Bioconcentration of nonylphenol in fathead minnows (Pimephales promelas)

Bioconcentration of p-nonylphenol (NP) by fathead minnows was determined under laboratory conditions. Fish were exposed continuously for 42 days to 0.33, 0.93 and 2.36 μgNP/l in a flow-through system. NP was Soxhlet extracted from whole fish homogenates with dichloromethane (DCM). The resulting extract was concentrated and bulk lipids removed by gel permeation and silica-gel chromatography. Compounds were identified and quantified by reverse-phase high-pressure liquid chromatography (RP-HPLC) with fluorescence detection. Mass spectrometry was used for verification of peak assignments. Bioconcentration factors (BCFs) ranged from 245 to 380.     

Quantification of bisphenol A, 353-nonylphenol and their chlorinated derivatives in drinking water treatment plants

Bisphenol A (BPA) and nonylphenols (NP) are of major concern to public health due to their high potential for human exposure and to their demonstrated toxicity (endocrine disruptor effect). A limited number of studies have shown that BPA and NP are present in drinking water. The chlorinated derivatives that may be formed during the chlorination step in drinking water treatment plants (DWTP) exhibit a higher level of estrogenic activity than their parent compounds. The aim of this study was to investigate BPA, 353NP, and their chlorinated derivative concentrations using an accurate and reproducible method of quantification. This method was applied to both surface and treated water samples from eight French DWTPs producing from surface water. Solid-phase extraction followed by liquid chromatography?Ctandem mass spectrometry was developed in order to quantify target compounds from water samples. The limits of detection ranged from 0.3 to 2.3?ng/L for BPA and chlorinated BPA and from 1.4 to 63.0?ng/L for 353NP and chlorinated 353NP. BPA and 353NP were found in most analyzed water samples, at a level ranging from 2.0 to 29.7?ng/L and from 0 to 124.9?ng/L, respectively. In most of DWTPs a decrease of BPA and 353NP was observed between surface water and treated water (36.6 to 78.9?% and 2.2 to 100.0?% for BPA and 353NP, respectively). Neither chlorinated BPA nor chlorinated 353NP was detected. Even though BPA and 353NP have been largely removed in the DWTPs studied, they have not been completely eliminated, and drinking water may consequently remain a source of human exposure.     

Determination of nonylphenol ethoxylates and octylphenol ethoxylates in environmental samples using 13C-labeled surrogate compounds

Alkylphenol polyethoxylates (APEOs) have been widely used as nonionic surfactants in a variety of industrial and commercial products. Typical compounds are nonylphenol polyethoxylates (NPEOs) and octylphenol polyethoxylates (OPEOs), which serve as precursors to nonylphenol (NP) and octylphenol (OP), respectively. NP and 4-t-OP are known to have endocrine disrupting effects on fish (medaka, Oryzias latipes), so it is important to know the concentrations of APEOs in the environment. Because the analytical characteristics of these compounds depend on the length of the ethoxy chain, it is necessary to use appropriate compounds as internal standards or surrogates. We synthesized two 13C-labeled surrogate compounds and used these compounds as internal standards to determine NPEOs and OPEOs by high-performance liquid chromatography (LC)-mass spectrometry. Method detection limits were 0.015 microg/L for NP (2)EO to 0.037 microg/L for NP(12)EO, and 0.011 microg/L for OP(3,6)EO to 0.024 microg/L for OP (4)EO. NPEO concentrations in water from a sewage treatment plant were less than 0.05-0.52 microg/L for final effluent and 1.2-15 microg/L for influent. OPEO concentrations were less than 0.05-0.15 microg/L for the final effluent and less than 0.05-1.1 microg/L for influent.     

