Humic substances can modulate the allelopathic potential of caffeic, ferulic, and salicylic acids for seedlings of lettuce (Lactuca sativa L.) and tomato (Lycopersicon esculentum Mill.)

The capacity of a leonardite humic acid (LHA), a soil humic acid (SHA), and a soil fulvic acid (SFA) in modulating the allelopathic potential of caffeic acid (CA), ferulic acid (FA), and salicylic acid (SA) on seedlings of lettuce (Lactuca sativa L.) and tomato (Lycopersicon esculentum Mill.) was investigated. Lettuce showed a sensitivity greater than that of tomato to CA, FA, and SA phytotoxicity, which was significantly reduced or even suppressed in the presence of SHA or SFA, especially at the highest dose, but not LHA. In general, SFA was slightly more active than SHA, and the efficiency of the action depended on their concentration, the plant species and the organ examined, and the allelochemical. The daily measured residual concentration of CA and FA decreased drastically and that of SA slightly in the presence of germinating seeds of lettuce, which were thus able to absorb and/or enhance the degradation of CA and FA. The adsorption capacity of SHA for the three allelochemicals was small and decreased in the order FA > CA > SA, thus suggesting that adsorption could be a relevant mechanism, but not the only one, involved in the "antiallelopathic" action.     

土壤腐殖质分组研究

采用Pallo法对两种耕地土壤腐殖质进行分组,初步研究了各组分的数量和结构特征。结果表明:焦磷酸钠提取的胡敏酸(HAp)是胡敏酸(HA)的主要组分;游离富里酸(FAf)是富里酸(FA)的主要组分;胡敏素(HM)的主要组分为非溶解性胡敏素(ISHM),而溶解性胡敏素(SHM)含量较低。通常随土层深度增加,HA、HM各组分的绝对数量和相对数量都下降;FA各组分的绝对数量也下降,而相对数量增加;HA/FA比值下降。一般来看,焦磷酸钠提取的富里酸(FAp)、氢氧化钠提取的富里酸(FAs)比相应的HAp、氢氧化钠提取的胡敏酸(HAs)的分子结构简单,铁结合胡敏素(HMi)比粘粒结合胡敏素(HMc)的分子结构简单;随土层深度增加,HAp、HAs的分子结构变简单,而HMi、HMc的分子结构变复杂。     

Anticlastogenic, antitoxic and sorption effects of humic substances on the mutagen maleic hydrazide tested in leguminous plants

The potential anticlastogenic and antitoxic effects of a soil humic acid (HA), a peat HA and a peat fulvic acid (FA) on the mutagen maleic hydrazide (MH) have been investigated in two legume species, Vicia faba and Pisum sativum. Both HAs and FA were tested at two different concentrations, 20 and 200 mg l^?1, either alone or after 24‐hour interaction with 10 mg l^?1 of MH before addition to the legume seeds. Anticlastogenicity, i.e. an antimutagenic action defined as the capacity for minimizing chromosome breakages, was evaluated by counting both micronuclei (MN) and aberrant anatelophases (AAT) in root‐tip cells. Length and dry weight of the seedling primary root were measured to test the antitoxic activity of HA and FA on MH. The possible occurrence and extent of adsorption or desorption of MH onto or from HA were also investigated. The two species responded differently to the anticlastogenic tests, with V. faba showing a greater number of MN and AAT anomalies than P. sativum. Peat HA and FA exhibited anticlastogenic and antitoxic activities of similar intensity and greater than those of soil HA. The adsorption capacity of both HAs for MH was small, thus suggesting that adsorption is not a major mechanism responsible for the reduction of clastogenicity and antitoxicity of MH by HA.     

