Comparative Evaluation of Humic Substances: Effect at Cell Level and Chlorophyll Retention during Accelerated Senescence

The influence of humic substances (HS) formulations derived from sedimentary and compost sources was studied on plant growth at cell level and chlorophyll retention during accelerated senescence of leaf tissue. The direct effect of HS formulations was studied on cell expansion using cucumber and radish cotyledon expansion test. The cucumber hypocotyl elongation test was used to study the effect on cell elongation. Chlorophyll pigment retention in excised leaf tissue incubated in dark with high temperature was assessed to study the effect on leaf senescence. Explant tissues were incubated directly in the solutions of the formulations at the concentration recommended for foliar application to the crop plants. HS formulations showed significant variations in their direct bio-stimulatory effects. Formulations derived from compost sources were found superior in terms of inducing a direct stimulatory effect on cell expansion and cell elongation and in maintaining chlorophyll pigment retention during accelerated senescence. HS from sedimentary sources stimulated cell expansion and delayed chlorophyll degradation to a lesser extent compared to HS from compost. However, HS formulations derived from sedimentary sources used in this study were not effective in inducing cell elongation in the cucumber hypocotyl elongation test. The direct bio-stimulatory effect of HS formulations differed significantly between the formulations that were evaluated.

Effect of humic substances on the precipitation of calcium phosphate

For phosphorus （P） recovery from wastewater, the effect of humic substances （HS） on the precipitation of calcium phosphate was studied. Batch experiments of calcium phosphate precipitation were undertaken with synthetic water that contained 20 mg/L phosphate （as P） and 20 mg/L HS （as dissolved organic carbon, DOC） at a constant pH value in the range of 8.0-10.0. The concentration variations of phosphate, calcium （Ca） and HS were measured in the precipitation process; the crystalline state and compositions of the precipitates were analysed by powder X-ray diffraction （XRD） and chemical methods, respectively. It showed that at solution pH 8.0, the precipitation rate and removal efficiency of phosphate were greatly reduced by HS, but at solution pH ≥9.0, the effect of HS was very small. The Ca consumption for the precipitation of phosphate increased when HS was added; HS was also removed from solution with the precipitation of calcium phosphate. At solution pH 8.0 and HS concentrations ≤3.5 mg/L, and at pH ≥ 9.0 and HS concentrations ≤ 10 mg/L, the final precipitates were proved to be hydroxyapatite （HAP） by XRD. The increases of solution pH value and initial Ca/P ratio helped reduce the influence of HS on the precipitation of phosphate.

Interaction of humic substances and hematite:FTIR study

The present work extended the knowledge on the binding and complexation of humic substances(humic acid or fulvic acid) and hematite by Fourier transform infrared spectroscopy(FTIR). The FTIR data gained gave the consist evidences by two different sampling preparation methods that the interaction mechanism between humic substances and hematite was mainly conform to the ligand exchange involving carboxylic functional groups of humic substances and the surfaces sites of hematite. The present method, although associated with some uncertainties, provided an opportunity to increase the knowledge in this field.

Competitive Complexation of Copper and Zinc by Sequentially Extracted Humic Substances from Manure Compost

