The chemical nature of components of PG (Green fraction of P type humic acid)

P type humic acids showing characteristic absorption maxima at 615, 575, and 450 nm have been found in various types of soils around the world (3, 5, 7,8). P type humic acid was fractionated into a brown fraction (Pb) and a green fraction (Pg) by column chromatography using cellulose powder (4) or Sephadex gel (3). The Pg fraction which has a strong absorption maxima at 615, 575, and 450 nm causes the characteristic absorption pattern of P type humic acid.     

The distribution of PG (green fraction of P type humic acid) and the degree of humification of PB (brown fraction of P type humic acid) in soils of central Japan

Abstract

Pg contents of soils chosen from the major soil groups occurring in the mountain area of Central Japan were measured by the method presented by the author in a previous paper. The Pg contents of the soils ranged from 0 to 0.58 per cent of the dry soils. In podzolic soils, Pg contents were high in A horizons and gradually decreased with depth and in some cases of brown forest soils, Pg contents were highest in B horizons. The percentage of Pg in humic acid ranged from 0 to 8.6 per cent, and the highest value was observed in the C horizon of Dando Bo soil, Pg contents were high in humid and acidic soils containing much humic acid. Humidity, soil pH and humus content may affect the Pg accumulation by controlling the activity of Pg-producing fungi. ? log K values of Pb were calculated to estimate the degree of humification of P type humic acids and indicated that Pb's were relatively immature.     

Characterization of the perylenequinone pigments in Japanese Andosols and Cambisol

The green fraction of humic acid (Pg) and the chloroform-extractable green fraction (CEGF) are characteristic soil organic matter (SOM) components. These alkaline solutions are green-colored due to the presence of 4,9-dihydroxyperylene-3,10-quinone (DHPQ) chromophore. While both of which are potential indicators for the effect of land use and paleoclimatic environment in the fields of soil science as well as geochemistry, CEGF as well as its relationship with Pg in soils are not yet fully documented. In this study, we firstly investigated the chemical properties of soil CEGF fractions by ultraviolet–visible (UV–Vis) and infrared (IR) method. Two CEGF components were separated by sequential liquid-liquid extraction using aqueous ammonium hydroxide (NH₄OH) followed by aqueous sodium hydroxide (NaOH). Results showed that the UV–Vis spectral shape of NH₄OH-extractable component is very similar to that of DHPQ, except that it is red-shifted. The solubility and UV–Vis spectrum of the NaOH-extractable fraction were completely identical with those of synthesized DHPQ. Their IR spectral shapes were also almost the same. Subsequently, the distribution of CEGF in humic acid (HA), fulvic acid (FA) and humin (HN) from Japanese Andosols and Cambisol was quantitatively evaluated by sequential extraction. Most of CEGF was detected in the HA (60–78%) and HN (22–40%), but not in the FA. While the UV–Vis spectral shape of CEGF extracted from Andosols HAs showed a relatively higher proportion of DHPQ than its derivative, the opposite was observed in Cambisol HA, whose CEGF is similar to that of sclerotium grain (one of the possible origin of CEGF). These results suggest the diversity of CEGF-producing soil fungi. Quantitative data also indicated that 35–49% of Pg consisted of a chloroform-soluble fraction (i.e., CEGF) and the remaining 51–65% of Pg was chloroform-insoluble. Based on these results, we propose that CEGF is composed of DHPQ and DHPQ-derivatives and that CEGF is one of the major fractions of Pg.     

Methods for estimating Pg content in P type humic acid and for calculating Δlog K of its Pb fraction

Based on the assumption that P type humic acid is composed of green fraction (Pg) and brown fraction (Pb),. methods for estimating Pg content and calculatlng Δlog K of Pb by analyzing the absorption curve of P type humic acid were presented. A green fraction of P type humic acid obtained from an alpine meadow soil by gel filtration was used as the Pg standard (λmax 450, 575, and 615 nm, E 1% 290, 126, and 168).

Alkaline solutions of standard Pg obeyed the Beer-Lambert law within the concentration range of 1.6 to 31 ppm. To verify the methods, Pg content in the mixed solutions of Pg and A, B, or Rp type humic acid was estimated. It was found that the recovery of Pg would be 95±14 (2S) per cent with a probability of 0.99. Also, the calculation of Δlog K of humic acid in the mixed solutions gave satisfactory results.

It was distinctly shown that the shape of absorption spectra of P type humic acid did not only depend on Pg content, but also on Δlog K of Pb.     

Chromatographic Separation of Green Humic Acid from Podzol Humus

One of the authors, Kumada ¹⁾ has found that absorption spectra of podzol humic acids, remarkably different from the other soil humic acids, have fairly distinct absorption bands near 620, 570 and 450 mµ, and he tentatively named them. P type humic acid. P type humic acid seems specific not only in its visible absorption spectra, but also in infra. red absorption spectra, in susceptible to electrolytes, etc. The authors are now engaged in the study of the nature and properties of P type humic acid, and recently they have found that it is composed of at least two main fractions, the one is reddish brown, and the other green, in alkaline solution. The experimental results are briefly presented.     

