Impact of ionic strength on Cd(II) partitioning between alginate gel and aqueous media.

Alginate gel is representative of polysaccharide-based components of cell walls which contain a large number of negatively charged functional groups. The structural charge gives rise to a Donnan potential in the gel, which impacts significantly on the partitioning of ions between the aqueous medium and the gel. We measured the Donnan potential and partitioning of Cd2+ in alginate gel as a function of ionic strength in the range 1-100 mM. The Cd2+ partition coefficient between gel and medium, as measured by in situ microelectrode voltammetry, reaches values between 10 and 100 in the 0.1-1 mM ionic strength range, and agrees well with Donnan partition calculations based on the charge density of the gels. The total Cd(II) concentration in the gel correlates approximately linearly with the free [Cd2+]gel. The results imply that metal ion activities in the biopolymer gel phase may generally differ drastically from those in the bulk medium to an extent that strongly depends on ionic strength. This feature must be taken into account in estimations of exposure conditions for predictions of bioavailability.

Relationship between metal speciation in soil solution and metal adsorption at the root surface of ryegrass.

The total metal content of the soil or total metal concentration in the soil solution is not always a good indicator for metal availability to plants. Therefore, several speciation techniques have been developed that measure a defined fraction of the total metal concentration in the soil solution. In this study the Donnan Membrane Technique (DMT) was used to measure free metal ion concentrations in CaCl(2) extractions (to mimic the soil solution, and to work under standardized conditions) of 10 different soils, whereas diffusive gradients in thin-films (DGT) and scanning chronopotentiometry (SCP) were used to measure the sum of free and labile metal concentrations in the CaCl(2) extracts. The DGT device was also exposed directly to the (wetted) soil (soil-DGT). The metal concentrations measured with the speciation techniques are related to the metal adsorption at the root surface of ryegrass (Lolium perenne L.), to be able to subsequently predict metal uptake. In most cases the metal adsorption related pH-dependently to the metal concentrations measured by DMT, SCP, and DGT in the CaCl(2) extract. However, the relationship between metal adsorption at the root surface and the metal concentrations measured by the soil-DGT was not-or only slightly-pH dependent. The correlations between metal adsorption at the root surface and metal speciation detected by different speciation techniques allow discussion about rate limiting steps in biouptake and the contribution of metal complexes to metal bioavailability.

Cd(II) speciation in alginate gels.

Polysaccharides, such as those occurring in cell walls and biofilms, play an important role in metal speciation in natural aqueous systems. This work describes the speciation of Cd(II) in alginate gels chosen as a model system for biogels. The gels are formed by bridging calcium ions at junction zones present along adjacent homopolymeric guluronic acid chain sequences. The free Cd2+ concentration in the gel phase is measured by a novel in situ microelectrode voltammetric technique that monitors the electroactive probe cation Cd2+ by its reduction at a Au-amalgam microelectrode. In situ voltammetric measurement coupled with total Ca(II) and Cd(II) determinations, as well as potentiometric titration, permits the full reconstruction of the charging environment and the cation binding forthe gel phase. Three independent combinations of measuring and modeling the charged gel layer thereby permit accurate prediction of the Donnan potential, psiD, and the Donnan enrichment coefficient, piD. At an ionic strength of 10 mM, Donnan potentials in the gel ranged from approximately -10 to -20 mV, corresponding to an enhancement in the level of free Cd2+ ions in the gel phase relative to the bulk solution by a factor of approximately 3. In contrast, the total level of Cd(II) was found to be enhanced by a factor of approximately 60, resulting predominantly from the specific binding of the Cd bythe uronic acids of the alginate gel. These results emphasize that large differences in Cd(II) speciation can arise due to the combination of specific and electrostatic modes of binding. The results of this speciation analysis, for charged biological gels, have important consequences for mechanistic interpretation of metal biouptake processes involved in complex media.

Metal uptake by Lolium perenne in contaminated soils using a four-step approach.

