Humic substances as catalysts in condensation reactions

Humic substances (HS) demonstrate appreciable impact on the rate of the condensation reactions as shown in the example of the reaction between hydrazine and 4-(dimethylamino)-benzaldhyde in an aquatic environment. The catalytic activity of HS has also been demonstrated in Knoevenagel and Claisen-Schmidt reactions for condensation of carbonyl compounds with CH acids. The aquatic fulvic acids are the most active in these reactions. The velocity of the studied reactions also depends on pH, temperature, the concentration and origin of HS used. A possible micellar and acid-base catalysis mechanism in aquatic media has been suggested.     

Influence of chemical characteristics of humic substances on the partition coefficient of a chlorinated dioxin

The partition coefficients (Koc) of 1,2,3,4,6,7,8-heptachlorodibenzo-p-dioxin (HpCDD) with respect to a variety of humic substances (HSs) were evaluated by a method involving solid-phase microextraction and gas chromatography-electron capture detection. The log Koc values for each of the HS samples were in the range of 6.4-7.7. The log Koc values for HAs from tropical peat, brown forest and ando soils were in the range of 7.3-7.6, similar to the calculated value for the octanol-water partition coefficient (log Koc=7.56). In contrast, the log Koc values for FAs and peat HAs were 0.5-1 unit lower than the calculated value. The parameters for the polarity of HSs, as calculated from (N+O)/C, O/C atomic ratios and the carboxyl group content, were numerically similar related to the log Koc for HpCDD. These results show that the Koc values for HpCDD are significantly influenced by the polarity of HSs.     

Does quinone or phenol enrichment of humic substances alter the primary compound from a non-algicidal to an algicidal preparation?

Dissolved organic matter (DOM) has been shown to affect phytoplankton species directly. These interactions largely depend on the origin and molecular size of DOM and are different in prokaryotes and eukaryotes. In a preceding study, however, two humic substance preparations did not adversely affect coccal green algae or cyanobacterial growth even at high concentrations of dissolved organic carbon (DOC). These results contradicted previous findings, showing a clear, negative response of different phototrophs to much lower DOC concentrations. To test whether or not at least defined building blocks of humic substances (HSs) are effective algicidal structures, we enriched two humic preparations with hydroquinone and p-benzoquinone, respectively, and exposed two different green algae, Pseudokirchneriella subcapitata and Monoraphidium braunii, and two cyanobacterial species, Synechocystis sp. and Microcystis aeruginosa, to the unmodified and enriched HSs. As response variables, growth rates in terms of biomass increase, chlorophyll-a content, and photosynthetic yield were measured. The highest concentration (4.17 mM DOC) of the modified HSs clearly inhibited growth; the cyanobacterial species were much more sensitive than the green algal species. However, realistic ecological concentrations did not adversely affect growth. Aerating the exposure solution for 24 h strongly reduced the inhibitory effect of the modified HSs. The algicidal effect was obviously caused by monomers and not by polymerised high molecular weight HSs themselves. Furthermore, the maximum quantum yield (Φ PSII max) was stimulated in the green algal species by low and medium DOC concentrations, but reduced in the cyanobacterial species upon exposure to higher HS concentrations. The quinone- and phenol-enriched HSs only showed algicidal activity at high concentrations of 4.17 mM DOC and lost their effects over time, presumably by oxidation and subsequent polymerisation. This study confirms that the applied humic substances themselves are not effective algicides even if enriched in effective structures.     

Release of substituents from phenolic compounds during oxidative coupling reactions

Phenolic compounds originating from plant residue decomposition or microbial metabolism form humic-like polymers during oxidative coupling reactions mediated by various phenoloxidases or metal oxides. Xenobiotic phenols participating in these reactions undergo either polymerization or binding to soil organic matter. Another effect of oxidative coupling is dehalogenation, decarboxylation or demethoxylation of the substrates. To investigate these phenomena, several naturally occurring and xenobiotic phenols were incubated with various phenoloxidases (peroxidase, laccase, tyrosinase) or with birnessite (delta-MnO(2)), and monitored for chloride release, CO(2) evolution, and methanol or methane production. The release of chloride ions during polymerization and binding ranged between 0.2% and 41.4%. Using the test compounds labeled with 14C in three different locations (carboxyl group, aromatic ring, or aliphatic chain), it was demonstrated that 14CO(2) evolution was mainly associated with the release of carboxyl groups (17.8-54.8% of the initial radioactivity). Little mineralization of 14C-labeled aromatic rings or aliphatic carbons occurred in catechol, ferulic or p-coumaric acids (0.1-0.7%). Demethoxylation ranged from 0.5% to 13.9% for 2,6-dimethoxyphenol and syringic acid, respectively. Methylphenols showed no demethylation. In conclusion, dehalogenation, decarboxylation and demethoxylation of phenolic substrates appear to be controlled by a common mechanism, in which various substituents are released if they are attached to carbon atoms involved in coupling. Electron-withdrawing substituents, such as -COOH and -Cl, are more susceptible to release than electron-donating ones, such as -OCH(3) and -CH(3). The release of organic substituents during polymerization and binding of phenols may add to CO(2) production in soil.     

