Humic Substances Fractions Research Articles

Effect of Natural Organic Substances on the Surface and Adsorptive Properties of Environmental Black Carbon (Char): Pseudo Pore Blockage by Model Lipid Components and Its Implications for N2-Probed Surface Properties of Natural Sorbents

Black carbon (BC; char and soot) particles emitted to the environment typically are formed with high microporosity and surface area, properties that are responsible for their presumed important role in adsorption of anthropogenic organic compounds in soils and sediments. An issue that has received little direct attention is the possibility that naturally occurring organic matter attenuates the surface activity of BC. We found that simulated "aging" of prepared wood char particles in a soil-water suspension leads to a strong decline in char total surface area (TSA) by N2 adsorption at 77 K with BET analysis and a more modest decline in affinity for dissolved benzene. To help determine the underlying cause, we measured the effects of adsorbed natural lipids or lipid fractions of humic substances, modeled by triglycerides of a commercial vegetable oil. With increasing lipid loading (up to 40% by char weight) from aqueous mixtures, N2 TSA was strongly suppressed (up to 100-fold), while CO2 cumulative surface area (CSA, 0-1.4 nm) at 273 K and benzene adsorption at 293 K were hardly affected. In addition, the rate of CO2 adsorption was retarded. We propose that externally adsorbed lipid molecules occupy pore throats with access to interior pore networks. At 77 K, as opposed to the higher temperatures, lipid chains are too inflexible to allow passage of adsorbate. It is concluded that benzene adsorption to char predominates at interior pore sites and does not correlate with N2-probed micropore properties when the char accrues pore-blocking substances from the surroundings. The findings question the suitability of N2 for probing hydrophobic microporosity of BC in soils and sediments.

Reversed-phase liquid chromatographic characterization and analysis of air particulates humic (-like) substances in presence of pollens

Newly developed method for characterisation and analysis of humic substances (HS) and humic-like substances (HULIS) in air dust particles was tested for potential interferences caused by abundant co-sampled pollens (common dandelion, common wormwood-absinth, apple tree). RP-HPLC using 10-step gradient of dimethylformamide (DMF) in a buffered aqueous mobile phase and a wide-pore (30 nm) octadecylsilica column has been applied to the analysis of HS and HULIS using tandem of spectrophotometric (DAD) and fluorimetric detection (FLD). Achieved results suggest that the devised method is reliable for characterisation, fractionation and analysis of terrestrial HS, air dust HS and air particulate HULIS in liquid extracts at a trace concentration levels. Fluorimetric detection (ex. 470 nm/em. 530 nm; LOD, 3.1 μg/ml) enables sensitive, highly selective and interference free determination of HS and HULIS regardless the presence of pollen constituents, whereas spectrophotometric detection is susceptible to interferences in UV region above 260 nm. However, even in this case, the interfering substances can be revealed by both different pattern and shape of their peaks, as well as by spectral features different from HS or HULIS. Analytical procedure based on air sampling, extraction and above-mentioned HPLC method enables characterisation and analysis of HS and/or HULIS at relative concentration levels down to 0.1% (m/m) in 10 mg mass scale of sampled air dust and particulate material.

Influence of alkaline extraction on the characteristics of humic substances in Brazilian soils

An investigation was made of the influence of alkaline extraction on the characteristics of humic substances extracted from Brazilian soil samples. Humic substances (HS) from seven different soils samples collected in Brazil were extracted using the procedure recommended by the International Humic Substances Society (IHSS). Soils, HS and humins were characterized by thermogravimetry and differential thermal analysis. About 8 mg of each material (soil, HS and humin) were placed in a platinum crucible and heated continuously from 20 to 750 °C at a heating rate of 10 °C min −1 in an atmosphere of synthetic air (100 ml min −1). A thermal analysis revealed a difference between the content and structural characteristics of organic matter present in HS and humin fractions in relation to their soils. The results indicated that alkaline extraction alters the characteristics of humic substances during the extraction process, underlining the importance of developing methodologies and analytical procedures that allow organic matter in soils to be studied without extracting it.

