International Humic Substances Society Research Articles

Role of the molecular structure of humified organic matter in rice plant response to environmental lead pollution.

Humified organic matter has been shown to decrease Pb toxicity in plants. However, there are still gaps in our understanding of the mechanism by which this phenomenon occurs. In this study, we aimed to assess the ability of humic substances (HSs), humic acids (HAs), and fulvic acids (FAs) to enhance defense mechanisms in rice plants under lead (Pb)-stressed conditions. HS fractions were isolated from vermicompost using the chemical fractionation methodology established by the International Humic Substances Society. These fractions were characterized by solid-state NMR and FTIR. Chemometric analysis was used to compare humic structures and correlate them with bioactivity. Three treatments were tested to evaluate the protective effect of humic fractions on rice plants. The first experiment involved the application of humic fractions along with Pb. The second comprised pretreatment with humic fractions followed by subsequent exposure to Pb stress. The third experiment involved Pb stress and subsequent treatment with humic fractions. The root morphology and components of the antioxidative defense system were evaluated and quantified. The results showed that HS + Pb, HA + Pb, and FA + Pb treatment preserved root growth and reduced the levels of O2- and malondialdehyde (MDA) in the roots by up to 5% and 2%, respectively. Pretreatment of the plants with humic fractions promoted the maintenance of root growth and reduced the contents of O2-, H2O2, and MDA by up to 48%, 22%, and 20%, respectively. Combined application of humic fractions and Pb reduced the Pb content in plant tissues by up to 60%, while pretreatment reduced it by up to 80%. The protective capacity of humic fractions is related to the presence of peptides, lignin, and carbohydrate fragments in their molecular structures. These results suggest that products could be developed that can mitigate the adverse effects of heavy metals on agricultural crops.

Metal Binding Studies of Soil Extracted Humic Acid with K (I), Zn (II) and Fe (III) ions

: Physico chemical parameters of soil samples collected from agriculture lands of Taluka Sobhodaro District Khairpur, Sindh, Pakistan were examined to check the quality of soil. Moisture contents percentage, pH and EC values indicated that texture of soils is mostly silt- loam type having water holding ability hence can be considered as good fertile land, beneficial for crops. Humic acids (HAs) were extracted from soil samples by International Humic Substance Society(IHSS) method. HAs form biogeochemical cycle due to presence of electron clouds resultant macro and micro nutrients, transferred to the plants. HAs obtained were characterized by UV-Visible spectroscopy. The optical parameter E4/E6 values illustrated that HAs possess high molecular condensation and proved to be hydrophilic and aromatic in nature. To analyze the binding ability with metal ions, isolated HAs were complexed with potassium, zinc and ferric ions under optimized conditions. The concentrations of selected metal ions were detected in complexed HAs samples by Atomic Absorption Spectrophotometer (AAS). The data obtained indicated that the maximum concentration of complexation of metal ions was found for K1+at pH 8, Zn2+7 and Fe3+6. The data was correlated and compared with literature.

Adsorption of Congo Red onto Humic Acid Isolated from Peat Soil Gambut Regency, South Kalimantan

Humic acid is one of the green materials for wastewater treatment including for removal dyes as an adsorbent. Humic acid was isolated from peat soil, Gambut Regency, South Kalimantan following International Humic Substances Society method, and used to adsorb Congo red. The adsorption process was carried out in a batch system and the effect of pH, contact time, and adsorption capacity of Congo red on humic acid were studied. The adsorbent characterization by using FTIR. The results showed that the adsorption of Congo red on humic acid occurred at the optimum pH of pH 6, the optimum contact time was 120 minutes. The adsorption capacity of Congo red onto humic acid of 33.33 mg/g and follows the Langmuir model with R2 = 0.9926. The characterization of humic acid functional groups before and after adsorption of Congo red showed that the signal at 1,712.79 cm-1 were shifted to 1,705.07 cm-1 and 1,273.02 cm-1 were shifted to 1,265.30 cm-1. These suggested that the mechanism interaction was the electrostatic interaction between -NH3+ functional group of Congo red group and -COO- functional group of humic acid. It means that humic acid isolated from peat soil could be used as an adsorbent for the removal Congo red.