Role of dissolved humic acids in the biodegradation of a single isomer of nonylphenol by Sphingomonas sp

This study shows the important role of humic acids in the degradation of ¹⁴C and ¹³C labeled isomer of NP by Sphingomonas sp. strain TTNP3 and the detoxification of the resulting metabolites. Due to the association of NP with humic acids, its solubility in the medium was enhanced and the extent of mineralization of nonylphenol increased from 20% to above 35%. This was accompanied by the formation of significant amounts of NP residues bound to the humic acids, which also occurred via abiotic reactions of the major NP metabolite hydroquinone with the humic acids. Gel permeation chromatography showed a non-homogenous distribution of NP residues with humic acids molecules, with preference towards molecules with high-molecular-weight. Solid state ¹³C nuclear magnetic resonance spectroscopy indicated that the nonextractable residues resulted exclusively from the metabolites. The chemical shifts of the labeled carbon indicated the possible covalent binding of hydroquinone to the humic acids via ester and possibly ether bonds, and the incorporation of degradation products of hydroquinone into the humic acids. This study provided evidences for the mediatory role of humic acids in the fate of NP as a sink for bacterial degradation intermediates of this compound.     

Removal of estrogenic activities of bisphenol A and nonylphenol by oxidative enzymes from lignin-degrading basidiomycetes

Bisphenol A (BPA) and nonylphenol (NP) were treated with manganese peroxidase (MnP) and laccase prepared from the culture of lignin-degrading fungi. Laccase in the presence of 1-hydroxybenzotriazole (HBT), the so-called laccase-mediator system, was also applied to remove the estrogenic activity. Both chemicals disappeared in the reaction mixture within a 1-h treatment with MnP but the estrogenic activities of BPA and NP still remained 40% and 60% in the reaction mixtures after a 1-h and a 3-h treatment, respectively. Extension of the treatment time to 12 h completed the removal of estrogenic activities of BPA and NP. The laccase has less ability to remove these activities than MnP, but the laccase-HBT system was able to remove the activities in 6 h. A gel permeation chromatography (GPC) analysis revealed that main reaction products of BPA and NP may be oligomers formed by the action of enzymes. Enzymatic treatments extended to 48 h did not regenerate the estrogenic activities, suggesting that the ligninolytic enzymes are effective for the removal of the estrogenic activities of BPA and NP.     

Variation in estrogenic activity among fractions of a commercial nonylphenol by high performance liquid chromatography

Estrogenic activity by recombinant yeast screen assay of the commercial NP was considerably higher when compared with that of n-nonylphenol (n-NP). Fractionation of the commercial NP by high performance liquid chromatography (HPLC) afforded seven isomers: 4-(1,3-dimethyl-1-propyl-butyl)-phenol, 4-(1,1,3-trimethyl-hexyl)-phenol, 4-(1,1-dimethyl-3-ethyl-pentyl)-phenol, 4-(1,1,4-trimethyl-hexyl)-phenol, 4-(1-methyl-1-propyl-pentyl)-phenol, 4-(1,1,2-trimethyl-hexyl)-phenol and 4-(1-ethyl-1-methyl-hexyl)-phenol. The structures of these isomers were determined by GC-MS and nuclear magnetic resonance spectroscopy (NMR). All of these isomers possessed tertiary alpha-carbon in their chemical structures. Another tertiary NP, 4-(1,1-dimethyl-heptyl)-phenol was synthesized in the present study and this synthetic NP also exhibited the estrogenic activity. One fractionated compound was identified as one of decylphenol, 4-(1-ethyl-1,4,4-trimethyl-pentyl)-phenol. The isomer, 4-(1,1,4-trimethyl-hexyl)-phenol exhibited the highest estrogenic activity corresponding to 1/10000 that of 17beta-estradiol (E2). The activity of n-NP was the least. This suggests that it may be possible to develop a technical NP mixture with relatively low estrogenic activity.     

Environmental transformation of 1-nitropyrene on glass surfaces

We have investigated the photochemical/chemical transformation of solid 1-nitropyrene (NP) in natural light to gain some understanding of the fate of nitroaromatic compounds released into the atmosphere. Glass plates coated with ¹⁴C-labeled NP were exposed continuously outdoors for periods of 12–670 h. Samples kept in the dark at ambient temperatures and at −20°C were also studied. Breakdown products were separated from the parent compound by high-pressure liquid chromatography, quantitated by liquid scintillation spectroscopy and identified by standard spectroscopic techniques. The rate of NP breakdown could be fitted with a two-component exponential. Half-times of breakdown for the rapid and slower components of breakdown are 14 and 533 h, respectively. Breakdown appears to have resulted in loss of the nitro group with formation of hydroxypyrene, possibly pyrene dione and dihydroxy pyrene, and several as yet unidentified compounds. Mutagenic activities of nitropyrene and its breakdown products were determined in Salmonella strain TA-98, with and without the addition of rat liver S-9. Mutagenic activities of breakdown products were significantly less than that of NP, probably because of loss of the nitro group. Further work is needed to determine if these results also apply to particle-associated NP that is airborne or deposited on surfaces.     

