Structural investigations of Australian coals—I. A characterization of Victorian brown coal lithotypes and their kerogen and humic acid fractions by i.r. spectroscopy

The functional groups of brown coal lithotypes, their humic acids and kerogens were investigated by elemental analysis and quantitative infrared spectroscopy using absorption coefficients.An inverse relationship is reported between the concentrations of kerogen and humic acids for each lithotype and significant variation in the levels of humic acids with lithotype are attributed to changing higher plant input and depositional environments.The structural implications of observed trends in absorption coefficients and their usefulness as a rapid method of functional group analysis has been investigated. Using this infrared technique, kerogen structures as reflected in functional group analyses were found to exhibit a dependance on lithotype, although no significant variation in the functionality of the humic acid structures was observed.     

Oxygen-containing functional groups in land-derived humic acids—I. Evaluation by derivatization methods

Determinations of reactive oxygen-containing functional groups were performed by usual chemical derivatization methods of four humic acids isolated from soils and deltaic sediments. Statistical tests and calculations based on elemental analysis demonstrated the validity and limits of these techniques and allowed us to identify from 60 to 71% of the total humic oxygen. Oxygen budgets of humic acids are discussed.     

Sources of sedimentary humic substances: vascular plant debris

A modern Washington continental shelf sediment was fractionated densimetrically using either an organic solvent, CBrCl₃, or aqueous ZnCl₂. The resulting low density materials (<2.06 g/ml) account for only 1% of the sediment mass but contain 25% of the sedimentary organic carbon and 53% of the lignin. The C/N ratios (30–40) and lignin phenol yields ($\text{Λ = 8}$) and compositions indicate that the low density materials are essentially pure vascular plant debris which is slightly enriched in woody (versus nonwoody) tissues compared to the bulk sediment. The low density materials yield approximately one-third of their organic carbon as humic substances and contribute 23% and 14% of the total sedimentary humic and fulvic acids, respectively. Assuming that the lignin remaining in the sedimentary fraction is also contained in plant fragments that yield similar levels of humic substances, then 50% and 30% of the total humic and fulvic acids, respectively, arise directly from plant debris.Base-extraction of fresh and naturally degraded vascular plant materials reveals that significant levels of humic and fulvic acids are obtained using classical extraction techniques. Approximately 1–2% of the carbon from fresh woods and 10–25% from leaves and bark were isolated as humic acids and 2–4 times those levels as fulvic acids. A highly degraded hardwood yielded up to 44% of its carbon as humic and fulvic acids. The humic acids from fresh plants are generally enriched in lignin components relative to carbohydrates and recognizable biochemicals account for up to 50% of the total carbon. Humic and fulvic acids extracted directly from sedimentary plant debris could be responsible for a major fraction of the biochemical component of humic substances.     

Stability, solubility and maximum metal binding capacity in metal–humic complexes involving humic substances extracted from peat and organic compost

The aim of this work is to investigate the influence of pH and the metal:humic substances (HS) ratio on HS complexing capacity and the stability and solubility of metal–HS complexes in solution. We selected four HS with different physicochemical properties and studied their interaction with Cu(II), Zn(II) and Fe(II) at different pH and metal:HS ratios. The selected HS were a humic acid and a whole humic extract (containing the humic and fulvic acids) extracted from black peat, and a fulvic acid and a whole humic extract extracted from a compost of grape solid wastes.Our results showed that HS complexing capacity significantly varied as a function of pH, thus indicating the influence of both functional group ionisation and molecular conformation on this property. As was expected, total acidity affected the complexing capacity of the selected HS.The results related to stability and complexing capacity indicated the possible presence of two binding patterns, one at acid-neutral pH probably involving carboxylates, and another at alkaline pH probably involving carboxylates and phenolic groups. The relationship between these binding patterns and the strength of the binding process varied according to the complexed metal.Complex solubility was greatly affected by the ratio between the concentration of free ionised functional groups and the molecular weight in the HS studied.     

