Comparative evaluation of compost humic acids and their effects on the germination of switchgrass (Panicum vigatum L.)

Purpose

This study aimed to investigate comparatively the main chemical and physico-chemical properties of the humic acid (HA) fraction of three different composts and to evaluate the bioactive effects of these HAs on the germination and early growth of four populations of switchgrass.

Materials and methods

Three compost HAs isolated from a green compost (HA_GC), a mixed compost (HA_MC), and a coffee compost (HA_CC) were characterized for some chemical and physico-chemical properties, such as ash content, elemental composition, total acidity, carboxylic and phenolic OH group contents, E₄/E₆ ratio, Fourier Transform infrared (FT IR), and fluorescence spectroscopies. In subsequent experiments conducted in vitro in a climatic chamber under controlled conditions, the bioactive effects of the three HAs at concentrations of 10, 50, and 200 mg?L^?1 were tested on the germination and early growth of four switchgrass (Panicum virgatum L.) populations, the octaploids Shelter, Shawnee and Dacotah, and the tetraploid Alamo.

Results and discussion

The ash content and the E₄/E₆ ratio were, respectively, much higher or slightly higher for HA_CC than for the other two HAs. HA_MC showed the lowest C and H contents and the highest O content, whereas HA_GC had the highest N content. The total acidity and phenolic OH group content followed the order: HA_MC?>?HA_CC?>?HA_GC. The fluorescence analysis of the three HAs evidenced a common fluorophore unit possibly associated to simple aromatic structures, such as phenolic-like, hydroxy-substituted benzoic and cinnamic acid derivatives. The FT IR spectra of all HA samples indicated the presence of aromatic phenolic structures. Significant beneficial effects were produced by any HA on switchgrass germination and early growth as a function of the population tested and the HA dose.

Conclusions

Results of this study demonstrated that the addition of compost HAs to the germination medium of four switchgrass populations positively influenced the germination process and the growth of primary root and shoot. Significant correlations were found between HA bioactivity and some HA properties. These results suggest a possible use of compost as soil amendment in areas where switchgrass grows naturally or is cultivated.     

Humus Characterization of Humid Tropical Forest,Tea Garden,and Field Crop Soils of West Bengal,India: Chemical,Potentiometric, and Spectroscopic Methods

Humic acids (HA) and fulvic acids (FA) were extracted from tropical humid forest, tea garden, and field crop soils, and their chemical, potentiometric, and spectrophotometric properties were measured. There was less HA than FA in the cultivated soils. The HAs contained more carbon (C) and nitrogen (N) and had greater C/H ratios but lower O (oxygen)/H (hydrogen) ratios than FAs. Cultivated soils had greater total acidity than the forest soil generally because of both —COOH and phenolic-OH. Molecular weights of HA an FA, estimated from intrinsic viscosities, revealed that cultivation had reasonably reduced the molecular weights. The difference in pH (ΔpH) values, corresponding to three-fourths and one-fourth of the pH at final inflexion point showed that they were polyprotic. The longer time required for stability of greater pH of HAs was related to greater degree of stable coiling. The (E₄/E₆), in general, revealed a greater amount of aliphatic moiety rather than aromatic moiety.     

Long term effects of a brown coal-based amendment on the properties of soil humic acids

In this study a typical grey-brown podzolic soil was amended with different doses of a brown coal-based preparation called Rekulter (R) largely used in Poland. After seven years, soils were analyzed and humic acids (HAs) were extracted both from the control soil and from the amended soils. All HAs were characterized by Fourier transform infrared spectroscopy and fluorescence spectroscopy both in emission, excitation and synchronous-scan mode and as Excitation-Emission-Matrix (EEM) contour maps. A higher carbon content was observed in the amended soils whereas significant differences were highlighted between the unamended and the amended soil HAs. HAs from amended soils showed a higher content of carboxyl groups and a more aromatic character, particularly HA extracted from the soil amended with the highest dose of R.     

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.     