Degradation of nonylphenol by anaerobic microorganisms from river sediment

We investigated the degradation of nonylphenol monoethoxylate (NP1EO) and nonylphenol (NP) by anaerobic microbes in sediment samples collected at four sites along the Erren River in southern Taiwan. Anaerobic degradation rate constants (k1) and half-lives (t1/2) for NP (2 microg/g) ranged from 0.010 to 0.015 1/day and 46.2 to 69.3 days respectively. For NP1EO (2 microg/g), the ranges were 0.009-0.014 1/day and 49.5-77.0 days respectively. Degradation rates for NP and NP1EO were enhanced by increasing temperature and inhibited by the addition of acetate, pyruvate, lactate, manganese dioxide, ferric chloride, sodium chloride, heavy metals, and phthalic acid esters. Degradation was also measured under three anaerobic conditions. Results show the high-to-low order of degradation rates to be sulfate-reducing conditions > methanogenic conditions > nitrate-reducing conditions. The results show that sulfate-reducing bacteria, methanogen, and eubacteria are involved in the degradation of NP and NP1EO, with sulfate-reducing bacteria being a major component of the river sediment.     

Biodegradation of nonylphenol in river sediment

We investigated the biodegradation of nonylphenol monoethoxylate (NP1EO) and nonylphenol (NP) by aerobic microbes in sediment samples collected at four sites along the Erren River in southern Taiwan. Aerobic degradation rate constants (k1) and half-lives (t1/2) for NP (2 microg g(-1)) ranged from 0.007 to 0.051 day(-1) and 13.6 to 99.0 days, respectively; for NP1EO (2 microg g(-1)) the ranges were 0.006 to 0.010 day(-1) and 69.3 to 115.5 days. Aerobic degradation rates for NP and NP1EO were enhanced by shaking and increased temperature, and delayed by the addition of Pb, Cd, Cu, Zn, phthalic acid esters (PAEs), and NaCl, as well as by reduced levels of ammonium, phosphate, and sulfate. Of the microorganism strains isolated from the sediment samples, we found that strain JC1 (identified as Pseudomonas sp.) expressed the best biodegrading ability. Also noted was the presence of 4'-amino-acetophenone, an intermediate product resulting from the aerobic degradation of NP by Pseudomonas sp.     

Determination of 4-tert-octylphenol, 4-nonylphenol and bisphenol A in surface waters from the Haihe River in Tianjin by gas chromatography-mass spectrometry with selected ion monitoring

The estrogenic pollutants 4-tert-octylphenol (OP), 4-nonylphenol (NP) and bisphenol A (BPA) were determined in surface water samples from the Haihe River, Tianjin, China. The analytes were extracted and concentrated from 300 ml acidified water samples by liquid–liquid extractions using dichloromethane, derivatized with trifluoroacetic anhydride, and quantified by gas chromatography–mass spectrometry (GC–MS) with selected ion monitoring (SIM). Among the samples collected from 14 sampling sites, only one sample was found to have a relatively high concentration of BPA (8.30 μg l⁻¹) and NP (0.55 μg l⁻¹). The concentrations of OP, NP and BPA in the other samples were in the range of 18.0–20.2, 106–296 and 19.1–106 ng l⁻¹, respectively. Recoveries for OP, NP and BPA in the spiked water samples were all over 80%.     

