Phenanthrene sorption to sequentially extracted soil humic acids and humins

Humic substances strongly influence the environmental fate of hydrophobic organic chemicals in soils and sediments. In this study, the sorption of phenanthrene by humic acids (HAs) and humins was examined. HAs were obtained from progressively extracting a soil, eight times with 0.1 M Na4P207 and two times with 0.1 M NaOH solution, and then the residue was separated into two humin fractions by their organic carbon contents. The chemical and structural heterogeneity of the HAs and humins were characterized by elemental analysis, ultraviolet-visible spectroscopy, diffuse reflectance infrared Fourier transform spectroscopy, and solid-state 13C NMR. There were significant chemical and structural differences among the HA fractions and humins; the later extracted HAs had relatively high aliphatic carbons content. All sorption data were fitted to a Freundlich equation, S = K(F)C(N), where S and C are the sorbed and solution-phase concentrations, respectively, and K(F) and N are constants. All of the phenanthrene sorptions were nonlinear, and the nonlinearity decreased with further extractions from 0.90 (first extracted HA) to 0.96 (ninth HA) and was the lowest (0.88) for the higher organic carbon content humin. Phenanthrene sorption coefficient by HAs significantly increased with progressive extractions, being the highest for the humins. For HAs isotherms, a positive trend was observed between the sorption coefficient and the aliphaticity, but a negative relation was shown between the nonlinearity and the aliphaticity and between the sorption capacity and polarity of HAs. Phenanthrene sorption was greatly affected by chemical structure and composition of humic substances, even from a same soil. In addition, polarity of humic substances seems to mainly regulate the magnitude of phenanthrene sorption rather than structure.     

Identification and characterization of sorption domains in soil organic matter using strucuturally modified humic acids

The sorption of phenanthrene was examined in humic acids (HAs) from different sources: a compost, a peat soil, and a mineral soil. Sub-samples of each HA were subjected to bleaching or hydrolysis to remove predetermined chemical groups from their structures. Bleaching successfully removed a large percentage of rigid, aromatic moieties, whereas hydrolysis removed the mobile, carbohydrate components. Phenanthrene sorption by all HAs was nonlinear (N < 1). However, the phenanthrene isotherms of the bleached HAs were more linear than those of the untreated HAs, whereas the removal of the carbohydrate components by hydrolysis produced more nonlinear isotherms. The introduction of pyrene to the phenanthrene sorption system yielded more linear isotherms for all the HAs, indicative of competitive sorption. Proton spin-spin (1H T2) relaxation determined by nuclear magnetic resonance (NMR) was used to identify separate rigid (condensed) and flexible (expanded) 1H populations and to determine their distribution. These 1H domains were highly sensitive to temperature and correlated well with reported glass transition temperatures for HAs. In combination with the chemical treatments, sorption, and spectroscopic data, we were able to observe some significant relationships among chemical groups, sorption behavior, and structural characteristics.     

Key role of the low molecular size fraction of soil humic acids for fluorescence and photoinductive activity

The IHSS soil humic acid (HA) standard and two HAs from soils of very different origin (Chernozem and Ranker) were fractionated by tandem size-exclusion chromatography-polyacrylamide gel electrophoresis. From each HA, three fractions with different molecular sizes (MSs) and electrophoretic mobilities were obtained and investigated for their fluorescence properties and abilityto photoinduce the transformation of 2,4,6-trimethylphenol and herbicide fenuron. Regardless of the source of the HA, the two high MS fractions were found to be very weakly fluorescent. They photoinduced the degradations of fenuron and 2,4,6-trimethylphenol less efficiently than the bulk HA (10-50-fold and 1.4-5.3-fold, respectively). In contrast, the low MS fraction was proved to be fluorescent and to photoinduce the transformation of probes as least as efficiently than the bulk HA. These results show that (i) most of fluorophores and a great part of photoinductive chromophores are located in the low MS fractions of soil HAs and (ii) this distribution of photochemically active constituents may be characteristic across broad soil types.     

