蒙脱土及腐殖酸的界面动电性质

自然界广泛存在的粘土和腐殖酸经常结合在一起,它们直接影响和决定着土壤结构和水/土界面的其它性质^[4-6].关于土壤胶体动电性质的研究五十年代比较活跃,但在以后却被大大地忽视了^[1].本工作研究了钠蒙脱土以及蒙脱土结合腐殖酸以后的粒子在水中的界面动电性质，考察了动电性质随pH的变化.这种纯粘土及其和腐殖酸结合体系的研究对于土壤胶体性质的认识是必要的.     

腐殖酸对滨海盐碱土水盐运移特征的影响

为了探索腐殖酸对土壤水盐运移特征及入渗参数的影响，进行了5种施量（0，1，3，5，7 g/kg）的一维垂直入渗试验。结果表明，入渗时间相同，累积入渗量和湿润锋均随腐殖酸施量的增加而减小；相同入渗深度随腐殖酸施量的增加所需的入渗时间、累积入渗量、剖面平均含水量均不断增加。入渗完成时相较于CK，施量为7 g/kg时累积入渗量、含水量分别增加8.0%，10.0%；施加腐殖酸可促进土壤剖面盐分向土壤深层运移并有效降低土壤pH，相较于CK施量为7 g/kg时0—20 cm土层平均脱盐率增加10.2%；Philip模型和代数模型能较好地描述施加腐殖酸条件下水分入渗特征。吸渗率S随腐殖酸施加量的增加而呈线性减小，综合形状系数α则随着腐殖酸施量的增加而线性增加。另外，均方根误差、平均绝对误差分别为0.005~0.018，0.004~0.014 cm³/cm³，表明代数模型可以较好地描述含水量的分布。该研究结果可为应用改良剂改良滨海盐碱地的提供一定参考。     

改良剂对镉污染酸性水稻土的修复效应与机理研究

为探明田间条件下施用石灰、钙镁磷肥、海泡石和腐殖酸等改良剂对Cd污染酸性水稻土的修复效应和作用机理, 通过在Cd污染区建立田间小区试验, 研究了改良剂单施和与石灰配施对Cd污染酸性水稻土中Cd作物有效性的影响。结果表明, 施用改良剂有效地改变了土壤中Cd的存在形态, 除腐殖酸外, 其他改良剂均使土壤酸提取态Cd不同程度地转化为可还原态Cd和残渣态Cd; 施用改良剂可使0.1 mol·L^-1 NaNO₃和 0.01 mol·L^-1CaCl₂提取态Cd 降低26%~97%, 降低效果为石灰+海泡石>海泡石>石灰+钙镁磷肥>钙镁磷肥>石灰>石灰+腐殖酸>腐殖酸; 改良剂使水稻地上部分的Cd吸收量降低6%~49%。试验结果还显示, 施用改良剂提高土壤pH是引起土壤中Cd作物有效性降低的主要原因之一。根据田间试验的结果, 海泡石可推荐作为Cd污染酸性水稻土的改良剂, 而腐殖酸则不宜使用。     

不同分子量风化煤腐殖酸对玉米植株主要代谢物的影响

【目的】腐殖酸的分子量决定了其功能特性,研究不同分子量腐殖酸对玉米生长发育的影响,可为腐殖酸资源的高效利用提供理论依据。【方法】选用‘郑单958’为供试玉米品种,以霍格兰营养液为基础溶液进行水培试验。选择分子量为> 50 kDa (HA_H)、10 kDa～50 kDa (HA_M)和<10 kDa (HA_L)三种腐殖酸,添加量均为碳10 mg/L,以不添加腐殖酸为对照(CK),共四个处理。幼苗生长20 d后收获,分为根、茎和叶样品,烘干称重,采用傅里叶变换红外光谱仪(型号VERTEX 70,德国Bruker公司)检测玉米植株根、茎和叶的光谱特征。【结果】腐殖酸处理玉米根、茎和叶的干物质重较对照分别显著提高了91.07%、89.27%和88.53%。三种腐殖酸处理,以小于10 kDa分子量腐殖酸(HAL)对玉米植株生长的促进作用最大,根、茎和叶的干物质重分别提高了143.14%、123.41%和150.54%,其次为10～50 kDa分子量腐殖酸(HA_M),大于50 kDa分子量腐殖酸(HAH)效果又次之。腐殖酸处理的玉米根系FTIR光谱在3420 cm^–1和1655 cm^–1波数附近的透射率均低于对照,茎FTIR光谱在3420 cm^–1和1655 cm^–1波数附近的透射率均高于对照,表明腐殖酸处理能增加玉米根系碳水化合物、蛋白质、多肽和氨基酸类物质的含量,减少其在茎中的积累,腐殖酸各处理玉米叶片在3420、2920、1735、1655、1518、1380、1250和1050 cm^–1波数附近的透射率均低于对照,小于10 kDa分子量腐殖酸表现尤为明显。这表明不同分子量(尤其是小于10 kDa分子量)腐殖酸能够增加玉米叶片碳水化合物、脂类物质、蛋白质、多肽、氨基酸类物质和核酸等的含量。【结论】外源添加腐殖酸能够增加玉米干物质重,引起玉米植株不同器官中碳水化合物、脂类物质、蛋白质、多肽、氨基酸类物质及核酸发生变化,小分子量(<10 kDa)的腐殖酸对玉米植株生长的促进作用大于大分子量的,在生产中应尽量选择小分子腐殖酸产品,以充分发挥其促生作用。     

