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Capillary electrophoretic behaviour of humic substances in physical gels.

We investigated the principles of the capillary electrophoretic behaviour of humic substances (HSs) in physical gels. Long chain (Mr 4000, 6000 and 20,000) polyethylene glycols (PEGs) at concentrations above their entanglement threshold caused the separation of HS fractions according to molecular size differences. Close linear relationships between effective mobilities and mean apparent molecular masses were observed at PEG concentrations between 2.5 and 15%. The efficiency of the separation does not increase in gels of increasing polymer concentrations. The possibility of interactions between HSs and gel-forming polymers was also investigated. Short chain (Mr 400) PEGs, added to the buffer at concentrations from 2.5 to 12.5%, increased the migration times of all HS fractions, but no separation was obtained even at large polymer concentrations, showing that gel formation was essential for the separation. In 2.5% polyvinyl alcohol (PVA) 49 000 all fractions show two unresolved, but well defined peaks. This separation is probably artefactual and depends on the relative concentration of HSs and PVA, as the relative abundance of the peaks changes with the sample concentration.

Electrophoresis, Capillary↗

Influence of chemical characteristics of humic substances on the partition coefficient of a chlorinated dioxin.

The partition coefficients (Koc) of 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD) with respect to a variety of humic substances (HSs) were evaluated by a method involving solid-phase microextraction and gas chromatography-electron capture detection. The log Koc values for each of the HS samples were in the range of 6.4-7.7. The log Koc values for HAs from tropical peat, brown forest and ando soils were in the range of 7.3-7.6, similar to the calculated value for the octanol-water partition coefficient (log Koc=7.56). In contrast, the log Koc values for FAs and peat HAs were 0.5-1 unit lower than the calculated value. The parameters for the polarity of HSs, as calculated from (N+O)/C, O/C atomic ratios and the carboxyl group content, were numerically similar related to the log Koc for HpCDD. These results show that the Koc values for HpCDD are significantly influenced by the polarity of HSs.

Carbon Isotopes↗

Investigations of cluster ions formed between cesium cations and benzoic, salicylic and phthalic acids by electrospray mass spectrometry and density-functional theory calculations. Toward a modeling of the interaction of Cs+ with humic substances.

A concerted theoretical (density-functional theory) and experimental electrospray mass spectrometry study was conducted on the formation of cesium cation adducts with small molecules taken as models of specific interactions sites in humic substances. Electrospray experiments with phenol, benzoic acid, salicylic acid, and phthalic acid, in methanolic solution containing cesium nitrate, were performed using a quadrupole ion trap. The formation of positively charged mixed clusters, [Cs(CsNO3)(n)(CsA1)(m)(Cs2A2)(p)]+ (A1 = benzoate, salicylate, and hydrogenophthalate, A2 = phthalate), was observed. Calculations of structures and bonding energetics of Cs+ in simple adducts formed with NO3-, CsNO3, A-, AH, and CsA are reported. The observation of variable cluster stoichiometry (n, m and p values) was interpreted in terms of more or less favorable interaction energies between Cs+ and the neutral species constituting the clusters. Phenol did not form clusters in significant abundances, despite a strong calculated interaction between Cs+ and cesium phenolate. This was attributed to its weak acid dissociation in the electrospray solution.

Acids, Carbocyclic↗

Influence of humic substances on the removal of pentachlorophenol by a biomimetic catalytic system with a water-soluble iron(III)-porphyrin complex.