Chicken manure with similar content of copper and zinc was changes of organic carbon and humus substance complexed chosen to conduct a composting experiment to investigate the copper （HS-Cu） and zinc （HS-Zn）, which were extracted by water （H2O）, sodium hydroxide （NaOH）, and sodium pyrophate-NaOH mixture （Na4P2O7-NaOH）, sequentially. Distributions of copper and zinc in fulvic acids （FA） and humic acids （HA） in the three extracts were studied. During manure composting, the concentrations of copper and zinc increased from about 500 mg kg^-1 in the raw material to 1 100 mg kg^-1 in the final products. HS-Cu in H2O, NaOH, and Na4P2O7-NaOH extracts occupied 6.7, 26.7, and 19% averagely of total copper and HS-Zn represented 2.7, 13.7, and 17% averagely of total zinc in compost, respectively. In water extracts, both HA and FA mainly complexed with Cu and the mole ratio of Cu to Zn was 2.8 in HA fractions and was 2.6 in FA fractions, respectively. HA mainly complexed with copper, so that the ratios of HA-Cu to HA-Zn averaged 3.4 in NaOH extracts. FA had a similar potential to complex with copper and zinc, so that the ratio of FA-Cu to FA-Zn was close to 1. In Na4P2O7-NaOH extracts, HA or FA had a similar potential to complex with copper and zinc. The ratio of HS-Cu to HS-Zn was close to 1. With manure composting, Na4P2O7-NaOH extractable HS-Zn increased to a level as high as HS-Cu. This indicated that more and more stable complexes of HS-Zn were formed in the late decomposition period. The competition between copper and zinc to be complexed with humic substance became weaker and weaker with the decomposition process.

Enhancement of Phosphorus Solubility by Humic Substances in Ferrosols

An investigation was conducted to study the effect of humic substance (HS) on the phosphorus (P) solubility in acidic soil. The soil (2.5 g), HS (0, 0.5, and 2.5 g), and P as monocalcium phosphate (0.31 and 1.25 g P kg-1 soil) were mixed with 50 mL distilled water and two different sequences of adding HS and P were used. The results indicated that the P concentration in water and 0.01 mol L-1 CaCl2 solution increased with increasing amounts of humic substance. The concentrations of Fe and Al were also increased. However, Olsen P decreased with increasing amount of humic substance. Water-soluble P concentrations from P rates at 0.31 and 1.25 g P kg-1 soil in the treatment with 0.5 g (2.5 g) humic substance addition were 360% and 70% (500% and 90%) higher, respectively, than those in the treatment with no humic substance addition. P extracted by 0.01 mol L-1 CaCl2 in the treatments with 0.5 and 2.5 g humic substance addition was increased by 400% and 540%, respectively, compared with that in the treatment without humic substance at the rate of 0.31 g P kg-1 soil, while the corresponding P concentrations were increased by 80% and 90% at the rate of 1.25 g P kg-1 soil. The order of mixing humic substance and phosphate did not significantly affect desorbed P and labile P extracted with CaCl2.

Removal of humic substances by biosorption

Fungal pellets of Aspergillus niger 405,Aspergillus ustus 326,and Stachybotrys sp.1103 were used for the removal of humic substances from aqueous solutions.Batchwise biosorption,carried out at pH 6 and 25°C,was monitored spectrophotometrically and the process described with Freundlich’s model.Calculated sorption coeffcients Kf and n showed that A.niger exhibited the highest effciency.A good match between the model and experimental data and a high correlation coeffcient(R2)pointed out to judicious choice of ...

Partitioning of water soluble organic carbon in three sediment size fractions:Effect of the humic substances

Water soluble organic carbon （WSOC） in sediments plays an important role in transference and transformation of aquatic pollutants. This article investigated the inherent mechanisms of how sediemnt grain size affect the partitioning coeffcient （k） of WSOC. Influences of NaOH extracted humic substances were particularly focused on. Sediments were sampled from two cross-sections of the middle Yellow River and sieved into three size fractions （〈 63 μm, 63-100 μm, and 100-300 μm）. The total concentration of WSOC in sediments （Cwsoc） and k were estimated using multiple water-sediment ratio experiments. Results showed that Cwsoc ranged from 0.012 to 0.022 mg/g, while k ranged from 0.8 to 3.9 L/kg. Correlations between the spectrum characteristics of NaOH extracted humic substances and k were analyzed. Strong positive correlations are determined between k and the aromaticity indicators of NaOH extracted humic substances in different sediment size fractions. Comparing with finer fractions （〈 63 μm）, k is higher in larger size fractions （63- 100 μm and 100-300 μm） related to higher aromaticity degree of NaOH extracted humic substances mostly. While negative relationship between k and the area ratio of fourier transform infrared spectroscopy （FT-IR） at 3400 and 1430 cm^-1 implied that the lowest k was related to the highest concentration of acidic humic groups in particles 〈 63 μm. WSOC in finer fractions （〈 63 μm） is likely to enter into pore water, which may further accelerate the transportation of aquatic contaminants from sediment to water.

