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Ferric ions can bind strongly with dissolved organic matter (DOM), including humic acids (HA), fulvic acids (FA), and protein-like substances, whereas isolation of Fe-DOM precipitates (Fe-DOMP) and their biochemical characteristics remain unclear. In this work FeCl3 was used to isolate DOM components from various sources, including river, lake, soil, cow dung, and standard tryptophan and tyrosine, through precipitation at pH 7.5-8.5. The Fe-DOMP contribute to total DOM by approximately 38.6-93.8% of FA, 76.2% of HA and 25.0-30.4% of tryptophan and tyrosine, whilst fluorescence spectra allowed to monitor/discriminate the various DOM fractions in the samples. The relative intensity of the main infrared peaks such as 3406‒3383 cm-1 (aromatic OH), 1689‒1635 cm-1 (‒COOH), 1523-1504 cm-1 (amide) and 1176-1033 cm-1 (‒S=O) show either to decline or disappear in Fe‒DOMP. These results suggest the occurrence of Fe bonds with various functional groups of DOM, indicating the formation of π-d electron bonding systems of different strengths in Fe‒DOMP. The novel method used for isolation of Fe-DOMP shows promising in opening a new frontier both at laboratory and industrial purposes. Furthermore, results obtained may provide a better understanding of metal-organic complexes involved in the regulation of the long-term stabilization/sequestration of DOM in soils and waters.

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The production of fluorescent dissolved organic matter (FDOM) by phytoplankton and its subsequent degradation, both of which occur constantly under diurnal-day time sunlight and by night time dark-microbial respiration processes in the upper layer of surface waters, influence markedly several biogeochemical processes and functions in aquatic environments and can be feasibly related to global warming (GW). In this work sunlight-mediated high-temperature was shown to accelerate the production of FDOM, but also its complete disappearance over a 24-h diurnal period in July at the highest air and water temperatures (respectively, 41.1 and 33.5 °C), differently from lower temperature months. Extracellular polymeric substances (EPS), an early-state DOM, were produced by phytoplankton in July in the early morning (6:00-9:00), then they were degraded into four FDOM components over midday (10:00-15:00), which was followed by simultaneous production and almost complete degradation of FDOM with reformation of EPS during the night (2:00-6:00). Such transformations occurred simultaneously with the fluctuating production of nutrients, dissolved organic carbon (DOC), dissolved organic nitrogen (DON) and the two isotopes (δ15N and δ18O) of NO3-. It was estimated that complete degradation of FDOM in July was associated with mineralization of approximately 15% of the initial DOC, which showed a nighttime minimum (00:00) in comparison to a maximum at 13:00. FDOM identified by excitation-emission matrix spectroscopy combined with parallel factor analysis consisted of EPS, autochthonous humic-like substances (AHLS) of C- and M-types, a combined form of C- and M-types of AHLS, protein-like substances (PLS), newly-released PLS, tryptophan-like substances, tyrosine-like substances (TYLS), a combined form of TYLS and phenylalanine-like substances (PALS), and their degradation products. Finally, stepwise degradation and production processes are synthesized in a pathway for FDOM components production and their subsequent transformation under different diurnal temperature conditions, which provided a broader paradigm for future impacts on GW-mediated DOM dynamics in lake water.

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Soil humic substances (HS) are involved in almost all biogeochemical processes and functions in soils, thus their extraction from aiming to their characterization is very important. However, many factors that influence HS extraction from soil still need further studies. The aim of this work was to assess and quantify comparatively the solubility of soil HS as a function of extraction time, various extractants, solid to liquid ratio and sequential extraction. In this work three different soils, i.e. a forest, a maize and a paddy soil, were examined to assess the solubility of soil HS based on their fluorescence (excitation-emission matrix, EEM) features and changes in nutrient (NO3--N, PO43--P and dissolved Si) contents using multiple extraction approaches (time-dependent, various extractants, solid to liquid ratio, and sequential extraction). Three fluorescent components, i.e. humic acids-like (HA-like), fulvic acids-like (FA-like), and protein-like fluorophores (PLF), were identified by parallel factor (PARAFAC) analysis of EEM spectra of the various soil extracts. The solubility of HS, dissolved organic carbon (DOC) and nutrients were shown to increase with extraction time, except for PLF. The FA-like fraction disappeared completely in KCl extracts of all three soils, suggesting the inefficiency of salt extraction. Conversely, HS and nutrients solubility substantially increased in alkaline extracts, and dissolved Si was correlated significantly with the fluorescent intensities of HA-like and FA-like, thus confirming the well-known typical process of alkaline dissolution of HS bound to phytolith and silicate minerals. The relative solubility of HS and nutrients was higher at lower solid to liquid ratio (1:250-1:100), whereas their maximum yields was achieved at high solid to liquid ratio (1:10) for all three soils. Sequential extraction results showed that the first water extraction step contributed 42-55% of HS, which suggested that a single extraction was insufficient to recover HS. In conclusion, water and alkaline extraction could provide, respectively, the labile and insoluble complexed HS existing in soil.