To investigate some basic aspects of soil remediation using biomimetic catalysts, the effects of humic substances (HSs) on the removal of xenobiotics, such as pentachlorophenol (PCP), were investigated. The use of a biomimetic catalytic system using tetra(p-sulfophenyl)porphine-iron(III) (Fe(III)-TPPS) and potassium monopersulfate (KHSO5) resulted in the disappearance of PCP, accompanied by dechlorination. In addition, this process was enhanced by the presence of several types of HSs. The degrees of enhancement (% delta(PCP)60) achieved by the presence of HSs from peat and compost soils were larger than those in the presence of other types of HSs (tropical peat, brown forest, and ando soils). In control experiments, no PCP disappearance and dechlorination were observed in the presence of only KHSO5, only Fe(III)-TPPS, or combinations of HSs and either KHSO5 or Fe(III)-TPPS. To better understand the role of added HS in enhancing or inhibiting PCP disappearance, correlations between the chemical parameters of the HSs and % delta(PCP)60 were investigated. The most effective HSs had lower carboxylic acid contents and lower degrees of unsaturation. The carboxylic acid content and degree of unsaturation increase with the extent of humification. Therefore, HSs of a lower degree of humification would be predicted to be more useful in enhancing the disappearance of PCP in an Fe(III)-TPPS/KHSO5 system.

Catalysis↗

Effect of humic substances on Cu(II) solubility in kaolin-sand soil.

The type and amount of organic matter present in industrially contaminated soils will influence the risk they pose. Previous studies have shown the importance of humic and fulvic acids (FAs) (important components of soil organic matter) in increasing the solubility of toxic metals but were not carried out using toxic metal levels and the pH range typical of industrially contaminated soils. This study investigated the influence of three humic substances (HSs: humates, fulvates and humins) on the solubility of copper(II) ions in kaolinitic soil spiked with Cu at levels representative of industrially contaminated soil. Humates, fulvates and humin were extracted from Irish moss peat, and controlled pH batch leaching tests were conducted on an artificial kaolin-sand soil that was spiked with each. Further leaching tests were conducted on soil spiked with each HS and copper nitrate. Dissolved organic contents were determined by titration and total and free aqueous copper concentrations in the leachate were measured using AAS and ion selective electrode (ISE) potentiometry respectively (dissolved complexed copper levels were determined by difference). It was found that humates and fulvates are partially sorbed by the soil, probably by chemisorption on positively charged gibbsite (Al-hydroxide) sites in the kaolinite. The addition of 340 mg/kg Cu(II) ions did not significantly affect the amount of humate or fulvate sorbed. Dissolved humates and fulvates form soluble complexes with copper over the pH range 3-11. However, in the presence of kaolinite, soluble copper humates and fulvates are unable to compete with the kaolinite for Cu ions at pH 6-7. Above pH 8, humate and fulvate complexes are the only forms of dissolved Cu. Humin is largely insoluble and has little effect on Cu mobility between pH 2 and 12. The implication of this study is that measurement of total soil organic content and water leaching tests should be a standard part of contaminated site investigation.

Adsorption↗

Effects of humic substances on the oxidation of pentachlorophenol by peroxosulfate catalyzed by iron(III)-phthalocyanine-tetrasulfonic acid.

In an attempt to enhance the oxidation of pentachlorophenol (PCP) in the Fe(III)-PcTS/KHSO5 system, the presence of added humic substances (HSs) was studied, investigating the chemical properties of HSs related to the enhancement in PCP oxidation by correlations with the degree of enhancement in PCP oxidation (%delta(PCP)60). The %delta(PCP)60 value increased with a decrease in the content of oxygen-containing functional groups, such as carboxylic acids. This indicated that HSs with a lower content of oxygen-containing functional groups would be useful for enhancing the oxidation of PCP. A negative correlation between %delta(PCP)60 and the kinetic constants of Fe(III)-PcTS self-oxidation indicated that the enhancement by added HSs could be attributed to the suppression of Fe(III)-PcTS deactivation by self-oxidation. Such a stabilization of Fe(III)-PcTS could be attributed to hydrophobic interactions between the catalyst and HSs.

Biodegradation, Environmental↗

[The effect of a low molecular weight synthetic humic substance on pre- and postnatal development in rats].

