Time-resolved Fluorescence Measurements of Aquatic Natural Organic Matter (NOM)

The fluorescence decay of aquatic natural organic matter (NOM) samples was investigated using the time-correlated single photon counting technique (TCSPC). Two different approaches for the data analysis are presented: the discrete component approach (DCA) and the exponential series method (ESM). The parameter set obtained in the DCA is discussed in terms of characterization for NOM of different origins. However, the obtained parameter set can only be interpreted as operationally defined. Using the ESM for a fluorescence decay time distribution analysis no a priori assumption about the number of fluorescing components was introduced into the data analysis. The interpretation of fluorescence decay time data for samples before and after ozonation is in good agreement with results of other analytical methods.     

The Effects of Nutrients on Natural Organic Matter (NOM) Removal in Biological Activated Carbon (BAC) Filtration

Effective biodegradation of organic compounds is one of the major objectives while optimizing biological drinking water treatment processes. Enhancing the biological activated carbon (BAC) filter performance with nutrient addition was studied using chemically pre-treated and ozonated lake water. Three parallel pilot-scale biofilters were operated: one with phosphorus addition, one with a mixture of inorganic nutrients addition, and one as a reference. The addition of nutrients had no statistically significant influence on the natural organic matter (NOM) removal when monitored by total organic carbon (TOC), UV absorbance, and assimilable organic carbon (AOC). However, the addition of nutrients significantly increased the heterotrophic plate count (HPC) bacteria of the filter effluent, while the adenosine triphosphate (ATP) analysis of the attached bacteria did not show any increase in BAC filters. It seemed that in BAC filters the bacterial growth was limited by phosphorus, but the increased bacteria could not attach themselves during the relatively short acclimatization period.     

Fluorescence as a Tool for Tracing the Organic Contamination from Pulp Mill Effluents in Surface Waters

Three‐dimensional fluorescence spectra of water samples from an eucalyptus bleached kraft pulp mill and from a river, upstream and downstream of the discharge of the effluent, revealed the existence of a peak at δ_exc = 280 nm and δ_em = 340 nm Δδ = 60 nm), characteristic of effluentπs organic matter. Humic substances were isolated from the effluent by sequential adsorption onto resins XAD‐8 and XAD‐4 in series. Their synchronous fluorescence spectra with Δδ = 60 nm do also exhibit an intense signal at δ_exc = 280 nm (≈ 300 nm in the humic acid fraction). The peak is absent in the spectra of humic substances isolated from a non‐polluted site of the river, but it is clearly seen in the spectra of the humic substances from a site downstream of the discharge of the effluent. Synchronous fluorescence spectra (Δδ = 60 nm) of water samples from the river and its lagoon were recorded and revealed to be an easy and fast way of tracing the organic contamination from the effluent.     

Characterization of Estuarine and Fluvial Dissolved Organic Matter by Thermochemolysis using Tetramethylammonium Hydroxide

Selected samples of dissolved organic matter (DOM) isolated by ultrafiltration (UDOM) have been analyzed by thermochemolysis in the presence of tetramethylammonium hydroxide (TMAH). This technique cleaves ester and ether bonds of bio‐ and geological macromolecules and releases monomer subunits and methylates them in situ as their methyl ethers and methyl esters. Compared with conventional pyrolysis, TMAH thermochemolysis avoids decarboxylation of preexisting carboxylic moieties and produces aromatic acids as their methyl esters. Various phenolic derivatives, which might originate from incorporated lignin‐derived structures, from the highly aliphatic and resistant biopolymer cutan and also from proteinaceous materials, were identified among the products produced from UDOM upon thermochemolysis. The presence of lignin derivatives in UDOM indicates input of organic matter derived from terrestrial sources. Various aromatic acids, perhaps representing the final steps in the oxidation of the side‐chain during microbial oxidation of lignin, were released upon TMAH thermochemolysis, suggesting they are structural constituents of the UDOM. Different ratios of lignin‐derived materials, commonly determined using the CuO oxidation method, such as the Δ value, indicative of the amount of lignin present, the acid/aldehyde ratio (Ad/Al)_G, indicative of the extent of oxidative degradation of the lignin component, and the syringyl/guaiacyl (S/G) and p‐hydroxyphenyl/guaiacyl (P/G) ratios, indicative of the contribution for the different types of lignin, were determined.     

Origin of Dissolved Organic Carbon Studied by UV‐vis Spectroscopy

Dissolved organic carbon (DOC) distributions in water from Lake Ipê, MS, Brazil, were investigated. The samplings were performed monthly (surface, 1 m depth, and bottom) from June 1999 to June 2000. Absorbance at 285 nm and DOC concentrations in mg dm^—3, p(DOC), were highly correlated for the three depths. 77% of the surface, 85% for 1 m and bottom samples presented a variation between 20 dm³ g^—1 cm^—1 and 50 dm³ g^—1 cm^—1 of A(285 nm)/p(DOC), that characterizes the dissolved organic matter in lake water as essentially fulvic. The ratio A(254 nm)/p(DOC) was also sensitive for fulvic matter, and an A(250 nm)/A(365 nm) = 4 ratio was characteristic of strongly colored waters. The ratios A(436 nm)/p(DOC) for the three depths also showed a significant correlation. The predominance of fulvic acid is explained by environmental characteristics such as the tropical climate, temperatures above 18 °C, and the lake environment. It was demonstrated that the variation in the water carbon content due to different compartments in the lake can be monitored by UV‐vis spectroscopy ratios.     