The double-edged sword of humic substances: contrasting their effect on respiratory stress in eastern rainbow fish exposed to low pH

High amounts of humic substances (HS) are commonly found in natural acidic waterways and have been suggested to offer some protection against low pH. This study investigated the ability of HS to decrease respiratory stress in eastern rainbowfish (Melanotaenia splendida splendida) exposed to decreases in pH (range of 7–3.5) in soft and hard water. Repeated measures ANOVA revealed a significant difference in respiration (time taken for ten operculum movements) between pH and HS treatments, with a significant interaction between pH and HS present in 5/6 trials. Respiratory stress was shown to increase with increasing acidity, but significantly decreased in treatments with HS (10 and 20 mg/L) compared to those without. The fish exposed to pH treatments without HS also displayed increased hyperactivity, larger operculum movements and increased mucous production. Increased morbidity was shown in HS treatments at pH 3.5 (soft water) and at pH 4 (hard water) compared to treatment without HS. This indicates that HS is helpful in ameliorating the effects of decreased pH on respiration at sublethal pH levels; however, as pH decreases further, it seems that HS increases the toxicity (morbidity) of the low pH.     

Effect of complexation with humic substances on diffusion of metal ions in water

Most studies on diffusion of metal ions in various water-rock systems have dealt with free ions (hydrated ions). However, it is often the case that metal ions are dissolved as complexed species such as with humic substances (HS) in natural waters. Hence, we need to study the diffusion behavior of these complexes in order to understand fully the diffusion phenomenon in natural. In this study, the diffusion coefficients of free metal ions (M(z+)) and their complexes with HS (M-HS) were compared to understand the effect of complexation with HS on the diffusion of metal ions such as Co(2+), Cd(2+), and rare earth elements (REE(3+)). Although the diffusion coefficients of free metal ions depend on ionic potential, such dependence was not observed in the presence of HS. Comparing the diffusion coefficients of metal complexes with ethylenediaminetetraacetate (EDTA), fulvic acid, and humic acid showed that the molecular weight (MW), or the size of the ligand, is of primary importance for the diffusion of M-HS. As a consequence, the diffusion coefficients of all REE(3+) were similar in the presence of HS, while they were different in the absence of HS due to the different size of each REE(3+). The similarity among the diffusion coefficients of REE-HS was caused by the much larger size of HS compared with each ion. However, the distribution coefficients of M-HS were not similar among REE(3+), Cd(2+), and Co(2+). REE(3+) and Cd(2+) which have higher affinities for larger MW fraction in HS diffused slower than Co(2+) which favors smaller MW fraction. The results show that the affinity for different MW fractions among HS controls the diffusion of M-HS, which must be important to predict precisely the diffusion behavior of metal ions bound to HS in natural systems.     

Natural Organic Matter Differently Modulates Growth of Two Closely Related Coccal Green Algal Species (6 pp)

Background

and Aim. Humic substances (HS) comprise the majority of dead and living organic carbon, including organisms. In the environment, they are considered to be chemically inert or at least refractory. Recent papers, however, show that HS (including natural organic matter – NOM, isolated by reverse osmosis) are natural chemicals which interact with aquatic organisms. They are taken up and cause a variety of stress defense reactions which are well known from man-made chemicals. These reactions include chaperon activation, induction and modulation of biotransformation enzymes, or induction of antioxidant defense enzymes. One specific reaction with freshwater plants is the reduction of photosynthetic oxygen release. In this contribution, we compare the susceptibilities (cell yield) of two closely related coccal green algae, Monoraphidium convolutum and M. minutum, towards various NOM isolates.