Paramagnetic Properties of Humic Substances in Taiga and Tundra Soils of the European Northeast of Russia

The study of paramagnetic activity of humic substances in taiga and tundra soils of the Komi Republic and the assessment of the influence of soil hydromorphism on concentrations of free radicals in the structure of humic acids (HAs) and fulvic acids (FAs) have been performed. The concentration of free radicals in HA specimens was up to 11 times higher than that in FA specimens due to a higher content of aromatic and other condensed structures in HA molecules. This fact attests to the high capacity of HAs to polymerization and complexation reactions with participation of radicals. The average value of g-factor is higher for FA specimens than for HA specimens, which attests to a greater electron density shift of unpaired electron to oxygen atom in the structure of FAs because of its spin-orbital interaction with oxygen-containing functional groups, the concentrations of which are significantly higher in FAs than in HAs. An increase in the concentration of free radicals in the molecular structure of HAs is observed in taiga soils with an increase in the degree of their hydromorphism (from automorphic to semihydromorphic soils), which is related to the biohydrothermal conditions of humus formation in bog-podzolic soils with retarded biochemical processes and low degree of plant litter humification. As a result, HAs with the high content of free radicals in their structure are formed. An opposite situation is observed for HAs in tundra soils with a decrease in the content of unpaired electrons under conditions of the increased hydromorphism. The difference in the character of changes in the paramagnetic activity of HAs in taiga and tundra soils with different degrees of hydromorphism may be related to different natures of plant residues participating in humification processes. A tendency for a decrease in the paramagnetic activity in both HAs and FAs from the south to the north is observed, which may be related to a general decrease in the content of poly-conjugated systems in the structure of humic substances in tundra soils.     

Inhibitory action of dissolved humic substances on the growth of soil bacteria degrading DDT

Abstract

Dissolved humic acid (HA) and fulvic acid (FA) prepared from a Dando brown forest soil (Typic Dystrochrept) inhibited the growth of soil bacteria degrading DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl)ethane) in the culture. The population of DDT-degrading Gram-variable rod Bll6 decreased by the application of both HA and FA, suggesting the presence of bactericidal effect. Such inhibitory effect was stronger for HA and resulted in a lower degrading activity of DDT in the culture of Bll6. No inhibitory effect was observed on the growth of DDT-degrading Bacillus sp. B75. The electron spin resonance spectra showed the presence of organic free radicals in both HA and FA. The relative concentration of the radicals was higher in HA. Storage of HA solution for 3 months at 4°C decreased the concentration of the radicals as well as the inhibitory action. The addition of catalase decreased the inhibitory effect of humic acid. It is suggested that a hydroxy radical, which is derived from free radicals of humic substances, is involved in the inhibition of bacterial growth and degradation of DDT.     

长期施肥对小麦-玉米作物系统土壤腐殖质组分碳和氮的影响

通过对华北平原小麦–玉米轮作农田生态系统18年田间施肥试验,研究了长期不同施肥处理对耕层（0—20 cm）土壤腐殖质及活性腐殖质组分碳和氮的影响。试验设化肥NPK不同组合（NPK、NP、NK、PK）,全部施用有机肥（OM）,一半有机肥+化肥NPK（1/2OMN）及不施肥（CK）共7个处理。结果表明,各施肥处理均能在不同程度上增加土壤腐殖质（胡敏酸、富里酸和胡敏素）及活性腐殖质（活性胡敏酸和活性富里酸）组分碳和氮含量,提高可浸提腐殖质（胡敏酸和富里酸）及活性腐殖质组分碳和氮分配比例;但施肥对土壤活性腐殖质组分碳和氮含量的增加率均分别高于腐殖质组分碳和氮。各处理土壤腐殖质及活性腐殖质组分碳和氮含量均为OM处理最高,且有机肥与化肥NPK配施高于单施化肥各处理;而化肥处理中NPK均衡施用效果最好。说明施用有机肥、有机肥与化肥NPK配施及化肥NPK均衡施用是增加土壤腐殖质及活性腐殖质组分碳和氮的关键;活性腐殖质组分碳和氮较腐殖质组分碳和氮对施肥措施的响应更灵敏。     

