Humic materials fractionation

Humic materials fractionated on the basis of hydrophobicity and proton affinity continue to exhibit two fluorophores as discussed in the section "Exciation-Emission Spectra. Strong evidence to establish the existence of at least two chromophores is seen in the phase-resolved spectra. These spectra are shown in Figures 4 a-f. They consist of the phase-resolved emission spectrum of each of the two fluorophores plotted separately and the normal emission spectrum of the humic fraction. If the nulling out of one fluorophore is exact then the sum of the two separate phase resolved spectra should be additive to equal the normal spectrum. In these figures the normal emission spectrum was measured separately from the two phase resolved emision spectra. The phase resolved spectra were then superimposed onto the scan of the normal emission spectrum. 

In this chapter, we present the theory and results of measurements on humic acid fractions using fluorescence techniques. The fluorescence techniques are attractive for this application because of the natural fluorescence of humic materials, the hi sensitivity of fluorescence detection, and the ability to directly observe the morphology of the molecule in aqueous solutions without the need for drying or applying harsh chemical conditions. Several interesting types of information are obtained from fluorescence measurements ... 

In a dialysis experiment, a dialysis bag containing the dissolved humic materials is placed in a solution of a pollutant (preferably radiolabeled). The dialysis tubing is chosen so the pollutant is free to diffuse through the bag while the humic materials are retained inside the bag. The solution is shaken at constant temperature until it comes to an equilibrium point. At equilibrium, the pollutant inside the dialysis bag consists of two fractions that truly dissolved and the bound to the humic materials. The concentration of pollutant on the outside of the dialysis bag consists only of the free, truly dissolved fraction. Any increase of the pollutant concentration on the inside of the dialysis bag is due to binding by dissolved humic materials. A series of dialysis experiments, therefore, can measure the bound fraction concentration as a function of the free concentration. 

In a solubility experiment the solubility of the compound of interest is measured in the presence and absence of dissolved humic materials. Two techniques were used to measure solubility a shake and filter method similar to that used by Yalkowsky, and a flow through column technique similar to that used by May et al. 9 The measured solubilities of a number of compounds in our experiments were always higher in the presence of humic materials. This increase in the solubility is due to the binding of the compound by humic materials. In the presence of humic materials the measured solubility consists of two fractions free and bound. The free concentration should be the same in the presence or absence of humic materials. The difference between the solubilities of the compound in the presence and absence of humic materials is therefore a measurement of the bound fraction. 

The third technique we used was a measurement of changes in the sorption behavior of a compound in the presence of humic materials. A thin film of OV-1, a methyl silicone gum used as a chromatographic stationary phase, was plated on the bottom of a 60 ml Hypo-Vial (Pierce Chemical Co., Rockford, 11.). A solution of radiolabeled pollutant was added to the vial in either buffered distilled water in a solution of humic materials. Again, it is assumed that the pollutant is solution consists of two fraction free and bound. It is also assumed... 

A fraction of organic C oxidizable with 333 mM KMn04 is another measure of labile organic matter (Blair et al. 1995). This fraction encompasses all those organic components that can be readily oxidized by KMnCL including labile humic material and polysaccharides (Conteh et al. 1999). It commonly accounts for 15-20% of total soil organic C (Blair et al. 1998 Conteh et al. 1998). 

Many researchers have attempted to unravel the mystery of the structure of humus. One approach has been to isolate fractions by extracting humus using various extraction procedures. These procedures result in the isolation of three or more fractions humic acid, fulvic acid, and humin. Humic material is isolated from soil by treating it with alkali. The insoluble material remaining after this treatment is called humin. The alkali solution is acidified to a pH of 1.0 and the precipitate is called humic acid, while the soluble... 

Japenga et al. [56] determined polychlorinated biphenyls and chlorinated insecticides in River Elbe estuary sediments by a procedure in which the sediments were pretreated with acetic acid, mixed with silica and Soxhlet-extracted with benzene/hexane. Humic material and elemental sulphur were removed by passing the extract through a chromatographic column containing basic alumina, on which sodium sulphite and sodium hydroxide were adsorbed. Silica fractionation was followed by gas chromatography to analyse chlorinated pesticides, polychlorinated biphenyls and polyaromatic hydrocarbons. Recovery experiments with standard solutions gave recoveries of 90-102%. 

