A comparative study of elemental composition of water-soluble humic substances,humic acids,and fulvic acids extracted from sod-podzolic soils

Elemental analysis of water-soluble humic substances extracted from three sod-podzolic soils was carried out. Data on elemental composition were compared to those of humic and fulvic acids extracted from the same soils.     

Chemical and physical characteristics of humic and fulvic acids extracted from soils of the Mediterranean region

Humic and fulvic acids were extracted from two soils from Israel and from four soils from southern Italy. Following purification, the humic substances were characterized by elementary and functional group analyses, spectrophotometric and spectroscopic methods, thermal analysis and by scanning electron microscopy.The analytical characteristics of the humic and fulvic acids extracted from the Israeli and Italian soils were generally similar to those of humic materials originating from soils formed under widely differing geographic and pedologic conditions. There were some differences in the content and composition of inorganics in the fulvic acids which were reflected in functional group analyses, IR and ESR spectra, DTA curves and SEM micrographs.     

Persulfate oxidation of humic acids extracted from three different soils

Three soil humic acids were degraded by a new mild chemical oxidation method using potassium persulfate at pH 2.0. The method degraded 30–40% of the starting material. The type and relative quantities of the products or fragments released as determined by GC/MS varied with the source of the polymers. Humic acid extracted from an Inceptisol yielded fatty, benzenecarboxylic and phenolic acids; humic acid from a Typic Chromoxerert produced largely dialkyl phthalates, and humic acid from an Humic Haplorthod gave only branched and straight-chain fatty acids. It is suggested that the method may be used to advantage as a first oxidant in a sequential degradation.     

Biological activity of humic substances extracted from soils under different vegetation cover

Abstract

Comparisons were made between the chemical compositions of humic substances extracted from three soils covered by different vegetation and their biological activities assayed using 15‐ and 30‐day‐old seedlings of Pinus sylvestris and Picea abies. The growth, “α‐amylase and invertase activity were affected by humic fractions and by gibberellic acid (GA), indicating that humic matter had a gibberellin‐like activity. The isoenzymatic polymorphism in the electrophoretic patterns of esterase was influenced in a similar way by all humic fractions and by indoleacetic acid (IAA), which also suggested that the humic fractions exhibited an auxin‐like activity. The humic fractions extracted from the grassland, exhibiting higher amounts of phenolic and a considerable amount of carboxyl carbon, showed the best metabolic effect. The forest humic fractions, characterized by lower phenolic carbon content, appeared less effective in influencing plant metabolism, whereas the grassland‐forest humic substances proved to be even more less effective. The auxin‐ and the gibberellin‐like activities were related to a high content of phenolic and carboxylic groups. These results presented evidence that the biological activity of the humic substances was attributed to their chemical structure and to their functional groups, which could interact with hormone‐binding proteins in the membrane systems, evoking a hormone‐like response.     

Microbial utilization and transformation of humic substances extracted from soils of long-term field experiments

Humic acids (HA)  extracted with alkali from unfertilized and fertilized (NPK + organic manure) plots of  long-term, i.e. 45 years, field experiments were added as a supplemental source of nutrients or as the sole sources of carbon and nitrogen to semi-aerobic cultures of complex microbial communities indigenous to the same individual soils. Depending on the experimental conditions, between 2.7% and 47.3% of the added HA preparations were utilized in 12 months. In full strength nutrient media these numbers seemed affected by some novel microbially produced HA-like substances. An average utilization rate was 33% for the HA from an unfertilized   Orthic-Luvisol plot, whereas HA preparations extracted from fertilized plots of the Orthic-Luvisol or  a Dystric-Cambisol soil site were utilized to 27% and 15% respectively. The HA preparations re-isolated after 12 months from microbial cultures exhibited differences in elemental and structural characteristics, corresponding mainly with the nutrient status of the individual  cultures. Those from nitrogen-deficient cultures, e.g., were partly depleted in N. The Fourier-transformed infrared (FTIR) spectra of  the re-isolated HA preparations indicated loss in aliphatic structures. Aromatic structures were strongly associated with  mineral moieties and  remained rather  unaffected. The results indicate in general that an increase in soil organic matter contents as frequently observed in long-term fertilized soils could be attributed to an enhanced resistance of humic acid fractions to microbial degradation.     