Fractionation of Humic Substances by Asymmetrical Flow Field-Flow Fractionation

Asymmetrical flow field-flow fractionation was used to investigate the effects of pH and ionic strength of the buffer, and the binding of polyaromatic hydrocarbons (PAHs) on the particle sizes of humic substances (HSs). Particle sizes were greater when HSs were present in acidic medium with phosphate buffer than when they were present in alkaline medium or in acidic medium with acetate buffer. The association of PAH did not lead to increase sizes of HSs. Bivalent calcium ion had a greater effect on the size than did monovalent sodium ion. Particle sizes in river water and seawater were mostly of the same magnitude of the standards HSs. Some larger particles (30–70 nm) were also found.

Biodegradability of Fractions of Dissolved Organic Carbon Leached from Decomposing Leaf Litter

Dissolved organic matter leached from decomposing organic matter is important in the leaching of nutrients from the root zone of ecosystems, eluviation of metals, and transport of hydrophobic pollutants. The objective of this study was to compare microbial mineralization rates in intact soil cores of various fractions of water-soluble dissolved organic matter. Uniformly 14C-labeled Populus fremontii leaf litter that had decomposed for 1 year was extracted in water and this extract was fractionated into phenolic, humic acid, fulvic acid, hydrophilic acid, and hydrophilic neutral fractions. Fulvic acid comprised 42.1% of C in dissolved organic carbon (DOC) extracted from the litter. These fractions were added to intact cores of soil or sand, and respired 14CO2 was collected. The percentage of labeled substrate C mineralized in soil at the end of 1 year was, in order from least to greatest, hydrophilic acid (30.5), fulvic acid (33.8), humic acid (39.0), whole, unfractionated DOC (43.5), unseparated hydrophilic acid and neutral (44.7), phenolic (63.3), glucose (66.4), and hydrophilic neutral (70.2). In acid-washed nutrient-amended sand that was inoculated with soil microbes, mineralization rates of fulvic acid and glucose were lower. The fractionation appeared to separate the DOC into components with widely different rates of mineralization. Results also supported the ideas that the dissolved humic substance and hydrophilic acid fractions are inherently difficult for microbes to mineralize, and this property can contribute to movement of refractory C in soil and into aquatic ecosystems.

Solubility and dissociation of lignitic humic acids in water suspension

Behavior of humic acids (HAs) extracted from lignite in water suspension was studied by means of potentiometry. Although humic acids are operationally defined as that fraction of humic substances, which is not soluble under acidic conditions, it does not mean that solid preparation easily dissolves in water. Presented results suggest that dissolving of solid humic acids in water environment is more complex than conventional solubility behaviour of sparingly soluble solids. Multi-step mechanism of solid humic acids interaction with water, including partial equilibrium dissolution as well as direct equilibrium dissociation from the solid state, was proposed and proved experimentally. Only very small part of solid humic acids can be dissolved in water and not giving necessarily saturation at the same time. In contrast to common sparingly soluble solids, solid humic phase thus possesses non-constant chemical potential even under constant temperature and pressure and its activity does not equal to unity, in general.

Heavy metal distribution between fractions of humic substances in heavily polluted soils

Soil organic matter is important in controlling heavy metal behaviour in soils. This study aims to determine the distribution of organically bound Cd, Pb, and Zn between humic and fulvic acids in 20 samples of heavily polluted Fluvisols. Traditional procedure of alkaline sodium hydroxide/pyrophosphate extraction of humic substances with consequent humic acids precipitation by acidification was used. All of the three metals were bound predominantly on fulvic acids (mean share was 98.4, 82.0, and 95.7% of total organically bound Cd, Pb, and Zn content, respectively). Mainly the level of pollution controlled the amount of metals bound on fulvic acids. On humic acids, a limited amount of specific sorption sites for the metals is expected. Lead content bound on humic acids was negatively correlated with soil pH. The results suggest relatively high potential mobility of organic fractions of heavy metals in soils under this study. The selectivity of alkaline extracting agent is briefly discussed.

Modeling the adsorption and precipitation processes of Cu(II) on humin.

