Nitrogen Forms in Synthetic Humic Acids Using Nitrogen-15 Nuclear Magnetic Resonance Technique

^15N-labelled phenolic polymers were synthesized by reactions of p-benzoquinone and 1,4-diphenol with ^15N-labelled glycine and were studied by using ^15N CP-MAS NMR technique in combination with chemical approaches.Results showed that the proportion of polymer nitrogen as N-phenyl amino acid N was not as great as expected,only accounting for 5%-15%;and most of N in polymers occurred in the forms of amide,pyrrole-and indole-like nitrogen,aliphatic amines and isonitrile.It seems that great differences existed between synthetic humic acids and soil humic acids in the type and distribution of nitrogen forms.     

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%).     

由木糖和15N标记的甘氨酸形成的美拉德氏聚合物的氮素形态

Water-soluble,nondialyzable Maillard polymers were prepared by reacting D-xylose with ^15N-glycine (and /or glycine)at 6８℃ and pH 8.0 at equimolar concentrations of 1,0.5 and 0.1 mol L^-1,respectively, for 13 days and partitioned into acid-insoluble(MHA) and acid-soluble(MFA) fractions.The nitrogen froms in these polymers were studied by using the ^15N cross polartisation-magic angle spinning nuclear magnetic resonance(CPMAS NMR) technique in combination with chemical methods .The ^15N nuclear magnetic resonance(NMR) data showed that while the yield,especially the MHA /MFA ratio,varied considerably with the concentrations of the reactants,the nitrogen distribution patterns of these polymers were quite similar. From 65% to 70% of nitrogen in them was in the secondary amide and /or indole form with 24%-25% present as aliphatic and /or aromatic amines and 5% to 11% as pyrrole and /or pyrrole-like nitrogen,More than half (50%-77%) of the N in these polymers were nonhydrolyzable,The role of Maillard reacion in the formation of nonhydrolyzable nitrogen in soil organic matter is discussed.     

固态13C和15N核磁共振法研究15N标记土壤的腐殖质组分

Five humic fractions were obtained from a uniformly ^15N-labelled soil by extraction with 0.1 mol L^-1 Na4P2O7,0.1mol L^-1 NaOH ,and HF/HCl-0.1 mol L^-1 NaOH,consecutively,and analyzed by ^13C and ^15N CPMAS NMR (cross polarization and magic angle spinning nuclear magnetic resonace).Compared with those of native soils humic fractions studied as a whole contained more alkyls ,methoxyls and O-alkyls,being 27%-36%,17%-21%and 36%-40%,respectively,but fewer aromatics and carboxyls(bein 14%-20% and 13%-90%,respectively),Among those humic fractions ,the humic acid(HA)and fulvic acid(FA) extracted by 0.1 mol L^-1 Na4P2O7 contained slightly more carboxyls than corresponding humic fractions extracted by 0.1 mol L^-1 NaOH ,and the HA extacted by 0.1 mol L^-1 NaOH after treatment with HF/HCl contained the least aromatics and carboxyls.The distribution of nitrogen functional groups of soil humic fractions studied was quite similar to each other and also quite similar to that of humic fraction from native soils.More than 75% of total N in each fraction was in amide from,with 9%-13% present as aromatic and /or aliphatic amines and the remainder as heerocyclic N.     

土壤腐殖质氮结构的X-光电能谱研究

X－ray photoelectron spectroscopy(XPS) was applied to examine the N structures of soil humic substances and some of their analogues.It was found that for soil humic substances XPS method gave similar results as those obtained by ^15N CPMAS NMR (cross-polarization magic-angle spinning nuclear magnetic resonance) method.70%-86% of total N in soil humic substances was in the form of amide,and 6%-13% was presented as ammes,with the remaining part as heterocyclic N.There was no difference in the distribution of the forms of N between the humic substances from soils formed over hundreds or thousands of years and the newly formed ones.For fulvic acid from weathered coal and benzoquinone-(NH4)2SO4 polymer the XPS results deviated significantly from the ^15N CPMAS NMR data.     