The influence of a low molecular synthetic humic substance (HS 1500) on pre- and postnatal development of rats was investigated. After oral application of 1000 mg/kg and 2000 mg/kg b. w. HS 1500 during the period of organogenesis (6th to 15th day p. c.) and from 16th day p. c. up to weaning neither adverse effects in the mothers nor in the fetuses were reported. The occurrence of a single malformation (gastroschisis) after application of 1000 mg/kg b. w. from day 6 to 15 of pregnancy, few ossifications of phalanges and differences in swimming behaviour are not regarded as the effect of the substance. In conclusion, the oral use of HS 1500 in the treatment of gastrointestinal diseases in animals during pregnancy is regarded to be riskless.

Administration, Oral↗

Using electrospray-assisted pyrolysis ionization/mass spectrometry for the rapid characterization of trace polar components in crude oil, amber, humic substances, and rubber samples.

We describe the use of electrospray-assisted pyrolysis ionization/mass spectrometry (ESA-Py/MS) to selectively ionize trace polar compounds that coexist with large amounts of nonpolar hydrocarbons in crude oil, amber, humic substances, and rubber samples. Samples of different origins are distinguished rapidly by their positive ion ESA-Py mass spectra without prior separation or chemical pretreatment. During ESA-Py analysis, the samples in their solid or liquid states were pyrolyzed at 590, 630 or 940 degrees C using a commercial Curie-point pyrolysis probe. The gaseous pyrolysates were transferred into a glass reaction cell. The polar compounds (M) in the pyrolysates were then ionized by electrospray ionization (ESI), yielding protonated molecules (MH+). Although the major components of the pyrolysates are nonpolar hydrocarbons, their lack of functional groups that can receive a proton in the ESA-Py source results in no hydrocarbon ion signals being produced; thus, the positive ions detected in ESA-Py mass spectra all result from trace polar components in the pyrolysates.

Amber↗

Strong copper-binding behavior of terrestrial humic substances in seawater.

In coastal areas, strong complexation of copper generally reduces its toxicity; our ability to monitor and regulate copper as a toxin therefore depends on our understanding of the sources and sinks of the copper-binding ligands. Terrestrial humic substances (HS) are well-recognized contributors to weak ligand concentrations in aquatic systems. In this work, we show that HS are likely contributors to both stronger and weaker ligand classes controlling copper speciation in coastal areas receiving typical inputs of terrestrial organic matter. We used competitive ligand exchange adsorptive cathodic stripping voltammetry (CLE-ACSV), with the added ligands benzoylacetone and salicylaldoxime, to examine copper binding by terrestrial HS in a seawater matrix, at HS and copper concentrations typical of coastal waters. Copper titration data of 1 mg/L Suwannee River humic acid (SRHA) in seawater could be modeled using conditional stability constants of 10(12.0) and 10(10.0) and total ligand concentrations of 10.4 and 199 nM for a stronger and weaker ligand, respectively. Similar results were obtained for Suwannee River fulvic acid (SRFA). Strong copper binding by SRFA in seawater was weaker than previously reported for a freshwater at similar pH, possibly indicating effects of Ca and Mg competition or ionic strength. Nevertheless,the concentrations and binding strengths of copper ligands we observed are comparable to the range reported in previous coastal speciation studies. In addition, we show that the weaker copper ligands cause internal calibration techniques to significantly underestimate the sensitivity of ACSV in the presence of HS concentrations typical of coastal waters. To address this issue, we demonstrate the use of "overload titrations", using a high enough concentration of added ligand to outcompete all natural ligands as an alternative calibration technique for analysis of coastal samples.

Adsorption↗

Characterization of aquatic humic substances to DBPs formation in advanced treatment processes for conventionally treated water.