Composition of humic acids extracted under air and nitrogen atmosphere

Abstract

Differences in nature and composition of humic matter, attributed to NaOH extraction under air or a N₂ gas atmosphere, were investigated. Samples from the Ap horizons of a Cecil and Onaway soil, and lignite were shaken with 0.1 M NaOH in the presence of air or under N₂ gas. The humic (HA) and fulvic acid (FA) separated were weighed, and analyzed for C, H, O, N, and S contents. Total acidity, carboxyl and phenolic‐OH group contents were determined by chemical analysis. Characterization was performed by ¹³C nuclear magnetic resonance (NMR) and infrared (IR) spectroscopy. The results indicated that differences in amounts of HA and FA, attributed to NaOH extraction under air and N₂ gas, were small and no definite trend was noted for more HA and FA extracted under air. No differences in C, H, O, N, and S contents of HA and FA were also noticed as a result of the influence of air or N₂ gas during extraction. However, HA and FA exhibited larger values for total acidities when extracted under a N₂ gas atmosphere than under air. The lower amounts of carboxyl and phenolic‐OH group contents in HA and FA as a result of NaOH extraction under air, however, do not support the idea of increased oxidation by NaOH. No differences were noticed in NMR and IR spectra of HA and FA attributable to the two extraction methods. The differences in NMR and IR features obtained were more the result of differences in origin than in methods of extraction.     

Soil humic acid aggregation by dynamic light scattering and laser Doppler electrophoresis

Humic acids (HAs), similar to other fractions of humic substances (HSs), have a large number of reactive functional groups enabling them to aggregate in solutions. Regardless of the origin of humic acid (aqueous or soil), this aggregation process is dependent on environmental conditions and strongly influences the mobility of soluble ionic and molecular pollutants. The aim of this work was to monitor the aggregation process of two humic acids isolated from different mineral soils (IHSS Elliot soil HA standard and Rendzic Leptosol HA) in the 2–11 pH range. Changes in aggregate size in HA sols were followed up using dynamic light scattering (DLS), while zeta potential (ZP) measurements in the same pH range were performed applying laser Doppler electrophoresis (LDE) technique. The effect of HA sol concentration and soil source on aggregation was examined as well. Besides, HA samples were characterized using Fourier transform infrared (FT‐IR) spectroscopy. By inspecting HA‐particle‐size dependence on pH, it can be concluded that both HAs in corresponding sols behave as molecular aggregates or supramolecular structures, formed from small individual moieties (sizes < 10 nm) at higher pH values. The ZP vs. pH curve for both HAs revealed the ZP minimum in the 5–7 pH range, caused most likely by dissociation of acidic functional groups prevailing at lower pH values and deaggregation predominating over dissociation at higher pH values.     

Effect of cultivated plants on physico‐chemical characteristics of humic substances of soils amended by sewage sludge

Abstract

Humic substances were extracted from raw sewage sludge samples as well as an alluvial slightly alkaline soil [Typic Xerofluvent (So)], a clay loam soil [Calcixerollic Xerochrept (M)] and the corresponding field plots amended with different rates of sewage sludge and cultivated with corn and cotton respectively, in a two‐year field experiment. These substances have been characterized by chemical and spectroscopic methods. The chemical analyses showed that humic acids (HAs) and fulvic acids (FAs), were effected by the cultivated plant. Humic substances extracted from field plots with cotton showed higher carbon, but less nitrogen (N), total acidity and carboxylic groups content, as compared with those under corn. Generally HAs and FAs extracted from the fields plots amended with sewage sludge had higher N content, lower values of total acidity, carboxylic groups and carbon contents than those from the unamended soils. The FT‐IR spectra of humic (HAs) and fulvic (FAs) acids extracted from sewage sludge indicated the presence of high percentage of aliphatic carbon, polysaccharides and proteinaceous materials. The spectra of the humic matter in the field plots showed less aliphatic but increased amide stretch in comparison with the sludge. The absorption due to C=O in carboxylic groups of the FAs was not pronounced, though the presence of polysaccharides and protein decomposition products was clearly indicated.     