腐殖酸-锌络合物稳定性的研究

腐殖酸-金属络合物在土壤形成和植物营养等方面具有重要的作用。六十年代以来,这方面的研究日益增多。锌是植物所需的微量元素之一,在土壤中常以络合态存在。因此,以络合态存在的锌的数量及其稳定性对土壤的供锌能力将有较大的影响。不少研究者虽曾研究了腐殖酸一锌的稳定性^[4,6,7,10],但所测得的稳定常数值差异甚大,有的可差几个甚至高达十个数量级。导致这种差异的原因很多,其中之一是各研究者所用的腐殖酸各人不同。一些研究者认为络合物的log K值因供试腐殖酸的腐殖化程度而异,另一些研究者认为log K值因腐殖酸分子大小不同而不同。本工作测定了不同来源的腐殖酸的某些性质及其与锌形成的络合物的稳定性,试图对它们之间的关系作一初步探讨。     

中低分子量腐殖酸提高冬小麦磷吸收和产量的机理

  【目的】  研究不同分子量腐殖酸与磷肥复合制备的腐殖酸磷肥对作物和土壤磷有效性的影响，为腐殖酸磷肥研发和磷素高效利用提供理论依据。  【方法】  利用超滤分级方法，将风化煤腐殖酸分子量分为 > 100 kDa、10～100 kDa和 < 10 kDa 3个部分，获得高 (HA_H)、中 (HA_M)、低 (HA_L) 不同分子量的腐殖酸，采用磷酸与KOH反应法制备普通磷肥 (P)、未分级腐殖酸磷肥 (PHA)、高分子量腐殖酸磷肥 (PHA_H)、中分子量腐殖酸磷肥 (PHA_M) 和低分子量腐殖酸磷肥 (PHA_L) 5种磷肥。采用深100 cm、内径25 cm的土柱进行冬小麦栽培试验，按等磷量原则，设置P、PHA、PHA_H、PHA_M、PHA_L 5个施磷处理，同时设置与4个施磷处理对应的等量腐殖酸处理 (HA、HA_H、HA_M、HA_L)，以不施磷肥为对照CK。测定小麦产量及产量构成因素、植株磷含量及不同层次土壤有效磷含量。  【结果】  1) 与CK相比，腐殖酸处理 (HA、HA_H、HA_M、HA_L) 小麦增产不明显。与普通磷肥相比，PHA、PHA_M和PHA_L处理产量显著提高了14.73%、18.84%、21.37% (P < 0.05)，3 个处理间产量差异不显著，PHA_H增产不明显。PHA_L处理千粒重显著高于普通磷肥处理，其余3个腐殖酸磷肥处理增幅未达显著水平。2) PHA、PHA_M和PHA_L处理籽粒吸磷量较普通磷肥处理分别显著提高14.97%、19.45%、22.68%，而PHA_H增幅未达显著水平；腐殖酸磷肥处理间秸秆吸磷量没有显著差异。3) 与普通磷肥相比，PHA、PHA_H、PHA_M、PHA_L磷肥偏生产力和农学效率分别提高14.71%、6.01%、18.82%、21.35%和14.95%、1.66%、20.18%、23.03%，磷肥表观利用率分别提高2.93、0.51、4.52、5.41个百分点，也以中、低分子量腐殖酸磷肥效果最为明显，腐殖酸中的氧烷基碳、羧基/酰胺基碳、烷基碳结构与小麦籽粒产量和磷肥利用率具有正相关性，与芳香碳、芳香C―O负相关。4) 4个腐殖酸磷肥处理间及其与普通磷肥处理间 0—20、20—40、40—60 cm 土层土壤有效磷含量差异不显著。  【结论】  田间土柱栽培条件下，单施腐殖酸对小麦没有表现出明显的增产效果。腐殖酸中的氧烷基碳、羧基/酰胺基碳、烷基碳结构与小麦磷素吸收具有正相关性，低分子量腐殖酸具有较多的烷基碳、氧烷基碳、羧基/酰胺基碳结构，因而低分子量腐殖酸提高磷肥中磷素利用率的作用好于中分子量腐殖酸，而高分子量腐殖酸的效果不显著。     