An advanced water treatment demonstration plant consisted of ozone/granular activated carbon processes was operated to study feasibility of the processes. Natural organic matter (NOM) from raw and process waters at the demonstration plant was isolated into humic and non-humic fractions by physicochemical fractionation method to investigate characteristics of humic fraction (i.e., humic substances, HS) as a predominant haloform reactant. Ozone did not significantly oxidize the carboxylic fraction (from 39.1 to 35.9%), while GAC removed some of the carboxylic fraction (from 35.9 to 29.1%). Formation potential of trihalomethanes (THMs) as compared to haloacetic acids formation potential (HAAFP) was highly influenced by HS. Higher yields of THMs resulted from chlorination of HS with a higher phenolic content and phenolic fraction in the HS gradually decreased from 60.5% to 15.8% through the water treatment. The structural and functional changes of HS were identified by elemental, Fourier-transform infrared (FT-IR) and proton nuclear magnetic resonance ((1)H NMR) analyses, and these results were mutually consistent. The functional distribution data obtained by using A-21 resin could be used to support the interpretation of data obtained from the spectroscopic analyses. Decreases in ratio of UV absorbance at 253 nm and 203 nm (A(253)/A(203)) and DBPFPs/DOC showed consistent trends, therefore, A(253)/A(203) ratio may be a good indicator for the disinfection by-product formation potentials (DBPFPs).

Chlorine↗

Retention behavior of humic substances in reversed phase HPLC.

Recovery as well as appearance and abundance (in percent) of different fractions of humic substances are found to depend on injected sample amounts in reversed phase HPLC. Sample amounts have been varied both by varying sample concentration and sample volume. In case of lowest amounts injected only two fractions were obtained for a commercial humic acid sodium salt, i.e. one for excluded molecules and one for hydrophobic components. The abundance of excluded molecules decreases upon increasing amounts injected. Another three fractions are obtained upon increasing amount injected: a hydrophilic fraction and two hydrophobic ones. This behavior is explained by auxiliary equilibria between excluded components and humic molecules previously adsorbed on the stationary phase.

Journal Article↗

Formation, adsorption and separation of high molecular weight disinfection byproducts resulting from chlorination of aquatic humic substances.

A significant portion of the unidentified disinfection byproducts (DBPs) in chlorinated drinking water can be attributed to high molecular weight (MW)-chlorinated DBPs (above 500 Da) that may have adverse health effects. In this work, issues on the formation, adsorption and separation of high MW-chlorinated DBPs were investigated by introducing radioactive (36)Cl into humic substance samples. The results show that the amount of high MW-chlorinated DBPs during chlorination decreased with the increase of contact time from 1 to 120 h, increased with the increase of pH from 5.5 to 9.5, and was less in the ultrafiltered samples from Suwannee River fulvic acid than from Suwannee River humic acid. The high MW-chlorinated DBPs were found to be effectively adsorbed by activated carbon and be possibly reduced to Cl(-) by activated carbon, but not to be readily desorbed from the activated carbon. Those high MW-chlorinated DBPs were demonstrated to be incapable of resolution into discrete peaks by any of the three liquid chromatography columns studied. The significant implications of these results are discussed.

Adsorption↗

Analytical fractionation of aquatic humic substances and their metal species by means of multistage ultrafiltration.