Cu and Fe associated with humic acids in sediments of a tropical coastal lagoon

The amount of Cu and Fe associated with humic acids was estimated in five sediment cores from a tropical coastal lagoon (Piratininga Lagoon, Rio de Janeiro, Brazil). Core samples were analysed for humic acid contents, total Fe and Cu content. Fe and Cu associated with humic acids were also measured. Results show amounts of humic acids ranging from 0.7 to 21.7% of the dry weight of sediment (average 4.6%, standard deviation 4.4%). Concentrations of Fe and Cu ranged from 0.3 to 6.0% (average 2.2%, S.D. 1.2%) and from <1.0 to 65.0 μg g⁻¹ (average 28.6 μg g⁻¹, S.D. 16.4 μg g⁻¹), respectively. The results of strongly bound metals show that while humic acids are the main carrier for Cu, Fe does not seems to be significantly associated with this organic matter.     

Methylation patterns of aquatic humic substances determined by13C NMR spectroscopy

¹³C NMR spectroscopy is used to examine the hydroxyl group functionality of a series of humic and fulvic acids from different aquatic environments. Samples first are methylated with¹³C-labeled diazomethane. The NMR spectra of the diazomethylated samples allow one to distinguish between methyl esters of car☐ylic acids, methyl ethers of phenolic hydroxyls, and methyl ethers of phenolic hydroxyls adjacent to two substituents. Samples are then permethylated with¹³C-labeled methyl iodide/NaH.¹³C NMR spectra of permethylated samples show that a significant fraction of the hydroxyl groups is not methylated with diazomethane alone. In these spectra methyl ethers of carbohydrate and aliphatic hydroxyls overlap with methyl ethers of phenolic hydroxyls. Side reactions of the methyltion procedure including carbon methylation in the CH₃I/NaH procedure, are also examined. Humic and fulvic acids from bog, swamp, groundwater, and lake waters showssome differences in their distribution of hydroxyl groups, mainly in the concentrations of phenolic hydroxyls, which may be attributed to their different biogeochemical origins.     

Nonaqueous titration of functional groups in humic acid

Humic acid was titrated by sodium methoxide in dimethylsulfoxide using platinum-calomel electrode systems. Adding benzoic acid and phenol as internal standards to humic acid yielded two inflections. The titer at the first inflection point was equivalent to the carboxyl groups whose pK_a (H₂O) values were less than 7. The difference between the titers at the two inflection points was equivalent to the phenolic hydroxyl groups whose pK_a (H₂O) values were 7–10. Calculated results for the carboxyl and phenolic hydroxyl groups in humic acid obtained by the nonaqueous titration method agreed closely with those obtained by conventional methods.     

Structural investigations of Australian coals—III. A 13C-NMR study on the effects of variation in rank on coal humic acids

A quantitative ¹³C-NMR technique was applied to humic acids, isolated from total solvent extracts obtained from coals, varying in rank from lignite to medium volatile bituminous. The results from elemental analysis and ¹³C-NMR structural ratios are interpreted in the form of three distinct structural phases in the maturation of coal humic acids. These phases reflect the rank of the parent coals. A decrease in polar functional group content and corresponding increase in aromaticity is observed with increasing maturation of humic acid chemical structure. Limits are postulated for the oxygen content and level of aromaticity for humic acids derived from higher rank coals.     

The lignin component of humic substances: Distribution among soil and sedimentary humic,fulvic, and base-insoluble fractions

Vanillyl, syringyl and cinnamyl phenols occur as CuO oxidation products of humic, fulvic and base-insoluble residual fractions from soils, peat and nearshore marine sediments. However, none of these lignin-derived phenols were released by CuO oxidation of deepsea sediment or its base-extractable organic fractions. Lignin analysis indicated that peat and coastal marine sediments contained significantly higher levels of recognizable vascular plant carbon (20–50%) than soils and offshore marine sediments (0–10%).Although accounting for less than 20% of the total sedimentary (bulk) lignin, lignin components of humic acid fractions compositionally and quantitatively resembled the corresponding bulk samples and baseinsoluble residues. Recognizable lignin, presumably present as intact phenylpropanoid units, accounted for up to 5% of the carbon in peat and coastal humic acids but less than 1% in soil humic acids. Fulvic acid fractions uniformly yielded less lignin-derived phenols in mixtures that were depleted in syringyl and cinnamyl phenols relative to the corresponding humic acid fractions.Within the vanillyl and syringyl families the relative distribution of acidic and aldehydic phenols is a sensitive measure of the degree of oxidative alteration of the lignin component The high acid/aldehyde ratios and the low phenol yields of soils and their humic fractions compared to peat and coastal sediments indicate extensive degradation of the lignin source material. Likewise, the progressively higher acid/aldehyde ratios and lower phenol yields along the sequence: plant tissues (plant debris)-humic acids-fulvic acids suggest that this pattern represents the diagenetic sequence for the aerobic degradation of lignin biopolymers.     