Immobilizing humic acid in a sol-gel matrix: a new tool to study humic-contaminants sorption interactions

Humic substances originated from aquatic, soil, or sediment environments are mixtures of humic compounds with various characteristics. Sorption interactions with isolated, well defined humic fractions can be studied either in an aqueous phase ("dissolved humic substances"), or in a solid-phase, by coating mineral particles with the humic materials, or simply by working with humic acid particles (powder) at low pH to minimize dissolution. Each attitude, by definition, can be studied by different experimental techniques and has a different meaning for understanding natural environmental processes. In this study, a new tool for studying sorption interactions is presented. Sol-gel was used as an inert matrix to immobilize (entrap) various humic acids (HAs), and then used to study the interactions of several polycyclic aromatic hydrocarbons (PAHs) with the entrapped HA. Linear and nonlinear sorption coefficients were highly correlated with contaminant hydrophobicity. Sorption of pyrene to immobilized HA was in the order of soil HA > Aldrich HA approximately = peat HA. It was concluded that the entrapped HAs retained their original properties in the gel matrix and were accessible to the external contaminant through the pore network. Additionally, binding coefficients of pyreneto dissolved humic substances and to dissolved organic matter (DOM) were determined from the reduction in pyrene sorption to immobilized HA in the presence of dissolved humic material or DOM in solution. Binding coefficients of pyrene were in the order of the following: dissolved Aldrich HA > dissolved peat fulvic acid (FA) > DOM derived from mature compost > DOM derived from fresh compost.     

Binding of atrazine to humic substances from soil,peat and coal related to their structure

Partition coefficients for the binding affinities of atrazine to 16 different humic materials were determined by the ultrafiltration HPLC technique. Sources included humic acids (HA), fulvic acids (FA), and combined humic and fulvic fractions (HF) from soil, peat, and coal humic acid. Each of the humic materials was characterized by elemental composition, molecular weight, and composition of main structural fragments determined by 13C solution-state NMR. The magnitude of K(OC) values varied from 87 to 575 L/kg of C, demonstrating relatively low binding affinity of humic substances (HS) for atrazine. On the basis of the measured K(OC) values, the humic materials can be arranged in the following order: coal HA approximately = gray wooded soil HA > chernozemic soil HA and HF > sod-podzolic soil HA approximately = peat HF > sod-podzolic soil FA > peat dissolved organic matter. The magnitude of the K(OC) values correlated strongly with the percentage of aromatic carbon in HS samples (r = 0.91). The hydrophobic binding was hypothesized as the key interaction underlying the binding of atrazine to HS.     

Impact of de-ashing humic Acid and humin on organic matter structural properties and sorption mechanisms of phenanthrene

Organic matter-mineral interactions greatly affect the fate of hydrophobic organic compounds (HOCs) in the environment. In the present study, the impact of organic matter-mineral interaction on sorption of phenanthrene (PHE) by the original and de-ashed humic acids (HAs) and humin (HM) was examined. After de-ashing treatment, the overall polarity of organic matter in HAs and HM consistently decreased. Differently, the surface polarity of HAs increased but that of HM decreased. No correlation between K(oc) values of PHE by all tested sorbents and their bulk polarity was observed due to inaccessibility of a portion of interior sorption domains. The inaccessibility of interior sorption domains in HAs and HM was partly due to the crystalline structure in organic matter as indicated by differential scanning calorimetric (DSC) and 13C NMR data and the interference from minerals. A good correlation between surface polarity of the original and de-ashed HAs and HMs and their K(oc) values for PHE indicated its importance in HOC sorption. Dissimilar changes in surface polarity of HAs and HM after de-ashing treatment can be ascribed to the distinct interactions between organic matter and minerals. The solid-state 13C NMR, XPS, and elemental composition data of all tested sorbents revealed that a larger fraction of O atoms in HAs were involved in organic matter-mineral interaction as compared to HM. Results of this work highlight the importance of soil organic matter (SOM)-mineral interactions, surface polarity, and microscaled domain arrangement of SOM in HOC sorption.     

Proton binding to humic acids from organic amendments and amended soils by the NICA-Donnan model