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The main aim of this work was to study the mechanisms of interaction between iron(II) ions and humic acids as a function of pH, iron concentration and various humic acids chemical properties, including the degree of humification, elemental composition, aromaticity and content of acidic functional groups. The results indicated that iron was bound by humic acids at pH 7 in amounts ∼2 times higher than at pH 5 (averaged capacities: 117 and 57 cmol/kg, respectively). Iron binding at pH 7 increased with increasing the total carboxylic and phenolic groups content and the degree of humification of humic acids (R-coefficients: 0.99 and 0.95, respectively). The stability of humic acid-iron complexes at pH 7 were only slightly lower than at pH 5 due to iron hydroxides formed at pH > 5 (averaged stability constants: 5.18 and 5.26, respectively). Iron coordination mode varied depending on pH: at pH 5, the bidentate (chelate) mode dominated, whereas at pH 7 the bridging mode appeared prevalent. The total amount of bound iron was much smaller than the content of the carboxylic and phenolic groups in humic acids, on average by ∼80 (pH 7) and ∼90.1% (pH 5) indicating the occurrence of steric effects in humic acid structure i.e. the reduction of the complexation capacity of free functional groups by adjacent groups occupied by iron and/or the formation of intramolecular aggregates with iron hindering the access of further metal ions. At pH 5 the complexes were soluble in the iron concentration range positively correlated to carboxylic and phenolic groups content, showing the protective nature of negatively charged functional groups on the stability of the solution. At this pH, the destabilization of the system was governed by the neutralization of humic acid charged structures by metal cations and the compression of the double electric layer. At pH 7 the stability of the humic acid-iron solution was largely determined by the form of iron, mainly by the precipitation of metal hydroxides acting as a flocculant destabilizing the solution by co-precipitation of humic acid-iron complexes.

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Soil organic carbon (OC) measurement is a crucial factor for quantifying soil C pools and inventories and monitoring the inherent temporal and spatial heterogeneity and changes of soil OC content. These are relevant issues in addressing sustainable management of terrestrial OC aiming to enhance C sequestration in soil, thus mitigating the impact of increasing CO2 concentration in the atmosphere and related effects on global climate change. Nowadays, dry combustion by an elemental analyzer or wet combustion by dichromate oxidation of the soil sample are the most recommended and commonly used methods for quantitative soil OC determination. However, the unanimously recognized uncertainties and limitations of these classical laboursome methods have prompted research efforts focusing on the development and application of more advanced and appealing techniques and methods for the measurement of soil OC in the laboratory and possibly in situ in the field. Among these laser-induced breakdown spectroscopy (LIBS) has raised the highest interest for its unique advantages. After an introduction and a highlight of the LIBS basic principles, instrumentation, methodologies and supporting chemometric methods, the main body of this review provides an historical and critical overview of the developments and results obtained up-to-now by the application of LIBS to the quantitative measurement of soil C and especially OC content. A brief critical summary of LIBS advantages and limitations/drawbacks including some final remarks and future perspectives concludes this review.

