Assessment of mercury(II) bioavailability using a bioluminescent bacterial biosensor: practical and theoretical challenges.

Critical methodological challenges in the microbial biosensor approach to assessing Hg(II) bioavailability were evaluated from the perspective of analytical chemists. The main challenge stems from the fact that the chemical speciation of Hg(II) in natural waters exerts a major control on its bioavailability, yet its natural complexation equilibria are extensively altered during conventional bioassays. New data, obtained using a bioluminescent Hg(II)-biosensor, that illustrate these challenges are presented and potential solutions proposed.

Characterization of methyl mercury in dental wastewater and correlation with sulfate-reducing bacterial DNA.

Dental wastewater (DWW) was collected over two months from a 12-chair clinic and a single-chair office to identify conditions that may affect Hg methylation. DWW was settled for 24 h and samples were collected from the top and bottom of the supernatant to simulate a range of particles that may escape in-line traps. Total Hg spanned 5 orders of magnitude (0.02-5000 microM), following a log-normal distribution with p10, p50, and p90 concentration values of 0.24, 31 and 4000 microM, respectively; typically well in excess of free aqueous Hg solubility. Methyl Hg was present in high levels (2-270 nM), also following a log-normal distribution with p10, p50, and p90 concentration values of 2.8, 17, and 100 nM, respectively. There were no statistically significant differences (90% CI) in p50 methyl Hg or total Hg between the clinic and office. Methyl Hg was predicted from total Hg data by (+/- 95% CI): Log (Me-Hg) = 0.33 (+/- 0.06) x Log (T-Hg) - 2.27 (+/- 0.13). Total methyl Hg from DWW to U.S. wastewater collection systems is estimated to be 2-5 kg yr(-1). Equilibrium speciation modeling predicted that DWW Hg was primarily in sulfide-Hg complexes, except at high total Hg levels where organo-Hg complexes become significant. DNA extracts amplified by quantitative polymerase chain reaction with primers for total eubacteria and sulfate-reducing bacteria (SRB) indicated that the total eubacterial DNA was composed primarily of SRB, and highly significant correlations were found between methyl Hg and both amplified Desulfobacteraceae (p < 0.0001) and Desulfovibrionacaea DNA (p < 0.00001). Both are known Hg methylators. In marked contrast, there was no significant correlation between methyl Hg and amplified Desulfobulbus DNA, a genus generally not known to methylate Hg at high rates. These results strongly suggest that SRB are implicated in DWW Hg methylation.

Thiourea catalysis of MeHg ligand exchange between natural dissolved organic matter and a thiol-functionalized resin: a novel method of matrix removal and MeHg preconcentration for ultratrace Hg speciation analysis in freshwaters.

Ultratrace analysis of dissolved MeHg in freshwaters requires both dissociation of MeHg from strong ligands in the sample matrix and preconcentration for detection. Existing solid phase extraction methods generally do not efficiently adsorb MeHg from samples containing high concentrations of natural dissolved organic matter. We demonstrate here that the addition of 10-60 mM thiourea (TU) quantitatively releases MeHg from the dissolved matrix of freshwater samples by forming a more labile complex (MeHgTU+) that quantitatively exchanges MeHg with thiol-functionalized resins at pH approximately 3.5 during column loading. The contents of these columns were efficiently eluted with acidified TU and MeHg was analyzed by Hg-TU complex ion chromatography with cold-vapor atomic fluorescence spectrometry detection. Routinely more than 90% of MeHg was recovered with good precision (average relative standard deviation of 6%) from natural waters-obtained from pools and saturated sediments of wetlands and from rivers-containing up to 68.7 mg C L-1 dissolved organic matter. With the preconcentration step, the method detection limit of 0.29 pg absolute or 0.007 ng L-1 in 40-mL samples is equivalent to that of the current state-of-the- art as practiced by skilled analysts. MeHg in 20-50-mL samples was completely trapped. On the basis of our knowledge of the chemistry of the process, breakthrough volume should depend on the concentrations of TU and H+. At a TU concentration of 12 mM breakthrough occurred between 50 and 100 mL, but overall adsorption efficiency was still 85% at 100 mL. Formation of artifactual MeHg is minimal; only about 0.7% of ambient MeHg is artifactual as estimated from samples spiked with 4 microg L-1 HgII.

Determination of MeHg in environmental sample matrices using Hg-thiourea complex ion chromatography with on-line cold vapor generation and atomic fluorescence spectrometric detection.