电荷辅助氢键对腐殖酸分子量和溶解性的影响

采用体积排阻色谱和总有机碳分析法,分别测定腐殖酸体系中加入盐酸、甲酸、乙酸和丙酸后其分子量和溶解性发生的变化;通过分析抗坏血酸、苯甲酸、苯酚和邻苯二酚4种模型化合物在甲酸影响下的紫外光谱,以验证电荷辅助氢键的存在。结果表明:小分子有机酸能够明显降低腐殖酸的分子量;此外,当腐殖酸体系pH接近小分子有机酸的pKa时(DpKa<0.5),腐殖酸溶解性明显增大。小分子有机酸可能与腐殖酸之间形成电荷辅助氢键,从而打破弱作用力维持的腐殖酸超分子的稳态结构,导致腐殖酸分子量的降低和溶解性的增加;且弱酸与有机物之间的pKa相差越小,形成的电荷辅助氢键能量越高,腐殖酸分子结构受到的扰动程度越大。一维紫外光谱和同步二维相关紫外光谱分析进一步表明,小分子有机酸可能与pKa接近的化合物之间形成电荷辅助氢键,整体跃迁所需能量提高,造成低波长吸收增大而高波长吸收减小的结果。     

土壤有机矿质复合体研究Ⅰ.土壤有机矿质复合体中腐殖质氧化稳定性的初步研究

土壤中有机矿质复合体的种类很多,广义地说,应当包括腐殖酸和各种金属离子所构成的盐类,腐殖质与含水铁绍氧化物的复合凝胶,腐殖质与粘土矿物直接结合或通过其他媒介结合的各种复合体等。很多工作者研究了各种有机矿质复合体的組成和特性^[1,6,7,8]。     

大米直链淀粉分子量分布及分子旋转半径的研究

为了从大米淀粉中有效的分离出直链淀粉，建立直链淀粉分子量的分析方法，从而得出确切的大米直链淀粉分子结构方面的信息，通过使用流变仪控制大米淀粉的升温糊化过程，用浸出法在不同温度下分离出大米直链淀粉，利用高效液相分子排阻色谱(HPSEC)与多角度光散射仪(MALLS)及折光检测器(RI)连用系统，分析了大米直链淀粉的分子量分布和分子旋转半径。得到不同品种的大米淀粉中分离出的直链淀粉的重均分子量范围为3.29×10⁵～2.75×10⁶。研究表明，90℃以上高温不利于用浸出法从直链淀粉含量低的大米淀粉中分离直链淀粉，当温度低于90℃而高于糊化温度时，各个温度下均可用浸出法分离出直链淀粉，得到的直链淀粉具有十分相似的分子量分布、重均分子量和分子旋转半径。该方法是一种简便、快速的分离大米直链淀粉，并测定其分子量的方法，具有较高的准确性。     

利用牧草与沼渣栽培毛木耳及其残渣改良土壤效果

以不同比例牧草与沼渣替代木屑栽培毛木耳对其产量、品质及氨基酸含量影响各异。30%草粉 +30%沼渣替代木屑效果较好,生物效率为107.5%,产量比对照高82.2%,粗蛋白、氨基酸、粗脂肪、可溶性糖含量分别比对照高3.7%、1.75%、0.13%、0.37%;以50%草粉或沼渣替代木屑其毛木耳产量分别比对照高55.1%和61.0%。山地果园连续3年施用5t/hm²沼渣、5t/hm²菌渣和5t/hm²鲜草处理区与对照相比,其红壤中腐殖酸碳含量分 别提高22.9%、26.5%和16.5%;红壤中团粒结构依次提高78.5%、79.9%和69.3%,改良土壤效果较好。     