Humins (HU) are the most insoluble fraction of humic substances. Chemically, they can be considered as humic macromolecules bonded to the mineral matter of soil. The HU have a marked colloidal character and they are extremely important in retention of pollutants in soils. The aim of this work is to combine adsorption data with spectroscopic techniques in order to study the adsorption and precipitation processes of Cu(II) on HU. Analysis of sorption isotherms by means of several single-adsorption-process-based models makes it possible to obtain the speciation diagrams of Cu(II) species on HU surfaces. Further, FTIR (which provides information about the changes in the surface groups of the HU) and DRX (which shows the formation of possible crystalline phases on the HU surface) were used to determine the specific interactions of Cu(II) cations with the surface reactive groups of HU. The shape of the isotherms at constant pH varies with pH from L1-type (pH 2-4) to L3-type (pH 5-6) and S-type (pH 8), which indicates a change in the retention mechanism. When pH is 2 the retention of Cu(II), as [Cu(H(2)O)(6)](2+), is the preferred retention mechanism. The retained quantity of Cu(II) as [Cu(OH)(H(2)O)(5)](+) increases with pH. Starting from pH 4 the Cu(II) begins its precipitation, which is the preferred retention mechanism at pH 8. The presence of HU has a great influence on the precipitation process of Cu(II), giving rise to botalackite, which reveals epitaxial growth of crystals.

Effect of low molecular size humic substances on nitrate uptake and expression of genes involved in nitrate transport in maize (Zea mays L.).

In this study, a detailed characterization of earthworm low molecular size humic substances (LMS) was performed and these substances were used to study their effect on the nitrate influx in roots, tissue nitrate content, and expression of maize genes putatively involved in nitrate uptake in maize (Zea mays L.). The results show that the humic fraction with low molecular size used in this study is endowed with the characteristic structural network described for most humic substances so far isolated and confirm the presence of IAA in this fraction. The results also show that the LMS fraction of humic substances stimulates the uptake of nitrate by roots and the accumulation of the anion at the leaf level. Moreover, the analysis of the expression of genes encoding two putative maize nitrate transporters (ZmNrt2.1 and ZmNrt1.1) and of two maize H(+)-ATPase isoforms (Mha1 and Mha2) show that these substances may exert direct effects on gene transcription in roots, as shown for the Mha2 gene, and long-distance effects in shoots, as observed for the ZmNrt2.1 gene.

Molecular Weight Fractions of Soil and Peat Humic Substances Analyzed by Isoelectric Focusing

Isoelectric focusing (IEF) has been used to study humic and humiclike substances. As a general interpretation, these substances are fractionated by IEF depending on their molecular weight. However, the nature of humic-substance fractionation is not certain when an IEF run is applied over them. Even if analysis and extraction methods have changed over the years, the above-mentioned interpretation is still the same. In order to check the actuality of the current interpretation, humic substances were separated to different molecular weight cutoff fractions and isoelectric focusing was applied to them. Results revealed that the dependence of IEF profiles on molecular weight is not a generally valid hypothesis, even if the lowest fraction (1,000–10,000 Da) of molecular weight showed that the smallest amount of material focused on the cathode side in over 50% of the samples studied.

Molecular Weight Fractions of Soil and Peat Humic Substances Analyzed by Isoelectric Focusing

Abstract Isoelectric focusing (IEF) has been used to study humic and humiclike substances. As a general interpretation, these substances are fractionated by IEF depending on their molecular weight. However, the nature of humic-substance fractionation is not certain when an IEF run is applied over them. Even if analysis and extraction methods have changed over the years, the above-mentioned interpretation is still the same. In order to check the actuality of the current interpretation, humic substances were separated to different molecular weight cutoff fractions and isoelectric focusing was applied to them. Results revealed that the dependence of IEF profiles on molecular weight is not a generally valid hypothesis, even if the lowest fraction (1,000–10,000 Da) of molecular weight showed that the smallest amount of material focused on the cathode side in over 50% of the samples studied.