The molecular-size fractionation of aquatic humic substances (HS) and their metal species by means of a novel sequential-stage ultrafiltration (UF) device equipped with five appropriate ultramembranes (1, 5, 10, 50 and 100 kD) is described. First of all, the concentration dynamics of macromolecules, particulary HS, during five-stage UF and its subsequent washing step has been modelled. Based on these results, the fractionation of aquatic HS (from ground and bog water) by means of multistage UF has been optimized for an analytical scale (10 ml sample, 1 mg/ml HS, 10 ml washing solution, pH 6.0). The molecular size-distribution of selected aquatic HS (BOC 1/2 from the "DFG-Versuchsfeld Bocholt", VM 5 from "Venner Moor", Germany) studied by five-stage UF exhibited strong systematic influences of the procedure used for their isolation. The molecular-size distribution of HS obtained by on-line UF and gel permeation chromatography (GPC) showed a satisfactory agreement in the range 1-50 kD. Moreover, when interrupting multistage UF for > 48 h a slow transformation in the HS samples has been found as gradually additional HS fractions of < 1 kD have been formed. Besides unloaded HS molecules, the molecular-size distribution of freshly formed metal species of HS (1.0 mg metal/g HS of Al(III), Cd(II), Cu(II), Fe(III), Mn(II), Ni(II), Pb(II), Zn(II), each) has been characterized by multistage UF as a function of pH-value, degree of loading and complexation time. Metal determinations as carried out by flame AAS, showed that considerable metal fractions in HS especially are present in molecules > 50 kD, which seemed to be rather acid-inert. With complexation times of < 2 days a transient shift of the molecular size distribution of both HS and their metal species (e.g., Al(III), Fe(III) to higher values (> 10 kD) has been found.

Journal Article↗

Influence of natural aquatic humic substances on the bioavailability of benzo(a)pyrene to Atlantic salmon.

Uptake and bioconcentration factors of benzo(a)pyrene (BaP) in Atlantic Salmon were determined in water containing natural aquatic humic substances (AHS) and control water with low humic content by measuring rate constants for uptake and depuration. The amounts of partition-like and strongly AHS bound BaP were determined by tube dialysis and cyclohexane extraction, respectively. Uptake and bioconcentration factors (BCF) were observed to decrease in the presence of AHS, the values of the BCFs being 2310 and 1160 for control water and AHS respectively. The rate constant for uptake of AHS bound BaP was found to be 30 percent of that of free dissolved BaP.

Animals↗

Molecular weight fractionation of humic substances by adsorption onto minerals.

Molecular weight (MW) fractionation of Suwannee River fulvic acid (SRFA) and purified Aldrich humic acid (PAHA) by adsorption onto kaolinite and hematite was investigated in equilibrium and rate experiments with a size-exclusion chromatography system using ultraviolet (UV) light detection. The extent of adsorptive fractionation based on UV detection was positively correlated with the percent carbon adsorption for both humic substances (HS), although the specific fractionation pattern observed depended on the particular HS and mineral used. Higher MW fractions of SRFA, an aquatic HS, were preferentially adsorbed to both kaolinite and hematite whereas the fractionation trends for PAHA, a terrestrial peat HS, differed for the two minerals. The contrasting fractionation patterns for SRFA versus PAHA can be explained reasonably well by the different structural trends that occur in their respective MW fractions and the underlying adsorption processes. Rate studies of adsorptive fractionation revealed an initial rapid uptake of smaller HS molecules by the mineral surfaces, followed by their replacement at the surface by a much slower uptake of the larger HS molecules present in aqueous solution.

Journal Article↗

Long-term effects of amendment with liquid swine manure on proton binding behavior of soil humic substances.

The acid-base properties of humic acids (HAs) and fulvic acids (FAs) isolated from liquid swine manure (LSM), soils amended with either 90 or 150 m(3)ha(-1)year(-1) of LSM for 7 years, and the corresponding unamended control soil were investigated by a current potentiometric titration method. The non-ideal competitive adsorption (NICA)-Donnan model for proton binding by two classes of binding sites (i.e., carboxylic- and phenolic-type groups) was fit to titration data, and a set of fitting parameters was obtained for each HA and FA sample. The NICA-Donnan model was shown to describe with a great degree of accuracy the behavior of experimental titration datasets, and highlighted important differences in the acid-base properties of the HAs and FAs examined. When compared to the unamended soil HA and FA, LSM-HA and LSM-FA, had smaller acidic functional group contents, larger proton binding affinities of both carboxylic- and phenolic-type groups, smaller heterogeneity of carboxylic-type groups, and smaller, in the case of HA, or similar, in the case of FA, heterogeneity of phenolic-type groups. Amendment with LSM caused a decrease of acidic functional group contents and a slight increase of proton binding affinities of carboxylic- and phenolic-type groups of soil HAs and FAs. Further, LSM application induced a decrease of the heterogeneity of carboxylic-type groups, whereas appeared not to affect substantially the heterogeneity of phenolic-type groups of LSM-amended soil HAs and FAs. These effects were more evident for HAs than for FAs and tended to slightly increase with increasing LSM amendment rate.