Characterisation of brown coal humic acids and modified humic acids using pyrolysis gcms and other techniques

It has been proposed that Victorian brown coal can be considered as a two-component structure — a lignocellulosic “host”, containing various amounts of weakly bound or entrapped “guest” material together with very small amounts of inorganic and/or mineral matter. The latter predominantly consists of wax esters and/or terpenoid material. In this paper we describe attempts to gain structural information regarding the more complex, “host” component of the coal. Our initial model compound has been humic acid that can be readily obtained from the coal by alkaline extraction. It has been found that “pure” humic acid, free from material associated with the “guest” components of the coal, can be obtained by a highly selective, low-yielding alkaline extraction. This humic acid has been studied by nmr spectroscopy and pyrolysis gas chromatography-mass spectroscopy (py-gc/ms). The products arising from py-gc/ms have been compared with those obtained from similar pyrolysis of whole coals. Alkylation of humic acids using alkyl halides in the presence of base has been successfully carried out and reactivity of the resulting materials compared with those of the parent coal and humic acid.     

Structural investigations of Australian coals II: A 13C-NMR study of the humic acids from Victorian brown coal lithotypes

The experimental conditions allowing quantitative interpretation of liquid state ¹³C-NMR spectra of the humic acids, derived from the five Victorian brown coal lithotypes are described. The structure of the two classes of humic acids investigated for each lithotype vary significantly in their level of aromaticity, the level of polar functional groups and aliphatic chains attached to their polyaromatic skeletons. Variation is also observed in the degree of aromaticity and oxygen-containing functions of humic acids with lithotype. These data are interpreted in the light of paleobotanical evidence, which suggests that the lithotypes represent variations in depositional environment and input to the coal seam.     

Quantifying the effect of humic matter on the emission of mercury from artificial soil surfaces

A set of surface samples was created using purified laboratory grade sand treated with 0.05 μg/g Hg as the HgCl₂ salt and various concentrations of purified humic and fulvic acids. Emissions of elemental Hg from these substrates to the atmosphere were inversely correlated with the organic content of the samples (99% confidence level). The greatest differences in Hg emissions were found between samples containing the lowest concentrations of humic matter (0% versus 0.01% humic, and 0.01% versus 0.1% humic), only small differences in Hg flux were found to exist for samples with higher concentrations of humic acid (1%, 5%, and 100%). This effect was independent of the type of humic substance used, with both humic and fulvic acids showing an inhibitory effect on surface Hg emissions.     

Aggregation of aquatic humic substances

Humic substances (HS) were isolated from Penwhirn Reservoir (PR) and Esthwaite Water (EW) and their removal from solution by centrifugation was studied as a function of pH, humic concentration and molecular weight, and CaCl₂ concentration. Large amounts (up to 50%) of PR HS could be removed but only small amounts (? 3%) of EW HS. At pH ? 5 removal of PR HS by Ca²⁺ can be explained satisfactorily in terms of decreases in humic solubilities induced by complexation with the cation. However, removal induced by protonation of the PR HS is unusual in that it decreases with increasing humic concentration.The results suggest that PR HS comprise a range of molecules differing in solubility, with the high-molecular-weight (40,000) components being the least soluble. The EW HS consist of molecules of weight-average molecular weight 5000 and resemble similarly sized PR HS in that they remain unaggregated in solution even when highly complexed with Ca²⁺.     