ADSORPTION OF HUMIC SUBSTANCES BY MACRO WEAKLY BASIC IONEXCHANGE RESIN AND THEIR EFFECTS ON REMOVAL OF Cu^(2+) AND Pb^(2+)

Adsorption of humic, tannic and gallic acids by a macro weakly basic ion-exchange resin JN-01 was studied. The adsorption capacity of this resin for gallic and tannic acids is much higher than that for humic acid, which can be explained on the basis of both their molecular size and ionization degree. Furthermore, humic acid is separated into different components with molecular weight in the range from 2000 Da to 100000 Da by ultra-filter, and their adsorption isotherms on resin JN-01 indicate that humic acid＇s molecular weight is an important factor which makes significant influence on adsorption. Finally, changes in the amount of Cu^2＋ and Pb^2＋ adsorbed on resin JN-01 as a function of the concentration of each of these three acids were studied. A large increase in the heavy metal ions uptake is observed in the presence of humic substance, such advantages are due to the interactions between the heavy metal ions and the unbound functional groups of the adsorbed organic acids.

Improving extraction yield of humic substances from lignite with anthraquinone in alkaline solution

Based on the characteristics of the lignite sample,effects of assistant anthraquinone(AQ) on extraction yield of humic substances(HS) and the action mechanisms of AQ in alkaline condition were studied by Fouvier transform infrared(FT-IR) spectroscopy.The results indicate that assistant AQ can not only increase the extraction yield of HS but also reduce the alkali dosage(NaOH) as well as the extraction temperature and extraction time.Under the optimal conditions of alkali dosage of 9%,AQ dosage of 0.75%,extraction temperature of 80 oC,extraction time of 30 min,stirring speed of 600 r/min and solid-to-liquid ratio of 1:3,the extraction yield of HS reaches 80.08%,which is increased by more than 20% compared with the conventional extraction.FT-IR spectra show that AQ is able to prevent dissolved HS from being destroyed into undissolved substance by alkali and HS obtained in the presence of AQ possesses more groups of -COOR and -COOH than that obtained without AQ.

^(15)N NMR Spectroscopic Study on Nitrogen Forms in Humic Substances of Soils

Nitrogen forms of humic substances from a subalpine meadow soil, a latentic red soil and a weathered coal 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 ammes, and the remaining intensities could not be assigned with certainty. Data on nonhydrolyzable residue of protein-sugar mixture and a 15N-labelled soil fulvic acid confirm the formation of nonhydrolyzable heterocyclic N during acid hydrolysis. Project (No. 39790100) supported by the National Natural Science Foundation of China.

Acid Dissociation Constants of Functional Groups in Humic Substances:I.Affinity Spectrum Model Analysis of Potentiometric Data of Fulvic and Humic Acids Solutions from Weathered Coal and Dark Loessial Soil

Potentiometric experiments were carried out on the proton binding equilibria of FA extracted from a weathered coal and HA and Fa extracted from a dark loessial soil.The affinity spectrum model was employed to treat the experimental data.The affinity spectrum model technique could“magnify” the heterogeneity of the proton binding equilibria.so it was useful for comparing and studying the characteristics of humic substances with similar properties.According to the affinity spectra,we also found that the direction of the titration could affect the properties of the equilibria of FA from the weathered coal,and the acidic functional groups contained in FA from the weathered coal were larger in quantity than those contained in HA and FA from the dark loessial soil.