A novel system for mercury speciation analysis using high-pressure ion chromatography (IC) has been developed and validated. Its chemistry permits separation of the two most abundant forms of Hg in natural waters, soils, sediments, and biota-monomethyl Hg (CH3Hg+) and mercuric Hg (Hg2+)-on the basis of the difference in charge of their respective thiourea (S=C(NH2)20) complexes. Once separated, both species are converted to Hg0 on-line and quantified by cold-vapor atomic fluorescence spectrometry (CVAFS). A column containing thiol-functionalized silica resin installed in the sample loop of the IC system traps Hg2+ and CH3Hg+ from prepared sample solutions without retaining interfering sample matrix components. The resulting matrix-independent chemistry permits external calibration of the system and a high sample throughput ( approximately 6 samples per hour). The system's accuracy has been validated with environmentally relevant reference materials. Figures of merit for the system, an average precision of approximately 2.5% and an absolute detection limit of <1 pg, are comparable to state-of-the-art gas chromatography approaches for Hg speciation analysis (ethylation/GC-CVAFS) and meet or exceed those of all extant LC systems. Low relative detection limits are attainable through preconcentration onto the thiol resin.

Modeling complexometric titrations of natural water samples.

Complexometric titrations are the primary source of metal speciation data for aquatic systems, yet their interpretation in waters containing humic and fulvic acids remains problematic. In particular, the accuracy of inferred ambient free metal ion concentrations and parameters quantifying metal complexation by natural ligands has been challenged because of the difficulties inherent in calibrating common analytical methods and in modeling the diverse array of ligands present. This work tests and applies a new method of modeling titration data that combines calibration of analytical sensitivity (S) and estimation of concentrations and stability constants for discrete natural ligand classes ([Li]T and Ki) into a single step using nonlinear regression and a new analytical solution to the one-metal/two-ligand equilibrium problem. When applied to jointly model data from multiple titrations conducted at different analytical windows, it yields accurate estimates of S, [Li]T, Ki, and [Cu2+] plus Monte Carlo-based estimates of the uncertainty in [Cu2+]. Jointly modeling titration data at low-and high-analytical windows leads to an efficient adaptation of the recently proposed "overload" approach to calibrating ACSV/CLE measurements. Application of the method to published data sets yields model results with greater accuracy and precision than originally obtained. The discrete ligand-class model is also re-parametrized, using humic and fulvic acids, L1 class (K1 = 10(13) M(-1)), and strong ligands (L(S)) with K(S) >> K1 as "natural components". This approach suggests that Cu complexation in NW Mediterranean Sea water can be well represented as 0.8 +/- 0.3/0.2 mg humic equiv/L, 13 +/- 1 nM L1, and 2.5 +/- 0.1 nM L(S) with [CU]T = 3 nM. In coastal seawater from Narragansett Bay, RI, Cu speciation can be modeled as 0.6 +/- 0.1 mg humic equiv/L and 22 +/- 1 nM L1 or approximately 12 nM L1 and approximately 9 nM L(S), with [CU]T = 13 nM. In both waters, the large excess (approximately 10 nM) of high-affinity, Cu-binding ligands over [CU]T results in low equilibrium [Cu2+] of 10(-14.5 +/- 0.2) M and 10(-13.3 +/- 0.4) M, respectively.

Effects of soil pH and soil water content on prosulfuron dissipation.

The sulfonylurea herbicide prosulfuron, 1-(4-methoxy-6-methyltriazin-2-yl)-3-[2-(3,3,3-trifluoropropyl)phenylsulfonyl]urea, is used for the selective control of broadleaf weeds in corn, sorghum, and cereal grains. To investigate its fate in soils, this study examined the effects of soil pH and water content on the rates of dissipation processes and the products formed under aerobic conditions. Radiometry and chromatography analyses were used to quantify the degradation products and bound residues formed in incubations of 10 different soils. The pH-dependent hydrolysis of the sulfonylurea bridge to form phenyl sulfonamide was the primary transformation process. Significant microbial degradation of prosulfuron occurred in 2 of the 10 soils, yielding (14)CO(2) and desmethyl prosulfuron among the major products. The time required for 50% dissipation of the herbicide (DT(50)) was determined for each soil and water content treatment. At equivalent water contents, prosulfuron DT(50) values were positively correlated with soil pH (P < 0.0001), varying from 6.5 days at pH 5.4 to 122.9 days at pH 7.9. Soil pH and water content strongly influence the fate of sulfonylurea herbicides in agricultural fields. Differences in the effect of soil water content on dissipation kinetics in a comparison of two soils were attributed to differences in soil pH, texture, and the ability of indigenous microorganisms to transform the herbicide.