Most research dealing with soil (solution) speciation and metal uptake by plants has focused on the relationships between a certain bioavailable fraction in the soil and metal uptake by aboveground parts of the plants. Here, a new approach to interpretation of metal uptake is presented that considers four steps: First, the metal concentration in the soil solution is related to the total metal content of the soil. Second, the metal adsorption to the root surface is related to the metal concentration in the soil solution. Third, the metal content in the roots is related to the adsorption of metal ions to the root surface. Fourth, the metal content in the shoots is related to the metal content in the roots. For grass grown on 10 different soils, it is shown that the metal adsorption to the root surface is pH-dependently related to the free or total metal concentration in the soil solution. The metal content in the roots depends linearly on the metal adsorption at the root surface, whereas the metal content in the shoots depends on the metal content in the roots, either linearly (Zn) or reaching a maximum (Cu, Pb, and Cd). For the Ni content in the shoots as a function of the root content, the relation is pH dependent, probably because of the competition effects of Ca. The pH of the soil has to be taken into account when CaCl2 extractions are used as a basis for risk assessment toward plants.

Measuring free metal ion concentrations in multicomponent solutions using the Donnan membrane technique.

Among speciation techniques that are able to measure free metal ion concentrations, the Donnan membrane technique (DMT) has the advantage that it can measure many different free metal ion concentrations simultaneously in a multicomponent sample. Even though the DMT has been applied to several systems, like surface waters, soil solutions, and manure slurry, basic features and calibrations with model calculations of the laboratory and field DMT have not been done sufficiently yet. Therefore, we tested the application of the DMT on metal complexation with several synthetic and natural ligands and the applicability of the dynamic mode of the DMT. The results show that there is a high agreement between the calculated and measured free metal ion concentrations in solutions containing synthetic (nitriloacetic acid, diglycolic acid) and natural organic ligands (fulvic acid, humic acid) at various pH values. Both the laboratory DMT and the field DMT give very similar results. In a solution containing labile ligands, equilibrium time is smaller than in a donor solution containing inert ligands or no ligands. Moreover, when labile ligands are present in the donor solution, a dynamic procedure can be used to decrease equilibrium time. This procedure cannot be applied when no ligands or only inert ligands are present.

Comparison of analytical techniques for dynamic trace metal speciation in natural freshwaters.

Several techniques for speciation analysis of Cu, Zn, Cd, Pb, and Ni are used in freshwater systems and compared with respect to their performance and to the metal species detected. The analytical techniques comprise the following: (i) diffusion gradients in thin-film gels (DGT); (ii) gel integrated microelectrodes combined to voltammetric in situ profiling system (GIME-VIP); (iii) stripping chronopotentiometry (SCP); (iv) flow-through and hollow fiber permeation liquid membranes (FTPLM and HFPLM); (v) Donnan membrane technique (DMT); (vi) competitive ligand-exchange/stripping voltammetry (CLE-SV). All methods could be used both under hardwater and under softwater conditions, although in some cases problems with detection limits were encountered at the low total concentrations. The detected Cu, Cd, and Pb concentrations decreased in the order DGT > or = GIME-VIP > or = FTPLM > or = HFPLM approximately = DMT (>CLE-SV for Cd), detected Zn decreased as DGT > or = GIME-VIP and Ni as DGT > DMT, in agreement with the known dynamic features of these techniques. Techniques involving in situ measurements (GIME-VIP) or in situ exposure (DGT, DMT, and HFPLM) appear to be appropriate in avoiding artifacts which may occur during sampling and sample handling.

Model predictions of metal speciation in freshwaters compared to measurements by in situ techniques.

Measurements of trace metal species in situ in a softwater river, a hardwater lake, and a hardwater stream were compared to the equilibrium distribution of species calculated using two models, WHAM 6, incorporating humic ion binding model VI and visual MINTEQ incorporating NICA-Donnan. Diffusive gradients in thin films (DGT) and voltammetry at a gel integrated microelectrode (GIME) were used to estimate dynamic species that are both labile and mobile. The Donnan membrane technique (DMT) and hollow fiber permeation liquid membrane (HFPLM) were used to measure free ion activities. Predictions of dominant metal species using the two models agreed reasonably well, even when colloidal oxide components were considered. Concentrations derived using GIME were generally lower than those from DGT, consistent with calculations of the lability criteria that take into account the smaller time window available forthe fluxto GIME. Model predictions of free ion activities generally did not agree with measurements, highlighting the need for further work and difficulties in obtaining appropriate input data.

