Molecular simulation of polycyclic aromatic hydrocarbon sorption to black carbon.

Strong sorption of hydrophobic organic contaminants to soot or black carbon (BC) is an important environmental process limiting the bioremediation potential of contaminated soils and sediments. Reliable methods to predict BC sorption coefficients for organic contaminants are therefore required. A computer simulation based on molecular mechanics using force field methods has been applied in this study to calculate BC sorption coefficients of polycyclic aromatic hydrocarbons (PAHs). The free energy difference between PAHs dissolved in water and in water containing a model structure of BC was calculated by thermodynamic integration of Monte Carlo simulated energies of transfer. The free energies were calculated with a hypothetical reference state that has equal free energies in both phases and is therefore cancelled in the calculated free energy difference. The calculated sorption coefficient of phenanthrene (log K(BC) = 5.17 +/- 0.54 L/kg C), fluoranthene (6.33 +/- 0.64 L/kg C) and benzo[a]pyrene (7.38 +/- 0.36 L/kg C) corresponded very well to experimental values available in the literature. Furthermore, an average spacing distance of 3.73 A between PAHs and BC was determined that is only slightly lower than an experimentally determined value of 4.1 A. The method applied in this study enables the calculation of the extent of PAH sorption to a soot surface for which no experimental values are available nor data for related compounds as required in quantitative structure-activity relationships.

Cytochrome P450cam-monoterpene interactions.

The use of the molecular mechanics AMBER force field (FF) to predict product profiles for the hydroxylation of the monoterpenes 1R-camphor, 1S-camphor, 1R-norcamphor, 1S-norcamphor and camphane by the enzyme cytochrome P450cam from the soil bacterium Pseudomonas putida was investigated. Predictions were carried out by applying multiple substrate (starting) orientations in the enzyme pocket in two procedures: a procedure based on molecular dynamics (MD) and a procedure based on short MD simulations followed by geometry optimisations. The latter (GO) procedure is faster and enabled the use of more monoterpene starting orientations. Monoterpene orientations were transformed into product profiles by applying both energetic and geometrical criteria appropriate for the (monoterpene) hydrogen abstraction reaction. Good predictions compared to experimental data were obtained for most compounds in both the MD and GO procedures. Prior to the product profile calculations, the FF was calibrated by reproducing the experimental data for the binding energy of 1R-camphor and 1S-camphor to P450cam and the energy of vaporisation of water. Focus of the calibration was on the value for the scaling factor for the electrostatic interactions.

Resistant sorption of in situ chlorobenzenes and a polychlorinated biphenyl in river Rhine suspended matter.

The desorption kinetics of in situ chlorobenzenes (dichlorobenzenes, pentachlorobenzene and hexachlorobenzene) and 2,4,4'-trichlorobiphenyl (PCB-28) were measured with a gas-purge technique for river Rhine suspended matter sampled in Lobith, The Netherlands. This suspended matter is the main source of sediment accumulation in lake Ketelmeer. In lake Ketelmeer sediment earlier observations showed that slow and very slow fractions dominate the desorption profile. For the river Rhine suspended matter, only for PCB-28 a fast desorbing fraction of around 1.6% could be detected. The observed rate constants were on the average 0.2 h(-1) for fast desorption, 0.004 h(-1) for slow desorption, and 0.00022 h(-1) for very slow desorption. These values are in agreement with previous findings for the sediment from lake Ketelmeer and with available literature data on fast, slow, and very slow desorption kinetics. The results from this study show the similarity of desorption profiles between river Rhine suspended matter, and the top layer sediment from lake Ketelmeer. This indicates that slow and very slow fractions are already present in material forming the top layer of lake Ketelmeer, and were not formed after deposition of this material in the lake. The absence of detectable fast fractions for most compounds could be caused by the absence of recent pollution of the suspended matter. But, the observations may also be explained by a rapid disappearance of compounds from the fast fraction due to a combination of a high affinity of very slow sites for these compounds, and their relatively high volatility.

Dissolved organic carbon--contaminant interaction descriptors found by 3D force field calculations.