Diffusion of sulphonamide antibiotics in agarose hydrogels enriched by humic acids

Many different pharmaceuticals can evince potentially dangerous behaviour in soil and aqueous environments. Their migration ability and toxicity can be significantly affected by interactions with organic matter. Sulfamethoxazole and sulphapyridine (used as representative commonly used sulphonamide antibiotics) are studied from the point of view of their transport in model hydrogels containing humic substances. Agarose hydrogel was used as a model medium for the transport experiments. Standard Elliot soil humic acid (4S102H) purchased from International Humic Substances Society was incorporated into the hydrogel as an active substance (and representant of organic matter) for interactions with antibiotics and potential suppression of their migration and toxicity in soil systems. Two different experimental arrangements were used: the diffusion from constant source, and the diffusion couple. Diffusion coefficients were determined for pure agarose hydrogels as well as for hydrogels containing humic acids and compared. It was found that the addition of humic acids into hydrogel resulted in the decrease in the effective diffusion coefficient of sulphapyridine and the increase in the effective diffusion coefficient of sulfamethoxazole. Humic acids can suppress the mobility of sulphapyridine and enhance the diffusion of sulfamethoxazole. Batch sorption experiments provided similar adsorption capacities of humic acids for both studied antibiotics. The ratio between adsorption and desorption rate constants were higher than 1 for sulphapyridine and lower than 1 for sulfamethoxazole. It indicated that adsorption predominated for sulphapyridine which resulted in observed lower diffusivity. Desorption experiments showed that a majority of sulphapyridine can be immobilized by interaction with humic acids whereas much more free movable sulfamethoxazole was determined in the comparison with immobilized one.

Chlorpyrifos fate in the Arctic: Importance of analyte structure in interactions with Arctic dissolved organic matter

The insecticide and current use pesticide chlorpyrifos (CLP) is transported via global distillation to the Arctic where it may pose a threat to this ecosystem. CLP is readily detected in Arctic environmental compartments, but current research has not studied its partitioning between water and dissolved organic matter (DOM) nor the role of photochemistry in CLP's fate in aquatic systems. Here, the partition coefficients of CLP were quantified with various types of DOM isolated from the Arctic and an International Humic Substances Society (IHSS) reference material Suwannee River natural organic matter (SRNOM). While CLP readily partitions to DOM, CLP exhibits a significantly higher binding constant with Arctic lacustrine DOM relative to fluvial DOM or SRNOM. The experimental partitioning coefficients (KDOC) were compared to a calculated value estimated using poly parameter linear free energy relationship (pp-LFER) and was found to be in good agreement with SRNOM, but none of the Arctic DOMs. We found that Arctic KDOC values decrease with increasing SUVA254, but no correlations were observed for the other DOM compositional parameters. DOM also mediates the photodegradation of CLP, with stark differences in photo-kinetics using Arctic DOM isolated over time and space. This work highlights the chemo-diversity of Arctic DOM relative to IHSS reference materials and highlights the need for in-depth characterization of DOM that transcends the current paradigm based upon terrestrial and microbial precursors.

Exploring the Macroscopic Properties of Humic Substances Using Modeling and Molecular Simulations