Kinetics and characteristics of humic acids produced from simple phenols

In the presence of H₂O₂ as donor, horseradish peroxidase was used to catalyze the polymerization of seven monomeric phenols. Yields of humic acid (HA) polymers from meta phenols—resorcinol and phloroglucinol—were insignificant. Of the five ortho and para phenols—phenol, catechol, hydroquinone, pyrogallol and 1,2,4-trihydroxybenzene—all except hydroquinone inhibited the enzyme at high concentration. The kinetics of polymerization of the ortho and para compounds were complex and dependent on the concentration of both electron acceptor and donor.The percentage yield of HA before dialysis was far greater from pyrogallol than from catechol or hydroquinone. After dialysis, the yield of the catechol HA was higher than those of the hydroquinone and pyrogallol HAs. A higher molecular weight for the catechol HA over those of the hydroquinone and pyrogallol HAs was also indicated by the lowest E₄/E₆ ratio and highest free radical content.All of the synthetic HAs were relatively rich in free radicals, suggesting that their synthesis occurred via free radicals, i.r. and ¹³C NMR spectra showed that the HAs were molecularly complex polymers or mixtures of complex aromatic structures rich in phenolic OH groups and to a lesser extent in CO₂H groups. The only HA which showed fine structure in the i.r. spectrum was the pyrogallol HA; the presence of aryl ethers was indicated. ¹³C NMR spectra showed that all synthetic HAs were highly aromatic, that aromatic rings of the initial phenols had been built into the HAs, but that molecular environments around phenolic OH groups had changed during the formation of the HAs.     

Humic matter isolated from soils and water by the XAD‐8 resin and conventional NaOH methods

Abstract

Differences were studied in humic (HA) and fulvic acid (FA) extracted from soils and streams in South Georgia by the Amberlite XAD‐8 resin and conventional NaOH method. Characterization analysis was performed by liquid ¹³C NMR, infrared (IR) spectroscopy, scanning electron microscopy (SEM), and chemical analysis. The NMR spectra indicated that the resin method yielded black water HA and FA with spectroscopic, chemical and elemental characteristics different from those isolated by the conventional NaOH method. Humic acids from both the resin and conventional NaOH methods were composed of aliphatic, aromatics and carboxyl groups, but the “resin”; HA contained more aliphatic groups. These differences were also noticed between the FA fractions obtained by the two methods. The differences corresponded to differences in IR spectra. The IR spectrum of “resin”; FA exhibited only a weak shoulder at 1625/cm for the COO”; stretching vibration, in contrast to that of FA isolated by the conventional NaOH procedure. Apparently, the high aliphatic‐CH₃ group content has blocked the vibration above, as evidenced by methylation of HA. Fulvic acid extracted by the resin method was also higher in total acidity, but considerably lower in N content than FA obtained by the conventional NaOH method. Both methods yielded black water FA which was less aromatic in nature than black water HA, or soil FA.     

Changes in forest soil organic matter quality affected by windstorm and wildfire

Purpose

Windthrows and fires are major natural disturbances in forest ecosystems, which can affect organic matter in the surface and the mineral layer of forest soil. The main aim of this study was to evaluate the changes occurring in the structure and properties of humic acid (HA) in the lands where windthrows and wildfires occurred.

Materials and methods

In November 2004, the forest in the area of 12,000 ha in the Tatra National Park, Slovakia, was seriously damaged by northern wind gusts exceeding 200 km/h. In July 2005, a wildfire broke out in a 220 ha of wind-damaged area. The HAs have been isolated from four research plots: (a) the area where the fallen trees were removed (EXT); (b) an area after windstorm covered by wood from struck trees (NEX), left for spontaneous succession; (c) an area after extracted timber, damaged by the surface wildfire (FIR); and (d) a reference intact spruce forest area (REF). Changes in the chemical structure of the HAs isolated from the research plots were determined on the basis of elemental analysis and UV-Vis, EPR, IR, and ¹³C NMR spectroscopy.

Results and discussion

All used analytical methods showed a decrease in the humification degree of the humic acids extracted from the soils where the spruce forest has been affected by a wildfire and a windthrow. In the case of the control sample HA (REF), the calculated atomic H/C and O/C ratios and the degree of aromaticity (α) calculated from the ¹³C NMR spectra were higher, indicating higher aromaticity of HA from the REF area. The more complex and developed structure of REF HA was confirmed by the higher value of E^1%₆ and the lower E4/E6 ratio obtained using UV-Vis spectroscopy. Also, the higher g-parameters determined from the EPR spectra of the stable radicals present in HA confirmed the lower aromaticity on the plots that have been subjected to the calamities. The ¹³C NMR spectra and the elemental analysis show that the structure of the HA extracted from the NEX plot is the closest to the REF.