Studies on the humus forms of forest soils

The results of humus composition analysis of each fraction separated physically from each horizon of the five forest soils used in the last paper (3) were presented.

Regular changes in humus composition were observed with the gradation of horizons and With the decrease in particle size. Most of the humic acids from plant residual fractions belonged to the Rp type; hurnic acids from the L layers showed a tannin-like character which disappeared with the progress of decomposition, and humic acids from the smaller particle fractions of the layers contained a Pg fraction.

As for humic acids from the A horizon, they all belonged to the P or B type and contained a Pg fraction. In addition, it was concluded that the humification process in the mineral layer Was clearly different from that in the organic layer.     

Humus composition of soils under forest, coffee and arable cultivation in hilly areas of south Sumatra, Indonesia

To understand the effect of land use changes on the composition of humus in tropical soils, samples from land under primary forest, secondary forest, coffee plantation, and arable crops were investigated at three sites in south Sumatra, Indonesia. Total carbon and total nitrogen contents were 1.7 to 4.3 times and 1.1 to 2.8 times greater in the topsoil under primary forest than under the other types of land use. Following change from primary forest to other uses, the proportion of humic acids in the organic matter of the topsoils decreased while that of the fulvic acid fraction increased. Within the range of land uses, differences in the yields of humic acids and fulvic acid fractions were, respectively, larger and smaller than those in total carbon content. The humic acids were classified into the low and middle classes in the degree of humification. Absorption due to the green fraction of humic acids, Pg, was detected in the UV‐visible spectra of almost all the humic acids. No relation was observed between the degree of humification of humic acids or the strength of Pg absorption in their spectra and land use change. The fulvic acid fractions were fractionated on insoluble polyvinylpyrrolidone (PVP) into the adsorbed fractions consisting of humic substances and the non‐adsorbed fractions consisting of non‐humic substances. A positive correlation between the amount of the fulvic acid fraction and the percentage of the PVP‐adsorbed fraction within it indicated that the variation in the amount of the fulvic acid fraction was attributable to acid‐soluble humic substances. The ionization difference spectra of solutions between pH 12 and pH 7 suggested that the chemical structures of the PVP‐adsorbed fulvic acids have been altered by land use change.     

Comparison of humus composition of charred Susuki (Eulalia,Miscanthus sinensis) plants before and after HNO3 treatment

Ando soils (Kurobokudo), which are characterized by the accumulation of large amounts of black humus, are widely distributed in Japan, and the black color is due to the presence of A type humic acids with a high degree of humification (Kumada 1987).     

Adsorption of Acrylonitrile on Some Soils and Minerals from Aqueous Solutions

Equilibrium and kinetic studies have been made on the adsorption of acrylonitrile(CH2=CHCN) on three soils and four minerals from aqueous solutions.It was shown that the organic matter was the major factor affecting the adsorption process in the soils.The conformity of the equilibrium data to linear type(one soil) and Langmuir type(two soils) isotherms indicated that different mechanisms were involved in the adsorption.This behavior appears bo be related to the hydrophobicity of soil organic matter due to their composition and E4/E6 ratio of humic acids.The adsorption kinetics were also different among the soils,indicating the difference in porosity of organic matter among the soils,and the kinetics strongly affected the adsorption capacity of soils for acrylonitrile.Acrylonitrile was slightly adsorbed from aqueous solutions on pyrophyllite with electrically neutral and hydrophobic nature,and practically not on montmorillonite and kaolinite saturated with Ca.However,much higher adsorption occurred on the zeolitized coal ash,probably caused by high organic carbon content(107g/kg).     

The involvement of iron and aluminum in the bonding of phosphorus to soil humic acid

Abstract

With a peat soil similar amounts of phosphorus (P) were coprecipitated with humic acid from alkali extracts over a limited range of strongly acidic pH, whereas with a mineral soil the amount was pH dependent. The difference between the two soils relates to the much greater total amounts of inorganic P and aluminum (Al) present in the extract of the mineral soil. In this acid mineral soil, Al rather than iron (Fe) may be involved in the formation of metal bridges in humic acid‐metal‐inorganic P complexes. Neither Al or Fe were implicated in binding of organic P to humic acid. The P species observed in humic acids was dependent on the pH at which they were precipitated from the alkali extracts. In the peat soil the inorganic P was an order of magnitude lower than the organic P.     