Alkaline Degradation of Dissolved Organic Matter

The degradation of dissolved organic matter (DOM) was studied in alkaline solution. The products were characterised using UV/vis spectroscopy, size‐exclusion chromatography (SEC), and by the analysis of low‐molecular‐weight organic acids (LMWOA). The degradation experiments were performed with water from a brown water lake or its isolated fulvic acid fraction and sodium hydroxide at different reaction times and temperatures. Depending on the wavelength and the reaction time, the UV/vis absorbance between 230 nm and 600 nm increased or decreased. The behaviour of model compounds during reactions in alkaline media was compared to the UV/vis spectroscopic behaviour of DOM. The release of LMWOA was described by kinetic data and compared to the data of model reactions. Evidence was given for the carboxylic esters playing a significant role in the release of LMWOA only during the beginning of the alkaline degradation. The results gained by SEC with on‐line UV and DOC detection showed that the average size of DOM was decreasing, and that a major part of the degradation products consisted of low‐molecular‐weight mono‐ and dicarboxylic acids.     

Association of Humic Substances: Verification of Lambert‐Beer Law

The precision UV/Vis‐absorption spectra of a naturally occurring aquatic humic substance were measured in the range of concentration 0.003 to 1.4 g/L DOC (dissolved organic carbon). Lambert‐Beer law is well obeyed at low concentrations whereas deviations have been observed for DOC concentrations above 0.1 g/L: they are interpreted in terms of self‐solvation and formation of micelles.     

Interaction of Aquatic Dissolved Organic Matter(DOM) with Amitrole: The Nature of the Bound Residues

In this study the binding behaviour of the herbicide amitrole with humic substances respectively dissolved organic matter (DOM) is presented. The results of the binding and release experiments using radio actively ¹⁴C-labelled amitrole ledto the conclusion that the binding of amitrole is very low. About 70% of amitrole bound to humic substances was not extractable. The releasable amitrole portion of the purified DOM-amitrolecomplexes led to a harmful effect in the cress test. Solid-state cross polarisation (CP) magic angle spinning (MAS) ¹⁵N-NMR spectroscopy is a powerful spectroscopic method to elucidate the structure of bound xenobiotics. In addition extraction experiments were used resulting in a binding scheme of amitrole to DOM, where above all the covalent bonding occurred via acid amide bonds.     

Dissolved Organic Carbon Fractions Formed during Composting of Municipal Solid Waste: Properties and Significance

The properties and transformation of dissolved organic matter (DOM) extracted (10 L water per kilogram compost) from municipal solid waste (MSW) compost at five stages (days 47, 77, 105, 126, and 187) of composting were investigated. The DOM was fractionated into hydrophobic or hydrophilic neutrals, acids, and bases. The unfractionated DOM, the hydrophobic acids and neutrals (HoA and HoN, respectively), and the hydrophilic neutrals (HiN) fractions were studied using solid-state ¹³C-NMR, FTIR, and DRIFT spectroscopy. The HoA fraction was found to be the dominant (percentage of total DOM) hydrophobic fraction, exhibiting a moderate increase during composting. The HoN fraction increased sharply from less than 1% to 18% of the total DOM during 187 days of composting, while the hydrophobic bases (HoB) exhibited the opposite trend. The HiN represented the major fraction of the hydrophiles up to 120 days of composting, decreasing thereafter by 38%. The relative concentration of the hydrophilic acids and bases (HiA and HiB, respectively) exhibited no consistent trend during composting. DRIFT spectra of the unfractionated DOM taken from the composting MSW revealed a decreasing level of polysaccharide structures with time. The ¹³C-NMR and FTIR spectra of the HoA fraction exhibited a polyphenol-humic structure, whereas the HoN spectra exhibited strong aliphatic features. The spectra of the HiN fraction confirmed its polysaccharide nature. During the final stage of composting, the DOM concentration was steady, while a relative decrease of HiN concomitant with an increase of HoA and HoN fractions was observed. These indicate that the DOM contained a low concentration of biodegradable organic matter and a higher content of macromolecules related to humic substances. The biological significance and heavy metal binding of these fractions are being studied based on earlier observations showing enhanced plant growth in the presence of DOM extracted from mature as opposed to immature compost.     