THE OVERLOOKED ECOSYSTEM DRIVING FORCE IN NON-EUTROPHICATED FRESHWATER SYSTEMS:DISSOLVED HUMIC SUBSTANCES-A SHORT REVIEW AND OUTLOOK

This review starts with the description of the quantitative significance of dissolved organic material in general and dissolved humic substances (HS) in particular in various ecosystems. Despite their high quantities, the knowledge about the role of HS is still very low and full of old, but still recycled paradigms. HS are thought to be inert or at least refractory and too large to be taken up by aquatic organisms. Instead, I present evidence that dissolved HS that mainly derives from the terrestrial environment, are taken up and directly and/or indirectly interfere with freshwater organisms and, thus, structure biocenoses. Relatively well known is in the meantime the fuelling function of allochthonous HS, which, upon irradiation, release fatty acids, which serve as substrates for microbial growth. This is an indirect effect of HS. Microbes, in turn, are food for mixotrophic algae and (heterotrophic) zooplankton. Thus, non-eutrophicated freshwaters are net-heterotrophic, meaning that respiration exceeds primary production. Furthermore, model calculations exemplify that only a very small portion of the terrestrial production is sufficient to cause net-heterotrophy in these freshwater bodies. But, recent papers show also that due to different stoichiometries the maximal plankton biomass production with algae or mixotrophs is higher than with bacteria. Very recently, several direct effects of HS have been elucidated. Among them are:induction of chaperons (stress shock proteins), induction and modulation of biotransformation enzymes, modulation (mainly inhibition) of photosynthetic oxygen release of aquatic plants, production of an internal oxidative stress, modulation of the offspring numbers in the nematode Caenorhabditis elegans[WTBZ], feminization of fish and amphibs, interference within the thyroid system, and action as chemical attractant to C. elegans. We are still in the phase of identifying the various physiological, biochemical, and molecular-biological effects. Hence, the ecological and ecophysiological significance of these HS-mediated effects still remain somewhat obscure. Nevertheless, HS appear generally to have an impact on the individual as well as on the community and even ecosystem level comparable to that of, for instance, nutrients.

Effects of pH value and coagulant dosage on contact filtration of humic substances

Humic substances (especially fulvic acid (FA)) are the major components of natural organic matter (NOM) that widely exist in drinking water source. Due to their potential effects on public health,the removal of FA was one of the main concerns during the water treatment. Therefore,the contact filtration of FA by using aluminum sulfate as coagulant on the basis of jar tests was carried out. The effects of pH and coagulant dosage on the FA removal and the development of head loss were investigated. The results show that the range of pH value during the FA contact filtration can be effectively influenced by the dosage of aluminum sulfate,and the high aluminum sulfate dosage is an important factor that can result in early filter breakthrough. The FA filtration by deep-bed filtration or by membrane filtration is sometimes disparate under the same coagulation conditions. The choice of aluminum sulfate dosage by the method of membrane filtration,i.e. the "true color measurement",may result in inappropriate filter run,whereas it can be determined with simple jar tests by observing the formation of micro flocs. Considering the effects of pH on aluminum sulfate dosage and FA removal,the optimal pH range of 5.5?6.0 is suggested.

CONTENTS OF DISSOLVED HUMIC SUBSTANCES IN THE RIVERS OF EASTERN CHINA

Concentrations of dissolved humic substances in the rivers of eastern China were found to be 0.9- 14.4 mg/L with a median value of 4.6.The contents of dissolved humic substances in river water generally decrease from north to south,a trend similar to that of aquatic sediments and soils.Results of multiple regression analysis and path analysis showed that the dissolved stream humic substances are of a combined autochthonous and allochthonous origin.