Soil organic matter (SOM) is composed of a complex and heterogeneous mixture of organic compounds, which poses a challenge in understanding it on an atomistic level. Based on the progress of molecular dynamics simulations and our efforts to create molecular systems that resemble SOM, in this work, we expanded our knowledge of SOM through the use of humic substances (HSs). Specifically, we studied the standardized samples of HS of the International Humic Substances Society (IHSS). This society provided the elemental and organic composition used as input parameters for our Vienna Soil Organic Matter Modeler 2 (VSOMM2). We modeled and simulated different HS samples from various sources, including soil, peat, leonardite, and blackwater river. In order to compare between different HS, we reduced the organic composition information to two principal components, which are associated principally with the amount of carboxyl and aromatic groups in the HS, denominated as PCacid and PCarom, respectively. We performed a plethora of analyses to characterize the structure and dynamics of the systems, including the total potential energy, density, diffusion, preferential solvation, hydrogen bonds, and salt bridges. In general terms, at the water content value of 0.2, we observed that most properties depend on the carboxyl group protonation state. The Coulombic interactions from this ionic specie and the interaction with cations determine the overall behavior of the studied systems. Furthermore, the type of cations and the pH influence those properties. This study exemplifies the importance of molecular dynamics to explain macroscopic properties from the structure and dynamics of the molecules modeled, such as the interaction network, i.e., hydrogen bonds or salt bridges of molecules presented in the system and their mobility.

Comparisons in molecular weight distributions and size-dependent optical properties among model and reference natural dissolved organic matter.

Humic acid (HA) and reference natural organic matter (NOM) have been widely used in environmental assessment, biogeochemistry, and ecotoxicity studies. Nevertheless, similarities and differences among the commonly used model/reference NOMs and bulk dissolved organic matter (DOM) have rarely been systematically evaluated. In this study, HA, SNOM (Suwannee River NOM) and MNOM (Mississippi River NOM), both from International Humic Substances Society, and freshly collected unfractionated NOM (FNOM) were concurrently characterized to evaluate their heterogeneous nature and size-dependent chemical properties. We found that molecular weight distributions, PARAFAC-derived fluorescent components, and size-dependent optical properties are NOM-specific and highly variable with pH. The < 1kDa DOM abundance followed the order of HA < SNOM < MNOM < FNOM. In addition, FNOM was more hydrophilic and contained more protein-like and autochthonous components with a higher UV-absorbance ratio index (URI) and biological fluorescenceindex, whereas HA and SNOM contained more allochthonous, humic-like components with a higher aromaticity and lower URI. Significant differences in molecular composition and size spectra between FNOM and model/reference NOMs suggest that environmental role of NOMs should be evaluated at the levels of molecular weight and functionalities under the same experimental conditions and that HA and SNOM may not represent bulkNOM in the environment. This study provides new information about similarities and differences in DOM size-spectra and chemical properties between reference NOMs and in-situ NOM and highlights the need to better understand the heterogenous roles of NOMs in regulating the toxicity/bioavailability and environmental fate of pollutants in aquatic environments.

Lead Ion Sorption Capacity and the Structural Properties of the Humic Acid Extracts from Evergreen Forests of Different Agro-ecological Zones of Kerala Western Ghats

Forest soils are enriched with soil carbon content. The vegetation and precipitation influences it’s chemical structure. The chemical structure directly contributed to its entire properties. Physico-chemical properties play vital role in its soil functioning. The sorption capacity of the humic acid can be applied in the nutrient uptake enhancement and waste water treatment. The study was conducted in the humic acid extract of evergreen forest of different agro-ecological zones of Kerala. Extraction was done by the acidic precipitation method recommended by International Humic Substance Society (IHSS). Determine the chemical structural variations and properties of the humic acid extracts followed by determining the sorption capacity of the material with lead ion. The study revealed the higher percentage of adsorption of lead ion with humic acid extract from evergreen forest of northern high hills –charnockite zone of western ghats of Kerala. This was the combined effect of enhanced cation exchange capacity and –COOH acidity of the material. These chemical structure and the functional properties to the sorption capacity of the humic acid extract to chelate the lead ion was evaluated .The higher cation exchange capacity and the –COOH acidity favoured the lead ion adsorption. Adsorption isotherm modeling was revealed the multilayer and monolayer adsorption of lead ion on humic acid depends with the heterogeneity of the HA surfaces. Maximum amount of lead ion molecule adsorbed on the HA surface was observed for humic acid from SHH-K agro-ecological zone. This method can be considered as economically viable green method for looking forward technology for waste water treatment.