Characterization of humic substances derived from swine manure-based compost and correlation of their characteristics with reactivities with heavy metals

Various composts contain a significant amount of humic substances including humic acid (HA) and fulvic acids (FAs). The FA fraction in soils is considered to be sensitive to agronomic and environmental factors. In this study, three fractions of humic substances, HA (MW > 1000 Da), FA (MW > 1000 Da), and FA (MW < 1000 Da) were extracted from swine manure-based compost and characterized, and then, their reactivities were correlated with heavy metals. Compositions of the three fractions of humic substances were characterized by elemental and total acidity analyses and electron spin resonance (ESR), Fourier transform infrared (FTIR), and 13C nuclear magnetic resonance with cross-polarization and magic-angle spinning spectroscopic techniques. Elemental analyses indicated that HA has higher contents of C, H, N, and S than those of FAs. However, FA (MW > 1000) and especially FA (MW < 1000) have higher contents of O than that of HA (MW > 1000). The g values of the ESR spectra of the three fractions showed that the organic free radical characteristics and the widths of the spectra and free radical concentrations of the three fractions are significantly different. The FTIR spectra indicated that HA (MW > 1000) is abundant in C=C bonds while FA (MW > 1000), especially FA (MW < 1000), are abundant in C=O bonds. In addition, 13C NMR spectra indicate that carboxyl contents of FA (MW > 1000), especially FA (MW < 1000), are higher than that of HA (MW > 1000). The sequence of the reactivity in terms of acidic functional groups was FA (MW < 1000) > FA (MW > 1000) > HA (MW > 1000). Elemental and functional group compositions of the three fractions significantly correlated with reported reactivities with heavy metals. The application of swine manure-based compost containing HA and FAs fractions to soil and associated environments may thus significantly affect the concerned reactions with organic and inorganic compounds including pollutants.     

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.     

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.     

Nitrogen Forms in Humic Substances

In this paper,the nitrogen forms in newly-formed humic substances,including humic acid (HA),fulvic acid (FA) and humic acid in humin (HAI),were studied by using the ^15N CP-MAS NMR technique in combination with chemical approaches.Results show that the majority of nitrogen in HA,FA and HAI was in the amide form with some presented as aliphatic and/ or aromatic amines and some as pyrrole type nitrogen,although the contents of nonhydrolyzable nitrogen in them differed greatly from each other (15-55%).     

Chemical and spectroscopic characterization of Humic acids from a soil toposequence in venezuela

Abstract

An investigation was conducted on physico‐chemical properties of humic acids (HAs) in Venezuelan soils. The HAs were extracted by the NaOH method from a Banco‐Bajio‐Estero soil toposequence (local names for soils located at high, intermediate and low topographic levels), in the Venezuelan plains (Mantecal, Apure State). The extracted HAs were analyzed for elemental composition and characterized by fluorescence, Fourier transform infrared (FT‐IR) and electron spin resonance (ESR) spectroscopies. The results showed that free radical concentration of HAs increased from soils at the highest to soils at the lowest topographic position. High carbon (C), nitrogen (N), and carboxyl group contents, E₄/E₆ ratio, aliphatic character and concentration of free radicals, and low oxygen (O) and phenolic hydroxyl group contents and total acidity were typical of HA from soils at the lower relief position. The FT‐IR spectra indicated that the HA from the soil at the lowest topographic position tended to have a slightly higher content of carboxyl groups than the HAs from soils at higher topographic levels. The observed fluorescence was attributed to the presence of condensed aromatic moieties and/or conjugated unsaturated systems of various complexity in the HA macromolecules.     