Conclusions

The results of the systematic research showed significant changes in the structure of HA taken from spruce forest soils that were subjected to windstorms and fires. An enrichment of the HAs in aliphatic carbon and so a lower humification degree of the organic matter in the areas calamity-affected were observed. The results clearly indicate that the HAs extracted from the disturbed plots of the spruce forest are not as stable as those extracted taken from the control plot.

     

The significance of solid‐state 13C NMR spectroscopy of whole soil in the characterization of humic matter

Abstract

Differences in characteristics of humic matter were investigated by solid‐state CP/MAS ¹³C NMR spectroscopy of whole (nontreated) materials and their extracted humic fractions. Samples used in the analysis were lignite, a commercial humate AG, and the Bh horizons of a Mascotte and a Lawnwood soil. Humic fractions were extracted by the 0.1 M NaOH or Na₄P₂O₇ (pH 9.8) method. The humic (HA) and fulvic acid (FA) obtained were weighed and analysed for total acidity, carboxyl and phenolic‐OH group contents. Whole lignite, humate AG and soil samples, and the HA and FA fractions were analyzed by solid state CP MAS ¹³C NMR and infrared spectroscopy. Carbon, H, and N contents were determined by chemical analysis. NMR spectra of the combined HA+FA extracts resembled the spectra of the whole materials. No additional signals were detected, indicating that alien compounds were not produced during the extraction. The best spectra were obtained with HA samples produced by the NaOH method. These spectra closely resembled those of the untreated materials. Spectral and chemical differences noticed between the HA (or FA) fractions were attributed more to differences in origin than to the extraction procedure. Aliphatic, aromatic and carboxyl groups were the major components of HA from lignite and humate AG. In contrast, HA from the two Haplaquods were characterized by four major components: the aliphatic, polysaccharide, aromatic, and carboxyl groups. Regardless of origin, all the HA fractions contained similar functional groups, as indicated by their close similarities in infrared spectra.     

Susceptibility of humic acids from soils with various contents of metals to microbial utilization and transformation

Humic acids (HAs) from a pseudogley soil with various metal contents were added as supplemental sources of nutrients, or as the sole sources of carbon or nitrogen, to aerobic cultures of complex microbial populations indigenous to the same individual soils. Depending on nutrient conditions in the individual cultures and origin of HAs, between 44% and 67% of the added HAs were utilized. The lowest utilization rate was obtained for HAs from soil heavily contaminated with Mg. The overall carbon mineralization in the microbial cultures was significantly reduced in the presence of HAs. Simultaneously, the formation of microbial biomass was enhanced up to 261%. Variovorax (Alcaligenes) paradoxus, Pseudomonas fluorescens, and a yeast Cryptococcus sp. have been identified as the dominant microbial species utilizing HAs. The individual HA preparations re-isolated from the microbial cultures exhibited distinct changes in elemental and structural characteristics. Diminished contents of ash and alterations in infra-red absorptions indicated a splitting of organic and mineral components in HAs exposed to microbial activities.     

Chemical structure of humic acids in biosolids-amended soils as revealed by NMR spectroscopy

We used NMR spectroscopy to characterize humid acids extracted from soils that had received long-term application of 2 levels of biosolids to evaluate the soil organic matter (SOM) stability in biosolids-amended soils. The study also quantified fulvic acids (FAs), humic acids (HAs) and Fe/Al oxides. The soils were collected in 2004 from 7 fields, in Fulton County, southwestern Illinois, which received biosolids at a cumulative rate of 0 (control), 554 (low biosolids) and 1,066 (high biosolids) Mg ha⁻¹. The application of biosolids increased both FA and HA contents, but biosolids-amended soil and control soil did not differ in FA/HA ratio. Biosolids application had no effect on water-soluble organic carbon content. Biosolids application increased the presence of Fe/Al in the SOM complex and lowered its C/Fe and C/Al ratios. ¹³C NMR spectra showed increased alkyl C and decreased aromatic C content in soil HAs with the application of biosolids, and the extent of such changes was higher with high than low biosolids treatment. Under biosolids application, the soil HAs’ C structure shifts from O-alkyl-dominant to alkyl-dominant. Biosolids application does not decrease SOM stability but rather increases the stability of soil humic substances.     