Changes of Acute Toxicity of Organic Chemicals to Daphnia magna in the Presence of Dissolved Humic Material (DHM)

With a series of substituted anilines, a series of substituted phenols as well as diazinon and tetrabromobisphenol-A alterations of short-term acute toxicity to the waterflea Daphnia magna have been studied in the presence of dissolved humic material (DHM). The results may be categorised as follows: No alteration in acute toxicity occurred with: tetrabromobisphenol-A, o-toluidine, 3,4-dichloroaniline, and pentachlorophenol. Toxicity decreases significantly with: diazinon, 4-chloroaniline, and 4-nitrophenol. Significant toxicity increases were observed with 2,4-dichlorophenol and 2,4,5-trichlorophenol. As demonstrated for diazinon, our study reveals that in the presence of DHM and light the chemical speciation of this chemical is altered and daughter products are produced most likely via photoactive species of DHM. Most probably the toxicity of both the parent and daughter compounds is changed as well. Alterations in acute toxicity of organic chemicals may thus be attributed to the adsorption to DHM as well as to the photoactive species mediated production of daughter products with toxicity different from that of the parent compounds.     

Approaches to the Immunochemical Analysis of Non-extractable Triazine Residues in Refractory Organic Substances (ROS) and Characterization of ROS

The tendency of several pesticides to form non-extractable residues requires simple and fast screening methods. Immunochemical methods can meet this demand. They are based on selective and highly sensitive antibody binding to pesticide residues exposed on refractory organic substances (ROS). s-Triazines were used as an example to show the feasibility of this approach. It was proven that under laboratory conditions atrazine was bound to ROS. Antibody recognition pointed to the availability of free ethyl and isopropyl groups exposed at the non-extractable s-triazine residue. Therefore, it is suggested on the basis of antibody selectivities that binding of atrazine to ROS may take place by substitution of the chlorine residue. In the next step, aquatic ROS isolated from surface waters located in an agricultural area were examined for non-extractable s-triazines. ROS from a lake were found to contain bound residues, whereas none were detected in the creek samples. As an outlook the feasibility of serological characterization of ROS is discussed. The novel concept of serotyping is based on the fractionation of ROS by electrophoretic techniques followed by an immunoreaction with polyclonal antibodies directed against different ROS. When immunoelectrophoresis is used, precipitation lines can be compared and checked for identity and cross-reactivity. This approach can form the basis for classifying ROS and determine similarities with unknown ROS.     

Spectroscopic Characterization of Organic Structures and Organic‐Inorganic Interactions in Paper Mill Sludge

The organic composition and organic‐inorganic interaction in paper mill sludge (PS) solvent extracts (hexane, ethyl acetate, acetone and ethanol) and humic fractions, humic acid (HA) and humin (HU) were studied by electron paramagnetic resonance spectroscopy (EPR), proton and carbon‐13 nuclear magnetic resonance spectroscopy (¹H NMR; ¹³C NMR), Fourier‐transformed infrared spectroscopy (FTIR), and ultraviolet‐visible spectroscopy (UV‐vis). The strategy of fractionating the PS, sequentially, with organic solvents of increasing polarity is a reliable analytical procedure for humic substance sample separation because it results in more purified fractions. FTIR, ¹H NMR and ¹³C NMR results showed that hexane extract consisted mainly of aliphatic hydrocarbon structures. Their contents in the extracts decreased as the polarity of the extracting solvent increased and the content of oxygen functional groups increased. Carboxylic and carboxylate functional groups were found in the acetone extract, and ester and ether functions were predominantly found in the ethanol extract. EPR spectra revealed some Fe³⁺ complexes with rhombic structure (g₁ = 4.3; g₂ = 9.0) in the humic fractions and in all solvent extracts, except hexane. Quasi‐octahedral Fe³⁺ complexes (g = 2.3; ΔH_pp ≤ 400 G) were found in the HU fraction and in the acetone extract. The organic free radical content in the HA fraction was higher than the non‐fractionated PS sample and HU fraction.     

Differences between Humic Substances from Riverine,Estuarine, and Marine Environments Observed by Fluorescence Spectroscopy

Sixteen samples of fulvic acids and XAD‐4 fractions of riverine, estuarine, coastal, and open ocean origin have been studied by emission and synchronous molecular fluorescence spectroscopy. Certain features of the molecular fluorescence are related to the nature, the content, and the origin of those aquatic humic substances (HS). Riverine HS appear several times richer in fluorophores than marine HS, which can be well observed by emission fluorescence spectroscopy. Synchronous‐scan spectra of fulvic acids and of XAD‐4 fractions from the aquatic environments studied, emphasized the quality differences of their fluorophores. These features are useful as tracers of humic substances related with their natural environment source or even with their anthropogenic origin.     

Biochemical Characterization of Refractory Organic Substances

The characterization of refractory organic substances (ROS) is very complicated because of their heterogeneous structure. Size-exclusion chromatography with continuous detection of dissolved nitrogen (LC-DN), dissolved organic carbon (LC-DOC), and UV-absorbance (LC-UV) is a very useful analytical tool for the characterization of changes of ROS in natural aquatic systems and in technological treatment. The effect of natural, oxidative, and biochemical processes on formation and removal of ROS is described. Additionally the role of hydrolysable carbohydrates in the composition of ROS is presented.