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The aim of this study was to elucidate the heterogeneous structural and functional composition of humic acids (HAs) and dissolved organic matter (DOM) isolated from two Alfisol profiles with different soil texture, in order to develop a better understanding of the organic matter dynamics. Soil samples were collected at different depths from three (Ap, 2AB and 2Bt) and eight (A1, A2, A3, E1, E2, 2Bt1, 2Bt2 and 2Bt3) soil horizons of two Alfisols located in the south (PR1) and north (PR2) of Italy, with a clay texture and a silt loam to loam ones, respectively. Chemical and spectroscopic methods were used to characterize the HAs and the DOM isolated from different soil horizons, including Ultraviolet-Visible (UV-Vis), Fourier Transform Infrared (FTIR), and Fluorescence spectroscopies. The HAs and the DOM isolated from the two Alfisols apparently showed significant differences in their compositional, structural and functional characteristics. In particular, the HAs isolated from the PR1 featured a higher degree of humification and molecular complexity with respect to those isolated from the PR2. On the contrary, the DOM samples isolated from the PR2 showed a more marked aromatic character and polycondensation degree. Both the HAs and the DOM obtained from the PR1 presented a greater qualitative homogeneity with respect to those obtained from the PR2. These results could be reasonably ascribed to the different texture and horizons of the two Alfisols, and to a greater pedogenesis occurred in the PR1.

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Sorption and biodegradation are the main mechanisms for the removal of endocrine disruptor compounds (EDs) from both solid and liquid matrices. There are recent evidences about the capacity of white-rot fungi to decontaminate water systems from phenolic EDs by means of their ligninolytic enzymes. Most of the available studies report the removal of EDs by biodegradation or adsorption separately. This study assessed the simultaneous removal of five EDs­the xenoestrogens bisphenol A (BPA), ethynilestradiol (EE2), and 4-n-nonylphenol (NP), and the herbicide linuron and the insecticide dimethoate­from a municipal landfill leachate (MLL) using a combined sorption/bioremoval approach. The adsorption matrices used were potato dextrose agar alone or added with each of the following adsorbent materials: ground almond shells, a coffee compost, a coconut fiber, and a river sediment. These matrices were either not inoculated or inoculated with the fungus Pleurotus ostreatus and superimposed on the MLL. The residual amount of each ED in the MLL was quantified after 4, 7, 12, and 20 days by HPLC analysis and UV detection. Preliminary experiments showed that (1) all EDs did not degrade significantly in the untreatedMLL for at least 28 days, (2) the mycelial growth of P. ostreatus was largely stimulated by components of the MLL, and (3) the enrichment of potato dextrose agar with any adsorbent material favored the fungal growth for 8 days after inoculation. A prompt relevant disappearance of EDs in the MLL occurred both without and, especially, with fungal activity, with the only exception of the very water soluble dimethoate that was poorly adsorbed and possibly degraded only during the first few days of experiments. An almost complete removal of phenolic EDs, especially EE2 and NP, occurred after 20 days or much earlier and was generally enhanced by the adsorbent materials used. Data obtained indicated that both adsorption and biodegradation mechanisms contribute significantly to MLL decontamination from the EDs studied and that the efficacy of the methodology adopted is directly related to the hydrophobicity of the contaminant.

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Bisphenol A (BPA) and 4-n-nonylphenol (NP) are two endocrine disruptor compounds dangerous to animals, especially aquatics, and humans. They can be leached from urban and industrial wastes and contaminate the environment. White rot fungi produce ligninolytic enzymes capable of biodegrading aromatic contaminants, including some endocrine disruptors. This investigation has evaluated the potential of three fungal species, Trametes versicolor, Stereum hirsutum and Pleurotus ostreatus, to remove BPA at a concentration of 4.6 mg L(-1) from two freshwaters, a lake and a river, and both BPA and NP each at a concentration of 10 mg kg(-1) from the corresponding sediments. A comparative assessment of mycelial growth during biodecontamination showed that, in general, the maximum fungal hyphae elongation was observed with T. versicolor in freshwaters and with P. ostreatus in sediments. The fungi T. versicolor and P. ostreatus exhibited a similar capacity for removing BPA from the two freshwaters, whereas S. hirsutum was much more effective in the decontamination of lake water than river water. A significant disappearance of both BPA and NP was shown in the two sediments inoculated with each fungus, especially of BPA in the lake sediment and of NP in the river sediment. The most effective removal of the two contaminants from sediments occurred during the first seven days after fungal inoculation.