Acids↗

Field-flow fractionation-inductively coupled plasma mass spectrometry: an alternative approach to investigate metal-humic substances interaction.

Interaction between metal ions and humic matter was investigated using a hyphenated technique, field-flow fractionation-inductively coupled plasma mass spectrometry (FFF-ICP-MS). Aggregation of a metal-spiked commercial Aldrich humic acid in an aqueous solution of calcium ion or in seawater was examined over time intervals of 0-4320 min. The aggregation was demonstrated by shifts in peak maximum of humic matter from smaller size (2.9 nm) to larger size (5.1 or 5.8 nm in Ca2+ solution or in seawater, respectively) and also by the broadening of size distribution profiles. With FFF, size distribution of humic aggregate was characterized. Further, dominant particle size (2.9 nm), mean particle size (3.8 nm), and diffusion coefficient (1.51 x 10(-6) cm2/s) of humic acid solution were determined. With FFF-ICP-MS, associations of Cd, Cu, and Pb with humic aggregates were examined. The mean diameters of Cd-, Cu-, and Pb-bound humic aggregates in the metal-spiked humic acid were 4.1, 4.5, and 5.8 nm, respectively. These diameters were shifted to 6.0, 6.0, and 6.9 nm, respectively, in the humic acid incubated with calcium solution, whereas they were shifted to 6.5, 5.7, and 7.4 nm, respectively, in the humic acid incubated with seawater for three days. Humic aggregate of small size showed more affinity for Cu than Cd and Pb, whereas the large aggregate showed more affinity for Pb than Cd and Cu, respectively.

Environmental Monitoring↗

Effects of humic substances on the bioavailability and aerobic biodegradation of polychlorinated biphenyls in a model soil.

The very high hydrophobicity of polychlorinated biphenyls (PCBs) strongly reduces their bioavailability in aged contaminated soils, thus limiting their bioremediation. The biodegradability of PCBs in heavily contaminated soils can be significantly enhanced by soil treatment with surface-active agents. In this work, the effects of naturally occurring surfactants such as humic substances (HS) on the aerobic biodegradation of PCBs in a model soil were studied. The soil was amended with biphenyl (4 g/kg), Fenclor 42 (1,000 mg/kg), the aerobic PCB-biodegrading bacterial co-culture ECO3 (inoculum: 10(8)CFU/mL), and treated in aerobic batch slurry-phase conditions (17.5% w/v) with and without the addition of HS at the rates of 1.5 and 3.0% (w/w). Low PCBs biodegradation and dechlorination yields were observed in the HS-free microcosms, probably as a result of the rapid disappearance of inoculated bacteria. The presence of HS influenced significantly the activity of the specialized biomass and the biodegradation of PCBs in the microcosms. The microcosms that received HS at the 1.5% rate showed a higher persistence of the specialized bacteria and yields of PCB biodegradation and dechlorination about 150 and 100%, respectively, larger than those found for the HS-free microcosms. Lower stimulating effects were observed in the microcosms added with the HS at 3.0% rate. These effects were attributed to an increased solubilization of PCBs in the hydrophobic domains of the humic supramolecular associations and to a different accessibility of PCBs by the specialized bacteria at the different rates of HS addition. Although the slurry-phase treatment generally showed a decrease of the original soil ecotoxicity, the addition of the originally non-toxic HS decreased soil ecotoxicity for the Collembola animal biomarker and increased that towards the Lepidium sativum vegetal biomarker.

Aerobiosis↗