Conformational changes of humic substances induced by some hydroxy-, keto-, and sulfonic acids

We studied the effect of organic acids of plant, microbial, or anthropic origin on the molecular size distribution of dissolved humic acids (HAs). High Performance Size Exclusion Chromatography (HPSEC) was used to evaluate size changes in four different HAs upon addition of hydroxy- (glycolic and malic), keto- (glyoxylic), and sulfonic (benzenesulfonic and methanesulfonic) acids. All humic substances showed a decrease of peaks absorbance when the pH of HAs dissolved in HPSEC mobile phase was lowered from 7 to 3.5 by acid addition before analysis. This effect, combined with an increase of peaks elution volumes in most cases, was interpreted as a disruption of supramolecular humic associations into smaller-size but energy-richer conformations brought about by the formation of mixed intermolecular hydrogen bonding upon acid treatment. The extent of size variation was related to the pKa of acids and also to the chemical and stereochemical affinity of humic components with the chemical structure of the acids. Dicarboxylic malic acid was the most effective in modifying humic conformations in all HAs whereas the aromatic-rich superstructure of HA from an oxidized coal was effectively disrupted by the relatively small methanesulfonic acid and the chemically akin benzenesulfonic acid. These results suggest that the conformational association of humus dissolved in the soil solution may be systematically altered by organic acids present in the rhizosphere and might have effects on plant and microbial activities.     

A comparative study on the chemical composition of humic acids from forest soil,agricultural soil and lignite deposit: Bound lipid,carbohydrate and amino acid distributions

Base- and acid-hydrolysable fractions of humic acids (HAs) isolated from a forest soil, an agricultural soil and a lignite deposit were analysed, and comparisons were made between the base hydrolysable lipid (bound lipid), carbohydrate and amino acid signatures of the different humic acids.Bound lipids differ depending on the humic acid origin. Their composition were rather similar for the two soil humic acids, with three main lipid classes identified: (i) aliphatic components, (ii) aromatic components and (iii) sterols and triterpenols. The aliphatic subfraction was the most abundant and consisted predominantly of cutin- and suberin-derived moieties some of which could be clearly related to the vegetation. A minor bacterial input was indicated by the presence of short chain α- and β-hydroxyalkanoic acids. Aromatic subfraction contributed to a low amount to the total base hydrolysates and consisted mainly of lignin-derived methoxyphenols. Present in trace amounts, sterols and triterpenols are mainly of higher plant origin. The base hydrolysate from lignite humic acid markedly differs. Bound lipids released from lignite HA comprised almost exclusively aliphatic components, largely dominated by long chain alkanoic acids. Lignin-derived moieties, hardly detected, consisted solely of vanillic and 4-hydroxybenzoic acids indicating a much higher degree of lignin alteration in lignite humic acid. Sterols and triterpenols were absent.Although the composition of monosaccharides released upon acid hydrolysis was rather uniform irrespective of the humic acid origin, the distribution changed with the degree of humification of the HAs. Ratios of (Galactose+Mannose) to (Xylose+Arabinose) increased from soil to lignite humic acids. The high values of the ratios indicate that carbohydrates are primarily of microbial origin.In all humic acids neutral and acidic protein amino acids dominated. Non protein amino acids were only minor components consisting mainly of hydroxy proline and ornithine. The amino acid distributions of both soil HAs were similar. The amino acid distribution of lignite HA resembled that of soil HAs except for the following differences: (1) the absence of hydroxy proline and the greater abundance of ornithine suggesting a higher microbial contribution to the amino acids as the degree of humification increases, (2) the higher contribution of polar amino acids suggesting a preferential preservation of these amino acids possibly by interaction with the humic acid surface through hydrogen bonds.     