Enthalpies of transfer at 300 K of various partitioning processes were calculated in order to study the suitability of 3D force fields for the calculation of partitioning constants. A 3D fulvic acid (FA) model of dissolved organic carbon (DOC) was built in a MM+ force field using AMI atomic charges and geometrical optimization (GO). 3,5-Dichlorobiphenyl (PCB14), 4,4'-dichlorobiphenyl (PCB15), 1,1,1-trichloro-2,2-bis-(4-chlorophenyl)-ethane (PPDDT) and 2-chloro-4-ethylamino-6-isopropylamino-s-triazine (Atrazine) were inserted into different sites and their interaction energies with FA were calculated. Energies of hydration were calculated and subtracted from FA-contaminant interactions of selected sites. The resulting values for the enthalpies of transfer from water to DOC were 2.8, -1.4, -6.4 and 0.0 kcal/mol for PCB 14, PCB15, PPDDT and Atrazine, respectively. The value of PPDDT compared favorably with the experimental value of -5.0 kcal/mol. Prior to this, the method was studied by the calculation of the enthalpies of vaporization and aqueous solution using various force fields. In the MM + force field GO predicted enthalpies of vaporization deviated by +0.7 (PCB14), +3.6 (PCB15) and -0.7 (PPDDT)kcal/mol from experimental data, whereas enthalpies of aqueous solution deviated by -3.6 (PCB14), +5.8 (PCB15) and +3.7 (PPDDT) kcal/mol. Only for PCB14 the wrong sign of this enthalpy value was predicted. Potential advantages and limitations of the approach were discussed.

Membrane-contaminant interaction found by 3D force field calculations.

Enthalpies of contaminant transfer from water to a 1,2-Dilauroyl-DL-phosphatidyl ethanolamine (DLPE) membrane were calculated in order to study the suitability of 3D force fields for the calculation of membrane-water partitioning constants (Kmw) and as potential descriptors for bio-concentration. A 3D DLPE membrane model was built in a MM+ force field using AM1 atomic charges. 3,5-Dichlorobiphenyl (PCB14), 4,4'-dichlorobiphenyl (PCB15), 1,1,1-trichloro-2,2-bis-(4-chlorophenyl)-ethane (PPDDT or p,p'-DDT) and 2-chloro-4-ethylamino-6-isopropylamino-s-triazine (atrazine) were inserted into apolar and polar sites and their interaction energies with the membrane were calculated by geometrical optimization (GO). Energies of hydratation were subtracted from membrane-contaminant interactions of selected sites. The resulting values for the enthalpies of transfer from water to the membrane were 4.7, -2.3, 11.5 and -9.2 kcal/mol for PCB14, PCB15, PPDDT and atrazine, respectively. In contrast to PCB15, the value of PCB14 compared favorably with the experimental values of similar membranes. PCB14, PCB15 and PPDDT had their lowest energies in the apolar sites of the membrane, whereas atrazine tended to accumulate in the polar membrane-bulk water boundary site. Potential advantages and limitations of the approach were discussed.

Determination and theoretical aspects of the equilibrium between dissolved organic matter and hydrophobic organic micropollutants in water (Kdoc).

Literature on the equilibrium constant for distribution between dissolved organic carbon (DOC) (Kdoc) data of strongly hydrophobic organic contaminants were collected and critically analyzed. About 900 Kdoc entries for experimental values were retrieved and tabulated, including those factors that can influence them. In addition, quantitative structure-activity relationship (QSAR) prediction equations were retrieved and tabulated. Whether a partition or association process between the contaminant and DOC takes place could not be fully established, but indications toward an association process are strong in several cases. Equilibrium between a contaminant and DOC in solution was shown to be achieved within a minute. When the equilibrium shifts in time, this was caused by either a physical or chemical change of the DOC, affecting the lighter fractions most. Adsorption isotherms turned out to be linear in the contaminant concentration for the relevant DOC concentration up to 100 mg of C/L. Eighteen experimental methods have been developed for the determination of the pertinent distribution constant. Experimental Kdoc values revealed the expected high correlation with partition coefficients over n-octanol and water (Kow) for all experimental methods, except for the HPLC and apparent solubility (AS) method. Only fluorescence quenching (FQ) and solid-phase microextraction (SPME) methods could quantify fast equilibration. Only 21% of the experimental values had a 95% confidence interval, which was statistically significantly different from zero. Variation in Kdoc values was shown to be high, caused mainly by the large variation of DOC in water samples. Even DOC from one sample gave different equilibrium constants for different DOC fractions. Measured Kdoc values should, therefore, be regarded as average values. Kdoc was shown to increase on increasing molecular mass, indicating that the molecular mass distribution is a proper normalization function for the average Kdoc at the current state of knowledge. The weakly bound fraction could easily be desorbed when other adsorbing media, such as a SepPak column or living organism, are present. The amount that moves from the DOC to the other medium will depend, among other reasons, on the size of the labile DOC fraction and the equilibrium constant of the other medium. Variation of Kdoc with temperature turned out to be small, probably caused by a small enthalpy of transfer from water to DOC. Ionic strength turned out to be more important, leading to changes of a factor of 2-5. The direction of this effect depends on the type of ion. With respect to QSAR relationships between Kdoc and macroscopic or molecular descriptors, it was concluded that only a small number of equations are available in the literature, for apolar compounds only, and with poor statistics and predictive power. Therefore, a first requirement is the improvement of the availability and quality of experimental data. Along with this, theoretical (mechanistic) models for the relationship between DOC plus contaminant descriptors on the one side and Kdoc on the other should be further developed. Correlations between Kdoc and Kow and those between the soil-water partition constant (Koc) and Kow were significantly different only in the case of natural aquatic DOC, pointing at substantial differences between these two types of organic material and at a high correspondence for other types of commercial and natural DOC.