Effects of humic acid and competing cations on metal uptake by Lolium perenne.

Within the biotic ligand model, which describes relationships between chemical speciation and metal binding at an organism's surface, multicomponent (long-term) metal uptake by plants has seldom been studied. In the present work, we exposed perennial ryegrass to nutrient solutions with two levels of Cd, Cu, Ni, Pb, and Zn (1 and 0.1 microM) and with or without 30 mg/L of humic acid. Iron and Mn concentrations were constant over all treatments. The hypothesis tested was that humic acid lowers the free and labile metal concentration and, therefore, reduces the metal uptake and, finally, the metal content of the plant. The free metal ion concentrations in the nutrient solutions were measured by the Donnan membrane technique and labile metal concentrations by diffusive gradients in thin-films. The metal content of the shoots depends on the metal content of the roots. The metal content of the roots is a function of the adsorption of metals on the root surface. In a multicomponent system at metal concentrations of 1 microM, humic acid decreased Cu, Pb, and Fe adsorption at the root surface, but it increased Cd, Zn, and Mn adsorption at the root surface. Complexation of cations such as Cu, Pb, and Fe with high affinity for (dissolved) organic matter may lead to increased uptake of cations with low affinity for organic matter (Ni, Zn, and Cd) because of competition between cations at the root surface. The results suggest that competition between metal ions can play a major role in multicomponent metal uptake, which has to be taken into account during risk assessments of metal-polluted soils.

Measuring free metal ion concentrations in situ in natural waters using the Donnan Membrane Technique.

Metal toxicity is not related to the total but rather to the free or labile metal ion concentration. One of the techniques that can be used to measure several free metal ion concentrations simultaneously is the Donnan Membrane Technique (DMT) in combination with the inductively coupled plasma-mass spectrometer (ICP-MS). However, free metal ion concentrations in natural waters are commonly below the detection limit of ICP-MS. We decreased the detection limit by making use of a ligand, and we developed a field DMT cell that can be applied in situ in natural waters. A kinetic approach can be used to calculate free metal ion concentrations when the equilibrium time becomes too large. The field DMT measured in situ in natural waters a free metal ion concentration ranging from 0.015% (Cu) to 13% (Zn) of a total metal concentration ranging from 0.06 nM (Cd) to 237 nM (Zn). The free metal ion concentrations were difficult to predict using an equilibrium speciation model, probably due to the uncertainty in the nature of the dissolved organic matter or the presence of other reactive colloids. It is shown that DMT can follow changes in the free metal ion concentration on times scales less than a day under certain conditions.

Applicability of the black slug Arion ater for monitoring exposure to polycyclic aromatic hydrocarbons and their subsequent bioactivation into DNA binding metabolites.

The applicability of terrestrial black slugs Arion ater (Mollusca, Gastropoda) was studied for biomonitoring environmental exposure to polycyclic aromatic hydrocarbons (PAHs). In laboratory experiments, slugs were orally exposed to benzo[a]pyrene (BaP) for a short term (3 days) or a long term (119 days) period. Test animals were collected in the field, or were reared under laboratory conditions to ensure that they had no history of PAH-exposure. Benzo[a]pyrene hydroxylase (BPH) activity was measured in the digestive gland as a biomarker for BaP exposure. Bulky DNA adduct formation in kidney was measured as an effect biomarker for BaP bioactivation into DNA-binding metabolites. Although success of clutching was relatively low (5 out of 18 slugs produced egg packages), sufficient number of slugs were obtained to perform exposure experiments due to high hatching (89%) and survival rates (79%). After a short exposure to a relatively high BaP doses of 20 and 200 microg/g fresh feed, a dose-dependent and significant increase of BPH activity and bulky DNA adduct levels could be demonstrated in A. ater. Induction factors were low (two times control level), but optimization of the test conditions yielded a higher BPH induction factor of 4.8 times control level. BPH activity and bulky DNA adduct levels, however, did not increase after a long-term exposure to environmentally relevant BaP doses (upto 0.25 microg/g fresh feed). Based on this lack of response after realistic exposure it is concluded that A. ater is not sensitive to BaP exposure and, therefore, not suitable for monitoring environmental exposure to PAHs.