Methods

Cultures of M. convolutum and M. minutum were obtained from the algal collection of the Leibniz Institute of Freshwater Ecology and Inland Fisheries, Berlin, and from the Culture Collection of Algae, Göttingen, and maintained in a common medium. The cultures were non-axenic. The algae were exposed to 5 mg L-1 DOC of each humic material, an environmentally realistic concentration. Cell numbers were counted microscopically in Neugebauer cuvettes in 5 replicates on days 1, 4, 7, 10, 14, and 21.

Results and Discussion

Almost all NOM isolates modulated the growth of the algae. Only the NOM of a Norwegian raised peat bog lake did not reveal any significant effect with M. convolutum. In general, the results with two algal species are by no means uniform. For instance, Suwannee River NOM causes a decrease in cell density with M. minutum, but temporarily stimulates the growth of M. convolutum. The opposite applies to Aurevann NOM: Growth increase in M. minutum, but a bi-phasic response in M. convolutum. Different responses of both Monoraphidium species must be attributed to intrinsic factors of the algae rather than only to chemical features of the exposed materials, because the exposures were identical with both algal species. The reduction in growth yields can be explained as a herbicide-like mode of action that affects the photosystem II most prevalently. The growth promoting effect remains somewhat obscure. It may be due to (1) an increase in bioavailability of some trace nutrients in the presence of HS, (2) the release of some growth promoting substances by microbial or photochemical processing of the humic materials, and (3) a hormetic effect upon the exposure of HS. Hormesis means stimulation of organisms or metabolic activities when exposed to noxes in low concentrations. However, it is still open to discussion why the growth promotion only applies to one or the other, but not simultaneously to both Monoraphidium species.

Conclusion

and Perspectives. Exposure of the closely related coccal green algal species to humic material changes their growth characteristics. Since the reactions are not consistent within the two species and the various humic materials, it seems that the less sensitive species is favored by HS exposure. The environmental relevance, however, is subject to future studies.     

Environmental signals: synthetic humic substances act as xeno-estrogen and affect the thyroid system of Xenopus laevis

According to outdated paradigms humic substances (HS) are considered to be refractory or inert that do not directly interact with aquatic organisms. However, they are taken up and induce biotransformation activities and may act as hormone-like substances. In the present study, we tested whether HS can interfere with endocrine regulation in the amphibian Xenopus laevis. In order to exclude contamination with phyto-hormones, which may occur in environmental isolates, the artificial HS1500 was applied. The in vivo results showed that HS1500 causes significant estrogenic effects on X. laevis during its larval development and results of semi-quantitative RT-PCR revealed a marked increase of the estrogenic biomarker estrogen receptor mRNA (ER-mRNA). Furthermore, preliminary RT-PCR results showed that the thyroid-stimulating hormone (TSHbeta-mRNA) is enhanced after exposure to HS1500, indicating a weak adverse effect on T3/T4 availability. Hence, HS may have estrogenic and anti-thyroidal effects on aquatic animals, and therefore may influence the structure of aquatic communities and they may be considered environmental signaling chemicals.     

Humic substances

GOALS, SCOPE AND BACKGROUND: Changing environmental conditions and handling stress are well known to cause chronic or acute stress situations in fish with subsequent infections. These requires a therapy by means of antibiotics and chemicals. In contrast to the huge number of pathogens, only a few substances are permitted for application as therapeutics in German aquaculture. Hence, there emerges an urgent need for highly effective and residueless alternatives. MAIN FEATURES: The prophylactic stimulation and training of the defense system of fish by alternative approaches becomes increasingly necessary. One approach is the application of dissolved humic substances (HS) of natural or artificial origin. For example, there exist several reports on the positive effect of HS to fishes. These effects shall be considered in detail. Furthermore, the impact of HS on the constitution of parasites and pathogens will be displayed. The reports on this issue are diverse, if not inconsistent. We try to shed some light on these discrepancies. The last aspect covered by this review is the outdated paradigm that calcium ions act as antidotes. In the presence of HS, even the opposite effect may occur. APPROACH: To overcome old paradigms on HS and their potential interactions with fish and fish parasites, we reviewed recent international literature, as well as 'grey' literature. We also include results from own former and ongoing studies. RESULTS AND DISCUSSION: HS are able to increase the physiological condition of the individuals and to reduce adverse physiological and histological consequences caused by stress; the mechanism behind remains obscure. HS detoxify heavy metals and organic pollutants. Damages caused by several fish pathogens, such as bacteria and parasites, can be repaired more quickly in the presence of HS. Some parasites--mainly fungi--appear to be directly affected by HS. Comparing the fungicidal effects of HS from various sources, evidence is increasing that the aliphatic moiety may be the effective structures. However, further research is necessary to relate more physiological and anti-pathogenic effects to the chemical characteristics of HS. CONCLUSIONS: HS are not real alternatives to strong traditional therapeutics. However, they show different advantages in repairing secondary, stress induced damages in fish. The ecophysiological relevance of HS in either aquatic systems or aquaculture is getting conspicuously. PERSPECTIVES: The lack of therapeutic and antiparasitic substances in aquaculture requires new strategies and ways of thinking. The search for alternatives to the 'traditional' chemical therapeutics calls for the intensive research. Inevitably, this search will lead to an intensive contemplation on HS as 'health promoting substances' and/or even therapeutics. Basic research is needed to detect the functional groups of the HS responsible for the effects observed. Health promoting effects of first investigations made in vitro to affect pathogens via application of HS and several field studies with HS raises hopes for a broader utilisation of HS to reduce stress consequences in fish and fish pathogens residuelessly.     