Effects of urease and nitrification inhibitors on the efficient use of urea for pastoral systems

Abstract

The humic substances contained in an animal organic waste were extracted and the total extract separated into three humic fractions with different molecular weights (low, F₁ <10³; medium, F₂, with molecular weights ranging from 10³ to 10⁴; and high, F₃ >10⁴). The C content was highest in F₂, the same fraction also showing the lowest N content. The molecular weight of the humic fractions influenced the electrical conductivity, the highest molecular weight resulting in the lowest degree of electrical conductivity. Membrane-controlled ultrafiltra-tion (the method used to separate the various fractions from the whole extract) was also suitable for purifying such enzymes as phosphatase and β-glucosidase: the total activity obtained from the three fractions was considerably greater than that determined in the whole extract, Pyrolysis-gas chromatography (Py-GC) applied to the whole extract and humic fractions showed that in the F₃ fraction (highest molecular weight) benzene was the major fragment while furfural was the major fragment of F₁ (lowest molecular weight). For this reason, the humification index benzene/toluene indicates that the fraction with the highest molecular weight was the most humified while the furfural/pyrrole ratio indicates that the fraction with the lowest molecular weight was the most degradable. The whole extract and the fraction F₁ had a negative effect on seed germination when the concentration was equivalent to 100 mg kg^?1 of C, while the germination index was higher than that of the control when only 10 mg kg^?1 were used. The F₂ fraction had a positive effect on germination regardless of the concentration used. When 10 mg kg^?1 of C of the humic substances studied were added to the nutrient solution for growth experiments with maize plants, F₃ led to increases in root weight and F₂ led to increases in shoot weight. An inhibitor effect was observed for fraction F₁.     

MOLECULAR WEIGHT AND SHAPE OF HUMIC ACID FROM SEDIMENTATION AND DIFFUSION MEASUREMENTS ON FRACTIONATED EXTRACTS

Whole humic acid extracts are usually too polydisperse for reliable molecular-weight measurement to be made in the ultracentrifuge by the sedimentation velocity technique. Consequently, the humic acid used in this study was fractionated with respect to molecular weight into fractions of low polydispersity by extensive use of gel-permeation chromatography and other fractionation techniques. The sedimentation and diffusion coefficients of the fractions were determined and molecular-weight values calculated. These values ranged approximately from 2 × 10³ to 1.5 × 10⁶, the higher figure not necessarily representing the upper limit for these substances. On the basis of the frictional parameters calculated from the experimental data, it is proposed that the molecule adopts the solution conformation of a randomly coiled polymer in which branching may be significant, particularly at higher molecular weights.     

Physiological action of humic substances on microbial cells

By incorporating molecular weight fractions of humic acids of various origins into selective substrates designed for the enumeration of physiological groups of microorganisms, it was found that the presence of humic acids at concentrations of up to 30mgl^?1 normally resulted in increased numbers of soil microbes active within a particular physiological group. Observed increases could be as much as 2000-fold. Microbes in an organic humus-rich soil were more stimulated by humic substances than organisms from a sandy soil.In certain microbes humic substances appeared to induce a change in metabolism, allowing the organisms to proliferate on substrates which previously they could not utilize. Indications were obtained that within the 10–30 mgl^?1 concentration range lower molecular weight humic fractions (approx. 5500 dallons) were more effective than higher molecular weight material. At higher concentrations the reverse was sometimes noticed. Similarly, fulvic acids at concentrations of up to approximately 50mgl^?1 would appear to have a more pronounced physiological effect than humic acids, whereas the latter might be more effective at higher concentrations.The response of certain physiological groups to humic products of natural origin appeared to be comparable to that of surfactants such as Tween and Brij. This would suggest that the physiological action of humic substances is, at least partly, the result of their surface activity, making the membrane one of the prime targets of the physiological action of humics on living cells.     