日本常规耕作和有机耕作(翻耕与免耕)农田的动物区系(螨和线蚓)比较

Nitrogen forms of humic substances from a subalpine meadow soil,a lateritic red soil and a weathered cola and the effect of acid hydrolysis on N structures of soil humic substances were studied by using ^15N cross-polarization magic angle spinning nuclear magnetic resonance(CPMAS NMR) spectroscopy,Of the detectable ^15N-signal intensity in the spectra of soil humic substances 71%-79% may be attributed to amide groups ,10%-18% to aromatic/aliphatic amines and 6%-11% to indole-and pyrrole-like N.Whereas in the spectrum of the fulvic acid from weathered coal 46%,at least,of the total ^15N-signal intensity might be assigned to pyrrole-like N,14% to aromatic/aliphatic amines,and the reamining intensities could not be assigned with certainty,Data on nonhydrolyzable reside of protein-sugar mixture and a ^15N-labelled soil fulvic acid confirm the formation of nonhydrolyzable heterocyclic N during acid hydrolysis.     

Composition and Structural Features of Calcium—Bound and Iron—and Aluminium—Bound Humus in Soils

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 ^13 C 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 aluminiumbound humus.The calcium-bound humic acid(HA1) showed higher E4/E6 ratios than iron-and aluminumboud,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 observerd for HA and FA from black soil.The 13C NMR spectroscopy revealed that HA2 was more aromatic than HA1.On the other ,FA1 exhibited a higher aromaticity than FA2.     

Enhanced anaerobic degradation of hexachlorobenzene in a Hydragric Acrisol using humic acid and urea

Humic substances acting as an electron shuttle and nitrogen transformation process influence remarkably the electron transfer in anaerobic reaction systems and thus may affect the reductive dechlorination of hexachlorobenzene(HCB). In order to develop an efficient agricultural strategy for the remediation of organochlorine-contaminated soils, a batch incubation experiment was conducted to study the effects of humic acid, urea, and their interaction on the reductive dechlorination of HCB in a Hydragric Acrisol with high iron oxide content. After 44 d of anaerobic incubation, the five treatments, sterile control,control, humic acid, urea, and humic acid + urea decreased HCB residues by 28.8%, 47.8%, 64.7%, 57.8%, and 71.3%, respectively. The amendment of humic acid or urea significantly decreased soil Eh values and accelerated Fe(Ⅲ) reduction to Fe(Ⅱ), thus promoting markedly reductive dechlorination of HCB. Humic acid had a larger dechlorination effect than urea. Since there was a synergistic interaction between humic acid and urea that accelerated HCB dechlorination, the treatment having both amendments together was the most efficient for HCB dechlorination. The results showed that the combination of NH_4~+-N supplied by a fertilizer and humic substance is a feasible strategy for the remediation of organochlorine-contaminated soils with abundant iron oxide.     

无定形锰氧化物在氮素损失中的作用

Studies have been made,by ^15N-tracer technique on nitrogen loss resulting from adding amorphous manganese oxide to NH4^ -N medium under anaerobic conditions.The fact that the total nitrogen recovery was decreased and that ^15NO2,^15N2O,^15N^14NO,^15NO,^15N2 and ^15N^14N were emitted has proved that,like amorphous iron oxide,amorphous manganese oxide can also act as an electron acceptor in the oxidation of NH4^ -N under anaerobic conditions and give rise to nitrogen loss.This once again illustrates another mechanism by which the loss of ammonium nitrogen in paddy soils is brought about by amorphous iron and manganese oxides.The quantity of nitrogen loss by amorphous manganese oxide increased with an increase in the amount of amorphous manganese oxide added and lessened with time of its aging.The nitrogen loss resulting from amorphous manganese oxide was less than that from amorphous iron oxide.And the nitrogen loss resulting from amorphous manganese oxide was less than that from amorphous iron oxide.And the nitrogen loss by cooperation of amorphous manganese oxide and microorganisms (soil suspension) was larger than that by amorphous manganese oxide alone.In the system,nitrogen loss was associated with the specific surface ares and oxidation-reduction of amorphous manganese oxide.However,their quantitative relationship and the exact reaction processes of nitrogen loss induced by amorphous manganese oxide remain to be further studied.     