Interaction of temperature and an environmental stressor: Moina macrocopa responds with increased body size,increased lifespan,and increased offspring numbers slightly above its temperature optimum

For organisms, temperature is one of the most important environmental factors and gains increasing importance due to global warming, since increasing temperatures may pose organisms close to their environmental tolerance limits and, thus, they may become more vulnerable to environmental stressors. We analyzed the temperature-dependence of the water-soluble antioxidant capacity of the cladoceran Moina macrocopa and evaluated its life trait variables with temperature (15, 20, 25, 30 °C) and humic substance (HS) concentrations (0, 0.18, 0.36, 0.90, 1.79 mM DOC) as stressors. Temperatures below and above the apparent optimum (20 °C) reduced the antioxidative capacity. Additions of HSs increased body length, but decreased mean lifespan at 15 and 20 °C. There was no clear HS-effect on offspring numbers at 15, 20, and 30 °C. At 25 °C with increasing HS-concentration, lifespan was extended and offspring numbers increased tremendously, reaching 250% of the control. Although the applied HS preparation possesses estrogenic and antiandrogenic activities, a xenohormone mechanism does not seem plausible for the reproductive increase, because comparable effects did not occur at other temperatures. A more convincing explanation appears to be the mitohormesis hypothesis which states that a certain increase of reactive oxygen production leads to improved health and longevity and, with Moina, also to increased offspring numbers. Our results suggest that at least with the eurythermic M. macrocopa, a temperature above the optimum can be beneficial for several life trait variables, even when combined with a chemical stressor. Temperatures approximately 10 °C above its optimum appear to adversely affect the lifespan and reproduction of M. macrocopa. This indicates that this cladoceran species seems to be able to utilize temperature as an ecological resource in a range slightly above its thermal optimum.     

Sorption of humic substances on aquifer material at artificial recharge of groundwater

Experiments in batch equilibrium system were carried out to evaluate the importance of physical and chemical factors determining the sorption efficiency of humic substances (HS) on aquifer material, which has been used for artificial recharge of groundwater (ARG) in drinking water production. Results showed that an increase of the amount of clay in the aquifer material and a decrease of pH in water increased the sorption efficiency. The sorption of higher molecular weight, more hydrophobic and aromatic HS (Aldrich and forest soil humic acids) were greater than the sorption of acidic HS (river fulvic acids), either on the aquifer material or to its representative sorbing phases, clay and organic matter. The sorption on the aquifer material was largely due to physical sorption (hydrophobic attractions). This study showed the importance of HS composition on their removal during ARG and contributed to an understanding of the HS sorption mechanisms in this process.     