土壤腐殖酸对毒死蜱水解的影响

土壤腐殖酸的VIS-UV光谱特征和E465/E665的测定结果表明,各种土壤腐殖酸的VIS-UV吸收曲线相似,但吸收峰强度和E465/E665差异较大,进而推得它们的腐殖化程度排序为:紫色潮土腐殖酸>中性紫色土腐殖酸>酸性紫色土腐殖酸>腐殖土腐殖酸,说明它们在组成上有共同的本性,但结构和性质差异较大。在此基础上,研究它们对毒死蜱水解的影响。结果表明,不同来源土壤腐殖酸均促进了毒死蜱的水解,但它们的影响程度有明显的差异。对于腐殖化程度最低的腐殖土腐殖酸,浓度为120mg L-1时,对毒死蜱水解的促进作用最大,而随着腐殖酸浓度的增大,促进作用却越来越弱;而对于其它三种样品,腐殖酸溶液对毒死蜱水解的促进作用随着腐殖酸浓度增大而加强,其中以腐殖化程度较低的酸性紫色土的促进作用较为明显。可见,毒死蜱的水解速率与土壤腐殖酸的浓度和腐殖化程度有关。其中,腐殖酸腐殖化程度的影响机理主要与腐殖酸的酸性和腐殖酸的吸附—催化作用有关,其具体作用机理有待于进一步从定性和定量方面去研究,以更好地指导毒死蜱农药的合理施用和污染土壤的修复。     

有机物料对不同作物根系土壤腐殖质组成和结构的影响

为探讨施用有机物料后不同作物根系土壤腐殖质各组分含量和胡敏酸元素组成的变化情况,以吉林农业大学试验田培肥2a的黑土为研究对象,试验选取3种作物(玉米、大豆、白菜),每种作物设4种施肥处理,包括化肥(CK)、玉米秸秆配施化肥、树叶配施化肥和鸡粪配施化肥。采用腐殖质修改法提取水溶性物质(WSS)、胡敏酸(HA)、富里酸(FA)、胡敏素(HM),利用重铬酸钾外加热法测定腐殖质各组分有机碳含量,采用分光光度计法测定HA和FA的光学性质,通过光密度E_4/E_6值和元素组成分析HA的结构变化。结果表明:土壤总有机碳(TOC)和腐殖质各组分有机碳含量均表现为白菜玉米大豆。相比CK,配施有机物料后作物根系土壤TOC、WSS、HA含量分别显著增加5%~8%,34%~55%,8%~20%,FA含量变化不明显。相比树叶和鸡粪,秸秆可显著提高TOC含量;WSS在各物料之间差异不明显;HM含量表现为秸秆树叶鸡粪CK。有机物料对PQ值的影响因作物而异,秸秆和鸡粪对玉米、大豆根系土壤PQ值(HA占腐殖酸的比率)的影响相似,而鸡粪对白菜根系土壤PQ值的影响显著高于玉米秸秆。施用有机物料后HA的E_4/E_6未发生明显改变,FA的E_4/E_6显著提高。有机物料使根系土壤HA缩合度升高,分子结构更加复杂,且有利于HA含氮基团的形成,以秸秆处理作用最好。由此可见,有机物料具有良好的培肥效果,其中鸡粪对白菜根系土壤培肥效果最显著,秸秆与鸡粪对玉米、大豆根系土壤培肥效果相似,均显著高于化肥。     

Phosphorus in humus acids

The distribution of phosphorus among humus acid groups in different soil types was studied. It was shown that the binding of P with acid-soluble and-insoluble organic components is determined by the acid-base status of soils, which controls the predominant interaction of P with Fe and Al or with Ca and Mg, and the solubility of the resulting organic phosphates in acid and alkali. The major part of P is associated with the group of humic acids (HAs) in the acid soils and with fulvic acids (FAs) in the neutral and calcareous soils. During the fractionation of organic matter from acid soils, the amount of P redistributed from the HA to the FA group can increase with an increasing acid or alkaline impact on the soil, which breaks the bonds of the phosphorus-bearing organic components with metals.     