Comparison of the structures of humic substances from aquatic and terrestrial sources by pyrolysis gas chromatography-mass spectrometry

Four humic extracts isolated from terrestrial, mahne, plankton and freshwater sources have been investigated by pyrolysis-gas chromatography-mass spectrometry. The results show that substantial quantities of phenols (some not derived from lignin), pyrroles and nitrites (derived from proteins, nucleic acids or porphyrins) are decomposition products. Small amounts of furans (derived from carbohydrates) and unsaturated ketones (derived from polycarboxylic aliphatic acids) are also present. The results illustrate how parent material, transport factors and biological activity affect the composition of humic extracts.     

Structural relationships between a brown coal and its kerogen and humic acid fractions

A South Australian Tertiary brown coal is fractionated into humic acid and kerogen fractions. These related samples are then subjected to a number of different analytical techniques including infrared and¹³C-Nuclear Magnetic Resonance spectroscopies and pertrifluoroacetic acid oxidation. Structural conclusions are drawn from an integrated consideration of the data. Brown coal aliphatic structure is concentrated in the kerogen and the solvent soluble polar acid fractions. The humic acids are the most aromatic and contain a high degree of hydrogen bonding. Only very minor amounts of long polymethylene chain structures are observed in the humic acids, in complete contrast to the kerogen fraction. Different organic detrital origins are proposed for the coal fractions.     

Pyrolysis-GC-MS and NMR studies of dissolved seawater humic substances and isolates of a marine diatom

¹³C- and ¹H-NMR spectra were obtained for humic substances isolated from a coastal marine environment and also for the intracellular and extracellular extracts of a marine diatom. Phaeodactylum tricornutum Bohlin. Highly branched alkyl chains constitute a large proportion of the structure of the marine humic material, whereas aromatic components are less important. Carbohydrate-type materials, possibly uronic acids, are also present in appreciable amounts. Furans (derived from carbohydrates). pyrroles and nitriles (derived from proteins, nucleic acids and/or tetrapyrroles) and phenols and methylphenols (non-lignin derived) are pyrolysis products derived from pyrolysis-gas chromatography-mass spectrometry of the extracts. The results indicate the similarities in chemical structure of P. tricornutum exudate and dissolved marine humic material.     

Proton-binding study of standard and reference fulvic acids, humic acids, and natural organic matter

The acid-base properties of 14 standard and reference materials from the International Humic Substances Society (IHSS) were investigated by potentiometric titration. Titrations were conducted in 0.1 M NaCl under a nitrogen atmosphere, averaging 30 min from start to finish. Concentrations of carboxyl groups and phenolic groups were estimated directly from titration curves. Titration data were also fit to a modified Henderson-Hasselbalch model for two classes of proton-binding sites to obtain “best fit” parameters that describe proton-binding curves for the samples. The model was chosen for its simplicity, its ease of implementation in computer spreadsheets, and its excellent ability to describe the shapes of the titration curves. The carboxyl contents of the IHSS samples are in the general order: terrestrial fulvic acids > aquatic fulvic acids > Suwannee River natural organic matter (NOM) > aquatic humic acids > terrestrial humic acids. Overall, fulvic acids and humic acids have similar phenolic contents; however, all of the aquatically derived samples have higher phenolic contents than the terrestrially derived samples. The acid-base properties of reference Suwannee River NOM are surprisingly similar to those of standard Suwannee River humic acid. Results from titrations in this study were compared with other published results from both direct and indirect titrations. Typically, carboxyl contents for the IHSS samples were in agreement with the results from both methods of titration. Phenolic contents for the IHSS samples were comparable to those determined by direct titrations, but were significantly less than estimates of phenolic content that were based on indirect titrations with Ba(OH)₂ and Ca(OAc)₂. The average phenolic-to-carboxylic ratio of the IHSS samples is approximately 1:4. Models that assume a 1:2 ratio of phenolic-to-carboxylic groups may overestimate the relative contribution of phenolic groups to the acid-base chemistry of humic substances.     