Isolation of bacterial DNA from soil

DNA has been isolated from the bacterial fraction of two soils. Numbers of bacteria, determined by fluorescence microscopy were 1.1 and 2.2·10¹⁰ cells g^?1 dry wt. The total amounts of bacterial DNA in these soils were 90 and 187 μg g^?1 dry wt respectively.A modification of Marmur's method was used to isolate DNA, but difficulties in separating DNA from humic substances gave low yields and impure DNA.DNA could be partly separated from humic material in the presence of 8 m urea by ion-exchange chromatography on DEAE-Sepharose CL-6B. Final purification was obtained by chromatography on a hydroxyapatite column. When lowering the EDTA concentration in the saline-EDTA solution used for lysis, the amount of humic substances in the cell-free lysate after centrifugation was considerably decreased. The lysate could then be chromatographed directly on hydroxyapatite. Quantities up to 1.5 mg DNA high purity was isolated from 90 g wet soil (37 g dry wt).The isolated DNA was characterized by treatment with DNAse and absorption spectra. No uncommon bases were revealed by thin layer chromatography of the DNA hydrolysates. Melting curves of the isolated DNA showed a relatively broad melting profile, with half maximum hyperchromicity (T_m) near 90°C. Sedimentation coefficients determined by analytical ultracentrifugation showed that the isolated DNA had a high molecular weight, ranging from 2.3 to 10.1·10⁵ daltons.     

1H NMR and i.r. spectroscopic investigations on soil organic fractions obtained by gel chromatography

Fulvic and humic acids fractionated by gel chromatography on Sephadex G-50 have been investigated by ¹H NMR and i.r. spectroscopic techniques. Fulvic acid gives rise to only one significant fraction (FA-I) whose spectrum does not substantially differ from that of the unfractionated sample. The HA sample is separated into a high (HA-I, nominal mol. wt > 10,000) and a low molecular weight fraction (HA-II, nominal mol. wt 500–10,000). The two fractions show worthwhile differences in the ¹H NMR spectra. Signals belonging to protons of long-chain aliphatic hydrocarbons or acids are present only in the HA-I fraction, which also displays a lower content of aromatic protons than the HA-II fraction. The pattern of the broad and intense absorption in the range 2.9 and 5.0 ppm of the two spectra is different. On the basis of the i.r. spectra, it has been suggested that this might be due to a different distribution of structures as polysaccharides. It seems likely that the non-humic substances are trapped in the voids of the high molecular wight polymer. The results obtained indicate that humic fractions narrower than the original material still show a high degree of polydispersity. The overall percentage distribution of chemical structures in humic fractions with varying molecular weights is different.     

Preparation and characterization of model humic polymers containing organic phosphorus

The nature of organic P in soil organic matter was studied by evaluating the incorporation of serine, phosphoserine, ethanolamine, phosphoethanolamine and glycerophosphate into model humic polymers prepared by chemical oxidation of polyphenols. Elemental and functional group analysis indicated that the composition of model humic polymers ranged as follows: organic C, 50.6–56.8%; total acidity, 7.86–11.87m-equiv g^?1; carboxyl, 1.42–2.00 m-equiv g^?1; total hydroxyl, 6.79-10.0 m-equiv g^?1; ash, 6.4–13.9%; E₄/E₆ ratio, 5.34–6.19; organic N, 0.70–1.65% and organic P, 0.254–0.942%. These values are within the ranges reported for soil humic substances. The only non-phenolic compounds incorporated into model humic polymers were those containing free amino groups. The P content of model polymers was not increased by the presence of KH₂PO₄, glycerophosphate, serine or ethanolamine whereas phosphoserine and phosphoethanolamine resulted in model polymers containing 0.254 and 0.942% P, respectively. Further characterization studies of the model polymer containing phosphoethanolamine (HA-PE) showed that most of the C (83.2%), N (79.8%) and P (75.3%) was in the humic acid fraction. Gel filtration of HA-PE showed that 0.5% of the polymer was present in high molecular weight (mol. wt) components (mol. wt > 100,000) and 74.8% of the polymer was in two components of mol. wt 10,000–50,000. The majority of the organic P in HA-PE was associated with the medium molecular weight fractions (79.2%) while 16.8% of the P was associated with materials possessing mol. wt < 10,000. Attempts to demonstrate the presence of organic P functional groups contained in HA-PE by infrared spectroscopy was limited by the relatively small amounts of organic P incorporated into the model humic polymers. The results obtained show that a portion of the unidentified organic P in soil humic substances may arise from the incorporation of organic compounds containing both amino and phosphate ester functional groups during oxidative polymerization of polyphenols.     