The acid-base properties of humic acids (HAs) are known to significantly affect the acid-base buffering capacity of soils, thus having a marked influence on the speciation of cations in the soil solid and liquid phases. Detailed information on the proton binding behavior of humic-like acids (HALs) from organic amendments and humic acids (HAs) from amended soils is, therefore, of intrinsic interest for the evaluation of the agronomic efficacy and environmental impact of soil amendment. In this work, the acid-base properties of HLAs isolated from sewage sludge (SS) and municipal solid waste compost (MSWC), and HAs isolated from soils amended with either SS or MSWC and the corresponding nonamended control soils were investigated by potentiometric titrations at various ionic strengths (0.01, 0.05, 0.1, and 0.3 M) over the pH range from 3.5 to 10.5. The nonideal competitive adsorption (NICA)-Donnan model that describes proton binding by two classes of binding sites with low and high proton affinity, i.e., carboxylic- and phenolic-type groups, was fit to titration data, and a set of fitting parameters was obtained for each HLA and HA sample. The NICA-Donnan model successfully described the shapes of the titration curves, and highlighted substantial differences in site density and proton-binding affinity between the HLAs and HAs examined. With respect to the nonamended control soil HAs, SS-HLA and MSWC-HLA were characterized by smaller carboxylic-type and phenolic-type group contents, larger affinities for proton binding by the carboxylic-type groups, and smaller affinities for proton binding by the phenolic-type groups. Amendment with SS and MSWC determined a number of modifications in soil HAs, including decrease of acidic functional group contents, slight increase of proton affinity of carboxylic-type groups, and slight decrease of the affinities for proton binding by phenolic-type groups. These effects were more evident in the HA fraction from the SS-amended soil than in the HA fraction from the MSWC-amended soil. Thus, both organic amendments examined can be a considered as a valuable source of organic matter for soil. However, MSWC appears to be an amendment of greater quality producing a smaller impact than SS on proton-binding behavior of soil HA.     

Infrared spectroscopic evidence supporting heterogeneous site binding models for humic substances

Infrared spectroscopy was used to corroborate predictions made by newly developed heterogeneous site binding models for humic substances. Experimental conditions to acquire the spectra of soil humic substances (humic and fulvic acid and a polysaccharide fraction) in an aqueous state using horizontal attenuated total reflectance Fourier transform infrared spectroscopy (HATR-FTIR) were established. Elimination of the water spectrum from that of the sample was achieved by spectral subtraction of the water peak at 2020 cm(-1). A KSCN internal standard with an absorption band at 2067 cm(-1) was used to verify the efficacy of the subtraction procedure. Spectral artifacts produced by the water spectrum subtraction and from contaminants within the humic materials have been identified. Three fulvic and one humic acid solution were examined in solutions of varying pH. Results show that the observed proportion of ionized carboxylate in relation to pH is consistent with models that assume electrostatic effects and a continuous distribution of proton association constants (log KH). The spectroscopic data were in accordance with calculations made using the generic humic and fulvic acid NICA-Donnan model parameters.     

Sorption of peat humic acids to multi-walled carbon nanotubes

Sorption of humic acids (HAs) from a peat soil by multiwalled carbon nanotubes (MWCNTs) was examined in this work. Sorption rate of HAs to MWCNTs was dominantly controlled by their diffusion from liquid-MWCNT boundary to MWCNT surfaces. Size exclusion chromatography analysis did not detect preferential sorption of HA fractions to MWCNTs at equilibrium, whereas the components with lower molecular weight in some HA fractions (e.g., HA1) would more preferentially be sorbed to MWCNTs at the initial sorption stage. Equilibrium sorption intensity of HAs by MWCNTs was dependent on their surface area and a sum of meso- and macropore volume. The surface area and sum of meso- and macroporosity-normalized sorption coefficient (K(d)) values of a given HA by MWCNTs increased with increasing outer diameter of MWCNTs, because MWCNTs with larger outer diameter were more strongly dispersed by HAs thereby making more sorption sites exposed for HA sorption. Van der Waals interaction between the alkyl components rather than the aromatic ones of HAs with MWCNTs was likely the key driving force for their sorption. This study highlights the sorption rate-controlling step of HAs from a same source to MWCNTs and the major factors affecting their sorption intensity at equilibrium.     

Quantitative evaluation of noncovalent interactions between glyphosate and dissolved humic substances by NMR spectroscopy

Interactions of glyphosate (N-phosphonomethylglycine) herbicide (GLY) with soluble fulvic acids (FAs) and humic acids (HAs) at pH 5.2 and 7 were studied by (1)H and (31)P NMR spectroscopy. Increasing concentrations of soluble humic matter determined broadening and chemical shift drifts of proton and phosphorus GLY signals, thereby indicating the occurrence of weak interactions between GLY and humic superstructures. Binding was larger for FAs and pH 5.2 than for HAs and pH 7, thus suggesting formation of hydrogen bonds between GLY carboxyl and phosphonate groups and protonated oxygen functions in humic matter. Changes in relaxation and correlation times of (1)H and (31)P signals and saturation transfer difference NMR experiments confirmed the noncovalent nature of GLY-humic interactions. Diffusion-ordered NMR spectra allowed calculation of the glyphosate fraction bound to humic superstructures and association constants (K(a)) and Gibbs free energies of transfer for GLY-humic complex formation at both pH values. These values showed that noncovalent interactions occurred most effectively with FAs and at pH 5.2. Our findings indicated that glyphosate may spontaneously and significantly bind to soluble humic matter by noncovalent interactions at slightly acidic pH and, thus, potentially pollute natural water bodies by moving through soil profiles in complexes with dissolved humus.     