Electrophoresis and size-exclusion chromatography of humic substances extracted from detritus and soils of different geneses

Electrophoresis in 10% polyacrylamide gel in the presence of denaturants and size-exclusion chromatography in Sephadex G-75 in 7 M urea were used for the comparative analysis of humic substances isolated from a typical chernozem, soddy-podzolic soil, and chestnut soil and from the easily decomposable organic matter (plant detritus) contained in these soils. After the electrophoresis, the gel with naturally colored bands of humic substances was further stained with a dye specific for proteins by immersing into a solution containing Coomassie Brilliant Blue R-250 and CuSO₄. The electrophoretic and chromatographic analyses showed that humic substances from the soils and the corresponding detritus fractions significantly differed in the intensity of the natural color of the electrophoretic fractions, the molecular-weight distribution, and the color of the electrophoretic fractions colored by the protein-specific dye (which was first discovered in this study). The hypothesis of Tyurin and Aleksandrova suggesting that the transformation of humus sources (plant detritus) into humic substances proceeds in the direction from the high-molecular compounds to the low-molecular compounds was experimentally confirmed.     

Fate of humic acids isolated from natural humic substances

Composition of humic acids (HA) is a function of plant-derived inputs, degradation processes regulated by microorganisms, organo-mineral interactions and age. Characterization of different origin humic substances is important for evaluation of their contribution to stabile and labile carbon pool in the environment. The relative abundance of chemical components in HA isolated from soils, compost, commercial lignohumates, alginite, acadiane and lignite was studied with aim to quantify content of important biomarkers such as amino acid, lipids and polyphenols. HA were considered as a heterogeneous complex and high concentration of peptides, polyphenols and lipids was determined in acadian-HA to compare with soil-HA. Compost-HA contained much more amino acids to compare with soil-HA samples. Alginite-HA and lignite-HA were similar in biomarkers content to soil-HA. Fourier transform infrared spectroscopy confirmed that chemical composition and functional groups content differs with the origin, humification degree and the age of studied samples. Soil-HA are typically composed of a variety of ?OH, COOH?, C–O, C–H₂, (aliphatic and aromatic) groups, quinines, lignin fragments, polysaccharide, monosaccharide and proteins fragments, which are linked together by ?O?, ?NH?, ?H=, >C=O, metal ions and –S? groups. ¹³C NMR spectroscopy showed that aromatic carbon content was the highest in lignite-HA and soil-HA.     

Aerobic short-term microbial utilization and degradation of humic acids extracted from soils of long-term field experiments

A humic acid (HA) fraction of the soil organic matter (SOM) was extracted with alkali from soil samples originated in non-fertilized and fertilized (NPK + organic manure) plots of long-term (45 years) field experiments. The HA preparations served as supplemental sources of nutrients or as sole source of either C or N for soil micro-organisms indigenous to the same soils. Under aerobic conditions (shake cultures) between 15% and 45% of HA were degraded in 21 days. The degradation was minimum if HAs were added supplementary, although the biomass formation was strongly enhanced. Preparations of HA from long-term fertilized soils appeared somewhat less susceptible to microbial degradation but they were capable of supporting microbial growth. Under copious nutritional conditions some novel HA-like substances were formed. The HA preparations re-isolated from individual cultures exhibited differences in elemental and structural characteristics. The FTIR spectra indicated an increasing proportion of aromatic structures that appeared as associated with mineral moieties. Conclusively, HAs from long-term fertilized and manured soils could be considered as more resistant to microbial activities than those from control soil, but under limited nutrient conditions their aliphatic constituents appear utilizable by micro-organisms.     

Chemical and physical characteristics of humic substances extracted from greek soils varying in classification order and natural drainage

Abstract

Humic and fulvic acids were extracted from the surface horizons of Alfisols, Vertisols, and Entisols with good and impeded natural drainage. These profiles are located in the temperate subhumid zone. The extracted humic substances were characterized by elementary and functional group analyses, IR‐spectroscopy, differential thermal analysis (DTA) and electron spin resonance spectroscopy (ESR). Some differences in the humic substances seem to be associated with differences in the natural drainage of the soils. When data from samples of the same soil order are compared, indicate that humic (HAs) and fulvic acids (FAs) extracted from soils with impeded drainage, have higher contents of carbon and spin concentration, but less total acidity and fewer carboxylic groups (COOH), than the respective humic substances extracted from well drained soils. The IR‐spectra of the HAs extracted from soils with impeded drainage showed more aliphatic groups than those of the well drained soils. No essential differences, which could be related to the drainage conditions of the studied soils, were observed in the IR‐spectra of FAs. DTA‐diagrams shows that the thermal stability of the HAs and the FAs are related to their carbon and ash contents. Iron was the dominant element in the ash of the humic acids, aluminum and silicon were the most abundant elements in the ash of the fulvic acids.     