Changes in Humic Substances and Phosphorus Fractions During Composting

The behavior of humic substances (HS) and phosphorus (P) fractions in livestock manure co-composted with rice straw was studied by chemical and spectroscopic analyses. Composting was accompanied by a decrease in the levels of carbon (C) and nitrogen (N) as well as HS and fulvic acid (FA) fractions. The elemental content (weight %) of humic acid (HA) extracted from the compost showed a high C content and low oxygen (O) content, possibly due to a low degree of oxidation. The N/C and H/C ratios of HA decreased as the compost matured, whereas the O/H and O/C increased. The decline in the H/C ratio from 1.24 at 0d to 1.0 at 195d indicated an increase in the aromatic structure of the HA. The absorbance curve for the HS showed a decrease in the slope with time, up to 42d. The optical density of HS increased with time of composting with a peak at 42d. Also the Δ log K values of FA increased appreciably with time of composting whereas those of HS and HA were little changed. Water-soluble and HCl-soluble-P were the dominant fractions of P in the compost, and ranged between 18 and 39% and 18 and 36%, respectively. However, the former declined progressively with time of composting, while the latter increased, indicating transformation of the more vulnerable water soluble P to the more recalcitrant HCl-extractable P.

Reduction of mercury(II) by tropical river humic substances (Rio Negro)—Part II. Influence of structural features (molecular size, aromaticity, phenolic groups, organically bound sulfur)

The influence of structural features of tropical river humic substances (HS) on their capability to reduce mercury(II) in aqueous solutions was studied. The HS investigated were conventionally isolated from Rio Negro water-Amazonas State/Brazil by means of the collector XAD 8. In addition, the isolated HS were on-line fractionated by tangential-flow multistage ultrafiltration (nominal molecular-weight cut-offs: 100, 50, 30, 10, 5 kDa) and characterized by potentiometry and UV/VIS spectroscopy. The reduction of Hg(II) ions to elemental Hg by size-fractions of Rio Negro HS was assessed by cold-vapor AAS (CVAAS). UV/VIS spectrometry revealed that the fractions of high molecular-size (F 1>100 kDa and F 2: 50–100 kDa) have a higher aromaticity compared to the fractions of small molecular-size (F 5: 5–10 kDa, F 6: <5 kDa). In contrast, the potentiometric study showed different concentration of functional groups in the studied HS fractions. The reduction of Hg(II) by aquatic HS fractions at pH 5 proceeded in two steps (I, II) of slow first order kinetics ( t 1/2 of I: 160 min, t 1/2 of II: 300 min) weakly influenced by the molecular-size, in contrast to the differing degree of Hg(II) reduction (F 5>F 2>>F 1>F 3>F 4>>F 6). Accordingly, Hg(II) ions were preferably reduced by HS molecules having a relatively high ratio of phenolic/carboxylic groups and a small concentration of sulfur. From these results a complex ‘competition’ between reduction and complexation of mercury(II) by aquatic HS occurring in tropical rivers such as the Rio Negro can be suggested.

Formation and Loss of Humic Substances During Decomposition in a Pine Forest Floor

Since twice as much C is sequestered in soils as is contained in the atmosphere, the factors controlling the decomposition rate of soil C are important to the assessment of the effects of climatic change. The formation of chemically resistant humic substances might be an important process controlling recycling of CO 2 to the atmosphere. Our objectives were to measure the rate of formation and loss of humic substances during 13 yr of litter decomposition. We placed nets on the floor of a white pine (Pinus strobus) forest to separate each annual layer of litter for 13 yr and measured humic substance concentration using NaOH extraction followed by chromatographic fractionation. The humic acid fraction increased from 2.1% of the C in litterfall to 15.7% after 1 yr. On a grams per square meter (g m -2 ) basis the humic substance fraction increased during the first year and then declined, with a half decay time (t 1/2 ) of 5.1 yr, which was significantly slower than the bulk litter (t 1/2 = 3.9 yr). The carboxylic C concentration estimated from 13 C nuclear magnetic resonance (NMR) increased in the litter over time, though total mass of carboxylic acid C in the forest floor also declined over the 13-yr period (t 1/2 = 4.6 yr). While humic substances in the forest floor decomposed at a somewhat slower rate than bulk litter during Years 1 to 13, they decomposed much faster than has been calculated from 14 C dating of the refractory fraction of organic matter in the mineral soil.