Separation and characterization of the constituents of compost and soil humic acids by two-dimensional electrophoresis

To investigate the chemical heterogeneity of humic acids (HAs), we applied two-dimensional (2-D) electrophoresis to HAs from a compost and two types of soils. In this method, HAs are first separated by isoelectric focusing (IEF) and then separated by polyacrylamide gel electrophoresis (PAGE). IEF and PAGE were carried out in the presence of 7?M urea. Upon 2-D electrophoresis of HAs, dark-colored substances were spread out across the gel mainly in the isoelectric point (pI) range of 3.0–4.5. Green fluorescence was observed in the smaller molecular size region of the gel, especially in the pI range of 3.0–4.5, and the most intense fluorescence was found at the moving front. The gels were divided into 36 sections, and then HA constituents were extracted from the individual sections and recovered by precipitation with acid. The distribution of organic carbon (C) among the gel sections coincided with that of the dark-colored substances on the gel. The total C recoveries were only 43–50%, suggesting that a considerable amount of HA constituents was lost during the extraction from the gels and purification. High-performance size-exclusion chromatography confirmed that the constituents of HAs were separated based on their molecular sizes by PAGE. The measurement of diffuse reflectance infrared Fourier transform (DRIFT) spectra indicated that the chemical properties of the HA constituents differed depending on the position on the gels and were affected by the molecular size rather than the pI. The fractions of the compost HA were characterized by higher proportions of aliphatic, proteinous and polysaccharide moieties and by the presence of lignin-derived structures. For the soil HAs, the fractions were characterized by a high proportion of the carboxyl group and a low proportion of aliphatic moieties. The proportion of proteinous and polysaccharide moieties in the fractions of soil HAs decreased with decreasing molecular size. The chemical properties of the green fluorescent substances remained unclear, since there was not enough of the substances to measure the DRIFT spectra. The present study showed that 2-D electrophoresis in the presence of concentrated urea offers an effective method for fractionating and isolating the constituents of HAs.     

土壤中钙结合和铁铝结合腐殖质的组成及结构特征

Calcium-bound and iron- and aluminium-bound humus extracted from different soils collected from north to south of China were characterized by chemical and spectroscopic methods. Meaningful differences in the composition and structure between them were revealed by ¹³C NMR, visible spectroscopy and elemental analysis. Results showed that the contents of carbon, hydrogen and nitrogen were higher in iron- and aluminium-bound humus than in calcium-bound humus while oxygen content in calcium-bound humus was shown to be higher. The calcium-bound humus had higher C/N and O/C ratios than iron- and aluminium-bound humus. The calcium-bound humic acid (HA1) showed higher E₄/E₆ ratios than iron- and aluminum-bound humic acid (HA2) while iron- and aluminum-bound fulvic acid (FA2) showed higher E₄/E₆ ratios than calcium-bound fulvic acid (FA1). An inverse relationship between E₄/E₆ ratios and aromaticity as determined by ¹³C NMR spectra was observed for HA and FA from black soil. The ¹³C NMR spectroscopy revealed that HA2 was more aromatic than HA1. On the other hand, FA1 exhibited a higher aromaticity than FA2.     

FTIR‐spectroscopic characterization of humic acids and humin fractions obtained by advanced NaOH,Na4P2O7, and Na2CO3 extraction procedures