Gel filtration chromatography of humic substances in soil solutions using HPLC-determination of the molecular weight distribution

The molecular weight distribution of humic substances in soil solutions from a dystric cambisol and an orthic podzol were determined by high-performance gel filtration chromatography (Toyo Soda TSKgel G2000 SWXL column). The retention volumes of humic substances were compared to those of narrow molecular weight distributed poly-styrenesulphonates with known molecular weight. A change in mobile phase pH or ionic strength usually resulted in an almost identical change in retention volume for the polystyrenesulphonates and the bulk of humic substances. The possibility of using the former to calibrate the column is discussed. The molecular weight at peak maximum was 1120–1190 and 1800 for the cambisol and podzol samples, respectively, using polystyrenesulphonates as molecular weight standards. No pretreatment of samples was used, but ion exchange is recommended for samples with high Al concentrations.     

Spatial variability of the molecular composition of humic acids from subtropical forest soils

Journal of Soils and Sediments - Humic acid (HA) plays vital roles in controlling the conservation of soil and water in the forest soils. The structure of HA is closely related to its stability. A...     

Particle and structure characterization of fulvic acids from agricultural soils

Purpose

Structural studies on fulvic acids (FAs) are of significant importance since FAs are involved in many environmentally important processes, such as adsorption and transportation of nutrients, trace elements, and organic pollutants. Interactions between suspended and dissolved soil components are controlled by a variety of attractive and repulsive interparticle forces influenced partially by FA properties. The aim of this paper is a detailed characterization of FAs derived from agricultural soils varied with physicochemical properties.

Materials and methods

Forty topsoils (pH_KCL?=?3.8–7.8, clay content?=?0–6%, and TOC?=?7.0–187.2 g kg^?1) were collected from rural area. Fulvic acids (FAs) were isolated according to the IHSS method. The overall FA solutions were purified by nonionic macroporous acrylic ester resin (DAX-8), and the organic carbon content in FA fraction (FA-OC) was determined by a liquid C–N analyzer. The particle size diameter (PSD) and polydispersity (PDI) were analyzed by a dynamic light scattering technique, while the zeta potential (ZP) was measured using an electrophoretic light scattering method. Spectroscopic properties of FAs, including occurrence and distribution of functional groups, were investigated by near-IR spectroscopy.

Results and discussion

Agricultural soils differed substantially, with FA-OC content ranging from 0.6 to 8.8 g kg^?1 that accounted for 0.5 to 22.6% of TOC. The PSD exhibited wide range of particle size (0.2 to 69.6 nm) and was characterized by different polydispersity (14–183.1%). The ZP described the behavior, and surface charge of FA particles varied from ??1.7 to +?3.3 mV. Low ZP characterized 77% of FAs and indicated the ease of aggregate formation and intermolecular connections. The measured ZP also showed that suspended organic particles of FAs had both positive and negative charges, which was confirmed by the spectroscopic analysis. The presence of negative charges on FA particle surfaces was connected with the occurrence of phenol and carboxyl groups while positive charge with amine.

Conclusions

Detailed characterization of FAs from agricultural soils confirms their heterogeneous and complex nature. The results indicate that FAs mainly exist as small molecules that form molecular aggregates or associations in solutions. FA in a solution of a similar ionic strength may be positively or negatively charged due to its chemical structure and aggregate behavior which affects their properties in the soil.

     

Binding of methylmercury compounds by humic and fulvic acids

Humic and fulvic acids were isolated from Fawn Lake in Ontario and investigated by membrane dialysis for their ability to bind CH₃Hg⁺. By measuring the distribution of methylmercury compounds in and outside of the dialysis membrane, for the first time, binding capacities and conditional stability constants for CH₃Hg⁺ and humic acids could be calculated. Equilibrium between inner and outer dialysis solution was reached within 24 hours without losses of CH₃Hg⁺ in humic solution. The ratio of bound to unbound CH₃Hg⁺ increased with decreasing total CH₃Hg⁺ levels in the system. The percentage of free methylmercury compounds increased with decreasing pH. Fawn Lake humic acid showed two different binding sites. The conditional stability constant of the stronger site was calculated as 1.3 × 10¹². This site was able to bind 0.2 ng CH₃Hg⁺ per mg of humic acid.     