Determination of DTNB-reactive sulphhydryl groups in soil humic acids: their enrichment in humic acids from volcanic acid soils

Thirteen acid soils were collected from typical volcanic regions in Japan (S content: 0.9–4.1, mean = 2.2g kg^?1; pH (H₂O): 2.81–3.93, mean = 3.33), as well as nine reference soils (S: 0.6–1.7, mean= 1.1 gkg^?1; pH(H₂O): 4.10–4.74, mean = 4.47). Humic acids were extracted from the soils separately with 0.1 M NaOH and precipitated by acidification (pH = 2, HCl). After purification, the humic acids were subjected to colorimetric analysis using a DTNB reagent [5,5′-dithiobis(2–nitrobenzoic acid] for the active -SH group. Since humic acids have significant absorption at 4I2 nm, the coloured compound (5–mercapto-2–nitrobenzoic acid) was separated from the humic acids by ultrafiltration or solvent extraction prior to the colorimetric measurement. Humic acids also caused discoloration of the coloured compound when they coexisted in the reaction solutions. Thus, the reproducible determination of -SH was accomplished by employing a standard addition technique (-SH standard: cysteine). Although -SH contents obtained by the ultrafiltration method were considerably higher than those by the solvent-extraction method, probably due to the denaturation of humic acids by the higher buffer concentration used in the ultrafiltration method, they correlated well with each other. The humic acids from acid soils contained apparently higher concentrations of -SH (120–510, mean = 270mg S kg^?1 by the ultrafiltration method; 8–110, mean = 38mg S kg^?1 by the solvent-extraction method) compared to those from reference soils [20–260, mean = 90mg S kg^?1 by the former; not detectable (<5)-34, mean = 11 mg S kg^?1 by the latter]. This -SH enrichment in the humic acids from acid soils may result from the degradation and subsequent humification of S-rich debris of plants and micro-organisms and/or direct incorporation of volcanic acid gas (H₂S) into the humic acids.     

The humus status of brown forest soils and its changes upon intensive tea growing in the subtropical zone of Russia

Changes in the humus status of acid brown forest soils used for tea growing have been studied in a long-term stationary experiment after 20 years of the application of mineral fertilizers in increasing doses. It is shown that long-term tea growing has resulted in a reliable increase in the humus content within the upper cultivated horizon (Ap, 0–40 cm) at the expense of the nonhydrolyzable fraction of humus (humin) and the first fractions of humic and fulvic acids. The fractional composition of humus has changed against the background of a stable fulvate type of humus (Cha/Cfa < 0.5). The portion of the first fraction of humic and fulvic acids in the total amount of humus has increased, as well as the portion of the first fraction of humic acids in the total amount of humic acids. These changes are due to the enhanced humification of plant residues with a parallel decrease in the contents of the second (Ca-bound) and third (bound with clay and stable sesquioxides) fractions of humic substances.     

ELECTRON-SPIN RESONANCE OF HUMIC ACIDS FROM CULTIVATED SOILS

An examination has been made of the electron-spin resonance spectra of humic acids from (i) two groups of cultivated soils from the north-east of Scotland, and the related natural soils from which the cultivated soils are considered to be derived, and (ii) some Rothamsted soils. With the Scottish soils the humic acids from the more acid natural soils (pH <4.6) show e.s.r. spectra with a four-peak structure (class I), and those of the less acid soils (pH >4.6) almost structureless spectra (class II). The cultivated soils contain humic acids with the same class of spectrum as that of the soils from which they are derived, even though they are now much less acid (_PH 5.9 –6.5). Humic acids from Rothamsted soils, sampled recently, were compared with those from the same site sampled 80 years previously. Where the soil has been allowed to revert towards a natural state but still contains free calcium carbonate the pH remained high (7.8–7.9) and the spectrum of the humic acid remains class II. Where the soil contained no free calcium carbonate the pH has fallen from 7.1 to 4.5 on reversion and the spectrum class of the humic acid has changed from II to I. The results support the hypothesis that the class of spectrum of the humic acid depends on the pH at which it is formed.     

Effects of prolonged irrigation on the humus of steppe soils in southwest Siberia

Changes in humus resulting from up to 20 years' irrigation with slightly saline water were studied in dry steppe soils in southwest Siberia. The amounts of brown humic acids and of the Pg fraction increased with the period of irrigation, and their maximum amounts occurred at greater depths in the profile. Other fractions of humus and humic acids fluctuated within the limits typical of dry steppe soils. The humus therefore remained essentially stable under irrigation for up to 20 years.     