Dissolution of Humic Substances from Highly Humic Volcanic Ash Soil as Affected by Anionic Surfactant, Electrolyte Concentration and pH

Dissolved humic substances separated from soils play an important role in the material cycle because they adsorb nutrients and contaminants and move with water. This study was conducted to investigate the influence of anionic surfactant, pH and electrolyte concentration on the dissolution of humic substances from a highly humic volcanic ash soil. The soil used in the experiment has a negative charge and the anionic surfactant, sodium dodecylbenzene sulfonate, has also the negative charge. The absorbance of supernatant of soil solution at different surfactant concentration and different electrolyte concentration (0.001 M, 0.01 M, 0.1 M & 0.5 M) of NaCl at pH 4.5 and 6.5 was measured at the wave-length of 400 nm;this corresponds to the relative concentration of dissolved humic substances. The surfactant adsorption and its equilibrium concentration under the same solution condition of the absorbance measurement were also measured in order to get their effect on dissolved humic substances. The zeta potential of soil particles was measured in order to evaluate the influence of electrostatic potential on dissolution of humic substances. The concentration of dissolved humic substances increased at higher surfactant concentration and adsorption, at higher pH and at lower electrolyte concentration, because the electrostatic repulsive force between the soil particles and the dissolving humic substances became larger. Therefore, surfactant concentration and adsorption, pH and electrolyte concentration are important when considering the fate of humic substances in soils.

Determination of Apparent Formation Constants of Eu(III) with Humic Substances by Ion Selective Liquid Membrane Electrode

This is the novel measurement of the apparent formation constants of trivalent lanthanide with humic substances by potentiometric titration using an ion selective electrode (ISE) consisting of bis(diphenylphosphoryl)methane as an ionophore. The ISE prepared exhibited a Nernstian response to the Eu3+ concentration in the test solutions in the presence of humic acid and fulvic acid. The effect of the solution conditions, including the pH, initial metal and humic substance concentrations, and ionic strength, on the formation constants was examined. The present formation constants were compared with the reported data using solvent extraction method.

APPLICATION OF SURFACE-ENHANCED RAMAN SPECTORSCOPY FOR CHARACTERIZING HUMIC SUBSTANCES

Surface-enhanced Raman Spectroscopy(SERS)has been used to characterize hurnic substances.By using HNO_3 -roughened silver foils as the enhancing surface,both fluorescence quenching and strong SERS signals have been observed at the same time.The results obtained herein show that SERS is a particularly sensitive and highly selective technique for the characterization of humic substances in the environment. Humic substances axe receiving ever-inereasing attention because of their importance in the environment. Very recently,we have reported the first successful application of Raman spectroscopy for characterizing humic substances.In this present paper,the application of surface-enhanced Raman spectroscopy(SERS)for humic substances has further been investigated.To our knowledge,no such studies have previously been reported.The results obtained herein dearly demonstrate that SERS has a great deal of promise as an analytical method for“In Situ”characterizing humic substances in the environment.

Structure characteristics and adhesive property of humic substances extracted with different methods

The structure characteristics and adhesive property of humic substance(HS) extracted with different methods were mainly studied by terms of elementary analysis,visible spectrum,FT-IR spectroscopy,viscosity,adsorption and pelletizing experiments.The results show that HSs extracted with new method(HS-a) own higher degree of aromatization and polymerization,larger relative molecular mass and more polar functional groups than HS extracted with usual method(HS-b).The viscosity of HS-b is about 30-40 mPa·s lower than that of HS-a.The maximum adsorption amounts of HS-a and HS-b onto iron concentrates are 9.11 mg/g and 8.08 mg/g,respectively.Meanwhile,HS-a has a better performance than HS-b in the practical application for pelletizing of iron concentrates.The difference in agglomeration behaviors with iron concentrates lies in the differences of the structure characteristics of HSs.With higher content of polar functional groups,larger relative molecular mass and viscosity of HSs,the agglomeration behavior is improved.

An XPS Study of Nitrogen Structures in Soil Humic Substances

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 amines, 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 benzoqu inone- (N H-4 )-2 S O-4 polymer the XPS results deviated significantly from the 15N CPMAS NMR data.