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Greenhouse and field studies were performed to examine the growth responses and possible phytoremediation capacity towards heavy metals of several Brassicaceae (Brassica alba, Brassica carinata, Brassica napus and Brassica nigra) and Poaceae (durum wheat and barley). Soils used featured total concentrations of Cr, Cu, Pb and Zn largely exceeding the maximum levels permitted by the Italian laws. Different organic amendments were tested such as a compost and the plant growth-promoting rhizobacterium Bacillus licheniformis. In the greenhouse experiment, plant length, leaf area index and shoots dry matter were evaluated periodically for the Brassicaceae examined. Whereas plant length, grains production, weight of 1,000 seeds, ear fertility and tiller density were determined under field conditions at the end of the crop cycle for wheat and barley. In general, the species tested appeared to be tolerant to high heavy metal concentrations in soil, and slightly significant differences were found for all parameters considered. A marked growth increase was shown to occur for Brassicaceae cultivated on compost- and bacillus-amended contaminated soils, with respect to non-amended contaminated soils. With some exception, higher growth parameters were measured for wheat and barley plants cropped from contaminated soils in comparison to non-contaminated soils. Further, bacillus amendment enhanced the length of wheat and barley plants in both non-contaminated and contaminated soils, while different effects were observed for the other parameters evaluated.

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Bisphenol A (BPA) is an endocrine disruptor compound of health concern in natural systems. In this study, BPA removal from solid and aqueous matrices by ligninolytic fungi was investigated. Three white rot fungi, Trametes versicolor (TRA), Stereum hirsutum (STE) and Pleurotus ostreatus (PLE) were evaluated for their capacity to remove BPA added at concentrations of 4.6 and 46 mg L(-1) from potato dextrose agar (PDA) growth medium and at 4.6 mg L(-1) from aqueous solutions. Further, the inhibition of mycelial growth exerted by BPA was evaluated in the experiments with PDA. Results obtained showed that BPA was toxic for TRA and STE only at the higher concentration in PDA. However, the efficiency of the three fungi for BPA removal was significant at either doses, with TRA showing the maximum removal efficiency. In the experiments in aqueous solutions BPA was removed efficiently only by TRA after 7 days and STE after 10 days.

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Bisphenol A (BPA) is an endocrine disruptor compound (EDC) of xenobiotic origin occurring in natural waters and wastewaters, especially in the most industrialized and urbanized areas. Recent investigations report the use of ligninolytic fungi for the removal of aromatic contaminants, including some EDCs, from different matrices. Humic acids (HA) are widely spread in all natural systems and their presence is ascertained to interfere with microbial growth and activity. The objective of this study was to assess the capacity of three ligninolytic fungi, Trametes versicolor, Stereum hirsutum and Pleurotus ostreatus, to remove BPA at the concentration of 4.6 mg L(-1) from water. Fungal growth on potato dextrose agar (PDA), in the absence and in the presence of a leonardite HA or a green compost HA, was evaluated during the biodecontamination process. The methodological approach adopted in this study excluded the presence of the mycelium in the contaminated water. Results obtained evidenced a relevant removal of BPA by any fungus when PDA only was used as growing medium. The addition of leonardite HA and compost HA stimulated the mycelial growth of any fungus, especially T. versicolor, and significantly enhanced the removal of the contaminant from water by, respectively, T. versicolor only and T. versicolor and S. hirsutum.

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The effects of grapemarc distillery effluents on the quality of soil organic matter is extremely important to ensure the environmentally-safe and agronomically efficient use of these materials as organic amendment. In this work, the effects of the application of untreated (UG) and anaerobically digested grapemarc distillery effluents, either added with (AGM) or without mycorrhiza (AG), on soil humic acid (HA) were investigated in field plot experiments in comparison to HAs from a control soil and an inorganic fertilized soil. The humic acid-like fractions (HALs) isolated from UG, AG and soils were characterized for compositional, structural and functional properties by the use of elemental and functional group analysis, and ultraviolet/visible, Fourier transform infrared and fluorescence spectroscopies. Results obtained indicated that anaerobic digestion of effluents produced an extended mineralization with loss of organic C and stabilization of residual organic matter by increasing the content of HALs in the effluent. With respect to control soil HA, HALs isolated from UG and AG were characterized by smaller acidic functional group contents, a prevalent aliphatic character and smaller aromatic polycondensation and humification degrees. The chemical and spectroscopic characteristics of native soil HA were not substantially modified by application of UG, AG and AGM to soil, which suggests the occurred incorporation of the effluent HAL into native soil HA. In conclusion, these results showed the possibility of a beneficial and safe recycling of grapemarc distillery effluents as soil amendment.