胡敏酸的酒精分级沉淀法分级

本文利用酒精分级沉淀法，将胡敏酸分成１２个级分，并对各级分的结构进行了表征。结果表明，随着溶液中乙醇浓度的增大，各级分胡敏酸的腐殖化程度大体上逐渐由强变弱，分子量逐渐变小，全氮，水解性总氮和氨基酸氮的含量逐渐增大，而氨基糖氮和铵态氮逐渐减少 ，同时还讨论了本法与凝胶分级法所获得的结果不同的可能原因。     

Aggregation of humic substances in aqueous media as determined by light-scattering methods

Samples of peat humic acid (PHA) and surface water humic (WBHA) and fulvic acids (WBFA) extracted from Whitray Beck in North Yorkshire, UK and previously studied by analytical (UV scanning) ultracentrifugation at low concentrations (10–20 mg dm^?3) (Reid et al., 1990) have been investigated by dynamic and static light-scattering methods at concentrations up to approximately 1700 mg dm^?3. Dynamic light-scattering gave very low diffusion coefficients for the humic acids (PHA and WBHA) suggesting the presence of aggregates. Static light-scattering from the fulvic acid (WBFA) showed that the second virial coefficients of this material in various media were negative, which is characteristic of reversible aggregation. The aggregates, which consisted of approximately 20 smaller molecules, could not be dissociated completely by the detergents sodium n-dodecylsulphate or Triton X-100, although limited dissociation by the latter may occur. Sedimentation velocity measurements in the same concentration range as the light scattering gave sedimentation coefficients which increased with concentration also consistent with reversible aggregation. The studies illustrate the problems inherent in the characterization of humic substances from natural sources and demonstrate that, at least for lower molecular weight fulvic acids, static light-scattering from solutions of relatively high concentration in combination with sedimentation equilibrium measurements at low concentrations distinguishes a system having a broad molecular weight distribution from one in which aggregation occurs.     

ADVERSE EFFECTS OF HUMIC SUBSTANCES FROM DIFFERENT ORIGIN ON LUPIN AS RELATED TO IRON SOURCES

Humic substances improve the efficiency of different iron (Fe) sources overcoming Fe deficiency chlorosis of plants. However, applied at high rates, they can promote negative effects on plants. The main objective of this work was to study the potential adverse effect of three humic acids from different origin when they were applied with two effective Fe sources for plants: Fe- ethylenediaminedihydroxyphenylacetic acid (EDDHA) and Vivianite. To this end, an experiment with lupin (Lupinus albus L.) was performed involving two factors: (i) Fe source, and (ii) humic substances from three different origin (composted cork, leonardite, and compost obtained from a mixture of olive husk with cotton gin trash) applied at 0, 0.1, and 0.5 g organic carbon (C) kg^?1 of growing media. At the rates used, humic substances promoted adverse effects on plant development, chlorophyll meter readings, and Fe content in lupin grown in calcareous media. Overall, the effect on dry matter and Fe content in plants was more relevant when Fe was supplied with Vivianite, the effect on chlorophyll meter readings being more significant when Fe was applied as Fe-EDDHA. Differences were also observed depending on the source of humic substances, those from leonardite promoting the greatest decrease in dry matter in roots and shoots. These humic substances possessed the highest values of spectroscopy index for aromaticity (A₂₅₄ ). On the other hand, the application of humic substances from olive husk compost, which exhibited the lower aromaticity index, resulted in the smallest decrease in dry matter production and chlorophyll meter readings. Dry matter in roots decreased logarithmically with increased values of the estimates of the amounts of aromatic compounds accumulated in the growing media (R² = 0.92; P < 0.01) with Vivianite as Fe source. Thus, the effects decreasing dry matter production, particularly in roots, and chlorophyll meter readings can be ascribed at least partially to the presence of phytotoxic aromatic compounds in humic substances.     

Complexometric gel permeation chromatography of soluble humic substances using Sephadex G-25

Sephadex G-25 was used with 0.01 m sodium acetate containing a concentration of 3×10^?5m Cu(II) as eluent to establish the molecular-size distribution (MSD) and the Cu complexation of molecular-weight fractions of organic solutes. The appropriateness of this modified gel permeation method for complexometric measurements on soluble humic Substances was investigated employing various types of soil solutions. The presence of Cu in the eluent had an effect on the MSD obtained by elimination of electrostatic repulsion between gel and solutes. No evidence was found that the presence of Cu in the chromatographic system induced aggregation of the organic substances. The Cu-binding capacity of the samples as obtained with this gel permeation method was found to increase linearly over a wide sample concentration range varying from 40–2000 mg C dm^?3. No interference was observed between the Cu-complexation by exclusion and inclusion peaks in the chromatograms. The analyses revealed that the fraction apparently low in molecular weight makes a large contribution to the complexation potential of forest soil solutions.