A fractionation procedure has been established and widely applied to studies of humic materials [42-44]. The procedure begins with natural OM (i.e., humus) and uses an aqueous basic solution (e.g., 0.1-0.5 mol/1 NaOH and Na2C03) to solubilize a fraction of the OM. The basic extract is then acidified which causes a precipitate to form, i.e., humic acids (HA). The fraction, which remains in solution, is called fulvic acids (FA). Humin is the name given to the insoluble organic fraction that remains after extraction of humic and fulvic acids. At nearneutral pH (pH 5 - 8), which is characteristic of most natural water, the FA are the most water soluble of these three fractions. HA are somewhat less soluble, with their solubility increasing as the pH increases. Humin is insoluble at all pH values. 

The difference in values of iCD0M for a solute with different types of fractionated humic materials has been explained in terms of the polarity, molecular size, and molecular configuration of the humic materials. It gives a reasonable estimate of the relative enhancing effects among humic extracts. 

The solubility of fuel oil no. 2, particularly the alkane and isoprenoid fractions, in seawater is increased by the presence of fulvic acid, although the solubilities of phenanthrene or anthracene, both polycyclic aromatic hydrocarbons, are unaffected by the presence of humic materials (Boehm and Quinn 1973). Unfiltered Narragansett Bay water was able to dissolve 1,560 g/L of fuel oil no. 2, although removal of... 

Environmental applications of FIFFF have been carefully collected in a review by Gimbert et al. [35]. Separations of nanoparticles belong to the FIFFF tradition and this sector has recently found new, fully deserved impulse for microparticle separations. The FIFFF technique has been applied to analyze humic material and submicron Fe colloids. Coupled with ICP-MS, FIFFF has been applied to detect the major and trace element chemistry of aquatic colloids in groundwaters and to determine the trace element distribution in soil and compost-derived humic and colloidal fractions in municipal wastewater. Recently, the ICP-AES has also been proposed as a specific detector for FIFFF to analyze inorganic nanoparticles (Figure 12.12). 

An extraction method for isolating humic substances from water by using XAD-8 has been proposed by Thurman and Malcolm (9) (see box). Humic substances in natural waters represent almost the entire hydrophobic acid fraction. This method has been used to isolate 4.25 g of humic substances from 24,500 L of ground water from the Fox-hills-Laramie aquifer and to obtain 500 g of humic material from 10,400 L of the Suwannee River (Table II). The sample from the Suwannee River was collected as a reference sample of aquatic humic substances by the International Humic Substances Society. In both of the examples cited, a fc cutoff of 100 was used. 

Humic material does not appear to be a prominent component of this HMW fraction. The principal components appear to be proteins and sugars, particularly N-acetylamino sugars, which probably originate from the degradation of microbial cell walls. 

The partitioning of Fluka humic material was studied at pH 3 and pH 7 to ascertain the amount of potential interference with subsequent GC analysis (19). Water-methylene chloride partition coefficients were determined by quantification of the humic concentration in pH 3 and pH 7 salt solutions by UV analysis at 254 nm of the humic material before and after methylene chloride extraction. The method followed the procedure of Suffet and Faust (7) in which p-values (fraction recovered in methylene chloride at 1 1 water methylene chloride) and E-values (fraction recovered in methylene chloride at any specified water-to-solvent ratio) were calculated. Watenmethylene chloride (10 1) was used for all E-value calculations. The values obtained were as follows for pH 3, p-value = 0.48 and 10 1 E-value = 0.07 for pH 7, p-value = 0.19 and 10 1 E-value = 0.02. 

A residual CP concentration is often observed after soil bioremediation. The leveling-off of degradation is not due to decreased microbial activity, since freshly added CPs are rapidly degraded (Harmsen, 1993 Salkinoja-Salonen et al., 1989). The residual concentrations are explained by the gradual diffusion of pollutants deep into micropores, as well as by their adsorption onto soil organic matter (Harmsen, 1993). Lagas (1988) observed that the nonextractable fraction of CPs in sterile soil increased according to the square root of time as a consequence of diffusion into humic material. 

Humic Materials. Humic materials are divided into three fractions based on their solubility in aqueous solutions as a function of pH humic acid, which is soluble in an alkaline aqueous solution fulvic acid, which is soluble in an aqueous solution regardless of pH and humin, which is insoluble in water at any pH value (and contains the OMN in soil organic matter). The chemical characteristics of humic acid and fulvic acid (e.g., Stevenson, 1994 Orlov, 1985 Rashid, 1985 Aiken et al., 1987 Hayes et al., 1987) and humin (Hatcher et al., 1985 Rice, 2001) are described in numerous reviews. 

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