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Several different plants are capable of removing and detoxifying the endocrine disruptor bisphenol A from water starting with initial concentrations of 4.6 mg L(-1) and 46 mg L(-1). Bisphenol A seems to be glycosylated, transformed to polar compounds, and bound as residue by five forage grasses, fescue, couch grass, perennial ryegrass, Siberian wheatgrass, and white clover, and three horticultural species, cucumber, marrow plant, and radish. Septic and axenic testing established that microbial degradation is possible for fescue and radish, and perennial ryegrass exudates seemed to contain enzymatic activity that transforms bisphenol A, but this activity is evidently deactivated by microorganisms. Although the grasses tested were more effective than the horticultural species, the optimal species of plants best suited for phytoremediation of bisphenol A was not determined. The limited plant testing during 16d does not define how nor which phytoremediation practices can be applied, but the removal efficiency and evident transformation of bisphenol A justify further feasibility, pilot, and treatability testing of different wastewaters.

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A spent mushroom substrate (SMS) was mixed with wheat straw (WS) in three proportions, C1 (2:1), C2 (4:1), and C3 (6:1), and composted for 90 days in static piles with periodic turning to ensure adequate aeration. Samples from each pile were collected periodically (after 0, 30, 60, and 90 days), and the humic acid-like fractions (HAs) were isolated to determine their elemental composition (C, H, N, S, and O), acidic functional group (carboxylic and phenolic) content, and structural and functional characteristics using spectroscopic methods including ultraviolet-visible, Fourier transform infrared (FTIR), and fluorescence. The results of elemental and functional group analyses show that, with increasing time of composting, the N, O, and acidic functional group contents of HAs increase, whereas their C and H contents and C/N ratio decrease. The analysis of FTIR and fluorescence spectra shows that, with increasing composting time, the presence of aliphatic and polysaccharide-like structures in HAs decreases, whereas oxygenation, polycondensation, and polymerization increase. These results suggest that the chemical and structural characteristics of the HA fractions in the final composts resemble those typical of native soil HAs, which indicate that an adequate degree of maturity and stability is achieved after the end of composting. The results of the present study confirm that composting is an appropriate treatment to transform fresh organic matter (OM) in SMS into humified forms, thus enhancing their quality, agronomic efficiency, and environmental safety as a soil OM resource for application as soil amendment.

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Binding of two model polycyclic aromatic hydrocarbons (PAHs), phenanthrene and pyrene, by humic acids (HAs) isolated from an organic substrate at different stages of composting and a soil was investigated using a batch fluorescence quenching method and the modified Freundlich model. With respect to soil HA, the organic substrate HA fractions were characterized by larger binding affinities for both phenanthrene and pyrene. Further, isotherm deviation from linearity was larger for soil HA than for organic substrate HAs, indicating a larger heterogeneity of binding sites in the former. The composting process decreased the binding affinity and increased the heterogeneity of binding sites of HAs. The changes undergone by the HA fraction during composting may be expected to contribute to facilitate microbial accessibility to PAHs. The results obtained also suggest that bioremediation of PAH-contaminated soils with matured compost, rather than with fresh organic amendments, may result in faster and more effective cleanup.

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The regulation capacity of four humic substance (HS) samples, a soil humic acid (HA) and two HAs and one fulvic acid (FA) isolated from a composting substrate, was evaluated at two concentrations on the growth in vitro of one plant pathogenic, Sclerotinia sclerotiorum, and two antagonistic, Trichoderma viride and T. harzianum, soil-borne fungi. The presence of any HS sample in the growing medium, especially those from the composting substrate, caused a relevant inhibition of the mycelial growth of S. sclerotiorum and a marked stimulation of sclerotial formation that was exhibited as early appearance and numerical increase. On the contrary, the same HS treatments generally did not inhibit the growth of the two Trichoderma species. In particular, T. viride was significantly stimulated by any HS sample at any concentration, with the only exception of HA from fresh-composting substrate, whereas T. harzianum appeared to be stimulated only slightly or not significantly influenced. Only S. sclerotiorum showed evident high correlations of both the extent of the inhibitory action on mycelial growth and the final number of sclerotia with some chemical and functional properties of HS, such as total acidity, COOH group content, and elemental composition.