Humic substances

GOAL, SCOPE AND BACKGROUND: Freshwater bodies which chemistry is dominated by dissolved humic substances (HS) seem to be the major type on Earth, due to huge non-calcareous geological formations in the Northern Hemisphere and in the tropics. Based on the paradigm of the inertness of being organic, direct interactions of dissolved HS with freshwater organisms are mostly neglected. However, dissolved organic carbon, the majority of which being HS, are natural environmental chemicals and should therefore directly interact with organisms. Major results that widened our perspective on humic substance ecology come from experiments with the compost nematode, Caenorhabditis elegans, which behaved contradictorily to textbook knowledge and provoked an in-depth re-consideration of some paradigms. APPROACH: To overcome old paradigms on HS and their potential interactions with organisms, we reviewed recent international literature, as well as 'grey' literature. We also include results from own ongoing studies. RESULTS: This review focuses on direct interactions of dissolved HS with freshwater organisms and disregards indirect effects, such as under-water light quenching. Instead we show with some macrophyte and algal species that HS adversely interfere with photosynthesis and growth, whereby closely related algal species show different response patterns. In addition to this, HS suppress cyanobacteria more than eukaryotic algae. Quinones in the HS appear to be the effective structure. Furthermore, HS can modulate the offspring numbers in the nematode C. elegans and cause feminization of fish and amphibians--they possess hormone-like properties. The ecological consequences of this potential remain obscure at present. HS also have the potential to act as chemical attractants as shown with C. elegans and exert a mild chemical stress upon aquatic organisms in many ways: induction of molecular chaperons (stress proteins), induction and modulation of biotransformation and anti-oxidant enzymes. Furthermore, they produce an oxidative stress with lipidperoxidation as one clear symptom or even stress defense strategy. Stronger chemical stresses by HS may even lead to teratogenic effects as shown with fish embryos; all physiological responses to HS-mediated stress require energy, which were compensated on the expense of yolk as shown with zebra fish embryos. One Finnish field survey supports the view of a strong chemical stress, as the weight yield in fish species decreases with increasing HS content in the lakes. DISCUSSION: HS exert a variety of stress symptoms in aquatic and compost organisms. According to current paradigms of ecotoxicology, these symptoms have to be considered adverse, because their compensation consumes energy which is deducted from the main metabolism. However, the nematode C. elegans looks actively for such stressful environments, and this behavior is only understandable in the light of new paradigms of aging mechanisms, particularly the Green Theory of Aging. In this respect, we discuss the mild HS-mediated stress to aquatic and compost organisms. New empirical findings with HS themselves and HS building blocks appear to be consistent with this emerging paradigm and show that the individual lifespan may be expanded. At present the ecological consequences of these findings remain obscure. However, a multiple-stress resistance may be acquired which improves the individual fitness in a fluctuating environment. CONCLUSIONS: It appears that dissolved HS have to be considered abiotic ecological driving forces, somewhat less obvious than temperature, nutrients, or light. PERSPECTIVES: The understanding of the ecological control by dissolved humic substances is still fragmentary and needs to be studied in more details.     

Comprehensive characterization of oil refinery effluent-derived humic substances using various spectroscopic approaches

Refinery effluent-derived humic substances (HS) are important for developing refinery effluent reclamation techniques and studying the environmental chemistry of wastewater effluents. In this study, dissolved organic matter (DOM) from refinery effluent was concentrated using a portable reverse osmosis (RO) system. HS were isolated from RO retentates with XAD-8 resin. A variety of approaches such as specific UV absorbance at 254nm (SUV(254)), elemental analysis, size exclusion chromatography (SEC), solid-state cross polarization magic angle spinning (13)C nuclear magnetic resonance spectrometry ((13)C CPMAS NMR), Fourier transform infrared spectrometry (FTIR), and electrospray ionization/ion trap/mass spectrometry (ESI/ion trap/MS) were employed for characterization of HS. The portable RO system exhibited high yield and recovery of DOM for concentrating refinery effluent. The concentration of dissolved organic carbon (DOC) in the refinery effluent was 9.9mg/l, in which humic acids (HA) and fulvic acids (FA) accounted for 2.3% and 34.6%, respectively. Elemental and SUV(254) analyses indicated relative high amounts of aliphatic structures and low amounts of aromatic structures in refinery effluent-derived HS. Refinery effluent-derived HS displayed lower molecular weight than natural HS. The number-average molecular weight (M(n)) and the weight-average molecular weight (M(w)) of HA were 1069 and 2934, and those of FA were 679 and 1212 by SEC, respectively. By ESI/ion trap/MS, the M(n) and the M(w) of FA were 330 and 383. Four kinds of carbon structures (aliphatic, aromatic, heteroaliphatic, and carboxylic carbons) were found in refinery effluent-derived HS by (13)C NMR analysis. The quantitative results support the interpretation that these HS are rich in aliphatic carbons and poor in aromatic carbons. Proteinaceous materials were identified by FTIR analysis in refinery effluent-derived HS.     