Influence of natural and synthetic humic substances on the activity of urease

The activity of a purified urease, obtained from Bacillus pasteurii, was inhibited by humic and fulvic acids obtained from an agricultural soil. Enzyme kinetic studies showed that the humic substances affected the affinity of the enzyme for its substrate (K_m) and the maximum velocity of the reaction (V_max). The V_max was inhibited to the same extent by both humic (HA) and fulvic (FA) acids, the precise effect depending on the pH and concentration of humic substance. At pH 4.0, HA concentrations of 25 pg cm^?3 and 10 μg cm^?3 inhibited the V_max by 38.5% and 20% respectively. HA and FA had similar effects on the K_m but in this case the lowering of the affinity of the enzyme for its substrate was not concentration dependent in the range 0–25 μg cm^?3 of humic substance. Typically, the affinity was decreased from a K_M of 50 mM in the control to 67 mM in the presence of HA and FA. The effects were not due primarily to the ash or N contents of the humic substances because de-ashed humic acid and synthetic model humic (made from catechol, guaiacol, pyrogallol, resorcinol and protocatechuic acid) and fulvic acid (made from polymaleic acid), containing virtually no ash or N, were equally as effective. The effect was not related to the phenolic monomers which, before polymerization, had no effect on urease activity.     

Phenanthrene sorption by compost humic acids

Nonlinear isotherms were observed for sorption of polycyclic aromatic hydrocarbons (PAHs) to humic materials, but the exact sorption mechanism is not clearly understood. This study aimed to investigate the sorption of PAHs by compost humic materials. Humic acids (HAs) were progressively extracted from two compost samples by a 0.1-M sodium pyrophosphate solution. Sorption of phenanthrene by the extracted HAs was studied with a batch equilibration technique. Competitive sorption between pyrene and phenanthrene in the HAs was also examined. Elemental analysis and solid-state ¹³C NMR were used to characterize HAs. All sorption isotherms were nonlinear although these samples contained little black carbons if any. Isotherm linearity increased with increasing number of extractions. Isotherm nonlinearity was negatively related to HA aliphaticity. Addition of pyrene to the phenanthrene-HA system increased isotherm linearity. Competition between phenanthrene and pyrene sorption was more pronounced in the first fraction of HAs with a higher content of aromatic moieties. This study showed that humic materials alone could lead to nonideal sorption for PAHs.     

施用猪粪对棕壤胡敏酸和富里酸热力学参数的影响

应用Vodyanitsky提出的方法，根据元素组成数据，计算了施用猪粪对棕壤胡敏酸(HA)、富里酸(FA)热力学参数的影响。结果表明，HA、FA的形成都是自发进行的放热反应；与HA相比，FA的能态较低，分子结构有序度和热稳定性较高；FA在土壤中比HA更容易形成。施用猪粪后，HA、FA的能态降低，分子结构有序度和热稳定性也降低，形成的自发性减弱且放热量减少，这些与它们元素组成的变化有关。     

Carbon and nitrogen in operationally defined soil organic matter pools

Humic substances [humic acid (HA), fulvic acid (FA), and insoluble humin], particulate organic matter (POM), and glomalin comprise the majority (ca 75%) of operationally defined extractable soil organic matter (SOM). The purpose of this work was to compare amounts of carbon (C) and nitrogen (N) in HA, FA, POM, and glomalin pools in six undisturbed soils. POM, glomalin, HA, and FA in POM, and glomalin, HA, and FA in POM-free soil were extracted in the following sequence: (1) POM fraction separation from the soil, (2) glomalin extraction from the POM fraction and POM-free soil, and (3) co-extraction of HA and FA from the POM fraction and POM-free soil. Only trace amounts of HA and FA were present in the POM fraction, while POM-associated glomalin (POM-glomalin) and POM alone contributed 2 and 12%, respectively, of the total C in the soil. Mean combined weights for chemically extracted pools from POM and from POM-free soil were 9.92 g glomalin, 1.12 g HA, and 0.88 g FA kg⁻¹ soil. Total protein and C, N, and H concentrations showed that glomalin and HA were, for the most part, separate pools, although protein was detected in HA extracts. Even though percentage carbon was higher in HA than in glomalin, glomalin was a larger (almost nine times) operationally defined pool of soil organic C. Glomalin was also the largest pool of soil N of all the pools isolated, but all pools combined only contained 31% of the total N in the soil.