Comparison of lignin peroxidase and horseradish peroxidase for catalyzing the removal of nonylphenol from water

Concentrations of aqueous-phase nonylphenol (NP), a well-known endocrine-disrupting chemical, are shown to be reduced effectively via reaction with lignin peroxidase (LiP) or horseradish peroxidase (HRP) and hydrogen peroxide. We systematically assessed their reaction efficiencies at varying conditions, and the results have confirmed that the catalytic performance of LiP toward NP was more efficient than that of HRP under experimental conditions. Mass spectrum analysis demonstrated that polymerization through radical–radical coupling mechanism was the pathway leading to NP transformation. Our molecular modeling with the assistance of ab initio suggested the coupling of NP likely proceeded via covalent bonding between two NP radicals at their unsubstituted carbons in phenolic rings. Data from acute immobilization tests with Daphnia confirm that NP toxicity is effectively eliminated by LiP/HRP-catalyzed NP removal. The findings in this study provide useful information for understanding LiP/HRP-mediated NP reactions, and comparison of enzymatic performance can present their advantages for up-scale applications in water/wastewater treatment.     

Effects of waterborne exposure to 4-nonylphenol on plasma sex steroid and vitellogenin concentrations in sexually mature male carp (Cyprinus carpio)

4-Nonylphenol (NP) has been shown to elicit estrogenic responses both in vivo and in vitro. The mechanism by which NP exerts estrogenic and other endocrine-modulating effects in vivo remains unclear, however. The goal of this study was to evaluate the ability of NP to elicit estrogenic responses through indirect mechanisms of action involving the modulation of endogenous steroid hormone concentrations. Sexually mature male common carp (Cyprinus carpio) were exposed to aqueous NP concentrations ranging from <0.05 to 5.4 microg NP/l for 28-31 d. Approximately 0.5-3.5 ppm of NP was detected in pooled plasma samples or tissue samples from the carp studied. NP exposure did not significantly increase plasma concentrations of 17beta-estradiol (E2), testosterone (T) or vitellogenin (VTG). Excluding outliers, plasma E2 concentrations ranged from <175 to 700 pg E2/ml. T concentrations ranged from 940 to 24,700 pg T/ml plasma. The greatest VTG concentration detected was 52 microg/ml. One-third of the plasma samples tested contained <1 microg VTG/ml. Overall, the results of this study did not support the hypothesis that exposure to waterborne NP can modulate concentrations of steroid hormones in the plasma of sexually mature male carp. The results did, however, raise a number of questions regarding the utility of estradiol equivalent (EEQ) estimates as a means of predicting in vivo effects of estrogenic substances. Furthermore, they provide information regarding the concentrations and variability of E2, T, and VTG in the plasma of sexually mature male carp, which may aid in design and interpretation of future studies.     

Seasonal and spatial distribution of nonylphenol in Lanzhou Reach of Yellow River in China

Nonylphenol (NP) is known as an endocrine disruptor and consequently has drawn much environmental concern. This study focused on seasonal variation and spatial distribution of NP in various matrices including water, suspended particles, and sediment taken from Lanzhou Reach of Yellow River in China. NP was measured in July and November in 2004. Concentrations of NP in water ranged from 34.2 to 599.0 ng/l, in suspended particles from 49.6 to 2835.2 ng/g dry wt, and in sediment from 38.4 to 863.0 ng/g dry wt. In terms of most water and suspended particles samples, concentrations were higher in warmer seasons than in colder seasons. Good linear correlations (R(2)=0.90 in July, R(2)=0.97 in November) were obtained for NP concentrations between water and suspended particles. In terms of sediment samples, concentrations were higher in November than in July, probably due to greater deposition of suspended particles. Reasonable linear correlations (R(2)=0.60 in July, R(2)=0.79 in November) were obtained for NP concentrations between water and sediment.     

Environmental dynamics of phosphate esters. II. Uptake and bioaccumulation of 2-ethylhexyl diphenyl phosphate and diphenyl phosphate by fish

Uptake, clearance and disposition of ¹⁴C-labelled 2-ethylhexyl diphenyl phosphate (EHDP) and ¹⁴C-diphenyl phosphate (DPP) by rainbow trout (Salmo gairdneri) were investigated using short-term static exposures. Bioconcentration factors of 1314 and 0.3 were calculated for EHDP and DPP, respectively. Highest levels of radioactivity were found in liver and intestine following exposure to 60 μg/L EHDP. A portion (up to 40%) of the radioactivity in each tissue was in the form of DPP while a similar portion was extractable with methanol but could not be identified.     