Dissolved organic matter: artefacts, definitions, and functions

In this concept paper, some basic problems in the obtaining of the dissolved organic matter (DOM) are described. Some of them are fairly obvious such as adsorption and desorption on the filters used to isolate DOM. However, some are subtler such as alteration in the tertiary physicochemical structure of DOM because of cavitation (filtration), concentration (ultrafiltration, reverse osmosis), or decomposition (electrophoresis). These structure changes may not have an influence on a variety of analytical techniques, but they may alter the ability of DOM to interact with certain ecosystem components such as the biota and pollutants. Therefore, the interpretation of the analyses done with such DOM cannot necessarily be extrapolated to in situ conditions. Fluorescence has been found to be a sensitive tool to follow such tertiary structure changes. In structured matrices with pore spaces such as soils and sediments, DOM may have different fates and functions depending on the pore spaces in which it is located. This again needs to be considered in sampling strategies. For example, DOM in macropores may play important roles in groundwater quality but presumably does not play a major role in the in situ production of the greenhouse gas N 2O via denitrification, since this pore space tends to be well aerated. A protocol for the separate sampling of the different pore spaces is presented. Another difficulty, which is shown here, is that minor changes in extraction methodology (such as the extractant–soil ratio) can have a serious effect on the apparent amount of DOM. DOM samples from a field can have considerable temporal and spatial variability, which is complicated by the fact that DOM from unsaturated systems can be strongly influenced by ambient water content. This again needs to be considered, especially when fields from different regions are sampled and compared. Another important point is that the results obtained from one DOM source cannot necessarily be extrapolated to the DOM from another source. For example, soil pore water DOM differs both from aquatic DOM and from the material, which can be extracted as the humic substance fraction. Finally, with the increasing awareness of the ecological functions of DOM, which are briefly presented here, there is also an increase in DOM research being done in different disciplines. This is laudable, but presentations of results need to define DOM more precisely to prevent misunderstandings.

Formation and Loss of Humic Substances During Decomposition in a Pine Forest Floor

Since twice as much C is sequestered in soils as is contained in the atmosphere, the factors controlling the decomposition rate of soil C are important to the assessment of the effects of climatic change. The formation of chemically resistant humic substances might be an important process controlling recycling of CO2 to the atmosphere. Our objectives were to measure the rate of formation and loss of humic substances during 13 yr of litter decomposition. We placed nets on the floor of a white pine (Pinus strobus) forest to separate each annual layer of litter for 13 yr and measured humic substance concentration using NaOH extraction followed by chromatographic fractionation. The humic acid fraction increased from 2.1% of the C in litterfall to 15.7% after 1 yr. On a grams per square meter (g m−2) basis the humic substance fraction increased during the first year and then declined, with a half decay time (t1/2) of 5.1 yr, which was significantly slower than the bulk litter (t1/2 = 3.9 yr). The carboxylic C concentration estimated from 13C nuclear magnetic resonance (NMR) increased in the litter over time, though total mass of carboxylic acid C in the forest floor also declined over the 13-yr period (t1/2 = 4.6 yr). While humic substances in the forest floor decomposed at a somewhat slower rate than bulk litter during Years 1 to 13, they decomposed much faster than has been calculated from 14C dating of the refractory fraction of organic matter in the mineral soil.

Effect of Humic Substances on Plutonium and Americium Speciation in Soils and Soil Solutions

The molecular-weight distribution of humic substances (HSs) of the organic and mineral soils and corresponding soil solutions was studied by gel chromatography, and their distribution over fractions of humus acids was evaluated by the Ponomareva-Plotnikova chemical fractionation technique. Distribution of 239,240Pu and 241Am over HS fractions of different dispersity was studied. In these fractions, HSs are associated with different mineral components and have different solubility and mobility. Plutonium and americium are more strongly associated with humic substances of organic soils as compared to those of mineral soils. Therefore, in passing from mineral to organic soils, the migration ability of radionuclides decreases. These effects are caused by qualitative difference in the HS chemical composition of the HSs of organic and mineral soils. As compared to plutonium, americium enters the composition of poorly soluble organomineral complexes to a smaller extent, forming to a greater extent mobile anionic complexes in the soil solutions. This difference can be responsible for enhanced americium mobility and its easier availability to plants.

Effects of humic substances on the pattern of oxidation products of pentachlorophenol induced by a biomimetic catalytic system using tetra(p-sulfophenyl)porphineiron(III) and KHSO5.