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The acid-base properties of humic acids (HAs) and fulvic acids (FAs) isolated from composted sewage sludge (CS), thermally-dried sewage sludge (TS), soils amended with either CS or TS at a rate of 80 t ha(-1)y(-1) for 3y and the corresponding unamended soil were investigated by use of potentiometric titrations. The non-ideal competitive adsorption (NICA)-Donnan model for a bimodal distribution of proton binding sites was fitted to titration data by use of a least-squares minimization method. The main fitting parameters of the NICA-Donnan model obtained for each HA and FA sample included site densities, median affinity constants and widths of affinity distributions for proton binding to low and high affinity sites, which were assumed to be, respectively, carboxylic- and phenolic-type groups. With respect to unamended soil HA and FA, the HAs and FAs from CS, and especially TS, were characterized by smaller acidic functional group contents, larger proton binding affinities of both carboxylic- and phenolic-type groups, and smaller heterogeneity of carboxylic and phenolic-type groups. Amendment with CS or TS led to 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. These effects were more evident in the HA and FA fractions from CS-amended soil than in those from TS-amended soil.

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Fulvic acids (FAs) were isolated by a conventional procedure from two mixtures of the sludge residue obtained from olive oil mill wastewater (OMW) evaporated in open-air pond and tree cuttings (TC) at different stages of the co-composting process. The FAs were analyzed for elemental (C, H, N, S, O) and acidic functional group (carboxylic and phenolic) composition, and by ultraviolet/visible, Fourier transform infrared and fluorescence spectroscopies. At the initial stage of composting, FAs from the OMW sludge-TC mixtures were characterized by a prevalent aliphatic character, large contents of C, S-containing groups, proteinaceous materials and polysaccharide components, extended molecular heterogeneity, small O and acidic functional group contents, and small degrees of aromatic ring polycondensation, polymerization and humification. As composting proceeded, C, H and S contents, C/N ratio, and aliphaticity decreased, whereas N, O, COOH and phenolic OH contents, C/H and O/C ratios, and aromaticity increased. These results suggested that, with increasing the composting time, the chemical and structural properties of the FA components of the two OMW sludge-TC mixtures approached the characteristics typical of native soil FAs. Thus, co-composting of OMW sludge mixed with TC may represent a suitable treatment for enhancing the quality of organic matter in these materials when used as soil amendments.

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Fluorescence spectroscopy has been used to probe the structural properties and Cu(II), Zn(II), Cd(II), and Pb(II)-binding behavior of humic acid (HA)-like fractions isolated from a municipal solid waste compost (MSWC) and HAs from unamended and MSWC-amended soils. The main feature of the fluorescence spectra, in the form of emission-excitation matrix (EEM) plots, was a broad peak with the maximum centered at an excitation/emission wavelength pair that was much shorter (340/437 nm) for MSWC-HA than for unamended and MSWC-amended soil HAs (455/513 and 455/512 nm, respectively). Fluorescence intensity for MSWC-amended soil HA was less than that for unamended soil HA. These results were indicative of more aromatic ring polycondensation and humification of soil HAs, and of partial incorporation of simple and low-humified components of MSWC-HA into native soil HA, as a result of MSWC amendment. Titrations of HAs with Cu(II), Zn(II), Cd(II), and Pb(II) ions at pH 6 and ionic strength 0.1 mol L(-1) resulted in a marked decrease of the fluorescence intensities of untreated HAs. By successfully fitting a single-site fluorescence-quenching model to titration data, the metal ion complexing capacities of each HA and the stability constants of metal ion-HA complexes were obtained. The binding capacities and stability constants of MSWC-HA were smaller than those of the unamended soil HA. Application of MSWC to soil slightly reduced the metal-ion-binding capacities and affinities of soil HAs.

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