Characteristics of Humic Substances in Paddy Soils

In the present paper,the composition of humus and the charateristics of humic acid from seven paddy soils were compared with those of upland (and/or natural) soils.Results show that:(1) in each group of the soil samples for comparison the HA/FA ratio of the humus of a paddy soil,in most cases,was appreciable higher than that of adjacent upland(and/or natural) soil derived from the same parent material;(2) the humic acid extracted from the paddy soils was characterized by a higher C/O ratio,a higher content of methoxyl groups,and a lower content of carboxyl groups than those from the corresponding upland (and/or natural) flooded soils,implying that the humic acid formed under rice cultivation is in a lower degree of humification than that formed under upland(and/or natural) conditions;and (3) the humic acid of paddy soils,however,was not always characterized by a lower aromaticity than that of the corresponding upland(and/or natural) soils.     

Temporal changes in distribution and composition of N from labeled fertilizer in soil organic matter fractions

Variations in the amount and composition of immobilized nitrogen (N) in major soil organic matter fractions were investigated in a 730-day soil incubation experiment using ¹⁵N-labeled urea and ¹⁵N nuclear magnetic resonance spectroscopy with the cross polarization/magic angle spinning (¹⁵N CPMAS NMR) method. After 730 days, 24.7% of the applied N was recovered from the soil as organic N. The urea-derived N recovered from humic acids and humin decreased from 11.2 and 33.8% of the applied amount after 14 days to 1.6 and 20.4% after 730 days, respectively. When these values were corrected for the microbial biomass (MB) N, they ranged from 9.0 to 1.2% and 28 to 18%, respectively. The proportion of urea-derived N recovered from fulvic acids was low, ranging between 0.4 and 5.8% (with MB N) or 5.6% (without MB N) of the applied amount, whereas that from water-soluble nonhumic substances (WS-NHS; NHS in the fulvic acid fraction) remained high, 28–33% of the applied amount after correction for the contribution of MB N up to day 365, and decreased to 0.9% thereafter. The ¹⁵N CPMAS NMR spectra of humic acids, fulvic acids, and humin showed the largest signal at −254 to −264 ppm, corresponding to peptide/amide N. The proportions of heterocyclic, peptide/amide, guanidine/aniline, and free amino N in the urea-derived humic acid N were 3–7, 83–90, 5–7, and 2–4%, respectively. More than 80% loss of the urea-derived humic acid N did not markedly alter their composition. No time-dependent variations were also observed for the proportions of respective N functional groups in humin N, which were 3–5, 71–78, 12–17, and 6–10% in the same order as above. These results suggest the greater importance of physical stability than structural variation for the initial accumulation of organic N in soil.     

Incorporation of nitrogen from urea fertilizer into soil organic matter in rice paddy and cassava upland fields in Indonesia

Incorporation of newly-immobilized N into major soil organic matter fractions during a cropping period under paddy and upland cropping systems in the tropics was investigated in Jawa paddy fields with and without fish cultivation and a Sumatra cassava field in Indonesia. ¹⁵N-labelled urea (¹⁵N urea) was applied as basal fertilizer, and the soil samples were collected after harvest. The percentage of distribution of the residual N in soil from ¹⁵N urea into the humic acids, fulvic acid fraction, and humin were 13.1–13.9, 19.0–20.5, and 53.4–54.3%, respectively, for the Jawa paddy soils, and 14.9, 27.4, and 52.4%, respectively, for the Sumatra cassava soil. These values were comparable to the reported ones for other climatic zones. The percentage of distribution of ¹⁵N urea-derived N into humic acids was larger than that of total N into the same fraction in all the soils. The distribution into the fulvic acid fraction was also larger for ¹⁵N urea-derived N than for total N in the Jawa soils. Humic and non-humic substances in the fulvic acid fraction were separated using insoluble polyvinylpyrrolidone (PVP) into the adsorbed and non-adsorbed fractions, respectively. Less than 5% of the ¹⁵N urea-derived N in fulvic acid fraction was detected in the PVP-adsorbed fraction (generic fulvic acids). The proportion of non-hydrolyzable N remained after boiling with 6 M HCl in the ¹⁵N urea-derived N was 9.4–13.5%, 17.3–26.7%, and 8.4–16.6% for the humic acids, generic fulvic acids, and humin, respectively. The significantly low resistance to acid hydrolysis suggested that the ¹⁵N urea-derived N was less stable than the total N in soil regardless of the fractions of humus.     