In the presence of humic substances (HSs), the oxidative conversion of pentachlorophenol (PCP) was found to be efficiently catalyzed by tetra(p-sulfophenyl)porphineiron(III) (Fe(III)-TPPS) using KHSO5 as an oxygen donor. Ortho-tetrachloroquinone (o-TeCQ), 2-hydroxyl-nonachlorodiphenyl ether (2H-NCDE), 4-hydroxyl-nonachlorodiphenyl ether (4H-NCDE), and octachlorodibenzo-p-dioxin (OCDD) were identified as the major byproducts of the reaction. Decreased amounts of these byproducts were produced in the presence of HS. In particular, the addition of HSs with a lower degree of humification resulted in a large decrease in the formation of dimers, such as 2H-NCDE, 4H-NCDE, and OCDD. More than 60% of the chlorine, which was released from PCP, was found in the HS fractions after the reaction. This suggests that chlorinated intermediates from PCP were incorporated into the HS. Pyrolysis-GC/MS and 13C NMR studies confirmed that the binding of the chlorinated intermediates was covalent in nature and that the intermediates were copolymerized with HS via oxidative coupling reactions. A Microtox test demonstrated that the toxicity of the HS fraction containing PCP-derived intermediates was much lower than that of the mixture of PCP and HS in the absence of a catalytic reaction.

Ozonation of Han River humic substances

This study investigated the characteristics of extracted humic substances (HS) which are ubiquitous in river waters and affect the treatment of the water. HS were extracted from the Han River and the samples obtained were analyzed before/after oxidation with the following analytical methods: DOC, THMFP, UV-abs, chemical composition, molecular weight distribution, IR. Ozonation experiments were conducted in batch-type reactors. Ozone dose was 1 mg/L and contact time was varied from 0 to 20 minutes. The fraction of hydrophobic matter was 56.0% of total organic carbon in the Han River and the ratio of humic/fulvic acids was 22.4/77.6, showing high fractions of aromatic and/or double carbon bond functional groups and high fractions of saturated aliphatic carbons. Oxygen fraction of Han HS was relatively high compared with other humic substances (Suwannee River humic and fulvic acids), which are consistent with the results of IR spectra. The IR spectra showed the increase in aliphatic groups by ozonation of the Han HS. Humic substances could not completely be oxidized to CO2, H2O etc., but were partly oxidized to low molecular weight matter (aliphatics, aldehydes, carboxylics, polysaccharides etc.), so low removal efficiencies in terms of DOC (31-35% with 5 minute contact) were observed. For UV254, 5 minute or shorter contact showed more than 80% removal efficiencies.

SEC-ICP-MS studies for elements binding to different molecular weight fractions of humic substances in compost extract obtained from urban solid waste.

In this work, the speciation of elements in compost was studied with emphasis on their binding to humic substances. In order to assess the distribution of As, Cd, Co, Cr, Cu, Mn, Mo, Ni, Pb, U, Th and Zn among molecular weight fractions of humic substances, the compost extract (extracted by 0.1 mol l(-1) sodium pyrophosphate) was analyzed by size exclusion chromatography coupled on-line with UV-Vis spectrophotometric and ICP-MS detection. Similar chromatograms were obtained for standard humic acid (Fluka) and for compost extract (254 nm, 400 nm) and three size fractions were operationally defined that corresponded to the apparent molecular weight ranges > 15 kDa, 1-15 kDa and < 1 kDa. The percentage of total element content in compost that was leached to the extract ranged from 30% up to 100% for different elements. The elution profiles of Co, Cr, Cu, Ni and Pb (ICP-MS) followed that of humic substances, while for other elements the bulk elution peak matched the retention time observed for the element in the absence of compost extract. Spiking experiments were carried out to confirm elements' binding and to estimate the affinity of individual elements for humic substances derived from compost. The results obtained indicated the following order of decreasing affinity: Cu > Ni > Co > Pb > Cd > (Cr, U, Th) >> (As, Mn, Mo, Zn). After standard addition, further binding of Cu, Ni and Co with the two molecular weight fractions of humic substances was observed, indicating that humic substances derived from compost were not saturated with these elements.