Humic Acid Extracted from Danty via Catalytic Oxidation Using H2O2/Birnessite: Characteristics and Agricultural Beneficial Effects.

Extraction optimization is very important for the quality of humic acid (HA). In this study, actived HA (HAb) was extracted from danty via catalytic oxidation using birnessite as a catalyst and H2O2 as an oxidant. Single-factor experiments and the response surface method were used to optimize the acidic functional group content of HAb. It was found that the maximum acidic functional group content of HAb can be achieved when danty-crushing time, H2O2 concentration, and birnessite dose were 105.7 min, 20, and 2%, respectively. Fourier transform infrared spectra showed that HAb had more surface functional groups than commercial HA (HAc) and HA extracted using the traditional method of the International Humic Substances Society (HAI). In addition, acidic functional group titration showed that HAb had 84.3% more acidic functional groups and 118.9% more carboxyl groups than HAI. Additionally, HAb had the greatest effect on promoting the dissolution of carbonate and bicarbonate, promoting the settlement of calcaline alkaline soil, and improving the germination rate of wheat seeds under saline and alkaline stress. This study provides a basis for the efficient extraction of active HA with rich functional groups and its application in agriculture and many other fields.

Humic Substances—Common Carriers of Micropollutants in Municipal Engineering

Research over several years has shown that municipal engineering is an important source of humic substance formation (HS). Effluents from municipal landfills and digested sludge, due to high concentrations of organic pollutants, including HS and inorganic pollutants, are discharged to a biological treatment plant, where their largely non-biodegradable pollutants constitute a burden on the activated sludge and generate problems related to wastewater treatment. To demonstrate that humic substances, primarily soluble acids, are the carriers of micro-pollutants, their isolation was carried out from samples taken at different municipal management sites. The following analyses were performed: elemental composition, and the content of inorganic micropollutants, including heavy metals. In addition, the sorption affinity of pure HS to selected pharmaceuticals was investigated. The sorbed pharmaceuticals were Diclofenac and Estrone. HS commonly found in municipal engineering have been shown to be carriers of inorganic and organic micropollutants, including pharmaceuticals. Based on the study data, surface water’s primary source of micropollutants is treated municipal wastewater and fulvic acids are the transfer agent. To demonstrate the locations of HS occurrence within municipal engineering and the micropollutants they carry, their isolation was carried out from samples taken at different municipal management sites. Fulvic acids were studied due to their high mobility. Extraction was carried out according to the method recommended by the International Humic Substances Society (IHSS) with the author’s modification. The following analyses were performed: elemental composition (N, C, O, and H); the content of inorganic micropollutants, including heavy metals; and the IR spectrum. In addition, the sorption affinity of pure HS to selected pharmaceuticals was investigated. The sorbed pharmaceuticals were Diclofenac and Estrone. The study confirmed the occurrence of HS in municipal landfill effluents, activated sludge, excess and digested sludge effluents, and treated wastewater. At the same time, HS commonly found in municipal engineering have been shown to be carriers of inorganic and organic micropollutants, including pharmaceuticals.

Screening of Humic Substances Extracted from Leonardite for Free Radical Scavenging Activity Using DPPH Method.

Humic substances (HSs) have been researched for a long time and still manage to surprise humanity today. According to the latest research, in addition to their previously well-known effects, they also have antioxidant properties. However, this previous research does not examine the difference in the antioxidant effect of the fractions extracted/produced in different processes; they do not consider the light absorption of the HSs, which falsifies analysis based on the measurement of color change over time. In the present work, HS fractions were obtained from leonardite, the extraction processes can also be implemented on an industrial scale. The fractions were characterized by elementary analysis, UV–Vis and FT-IR spectroscopies, to prove that our self-extracted samples have similar characteristics to the International Humic Substances Society (IHSS) standard samples. The different methods of HS fractionation affected the elemental composition, and the spectral characteristics. The antioxidant effect was investigated using the DPPH method to screen the antioxidant efficiency of humic, fulvic, and himatomelanic acids. In addition, we compared our results with the IHSS standard samples to obtain a more comprehensive picture of the antioxidant effect of HSs extracted in different ways according to the DPPH method. Based on our results, the extraction method affects not only the physico-chemical properties but also the free radical scavenging activity of the fractions.