Transformation of 15N-labelled leguminous plant material in three contrasting soils

Summary Two soils from Pakistan (Hafizabad silt loam and Khurrarianwala silt loam) and one from Illinois, USA (Drummer silty clay loam) were incubated with ¹⁵N-labelled soybean tops for up to 20 weeks at 30°C. Mineralization of soybean ¹⁵N was slightly more rapid in the Pakistani soils, and after 20 weeks of incubation, 50%, 53%, and 56% of the applied ¹⁵N was accounted for as (NH₄ ⁺+NO₃ ^–)-N in Drummer, Hafizabad, and Khurrarianwala soils, respectively. Potentially mineralizable N (determined by anaerobic incubation) varied between 1.5% and 10% of the applied ¹⁵N in the three soils at different stages of incubation; somewhat higher percentages were mineralizable in the Pakistani soils than in the Drummer soil. From 3.7% to 9% of the applied ¹⁵N was accounted for in the microbial biomass. From 10% to 32% of the applied N was recovered in the humic acid and fulvic acid fractions of the organic matter by sequential extraction with Na₄P₂O₇ and NaOH; from 12% to 49% was recovered in the humin fraction. Of the three soils, Drummer soil contained more ¹⁵N as humic and fulvic acids. In all cases, the ¹⁵N was approximately equally distributed between the humic and fulvic acid fractions. A significant percentage of the humin ¹⁵N (52%–78%, equivalent to 8%–34% of the applied ¹⁵N) occurred in non-hydrolyzable (6 N HCl) forms. Of the hydrolyzable ¹⁵N, 42%–51% was accounted for as amino acid-N followed in order by NH₃ (17%–30%), hydrolyzable unknown forms (20%–22%), and amino sugars (6%–2%). The recovery of applied ¹⁵N for the different incubation stages was 87±22%. Recovery was lowest with the Khurrarianwala soil, presumably because of NH₃ volatilization losses caused by the high pH of this soil.     

长期施肥条件下土壤养分的动态和平衡——Ⅱ.对土壤氮的有效性和腐殖质氮组成的影响

长期施用有机肥、化肥或二者配合施用均增加了土壤全氮、碱解氮、酸解氮和植物吸收氮量,提高了土壤氮的有效性,其后效可保持至第3年;土壤腐殖质的胡敏酸氮减少,富里酸氮增加。富里酸氮易被6mol/L HCl水解,水解氮占71.4％～74.2％;胡敏酸氮比较稳定,水解氮占40％～45％。在水解氮中,氨基氮的比例为2/3～3/4,铵态氮为1/4～1/3。胡敏酸中非水解氦的主要成分是杂环化合物氮(37.5％～52.5％),其次是苯系化合物氮(31％～35.2％)和非苯系化合物氮(10.6％～26.7％)。长期施用有机肥和化肥,杂环氮比例减少,苯系和非苯系化合物氮比例增加。胡敏酸表层氮占1/3左右,核心氮占2/3左右,施用有机肥和化肥,胡敏酸表层氮增加,核心氮降低。     

长期施肥条件下土壤养分的动态和平衡——Ⅱ.对土壤氮的有效性和腐殖质氮组成的影响

长期施用有机肥、化肥或二者配合施用均增加了土壤全氮、碱解氮、酸解氮和植物吸收氮量,提高了土壤氮的有效性,其后效可保持至第3年;土壤腐殖质的胡敏酸氮减少,富里酸氮增加。富里酸氮易被6mol/L HCl水解,水解氮占71.4％～74.2％;胡敏酸氮比较稳定,水解氮占40％～45％。在水解氮中,氨基氮的比例为2/3～3/4,铵态氮为1/4～1/3。胡敏酸中非水解氦的主要成分是杂环化合物氮(37.5％～52.5％),其次是苯系化合物氮(31％～35.2％)和非苯系化合物氮(10.6％～26.7％)。长期施用有机肥和化肥,杂环氮比例减少,苯系和非苯系化合物氮比例增加。胡敏酸表层氮占1/3左右,核心氮占2/3左右,施用有机肥和化肥,胡敏酸表层氮增加,核心氮降低。