Sorption and desorption of triadimefon by soils and model soil colloids

Sorption-desorption of the azole fungicide triadimefon [1-(4-chlorophenoxy)-3,3-dimethyl-1-(1H-1,2, 4-triazol-1-yl)-2-butanone] on eight soils and a series of single, binary, and ternary model soil colloids was determined using the batch equilibration technique. Regression analysis between Freundlich sorption coefficients (K(f)) and soil properties suggested that both clay and organic C (OC) were important in triadimefon sorption by soils, with increasing importance of clay for soils with high clay and relatively low OC contents. Triadimefon sorption coefficients on soil were not significantly affected by the concentration of electrolyte or the presence of soluble soil material in solution, but they were highly dependent on the soil:solution ratio due to the nonlinearity of triadimefon sorption on soil. Freundlich sorption isotherms slopes were very similar for all soils (0.75 +/- 0.02). Desorption did not greatly depend on the concentration at which it was determined and showed higher hysteresis for more sorptive soils. Results of triadimefon sorption on model sorbents supported that both humic acid and montmorillonite-type clay constituents contribute to triadimefon retention by soil colloids.     

Organic matter of technogenic soils and substrates on the dumps of sulfidic mine rock (carbonaceous shale) in Western Donets Basin

Data on the total carbon content and on the composition of humic substances extracted from the technogenic soils and substrates of dumped mine rocks are analyzed. It is shown that two types of humic substances are formed upon the oxidation of sulfidic rocks and their overgrowing with vegetation. The first type is represented by humic substances originating in the course of transformation of plant remains. The second type includes humic substances formed due to the oxidation of coal particles; these humic substances are characterized by a higher C_ha/C_fa ratio. The role of humic acids in decreasing the mobility of aluminum ions in the strongly acid mine substrate is experimentally confirmed.     

THE OCCURRENCE OF COPPER-PORPHYRIN COMPLEXES IN SOIL HUMIC ACIDS

Electron paramagnetic resonance (e.p.r.) spectroscopy has been used to show that humic acids, which were isolated from a range of soils and from which most of the iron was removed by boiling with hydrochloric acid, contain copper in the form of porphyrin complexes. The amounts of porphyrin-bound copper in the acid-boiled humic acids were estimated to be ≤ 20 mg/kg (equivalent to 0.05-0.9 mg/kg in the soils). The acid-boiled humic acids from cultivated mineral soils showed no great capacity for further uptake of copper as copper porphyrin when treated with CuSO₄, most of this additional copper being held by groups not involving N-donor groups. In contrast, copper treatment of the acid-boiled humic acid from a raised bog peat produced a large increase in copper-porphyrin signal strength, indicating that in the peat there are considerable amounts of porphyrin by which added copper could be complexed. It is suggested that porphyrin groups may be involved in the fixation of copper by organic matter in soils.     

Nitrogen forms in 1- to 7-year-old opencast lignite mine soils

We studied the distribution of different forms of N in very young (0- to 7-year-old) soils from the Meirama lignite mine in northwest Spain. Total N increased rapidly with soil age, largely in the humic acid-associated fraction. Acid hydrolysis indicated that amino acid N and a hydrolysable unknown form of N increased with soil age. The fractionation reported by Bettany et al. (1980) indicated that alkali/pyrophosphate-extractable humus increased with soil age. All these data suggest that there is rapid stabilization of organic N during the first few years of evolution of these mine soils, to the extent that distribution of N forms in the 7-year-old soil is very similar to that in native soils.     

Analysis of the green fraction of humic acids

The green fraction of humic acids (HAs), Pg, was fractionated by gel chromatography on Sephadex G-50. Repeated chromatography of the crude Pg obtained by the first chromatography of HA yielded a sharp peak (G₁) and two broad bands (G₂) of purified Pg. The recovery of Pg in G₁ and G₂ as percentages of the total Pg content of the HA were 16.2 and 14.5%, respectively. Distinct peaks were detected at 615, 570, 450 and 280 nm in the UV-visible spectra of both G₁ and G₂ in alkali solutions. Estimates of weight-average molecular weight were 9.7 * 10³ for G₁ and 1.23 * 10⁴ for G₂. In the ¹H-NMR spectra the percentage of hydroxyl groups of 4,9-dihydroxy-perylene-3, 10-quinone (DHPQ) nuclei, which are known to be the chromophore of Pg, was 4.1 and 4.5% of the total H, respectively. Based on these values and the H/C ratios, DHPQ-C was estimated as 28% of total C. ¹³C-NMR spectra obtained using inverse-gated decoupling yielded 40–41% aromatic C, which suggested the presence of aromatic rings other than DHPQ. All of the IR, ¹³C- and ¹H-NMR spectra indicated that the two purified Pg samples largely differed from each other in their contents of alkyl and polysaccharide components.     

Purification of humic acid by the use of ion exchange resin

Abstract

The humic acid fraction, extracted from soil by alkali or neutral salt solution and acid-recipitated, is generally contaminated with a considerable amount of inorganic matter. The presence of the inorganic matter is often undesirable for studies on humic acid. The inorganic matter is generally taken off by ultrafiltration or electrodialysis. But these methods take time and are unsuitable for the bulk preparation of humic acid.