Humic Acids Formation during Compositing of Plant Remnants in Presence of Calcium Carbonate and Biochar

The investigation of the mechanisms organic matter transformation in compost organic fertilizers is an urgent task of modern soil ecology and soil chemistry. The main components of such fertilizers are newly formed, weakly humified labile humic acids (HAs). The objective of the study is to determine the mechanism of converting the newly formed HAs into the forms with increased resistance to microbiological and biochemical influences. Obtained during the plant residues decomposition, HAs were studied in the incubation experiment (0, 30, 90 days). Calcium carbonate and biochar produced by rapid pyrolysis from birch and aspen wood, at 550 °C, were used as the composting mixture compounds. Decomposed plant residues—fresh aboveground mass of clover (Trifolium pratense L.), rye (Secale cereale L.), as well as dry oat straw (Avena sativa L.) were the material used for humification. To obtain Has, 0.1 M NaOH and 0.1 M Na4P2O7 were used. Then, HAs were separated from fulvic acids (FAs) using a 0.5 M H2SO4. The amount of labile HAs (HAs1) was estimated by their content in 0.1 M NaOH. The amount of stabilized HAs (HAs2) was calculated by the difference between the HAs content in 0.1 M Na4P2O7 and 0.1 M NaOH. Preparation of HAs for elemental composition and NMR analysis was performed according to the International Humic Substances Society’s recommendations. The possibility of converting newly formed HAs into stable forms (calcium humates), whose share in the HAs composition reaches 40–50%, has been shown. However, the mechanism of HAs transformation under the studied reagent’s influence was different. In the presence of calcium carbonate, it is caused by the physicochemical processes of newly formed HAs rearrangement. However, in the presence of biochar, this is due to the humification processes’ intensification and to the increase in the aromatization degree confirmed by the increase in the optical density, as well as by the increase in carbon and oxygen proportion, and by the decrease in hydrogen proportion in HAs molecules. The understanding of HAs formation and transformation mechanisms at the early humification stages can help to optimize the methods of obtaining organic fertilizers.

Deciphering the Composition of Fulvic Acids from Recirculating Aquaculture Systems by Quadrupole Time-of-Flight Mass Spectrometry

The constant reuse of waters in recirculating aquaculture systems (RAS) together with the continuous addition of organic matter from fish degradation components, faeces and non-consumed feed promote the accumulation of dissolved organic matter (DOM), particularly of fulvic acids (FA), leading to the yellow discoloration of their waters. The accumulation of these acids in RAS can have potential effects on its water quality, and consequently fish health and welfare. Thus, the characterization of FA is paramount for improving water quality, and subsequently fish productivity in RAS. In this study, a non-targeted analysis by quadrupole time-of-flight mass spectrometry (QTOF-MS) was used to characterize the recirculating aquaculture systems fulvic acids (RASFA) and then compare their molecular fingerprints with actual reference standards of fulvic acids (FA) (Suwannee River; SRFA, Elliott soil; ESFA and Pahokee Peat; PPFA) purchased from the International Humic Substance Society (IHSS). The results of this study demonstrated the applicability of QTOF-MS as a rapid and comprehensive screening technique to characterize the FA fraction of DOM from RAS and to monitor differences in their molecular fingerprints when compared with other FA samples (SRFA, ESFA and PPFA). The QTOF-MS data from SRFA and ESFA standards matched the list of formulas obtained by 17 high-resolution mass spectrometry (HRMS) instruments with 90 and 76% accuracy, respectively, which guaranteed the power of QTOF-MS without the need for further coupling to liquid chromatography (LC). RASFA was found to be rich in low- and high-oxygen unsaturated classes of compounds (lowOC: 61.73% and highOC: 19.28%) and was similar in composition to SRFA. On the contrary, the ESFA and PPFA soil standards consisted mainly of aliphatic compounds (36.77 and 55.74%, respectively) and differed significantly in composition from the RASFA and SRFA water samples. RASFA matched with 66% of the elemental compositions obtained from the DOM of makeup waters analyzed in a previous experiment, indicating freshwater and seawater origins with a high fraction of terrestrial-derived organic matter (Tanimoto score: 0.53 between RASFA and SRFA). The unique information obtained from the molecular-level analysis of FA samples by QTOF-MS highlights the importance of this technique to characterize and compare FA from different origins rendering it possible to track the FA compositional changes along the water treatment processes of RAS. To our knowledge, this is the first study that characterizes the molecular fingerprints of the RASFA by QTOF-MS and compares them with the available FA reference standards from IHSS.

