or perfectly labile. Lability criteria to predict the contribution of complexes in DGT measurements are reported. The key role of the resin thickness is highlighted. For complexes that are partially labile to the DGT measurement, their dissociation inside the resin domain is the main source of metal accumulation. This phenomenon explains the practical independence of the lability degree of a complex in a DGT device with respect to the ligand concentration. Transient DGT regimes, reflecting the times required to replenish the gel and resin domains up to the steady-state profile of the complex, are also examined. Low lability complexes (lability degree between 0.1 and 0.2) exhibit the longest transient regimes and therefore require longer deployment times to ensure accurate DGT measurements.

Contribution of partially labile complexes to the DGT metal flux.

Penetration of complexes into the resin layer can dramatically increase the contribution of complexes to the metal flux measured with a DGT (diffusive gradients in thin films) sensor, but equations to describe this phenomenon were not available. Here, simple approximate analytical expressions for the metal flux, the lability degree and the concentration profiles in a DGT experiment are reported. Together with the thickness of the reaction layer in the gel domain, the effective penetration distance into the resin layer that would be necessary for full dissociation of the complex (λ(ML)) plays a key role in determining the metal flux. An increase in the resin-layer thickness (r) effectively increases the metal flux and the lability degree until r ≈ 3λ(ML). For the usual DGT configuration, where the thickness of the gel layer exceeds that of the resin layer, the complex is labile if r > (D(ML)/k(d))½, where D(ML) is the diffusion coefficient of the metal complex and k(d) its dissociation rate constant. A general procedure for estimating the lability of any complex in a standard DGT configuration is provided.

Key role of the resin layer thickness in the lability of complexes measured by DGT.

Analysis of the dynamic features of diffusion gradients in thin film devices (DGT) indicates that the penetration of complexes into the resin layer dramatically increases their lability. This should be taken into account when interpreting DGT measurements in terms of the dynamics of solution speciation. The experimental accumulation of Cd by DGT sensors in Cd-NTA systems confirmed these theoretical analyses. A computational code, which allows a rigorous digital simulation of the diffusion-reaction processes in the gel and resin layers, was used to model the results and to demonstrate the effect of the complex penetration into the resin layer on the lability degree. These findings suggest that DGT renders all complexes much more labile than if the resin-diffusive gel interface was considered as a perfect planar sink, explaining why DGT often measures a high proportion of the metal in a natural water. This information is relevant since some studies have stressed the importance of labile complexes as a source of bioaccumulated metal.

Ligand mixture effects in metal complex lability.

The degree of lability of a given metal complex species is modified in the presence of a mixture of ligands. This modification is a consequence of the coupling of the association and dissociation processes of all of the complexes according to the competitive complexation reaction scheme. We show that, because of the mixture effect, the lability of a given complex usually increases when another more labile complex is added into the system, while it decreases upon addition of a less labile one. Typically, complexes tend to adapt to the global lability of the mixture. A quantitative evaluation of these effects for diffusion-limited conditions in a finite domain by rigorous numerical simulation in a system with two complexes indicates that the lability degree of a complex can change by more than 100% with respect to that in the single ligand system. The impact of the mixture effect on the metal flux depends at least on two main factors: the respective abundance of the metal species and the particular values of their lability degrees. Dominant complexes (i.e., those most abundant when these complexes have equal diffusion coefficients) undergo smaller changes in their own lability degree, but these changes have the greater impact on the overall metal flux. Partially labile complexes are more easily influenced by the mixture than labile or inert ones. Some mixture effects can be qualitatively predicted by an analytical expression for the lability index derived using the reaction layer approximation. For a mixture of many complexes, the change in the lability degree of a complex due to the mixture effect can be understood as a combination of the changes due to all of the complexes present.

Lability criteria for successive metal complexes in steady-state planar diffusion.

The lability of sequential metal complexes, ML, ML2, ML3, ... , up to a general 1:n metal/ligand stoichiometric ratio is considered for the case of metal ions (M) being accumulated at a surface (analytical sensor or organism). The analytical solution for the steady-state diffusion of M within a sequential complexation scheme allows quantification of the contribution from the dissociation of all of the complex species to the metal flux through the so-called lability degree, xi. A lability degree for each sequential complexation step is also defined which, due to the sequential character of the complexation scheme, depends not only on the proper kinetic constants of the given complexation step but also on the kinetics of the previous ones. When all contributions from the complexes are diffusion limited, the system is fully labile and xi=1. To provide simple lability criteria, the reaction layer approximation is extended to specifically deal with this sequential complexation scheme, so that a reaction layer thickness is defined when the existence of one particular rate-limiting step is assumed. Expressions for the classical lability parameter, L, are formulated using the reaction layer approximation. The change of the lability of the system as the diffusion layer thickness is modified is analyzed in detail. The contribution of the complex flux reflects the evolution of the system from labile to inert as the thickness of the sensor is appropriately decreased.