Acid properties of fulvic and humic acids isolated from two acid forest soils under different vegetation cover and soil depth

We studied the acid‐base properties of 16 fulvic acids and 16 humic acids isolated from the surface (3–15 cm) and subsurface (> 45 cm) horizons of two types of acid forest soils, derived respectively from amphibolite and granite rocks, under five different types of vegetation. The observed differences between the contents of humic substances in the two types of soils were related to the degree of Al‐saturation of the soil organic matter, as indicated by the molar ratio between pyrophosphate extractable Al and C. Humic fractions were characterized in terms of elemental composition, and CPMAS ¹³C NMR spectrometry. The contents of carboxylic and phenolic groups were estimated by potentiometric titrations conducted in 0.1 m KNO₃ in a nitrogen atmosphere. The fulvic acids contained more carboxylic groups but less phenolic groups than the humic acids: the ratio of phenolic to carboxylic groups in the humic acids was 0.48 ± 0.10 and in the fulvic acids 0.23 ± 0.05. The mean values of the protonation constants of each of the humic substance fractions can be used as generic parameters for describing the proton binding properties. The fulvic acids isolated from the subsurface horizon of the soil contained between 2.6 and 23% more carboxylic groups, and the humic acids between 8 and 43% more carboxylic groups than those isolated from the surface horizon of the same soil.     

Mobilization of Cu and Pb from multi-metal contaminated soils by dissolved humic substances extracted from leonardite and factors affecting the process

Journal of Soils and Sediments - The aim of the research was to determine the potential of dissolved humic substances extracted from leonardite to mobilize Cu and Pb from multi-metal contaminated...     

Effect of ph on precipitation of humic acid from peat and mineral soils on the distribution of phosphorus forms in humic and fulvic acid fractions

Abstract

Humic and fulvic acid fractions were isolated from a mineral soil and a peat by adjusting the pH of the alkali extracts to a range of values from 0.2 to 2.5. Total inorganic and organic forms of phosphorus (P) in the acids were measured by chemical analysis and by ³¹P NMR spectroscopy. As the pH of precipitation of the mineral soil humic acid increased, there was an increase in the total P of the humic acid which related to the inorganic P component. In contrast with the peat, the increases observed in the pH range 0.2 to 1.5 were the result of changes in organic P. Using ³¹P nuclear magnetic resonance spectroscopy, the ratio of inorganic to organic P as mono‐ and di‐esters in the peat humic acid was found to increase from 1:4.8 at pH 2 to 1:19 at pH 2.5. In contrast with mineral soil humic acid, the ratio decreased from 1:6.1 at pH 0.2 to 1:1.3 at pH 2.5. The mono‐ester to di‐ester ratio was about 3 in the peat and 10 in the mineral soil and varied little with pH of precipitation. Phosphonates were detected only in the peat humic acid precipitated in the pH range 1.0 to 2.0     

Systems of solutions,minerals, and humic substances in soils

The equilibrium composition of solutions, minerals, and humic substances of the humus-clay plasma from the humus horizon of a leached chernozem was calculated using the method of computerized physicochemical simulation based on Gibbs’ principle of minimum free energy in the equilibrium state of the system. It was shown that the plasma from the humus horizons of chernozems and soddy-deeply podzolic soil mainly consists of oxides, hydroxides, humic substances, kaolinite, and 2: 1 illite-smectite minerals. The stoichiometry of the humic substances corresponds to calcium-saturated humus acids with appreciable contents of iron and aluminum. The crystallochemical composition of the 2: 1 layered minerals classifies them among ferruginated mixed-layered illite-montmorillonites. Simulation at an increased concentration of carbon dioxide indicated the degradation of the humus-clay plasma in a leached chernozem under extensive soil use, which can be restored by application of potassium at rates 1-to ?1.5-fold its removal.     

Permanganate oxidation of methylated fulvic and humic acids in chloroform

Methylated fulvic and humic acids were oxidized by permanganate in chloroform in the presence of a cyclic polyether, 18-crown-6. The total amount of oxidation products identified was 6.5% for fulvic acid and 6.7% for humic acid. About 46% of the oxidation products found from both fractions were dicarboxylic acids (15 compounds), the rest were benzenecarboxylic acids and their methoxyl derivatives (15 compounds). Alkanes and fatty acids were also found.Nonandioic acid and 3,4-dimethoxy-benzenecarboxylic acid are found in almost equal amounts. Each accounts for 1.3% of the starting material, and together they represent 40% of the oxidation products. 3,4-Dimethoxy-benzoic acid and 4-methoxy-benzoic acid are believed to result from the oxidation of terminal groups in the humic polymer.Proton resonance spectra show that about 25% of the aliphatic protons are part of methylene chains. The most prominent chain lengths consist of 6–8 methylene groups as shown by the dicarboxylic acids formed by oxidation.