Effect of Humic Acids on Soybean Seedling Growth under Polyethylene-Glycol-6000-Induced Drought Stress

Humic substances (HS) are the most important natural biostimulant of plants. However, the relationship between their structure and biological activity in plants is still not well recognized. The objective of this paper was to assess the influence of molecular fractions of humic acids (HA) (HA &lt; 30 kDa and HA &gt; 30 kDa) on reducing negative effects of drought stress in soybean (Glycine max (L.) Merr.) seedlings of Progres and Nawiko cultivars. Drought stress was induced in laboratory conditions by the addition of polyethylene glycol 6000 (PEG 6000) to make a water potential of −0.5 MPa. HA were extracted according to the International Humic Substances Society procedure, and then were separated into two molecular fractions by membrane filtration. The following physiological and biometric parameters were determined: chlorophyll content, photosynthesis activity, electrical conductivity, fresh and dry mass of overground and roots, and plant length. The enzyme activity and ion contents were also measured. Differences in response to drought stress with the addition of HA &lt; 30 kDa and HA &gt; 30 kDa or not to the Hoagland’s solution were observed among studied cultivars. Drought stress caused a decrease in the most physiological parameters and increase in peroxidase activity in the case of both studied cultivars. However, the results of biometric measurements showed that the Progres cultivar appears to have better tolerance to drought stress. The significant influence of water deficit on most macroelement content in dry matter leaves of both studied cultivars was not observed, while its effect on microelement uptake by soybean plants was concluded. In the case of the Progres cultivar, the results showed a significant decrease in microelement content in the dry matter of leaves, whereas in the leaves of Nawiko cultivar there was a significant increase. The influence of HA &gt; 30 kDa and HA &lt; 30 KDa fractions on physiological features of both studied cultivars was varied. HA &gt; 30 kDa fraction better up-regulated the antioxidant defense system. Unfortunately, no effect of either HA fraction on the macro- and micronutrients uptake system of both studied cultivars was observed.

The Effect of Supramolecular Humic Acids on the Diffusivity of Metal Ions in Agarose Hydrogel.

Humic acids are known as natural substances of a supramolecular nature. Their self-assembly ability can affect the migration of heavy metals and other pollutants in nature. The formation of metal-humic complexes can decrease their mobility and bioavailability. This study focuses on metal ions diffusion and immobilization in humic hydrogels. Humic acids were purchased from International Humic Substances Society (isolated from different matrices—peat, soil, leonardite, water) and extracted from lignite mined in Czech Republic. Copper(II) ions were chosen as a model example of reactive metals for the diffusion experiments. The model of instantaneous planar source was used for experimental data obtained from monitoring the time development of copper(II) ions distribution in hydrogel. The effective diffusion coefficients of copper(II) ions showed the significant dependence on reaction ability of humic hydrogels. Lower amounts of the acidic functional groups caused an increase in the effective diffusion coefficient. In general, diffusion experiments seem to act as a valuable method for reactivity mapping studies on humic substances.