The lanthanum precipitation method. Part 2: quantification of the conditional interaction constant between technetium(IV) and humic substances

The interaction between colloidal Tc(IV) species and colloidal Gorleben humic substances (HS) was quantified after application of the La-precipitation method on supernatant solutions obtained under various experimental conditions but at constant ionic strength of the Gorleben groundwater (0.04M). The determined interaction constant LogK(HS) (2.3+/-0.3) remained unchanged over a large range of Tc(IV) and HS concentrations and was independent of the pH of the original supernatant solution (pH range 6-10), Tc(IV)-HS loading (10(-3)-10(-6)molTcg(-1) HS) and the nature of the reducing surface (Magnetite, Pyrite and Gorleben sand) used for the pertechnetate reduction. The LogK(HS) value determined by the La-precipitation method is lower than the LogK value obtained from a previous study where the interaction between colloidal Tc(IV) species and Gorleben humic substances was quantified using a modified Schubert approach (2.6+/-0.3). The La-precipitation method allows to accurately determine the amount of Tc(IV) associated with HS but leads to a (small) overestimation of the free inorganic Tc(IV) species.     

Effect of some heavy metals and soil humic substances on the phytochelatin production in wild plants from silver mine areas of Guanajuato, Mexico

Phytochelatins (PCs) were determined in the wild plants, focusing on their relationship with the levels of heavy metals and humic substances (HS) in soil. Ricinus communis and Tithonia diversifolia were collected from several sites in Guanajuato city (Mexico), which had long been the silver and gold mining center. The analysis of PCs in root extracts was carried out by liquid chromatography (derivatization with monobromobimane). Total Ag, Cd, Cu and Pb in plant roots and in soil samples, as well as soil HS were determined. The association of metals with HS in soils was evaluated by size exclusion chromatography (SEC) with UV and ICP-MS detection. The results obtained revealed the induction of PCs in R. communis but not in T. diversifolia. The levels of Cd and Pb in plant roots presented strong positive correlation with PC-2 (r = 0.9395, p = 0.005; r = 0.9573, p = 0.003, respectively), indicating that these two metals promote PCs induction in R. communis. On the other hand, the inverse correlation was found between soil HS and metal levels in roots of R. communis (Cu > Pb > Cd > Ag), in agreement with the decreasing affinity of these metals to HS. Importantly, the inverse correlation between soil HS and plant PC-2 was observed (r = −0.7825, p = 0.066). These results suggest that metals strongly bound to HS could be less bioavailable to plants, which in turn would limit their role in the induction of PCs. Indeed, the SEC elution profiles showed Pb but not Cd association with HS and the correlation between metal in soil and PC-2 in plant was statistically significant only for Cd (r = 0.7857, p = 0.064). Based on these results it is proposed that the role of heavy metals in PCs induction would depend on their uptake by R. communis, which apparently is controlled by the association of metals with soil HS. This work provides further evidence on the role of environmental conditions in the accumulation of heavy metals and phytochelatin production in plants.     

Fluorescence of sediment humic substance and its effect on the sorption of selected endocrine disruptors

Humic substances (HS) have a critical influence on the sorption of organic contaminants by soils and sediments. This paper describes investigations into the sorption behavior of three representative endocrine disruptors, bisphenol A (BPA), 17beta-estradiol (E2), and 17alpha-ethynylestradiol (EE2), onto sediments and HS extracted sediments using a batch technique. The organic carbon-normalized partition coefficients (K(oc)) for the extracted HS (K(oc)(hs)) were calculated, and the fluorescence spectra of the HS extraced from different sediment samples were gained using excitation/emission matrix (EEM). Particular attention was paid to the correlations between the fluorescence characteristics of HS and the log K(oc)(hs) of selected endocrine disruptors. The results show that the log K(oc)(hs) values range from 3.14 to 4.09 for BPA, from 3.47 to 4.33 for E2, and from 3.65 to 4.32 for EE2. Two characteristic excitation-emission peaks were observed for HS samples extracted from sediments. They are located at Ex/Em=250-260 nm/400-450 nm (peak alpha') and Ex/Em=310-330 nm/390-400 nm (peak alpha) respectively. The alpha' and alpha peak relative intensities I(alpha')/I(alpha) vary from 0.46 to 1.64 for different extracted HS samples. The similarity between fulvic acids (FA) Ex/Em pairs and those observed for HS indicates that FA is the predominant fraction of HS extracted from sediments. Moreover, the log K(oc)(hs) values of BPA, E2, and EE2 have a negative linear correlation to I(alpha')/I(alpha) values. Peak alpha is often attributed to relatively stable and high molecular weight aromatic fulvic-like matter. Therefore, the result presented here reveals that the abundance of aromatic rings in HS molecular structure plays a critical role in the sorption of selected endocrine disruptors.