Slow desorption of PCBs and chlorobenzenes from soils and sediments: relations with sorbent and sorbate characteristics.

The kinetics of slow desorption were studied for four soils and four sediments with widely varying characteristics [organic carbon (OC) content 0.5-50%, organic matter (OM) aromatic content (7-37%)] for three chlorobenzenes and five polychlorinated biphenyls (PCBs). Slowly and very slowly desorbing fractions ranged from 1 to 50% (slow) and 3 to 40% (very slow) of the total amount sorbed, and were observed for all compounds and all soils and sediments. In spite of the wide variations in sorbate K(OW) (factor 1000) and sorbent characteristics, the rate constants of slow (k(slow), around 10(-3) h(-1)) and very slow (k(very slow), 10(-5)-10(-4) h(-1)) desorption appeared to be rather constant among the sorbates and sorbents (both within a factor of 5). There was a good correlation (r(2) above 0.9) between the distribution over the slow, very slow and rapid sediment fractions and log K(OC), indicating that sorbate hydrophobicity may be important for this distribution. No correlation could be found between sorbent characteristics [OC, N, and O in the organic matter, polarity index C/(N+O), OC aromaticity as determined by CP-MAS (13)C-NMR] and slow desorption parameters (slowly/very slowly desorbing fractions+corresponding rate constants). The absence of (1) a correlation between k(slow) and k(very slow), respectively, and OC content, and (2) the narrow range of k(slow) and k(very slow) values, indicates that intra-OM diffusion is not the mechanism of slow or very slow desorption, because on the basis of this mechanism it would be expected that increasing OC content would lead to longer diffusion pathlengths and, consequently, to smaller rate constants. In addition, it was tested whether differential scanning calorimetry would reveal a glass transition in the soils/sediments. In spite of the sensitivity of the equipment used (changes in heat flow in the micro-Watt range were measurable), a glass transition was not observed. This means that activation enthalpies of slow desorption can be calculated from desorption measurements at various temperatures. In the present study these values ranged from 60 to 100 kJ/mol among the various soils and sediments studied.

Trace metals in populations of freshwater isopods: influence of biotic and abiotic variables.

Trace metal levels in water, sediments and freshwater isopods from 28 different water systems in the Netherlands were measured during the period of 1986 to 1989. Distinct element-specific internal distribution patterns were present, with Cd and Cu stored mainly in the hepatopancreas (30-60% of total body burden) and Pb and Zn in the hindgut and exoskeleton with hemolymph. Mean whole-body concentrations of the non-essential elements Cd and Pb in individually analyzed isopods varied over three orders of magnitude between populations. The variability of Zn and Cn were within one order of magnitude difference. The variability of trace metal levels between populations exceeded within-population variability. Within-population variability was related to seasonal and biological factors such as body weight, sex, reproductive state, and species abundance. The highest concentrations were found in small juvenile animals compared to adults, females compared to males, and Proasellus meridianus compared to Asellus aquaticus. However, after correction for size effects using a power-curve regression model no significant differences remained between sex and species. Seasonal fluctuations accounted for 33-79% of the within population variability. Trace metal levels in isopods were predicted from concentrations in water and sediments in combination with aqueous Ca, Cl-, DOC, and sediment characteristics (Org-C, clay, CaCO3) using a multiple regression model. With this predictive model 42-63% of the variance could be explained. In situ determined partitioning coefficients (apparent BCF, biota-sediment BSAF, and sediment-water distribution coefficient Kd) varied between locations and covaried with factors related to trace metal bioavailability (aqueous Ca, Cl- and DOC, sediment Org-C, clay, and CaCO3). Especially for Cd and Cu field-derived BCF values were in agreement with previously reported experimental studies. It is concluded, that A. aquaticus may be a suitable candidate-organism for biomonitoring available trace metal levels in littoral freshwater systems. Finally, some practical recommendations are given for future field surveys with freshwater isopods with respect to sample size, allometric standardization, period of sampling and statistical design.