Humic Acid-Modified Magnetite Nanoparticles for Removing [AuCl4]− in Aqueous Solutions

Humic acid-modified magnetite nanoparticle (MnP-HA) has been synthesized using the co-precipitation method and applied for removal of [AuCl4]−. Modifying of MnP-HA was prepared with the mass ratio of MnP-HA=10:1 and 10:3. The HA was extracted from peat soil of Sambutan Village, East Kalimantan, Indonesia, by the recommended procedure of the International Humic Substances Society (IHSS). The saturation magnetization of MnP-HA was decreased compared to unmodified MnP. The interaction between MnP and HA was occurred due to the chemical bond between Fe from MnP with the carboxylic group from HA. The coating HA on the surface of MnP unchanged the formation of the crystal structure of MnP and increased the particle size. The optimum removal of [AuCl4]− on MnP and MnP-HA materials was optimum at pH 3.0. The Langmuir isotherm model with sorption capacity was 0.23, 4.85, and 4.65 mol g–1 for MnP, MnP-HA=10:1, and 10:3, respectively. Using a pseudo-second-order equation, the degradation of the kinetics model of [AuCl4]− on MnP, MnP-HA=10:1 and 10:3 with adsorption rate constant (k) were 0.02, 0.07, and 0.06 g.mol min–1.

Optimized isolation method of humin fraction from mineral soil material

Humic substances, including humin fraction, play a key role in the fate of organic and inorganic xenobiotics contaminating the environment. Humin is an important fraction of humic substances, which has been the least studied to date. This is due to the difficulties connected with its isolation that pose a number of methodological problems. Methods of humin fraction isolation can be divided into following main groups: (1) digestion of mineral soil components with HF/HCl followed by alkali extraction of HA and FA; (2) alkali extraction of HA and FA followed by extraction of humin by different organic solvents; and (3) alkali extraction of HA and FA followed by HF/HCl digestion of mineral soil components. Nevertheless, each of these methods has different limitations. We described in detail a useful procedure of humin isolation, in which this fraction was not extracted, but isolated from the soil by removing its soluble organic and mineral components. A modified method of HA and FA extraction with 0.1 M NaOH, according to the International Humic Substances Society, was used in the first step. Then, the mineral components in the residue were digested with the 10% HF/HCl. Unlike the procedures oriented to increase the concentration of organic matter, samples were treated several times with the HF/HCl mixture until the mineral fraction was almost completely digested. The main assumption of the method modification was to obtain the highest yield with the lowest possible ash content, but without affecting humin chemical structure. The results showed that the proposed procedure is characterized by a high efficiency and recovery and, therefore, it can be used to isolate high amounts of humin from soil.

Spectral Characteristics Related to Chemical Substructures and Structures Indicative of Organic Precursors from Fulvic Acids in Sediments by NMR and HPLC-ESI-MS.

The aim of this work was to determine Fulvic Acids (FAs) in sediments to better know their composition at the molecular level and to propose substructures and structures of organic precursors. The sediment samples were obtained from a priority area for the conservation of ecosystems and biodiversity in Mexico. FAs were extracted and purified using modifications to the International Humic Substances Society method. The characterization was carried out by 1D and 2D nuclear magnetic resonance (NMR) and high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) in positive (ESI+) and negative (ESI−) modes. Twelve substructures were proposed by the COSY and HSQC experiments, correlating with compounds likely belonging to lignin derivatives obtained from soils as previously reported. The analysis of spectra obtained by HPLC-ESI-MS indicated likely presence of compounds chemically similar to that of the substructures elucidated by NMR. FAs studied are mainly constituted by carboxylic acids, hydroxyl, esters, vinyls, aliphatics, substituted aromatic rings, and amines, presenting structures related to organic precursors, such as lignin derivatives and polysaccharides.