Suspending multi-walled carbon nanotubes by humic acids from a peat soil

Suspension of the pristine and COOH-substituted multi-walled carbon nanotubes (P- and C-MWCNTs) with different outer diameters (ODs) by humic acids (HAs) from a peat soil was examined. Under shaking condition, MWCNTs were not suspended within 5 d. Without HAs, C-MWCNTs were slightly suspended by sonication within 16 h, but no suspension was observed for the pristine ones (P-MWCNTs). HAs greatly enhanced suspension of both P- and C-MWCNTs. The suspension enhancement was attributed to HA sorption, which increased electrostatic repulsion and steric hindrance between individual MWCNTs. Introduction of O-containing hydrophilic moieties to MWCNTs via HA sorption enhanced the interactions of their surfaces with water through H-bonding. Suspending capability of various MWCNTs on suspended mass concentration basis by four HAs showed inconsistent orders with the increasing or decreasing trend of their ODs. However, the suspended surface area concentrations of both P- and C-MWCNTs by individual HAs consistently followed an order of P8 > P30 > P50, and C8 > C30 > C50 (P and C, respectively, refer to P- and C-MWCNTs, and the numbers represent their ODs). These data implied that MWCNTs with smaller OD could be more strongly suspended by a given HA relative to those with larger OD under sonication condition.     

Birnessite-induced binding of phenolic monomers to soil humic substances and nature of the bound residues

The nature of the abiotic birnessite (δ-MnO(2))-catalyzed transformation products of phenolic compounds in the presence of soil organic matter is crucial for understanding the fate and stability of ubiquitous phenolic carbon in the environment. (14)C-radioactive and (13)C-stable-isotope tracers were used to study the mineralization and transformation by δ-MnO(2) of two typical humus and lignin phenolic monomers-catechol and p-coumaric acid-in the presence and absence of agricultural and forest soil humic acids (HAs) at pH 5-8. Mineralization decreased with increasing solution pH, and catechol was markedly more mineralized than p-coumaric acid. In the presence of HAs, the mineralization was strongly reduced, and considerable amounts of phenolic residues were bound to the HAs, independent of the solution pH. The HA-bound residues were homogeneously distributed within the humic molecules, and most still contained the unchanged aromatic ring as revealed by (13)C NMR analysis, indicating that the residues were probably bound via ester or ether bonds. The study provides important information on δ-MnO(2) stimulation of phenolic carbon binding to humic substances and the molecular distribution and chemical structure of the bound residues, which is essential for understanding the environmental fates of both naturally occurring and anthropogenic phenolic compounds.     

The identification of plant derived structures in humic materials using three-dimensional NMR spectroscopy

Here we demonstrate the application of 3-D NMR spectroscopy to structural studies of humic substances, the most abundant of organic compounds on earth. The increased spectral dispersion provided by the additional dimension is proven to be highly advantageous in separating the overlapping signals observed in 2-D spectra. Assignments of the major aliphatic structures and selected aromatic moieties are given as examples. We find that in a forest soil fulvic acid the major aliphatic materials are likely derived from leaf cuticles and further demonstrate that lignin signatures can be identified among the aromatic species. Once identified from the 3-D spectra, these structures can be assigned using the partial information available in 2-D, and in some cases, in the 1-D spectra. These signals are demonstrated to be characteristic to given samples of natural organic matter, and the case is made for their use as indicators of terrestrial biomarkers in mixtures of compounds with unknown origins.     

Molecular and quantitative analysis of metal ion binding to humic acids from sewage sludge and sludge-amended soils by fluorescence spectroscopy

Humic acids (HAs) isolated from sewage sludge (SS) and control and SS-amended soils were characterized by fluorescence spectroscopy. The main feature of fluorescence spectra was a broad band with the maximum centered at excitation/emission wavelengths that were much shorter for SS-HA (340/438 nm) than for any soil HA (440/510 nm). Titration with Cu(II), Zn(II), Cd(II), and Pb(II) ions decreased fluorescence intensities of HAs. Titration data were fitted to a single-site fluorescence quenching model, and metal ion complexing capacities of each HA sample and stability constants of metal ion-HA complexes were calculated. The binding capacities of HAs and strengths of metal ion-HA complexes followed the order Pb(II) > Cu(II) > Cd(II) > Zn(II). With respect to the control soil HA, SS-HA, which showed the smallest contents of acidic functional groups and the lowest humification degree, was characterized by much smaller binding capacities and stability constants. The binding capacities and affinities of SS-amended soil HA were intermediate between those of native soil HA and SS-HA but closer to the former, thus suggesting a partial incorporation of HA fractions of SS into native soil HAs. These effects are expected to have a great impact on the behavior of metals in SS-amended soils.     