Compositional and source characterization of base progressively extracted humic acids using pyrolytic gas chromatography mass spectrometry

Cyclic base extraction is a commonly used method for the isolation of humic acids from soils and sediments. However, every extract may differ in chemical composition due to the complex nature of humic acids. To better understand the chemical composition of each extract, the heterogeneous property of humic acids and their speciation in environmental samples, eight fractions of humic acids were obtained in the present work by progressive base-extraction of Pahokee peat, and their chemical composition was characterized using two complementary pyrolytic techniques, namely conventional pyrolysis and methylation pyrolysis (TMAH) GC/MS. These quick and effective procedures provide an insight into the structure of macromolecules. The work shows that the lignin-derived aromatic compounds are major components of pyrolysates in both pyrolytic techniques, while aliphatic compounds originating from microorganisms and plants are minor components. Other compounds derived from proteins and carbohydrates at lower concentrations were also detected. Fatty acids were found in the pyrolysis without methylation, indicating their association with humic acid in a free state. These compounds are different from those formed during pyrolysis with in situ methylation, where fatty acids are generally believed to be the cleavage products of carboxylic groups bound to humic acids. A relative decreasing abundance of aromatic components and increasing abundance of aliphatic components in the pyrolysates as the peat was progressively extracted was also observed in this work, suggesting that the extraction of more hydrophobic aliphatics may be delayed in comparison to the aromatic components. Speciation and origin differences may also be important particularly considering that the contribution from lignin organic matter decreased with extraction number, as the contribution of microbial organic matter increased. The observed change in chemical composition with the extracted fractions indicates again that the humic acid distribution and their speciation are complex, and complete extractions are necessary to obtain a representative humic acid sample.     

Covalent coupling of peptides to humic acids: Structural effects investigated using 2D NMR spectroscopy

Covalent and non-covalent interaction of proteinaceous materials in soils and sediments has been suggested as an important mechanism for immobilizing nitrogen in numerous types of environments. In a previous study (Hsu P.-H., Hatcher, P.G., 2005. New evidence for covalent coupling of peptides to humic acids based on 2D NMR spectroscopy: A means for preservation. Geochimica et Cosmochimica Acta 69, 4521–4533), we provided molecular evidence for covalent, as well as non-covalent, bonding between ¹⁵N-labeled peptides and humic acid molecules using the 2D HSQC (heteronuclear single quantum coherence) NMR technique. In this report, we examine the influence of aromaticity and aliphaticity of peptides and humic materials on these covalent and non-covalent interactions. We use 2D NMR techniques to evaluate bonding interactions of ¹⁵N labeled peptides, having different aromatic and aliphatic properties, with three humic acids that vary in degree of aromaticity. The peptide containing primarily aromatic amino acid residues is observed to form covalent and non-covalent bonds with mainly aromatic-rich humic acids. The peptide composed of aliphatic amino acid residues shows, on the other hand, only bonding interactions with aliphatic-rich humic acids. These observations provide the first direct molecular evidence that aliphatic functional groups are involved in bonding with proteinaceous materials. The process may play an important role in sequestration of proteinaceous materials in sedimentary systems such as marine systems where the humic materials are mainly aliphatic in nature.     

Phosphorus in marine and non-marine humic substances

The phosphorus content of marine humic acids (HA) is in the range of 0.1–0.2%. The C/P ratios of the HA are 300 to 400. Marine fulvic acids (FA) contain 0.4–0.8% P and have C/P ratios of 80 to 100. High molecular weight organic matter dissolved in pore waters (DOM) contains 0.5% P and has C/P of 90. The data suggest that during the formation sequence: Plankton → DOM → FA → HA → Kerogen, phosphorus is lost, mainly in the FA → HA (and possibly also in the HA → Kerogen) step. Diagenesis of sedimentary humic acids is accompanied by loss of phosphorus (as well as of nitrogen) to form HA with C/P ratios of 1000.Soil humic substances resemble marine humates in P content (0.3%) and soil FA's are about three to fivefold enriched in P relative to HA. C/P ratios are lower in soil HA (ca. 200) as compared with marine HA. Humic acids from diagenetic products such as peat and lignite are highly depleted in P. Rough calculations indicate that humate bound P may account for 20–50% of the organic phosphorus reservoir in sediments. The chemical speciation of this P is unknown, but lack of correlation with ash, Fe, Ca or Al content (in marine humates, at least) indicates that it is organically bound.