New possibility for characterization of dissociation behaviour of supramolecular electrolytes: results obtained for the International Humic Substances Society standard and reference samples by coulometry

BackgroundHumic substances can be considered as polyelectrolytes with supramolecular character and complicated behaviour in water environment. The fractions of humic substances dissolved in water are the most active ones and determinative for their functioning in nature, where the proton-binding and dissociation ability play a crucial role. The dissociation behaviour of humic and fulvic acids can be affected by different circumstances including their concentration which is directly connected with the molecular organization of humic particles in solution and the accessibility of their ionizable functional groups. This study is focused just on these active fractions and their dissociation behaviour in the dependence on their content in studied system.ResultsStandards and reference samples of International Humic Substances Society were used. Flow-through coulometry was used to determine the total content of acidic functional groups in fulvic solutions and humic leachates. The amount of dissociated acidic groups was determined on the basis of potentiometry. Several differences between the behaviour of humic and fulvic acids were found. While whole samples of fulvic acids including the weakest functional groups were analysed, only the active dissolved humic fractions containing stronger acidic functional groups were characterized. The fractions containing higher amounts of the weakest functional groups remained insoluble. The dissociation degree of fulvic acids decreased with their increasing content, but a maximum on its concentration dependence obtained for humic acids was observed.ConclusionsTwo different values of dissociation constants were determined for each sample. The first was determined on the basis of the extrapolation of infinite dilution, second was determined as their average value in the region of high concentrations in which it was constant. Obtained values characterize the functioning of dissolved humic substances from point of view of their dissociation ability. The results obtained by this approach can help to predict the acid–base behaviour of dissolved organic carbon in soil and generally in nature. The acid–base behaviour of dissolved organic carbon depends on its ratio to water. It behaves differently in soil which is dry and in soil which is wet or saturated by water.

Characterization of Soil Organic Matter Individual Fractions (Fulvic Acids, Humic Acids, and Humins) by Spectroscopic and Electrochemical Techniques in Agricultural Soils

The objective of this paper was to investigate the molecular characterization of soil organic matter fractions (humic substances (HS): fulvic acids-FAs, humic acids-HAs, and humins-HNs), which are the most reactive soil components. A wide spectrum of spectroscopic (UV–VIS and VIS–nearIR), as well as electrochemical (zeta potential, particle size diameter, and polydispersity index), methods were applied to find the relevant differences in the behavior, formation, composition, and sorption properties of HS fractions derived from various soils. Soil material (n = 30) used for the study were sampled from the surface layer (0–30 cm) of agricultural soils. FAs and HAs were isolated by sequential extraction in alkaline and acidic solutions, according to the International Humic Substances Society method, while HNs was determined in the soil residue (after FAs and HAs extraction) by mineral fraction digestion using a 0.1M HCL/0.3M HF mixture and DMSO. Our study showed that significant differences in the molecular structures of FAs, Has, and HNs occurred. Optical analysis confirmed the lower molecular weight of FAs with high amount of lignin-like compounds and the higher weighted aliphatic–aromatic structure of HAs. The HNs were characterized by a very pronounced and strong condensed structure associated with the highest molecular weight. HAs and HNs molecules exhibited an abundance of acidic, phenolic, and amine functional groups at the aromatic ring and aliphatic chains, while FAs mainly showed the presence of methyl, methylene, ethenyl, and carboxyl reactive groups. HS was characterized by high polydispersity related with their structure. FAs were characterized by ellipsoidal shape as being associated to the long aliphatic chains, while HAs and HNs revealed a smaller particle diameter and a more spherical shape caused by the higher intermolecular forcing between the particles. The observed trends directly indicate that individual HS fractions differ in behavior, formation, composition, and sorption properties, which reflects their binding potential to other molecules depending on soil properties resulting from their type. The determined properties of individual HS fractions are presented as averaged characteristics over the examined soils with different physico-chemical properties.