Monitoring the effect of three humic acids on a model membrane system using 31P NMR

The sorption of three humic acids to 1,2-dipalmitoyl-sn-glycero-3-phosphocholine multilamellar vesicle model membrane systems was studied by phosphorus nuclear magnetic resonance (31P NMR). The effects of temperature and pH were investigated. The gel --> bilayer transition did not appear to be affected by any of the humic acids at pH 7; however, all three humic acids induced a perturbation to this transition and to the bilayer structure at pH 4. On the basis of the findings from this and other work, a conceptual adsorption/absorption model for the sorption of humic acid (HA) to biomembranes has been put forward. The model requires an initial adsorption step initiated at an acidic pH by hydrogen bridging and electrostatic interactions between the functional groups of the HAs and the head groups of the phospholipids. Once the HA material is adsorbed, its hydrophobic domains can further seek a more thermodynamically favorable environment within the bilayer using hydrophobic interactions. These interactions lead to the HA being absorbed into the membrane, which subsequently induces the observed perturbation by disturbing the ordered packing of the phospholipid tail groups. This model is also related to other humic substances/biomembrane observations in the literature.     

Identification of black carbon derived structures in a volcanic ash soil humic acid by Fourier transform ion cyclotron resonance mass spectrometry

Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS), coupled with cross-polarization magic angle spinning 13C nuclear magnetic resonance (NMR) spectroscopy and Kendrick mass defect analysis, was used to study the molecular composition of an aromatic carbon-rich humic acid extracted from a dark black soil from Iwata, Japan. Black carbon, produced by the incomplete combustion of fossil fuels and organic matter, has been suggested as a major component of humic acids having intense peaks in the aromatic and carboxyl regions of the 13C NMR spectrum. Taking advantage of the high resolving power of FT-ICR MS to make precise formula assignments, three different types of highly carboxylated polycyclic aromatic compounds were identified in the sample: linearly fused aromatic structures, aromatic structures linked by carbon-carbon single bonds, and highly condensed aromatic structures. These carboxylated aromatic structures have a low mass defect in their mass spectra due to their abundance of oxygen and deficiency of hydrogen. This mass defect is observed in the vast majority of peaks present in the entire mass spectrum, differentiating them from structures that are hydrogen-rich (e.g., fatty acids, proteins, carbohydrates). Thus, we conclude that the bulk of the sample analyzed is comprised of these heavily carboxylated, hydrogen-deficient, condensed aromatic structures, features believed to be characteristic of black carbon-like material.     

Aggregation and disaggregation of humic supramolecular assemblies by NMR diffusion ordered spectroscopy (DOSY-NMR)

Diffusion ordered nuclear magnetic resonance spectroscopy (DOSY-NMR) was applied to a number of fulvic (FA) and humic (HA) acids of different origin. Spectral separation achieved by DOSY based on diffusion coefficients (D), and correlated to molecular sizes by calibration standards, showed that carbohydrates had the largest molecular size in FA, whereas alkyl or aromatic components were the most slowly diffusing moieties in HA. At increasing concentrations, these components had invariably lower D values in DOSY spectra for all humic samples,thereby indicating an aggregation into apparently larger associations, whose increased hydrodynamic radius was confirmed by viscosity measurements. When humic solutions were broughtfrom alkaline to acidic pH (3.6), components diffusivity detected by DOSY increased significantly, suggesting a decrease of aggregation and molecular size. A general comparison of HA and FA molecular sizes was achieved by multivariate statistical analysis. While a larger extent of aggregation and disaggregation was observed for HA than for FA, no aggregation was detected, under similar conditions, for a true macropolymeric standard. Such difference in diffusion between a polymeric molecule and humic samples, is in line with the supramolecular nature of humic matter. The possible formation of humic micelles was also investigated by both changes of diffusivity in DOSY spectra and shift of 1H NMR signals. Except for HA of peat and soil origin, revealing a self-assembling in micelle-like structures at the 4 mg mL(-1) concentration, no other humic sample showed evidence of critical micelle concentration (cmc) up to 20 mg mL(-1). These results indicated that DOSY-NMR spectroscopy is a useful technique to evaluate components of different molecular size in natural humic superstructures.