Aim of our study was the development of the methodological basis for the characterization of humic fractions of a long‐term field experiment. Humic acids (HAs) were extracted from three layers of a nontilled soil using three different extractants (1 M NaOH, 0.1 M Na₄P₂O₇, 1 M Na₂CO₃), and the humin fraction was enriched. NaOH as extractant for FTIR analysis of humic substances yields higher resolved IR spectra, especially in the important regions of stretching vibrations including aromatic and aliphatic groups and in the fingerprint area including amides, aliphats, and aromats than the other extractants. The NaOH extraction has lower extraction yields as compared to Na₄P₂O₇ and Na₂CO₃ and represents a different part of the soil organic matter (SOM). This is reflected by lower C : N ratios and higher E₄ : E₆ and fulvic acid–to–humic acid ratios as compared to the other extractants. The FTIR band areas of HA fraction obtained by NaOH showed an increase of the aromatic and carbonyl groups and a decrease of amide groups with increasing soil depth. Aliphatic groups showed contradicting results: The bands of the stretching vibrations increased, and the band of the bending vibrations decreased. We assume that band interactions in the bending vibrations were responsible for that phenomenon under the assumption of an increase of aliphatic groups with increasing soil depth. The IR bands of the enriched humin fraction showed a decreasing trend in case of both aliphatic bands deriving from stretching vibrations and an increase of aromatic characteristics with depth. Our study led to the conclusion that HA fractions obtained by 1 M NaOH represent a small and dynamic fraction indicated by the measured yields in combination with values of N_t, C : N, E₄ : E₆ ratios, and ratios of fulvic acids (FA) to HA. The humin fraction has a high contribution to the total organic C and represents a more stabilized fraction of SOM which still shows changes in its aromatic and aliphatic characteristics with soil depth.     

Soil organic matter alteration under biochar amendment: study in the incubation experiment on the Podzol soils of the Leningrad region (Russia)

Purpose

Biochar is one of the most widely used ameliorants for soil amendment, which is known as factor which rises crop yields and levels of soil biological activity. Nowadays, it is under investigated how biochar application affects the dynamics of the humic components and whole soil organic matter (SOM) and the processes of its alteration. This investigation is aimed to evaluate the influence of biochar on the content, composition, and transformation of humic acids (HAs) as the main component of the SOM.

Materials and methods

The incubation experiment was carried out on three Podzol Antric soils, with varying amounts of initial total organic carbon. The incubation time was 90 days, using biochar gravimetric doses of 0.1 and 1.0%. The biochar was produced by fast pyrolysis of birch and aspen wood at 550 °С. Humus composition was analyzed for the organic matter fractions extracted with 0.1 M NaOH (containing HAs 1 + fulvic acids (FAs) 1) and 0.1 M Na₄P₂O₇ (containing HAs 1 + FAs 1 + HAs 2 + FAs 2). Isolated HAs were characterized for their elemental composition (C, N, H, and S) and molecular composition with the use of solid-state 13C nuclear magnetic resonance (13C-NMR) techniques.

Results and discussion

We found that 0.1% of biochar amendment does not influence SOM mineralization, but 1.0% of biochar increases the mineralization by 15–18%. This process is accompanied by changes in the composition and properties of the HS. The increased proportion of HA aromatic fragments in biochar indicates an increasing of their stability. However, in soils with high humus content and a significant amount of insoluble matter, the processes of mineralization and the growth of HAs are taking place simultaneously. The replenishment of HAs could be the outcome of both the intensification of the transformation processes (mineralization and humification) of the more sustainable insoluble matter compounds and the humification of the biochar itself.

Conclusions

The influence of biochar on humification in Podzol Antric soils was revealed on the basis of incubation experiment. Both negative and positive changes under biochar in HS system were demonstrated. The active decrease of humus total contents and also the labile HS ought to qualify as negative changes. The increase of HA chemical maturity that leads to the stability of humus in whole as well as the intensive new HA formation thought to qualify as positive changes.

     

Quantitative and qualitative characterisation of humic products with spectral parameters

Purpose

The application of different humic products for the treatment of soils and plants has increased in recent years. The characteristics of humic products, such as the content and composition of organic carbon and the maturity, provide valuable information which is essential for an adequate application. Such information is crucial for manufacturers, business consultants and users involved in the production, distribution and implementation of humic products. This article presents the correlation between the quantitative indicators of commercial humic products and their spectral characteristics via measurements in the ultraviolet spectrum at 300 nm, in the visible area at 445 and 665 nm and in the near-infrared spectrum at 850 nm.

Materials and methods

We evaluated humic products (liquid and solid) of different origins. Via wet combustion, the content of total organic carbon in humic products can be determined. The precipitation of humic acids from the starting solution determines the composition of the humic products in terms of humic acids (HAs) and fulvic acids (FAs). The dissolution of HAs determines their concentration by titration, while the specific extinction can be assessed via spectrophotometry via measuring the absorption of HAs spectra at the following wavelengths: 300, 465, 665 and 850 nm. The degree of aromaticity and condensation of humic products determines the optical density of the HAs via the E₄/E₆ ratio.