Determination of bioconcentration factors of eight tetrachlorobenzyltoluenes in the zebra mussel Dreissena polymorpha.

Accurate duplicate bioconcentration factors were determined for tetrachlorobenzyltoluenes (TCBTs). This is the first study that quantifies bioconcentration factors for single TCBT isomers. A continuous flow uptake experiment was performed during 21 days with the zebra mussel (Dreissena polymorpha) as a test organism. Bioconcentration factors (BCFs) and rate constants were derived by an iterative integration method. With this method BCFs and rate constants can be derived from experimental data of a bio-concentration test, even if the concentration in the water is not constant and steady state has not been reached. Mean log BCF values of TCBTs ranged from 4.43 +/- 0.05 to 5.19 +/- 0.15 liter.kg-1 on a wet weight basis. The experimental log BCF values were compared with log BCF values of TCBTs, calculated according to linear and polynomial relationships between log BCF and log Kow.

Effect of fatty acids and the aqueous diffusion barrier on the uptake and transport of polychlorinated biphenyls in Caco-2 cells.

Polychlorinated biphenyls (PCBs) dissolved in dietary fat are absorbed in the gastrointestinal tract by the enterocytes in combination with the fatty acids proceeding from the lipid hydrolysis in the gut lumen. The effect of fatty acid absorption on the uptake and transport of 14 PCBs in enterocytes was studied using monolayers of the human intestinal Caco-2 cell line as a model system. The diffusive resistance of the unstirred water layer and the facilitating role of mixed bile salt micelles on the PCB uptake were examined by varying the thickness of the unstirred water layer adjacent to the apical membrane. In additional experiments, the polarity of the PCB uptake and transport in Caco-2 cells was determined. The solubility of PCBs in the mixed bile salt-fatty acid micelles was 2.7 to 4.8-fold higher than the solubility in plain bile salt micelles. Both the uptake and transport of PCBs in Caco-2 cells were significantly higher (up to 10-fold) when the PCBs were presented mixed with fatty acids. Reducing the thickness of the unstirred water layer resulted in an increased uptake of PCBs. The PCB uptake in Caco-2 cells exceeded the uptake as expected from monomer diffusion only, indicating that bile salt micelles facilitate the PCB transport over the unstirred water layer. Concentrations of dichlorobiphenyls accumulating in the basolateral medium stayed unexpectedly low, suggesting that Caco-2 cells might possess the capability of metabolizing lower chlorinated biphenyls. Uptake of PCBs into the Caco-2 cells was not significantly different whether the PCBs were presented at the apical side or at the basolateral side. However, transport of PCBs over the cell monolayer was significantly higher when the PCBs were presented at the apical side as compared to the basolateral side, suggesting that the unidirectional transport of lipids and lipoproteins affected the PCB transport as well. Our studies indicate that monolayers of the Caco-2 cell line offer a useful model system for studying the intestinal uptake and transport processes of hydrophobic xenobiotics such as polychlorinated biphenyls.

Ecological hazard assessment of dioxins: hazards to organisms at different levels of aquatic food webs (fish-eating birds and mammals, fish and invertebrates).

An ecological hazard assessment was performed for 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin in the aquatic environment, taking into account four different classes of species which differ with respect to their positions in aquatic food webs: invertebrates, fish, birds preying on aquatic organisms, and mammals preying on aquatic organisms. The most sensitive toxicological endpoints for each class were selected from the literature. These endpoints were extrapolated to no effect concentrations (NECs) in water with a two-level food chain model as indicator for secondary poisoning. From the results it is concluded that fish-eating birds and mammals experience larger hazards from dioxins and related compounds than fish. The limited data available appear to indicate that invertebrates are rather insensitive to dioxins. The study indicates that assessment of secondary poisoning affects environmental quality criteria setting and risk assessment.

Comparison of bioconcentration factors of tetrachlorobenzyltoluenes in the guppy (Poecilia reticulata) and zebra mussel (Dreissena polymorpha).

Preliminary bioconcentration factors (BCFs) were determined of six tetrachlorobenzyltoluenes (TCBTs) in guppies (Poecilia reticulata). BCFs and rate constants were derived by an iterative integration method. With this method BCFs and rate constants can be derived from experimental data of a bioconcentration test, even if the concentration in the water is not constant and steady state has not been reached. Log BCF values of TCBTs in guppies ranged from 1.67 to 2.68 L.kg-1 on a wet weight basis. These values are much lower than the reported log BCF values of TCBTs in zebra mussels (Dreissena polymorpha) determined with the same method. Biotransformation of TCBTs in guppies as an explanation for the higher log BCF values in zebramussel is discussed.