Results and discussion

The content of total organic carbon varied widely from 0.55 to 37.5% across all groups. The content of carbon in HAs, as a percentage of the total carbon in fulvic-type humic products, ranged from 1.29 to 16.00%, while in humic-type products, it ranged from 51.43 to 91.92%. The minimum value of the E₄/E₆ ratio was 2.97, while the maximum value was 6.35. We observed a direct relationship between the dominant type of acids in humic products and the E₄/E₆ ratio.

Conclusions

The optical density of HAs indicates their quality characteristics. The presented optical characteristics for humic products show that there is a direct relationship, especially between HAs/FAs and E₄/E₆ ratios. Measurement at 300 nm (E₃₀₀) in the near-ultraviolet area and at 850 nm (E₈₅₀) in the near-infrared area can increase the range of the spectral study.

     

Exploratory analysis of the structural variability of forest soil humic acids based on multivariate processing of infrared spectral data

Semi‐quantitative data of peak intensities in infrared (IR) spectra of humic acids (HAs) from semiarid soils under contrasting environmental conditions (vegetation type, geological substrate and local climate) were analysed by multivariate data treatments. Resolution‐enhanced IR spectra (applying a second derivative‐based subtractive operator) showed a typical lignin pattern, which was coded to obtain an index used to classify the degree of diagenetic alteration of the lignin moiety in the HA fraction. Partial least squares regression (PLS) was used in the exploratory screening for supervised data reduction previous to other multivariate data treatments as well as to identify IR peaks responsive to soil dependent variables. Regression models and multi‐dimensional scaling (MDS) were applied in order to classify individual IR peaks or sets of peaks associated with the degree of diagenetic alteration of organic matter, or inform on soils' potential for carbon (C) accumulation. Soil properties co‐varying with the intensities of these peaks were mainly related to soil texture and consequently to water holding capacity at different pressures. Principal component analysis (PCA) based on the IR peaks selected in the previous PLS treatments maximized differentiation in terms of the impact of environmental factors on HA characteristics: (i) vegetation type (angiosperms or gymnosperms), (ii) the effect of the geological substrate (granite or limestone) on soil organic matter dynamics and (iii) soil taxonomical differences reflected by independent clusters. The successful forecasting of several factors related to soil C sequestration indicated the validity of the semi‐quantitative information extracted from the IR spectra of the HAs and the potential of the multivariate data treatments used to identify biogeochemical proxies of the soil organic matter stabilization processes.     

Effects of pig slurry application on soils and soil humic acids

The effect of three annually consecutive additions of pig slurry at two rates (90 and 150 m3 x ha(-1) x year(-1) on soils and soil humic acids (HAs) was investigated in a field experiment under semiarid conditions. Soils and pig slurries were analyzed by standard methods. The HAs were isolated from soils and pig slurry by a conventional procedure based on alkaline extraction, acidic precipitation to pH 1, purification by repeated alkaline dissolutions and acidic precipitations, water washing, dialysis, and final freeze-drying. The HAs obtained were analyzed for elemental (C, H, N, S, and O) and acidic functional group (carboxylic and phenolic) composition, and by UV-vis, FT-IR, fluorescence, and ESR spectroscopies. With respect to the control soil, the pig slurry amended soils had greater pH and electrical conductivity, slightly larger total N content, and smaller values of C/N ratio. A decrease of total organic C was observed only in soils amended for 2 and 3 years at the higher slurry rate. With respect to control soil HA, pig slurry HA was characterized by larger contents of S- and N-containing groups, smaller acidic functional group and organic free radical contents, a prevalent aliphatic character, extended molecular heterogeneity, and smaller aromatic polycondensation and humification degrees. Amendment with pig slurry HA determines a number of modifications in soil HAs, including increase of C, S, and COOH contents, C/N ratios, and aliphaticity and decrease of extraction yields and N, O, phenolic OH, and organic free radical contents. These effects are generally more evident after the first year of slurry application and tend to disappear with increasing number of treatments. Most probably, over the years the slightly humified slurry HA is mineralized through extended microbial oxidation, whereas only the most recalcitrant components, such as S-containing, phenolic, and aliphatic structures, are partially accumulated by incorporation into soil HA.