Aerobic degradation of polychlorinated biphenyls by Alcaligenes sp. JB1: metabolites and enzymes.

In contrast to the degradation of penta- and hexachlorobiphenyls in chemostat cultures, the metabolism of PCBs by Alcaligenes sp. JB1 was shown to be restricted to PCBs with up to four chlorine substituents in resting-cell assays. Among these, the PCB congeners containing ortho chlorine substituents on both phenyl rings were found to be least degraded. Monochloro-benzoates and dichlorobenzoates were detected as metabolites. Resting cell assays with chlorobenzoates showed that JB1 could metabolize all three monochlorobenzoates and dichlorobenzoates containing only meta and para chlorine substituents, but not dichlorobenzoates possessing an ortho chlorine substituent. In enzyme activity assays, meta cleaving 2,3-dihydroxybiphenyl 1,2-dioxygenase and catechol 2,3-dioxygenase activities were constitutive, whereas benzoate dioxygenase and ortho cleaving catechol 1,2-dioxygenase activities were induced by their substrates. No activity was found for pyrocatechase II, the enzyme that is specific for chlorocatechols. The data suggest that complete mineralization of PCBs with three or more chlorine substituents by Alcaligenes sp. JB1 is unlikely.

Solubility and micelle-water partitioning of polychlorinated biphenyls in solutions of bile salt micelles.

Experimental data are presented on the enhanced solubilities and micelle/water partition coefficients of 14 polychlorinated biphenyls (PCBs) in solutions of the bile salts sodium taurocholate (TC) and sodium taurodeoxycholate (TDC) below and above the critical micellar concentration (CMC). Solubilities of PCBs in micellar solutions above the CMC depend linearly on bile salt concentration and the solubility enhancement is up to four orders of magnitude. PCB partition coefficients (log Kmw) are independent of bile salt concentrations and are systematically higher in solutions of TDC than in solutions of TC, ranging from 4.26 to 6.24 and from 3.89 to 5.89 respectively. The partitioning behavior of the pertinent extremely hydrophobics between the aqueous and micellar phases is not completely comparable with simple organic solvent/water systems and the correlation between log Kow and log Kmw is non-linear.

Descriptors for isomer resolution of (bio-) distribution of chlorinated aromatic compounds.

Both Solubility Parameter (SOLPAR) and Linear Solvation Energy Relationship (LSER) theory utilize molar volume and energies as descriptors for distribution phenomena. They provide powerful methods for the prediction of the GLC retention indices of chlorobenzenes (CBzs) and tetrachlorodibenzo[p]dioxins (TCDDs). Almost complete isomer resolution is obtained with correlation coefficients (r) of greater than or equal to 0.9984 (CBzs) and greater than or equal to 0.9839 (TCDDs). The n-octanol/water partition coefficient (log Kow) of CBzs can be calculated with a standard error of regression of 0.04, which compares favourably with standard deviations of 0.03-0.26 in mean values based on four different experimental methods. The HPLC capacity factor of TCDDs on C18 columns can be predicted by SOLPAR with moderate accuracy (r = 0.9470), allowing for a preliminary calculation of log Kow. A simplified LSER method fails.

QSARs and PARs for biodegradation of PCBs.

Relationships between the biodegradation rate constants of a number of polychlorinated biphenyls (PCBs) and hydrophobic and electronic structural parameters are compared. There is no simple relationship with octanol-water partition coefficients, indicating that the biodegradation rates of PCBs are probably not determined by their rates of permeation through the bacterial membranes. Biodegradation rate constants correlated much better with both the electronic and hydrophobic properties of the chlorine substituents, which suggests that the reactivity and possibly enzyme binding of PCBs control their biodegradation rates.

Quantitative structure-activity relationships for biodegradation.

Quantitative structure-activity relationships (QSARs) between biodegradation rates of organic compounds and chemical structure parameters are reviewed. Although a number of such relationships have been developed, they in general only apply to restricted ranges of compounds, limiting their value as predictors of biodegradation rates. For many of these classes of chemicals relationships have been reported with different structural descriptors, varying from macroscopic physical properties to molecular structure parameters. More information on the mechanism and rate-determining steps of biodegradation, which can lead to a better-founded choice of descriptors, and more biodegradation rate data are required to further develop QSARs for biodegradation.