Determination and distribution characteristics of degradation products of nonylphenol polyethoxylates in the rivers of Taiwan

Concentrations of degradation products of nonylphenol polyethoxylates (NPEOs) were analyzed in river water samples in order to determine the distribution characteristic of these alkylphenolic compounds in 18 major rivers of Taiwan. The degradation products of NPEOs were detected in all river samples, with the dicarboxylates alkylphenolic degradation products (CAPEC) being detected most frequently and at the highest concentrations. Concentrations of NP and NP1EO in rivers ranged from n.d. to 5.1 μg l⁻¹ and n.d. to 0.5 μg l⁻¹, respectively. The total concentrations of shortened carboxylates (i.e., NP1EC + NP2EC + NP3EC) and dicarboxylates alkylphenolic degradation products (CAP1EC + CAP2EC) ranged from n.d. to 63.6 μg l⁻¹ and n.d. to 94.6 μg l⁻¹, respectively. Concentrations of NP2EC, NP3EC and all CAPEC residues were determined semi-quantitatively by comparing with the internal standard. Significantly higher concentrations of CAPEC residues were detected in the river waters as compared to those of NP, NP1EO and NPEC degradation products and the average proportions of these compounds in the samples of the rivers were as follows: NP + NP1EO was 5 ± 2.5%, total NPEC was 25 ± 12%, and total CAPEC was 70 ± 12%. The high concentration ratios of CAPEC/NPEC illustrate that aerobic biodegradation plays a main route in the fate of NPEO in the rivers of Taiwan.     

An exploration of the relationship between adsorption and bioavailability of pesticides in soil to earthworm

A study was conducted to determine the adsorption/desorption of butachlor, myclobutanil and chlorpyrifos on five soils using a batch equilibration technique and to study the relationship between bioavailability to Allolobophora caliginosa and the adsorption/desorption of these three pesticides. The results showed that the adsorption/desorption processes of the tested compounds were mainly controlled by soil organic matter content (OM) and octanol/water-partitioning coefficient (K(ow)), and that the bioavailability of the pesticides was dependent on characteristics of pesticides, properties of soils, and uptake routes of earthworms. Bioconcentration of butachlor and myclobutanil was negatively correlated with Freundlich adsorption constant K(af) and K(df). However, only a slightly positive correlation between bioconcentration and K(af) and K(df) was observed for chlorpyrifos due to its high affinity onto soil.     

Comparison of biodegradation of surfactants in soils and sludge-soil mixtures by use of 14C-labelled compounds and automated respirometry

The biodegradability of dodecyl benzene sulphonate (LAS), nonylphenol-di-ethoxylate (NP2EO) and tridecyl-tetra-ethoxylate (LAE) in soil was examined by use of 14C experiments at two concentrations (10 and 400 mg/kg). Increasing the concentration of test chemical from 10 to 400 mg/kg resulted in a decrease in the relative maximum mineralization rate and an increase in the estimated lag times of a factor of approximately 3.5. In sludge-amended soil, the highest expected environmental concentration (just after sludge application) will be around 10 mg/kg for linear alkylbenzene sulphonate (LAS), while the concentration of NP2EO and linear alcohol ethoxylates (LAE) will be much lower. However, when using a respirometric method it is necessary to use a higher concentration of test substance in order to detect biodegradation. In our experiment, amendment with anaerobically digested sludge resulted in a decrease in the mineralization of LAS, NP2EO and LAE for all soils. Respirometric experiments were carried out at 400 mg/kg and could be used for estimation of biodegradation potential of LAS, NP2EO and LAE in soil and sludge-amended soil. For LAS, the results obtained from the respirometric experiments were similar to the results obtained in the 14C experiments, whereas NP2EO and LAE showed a faster degradation in the respirometric experiments.     

Trace metal accumulation by algae in Sepetiba Bay, Brazil

Trace metal concentrations (Zn, Cd, Pb, Cr and Cu) were determined in some benthic algae from Sepetiba Bay, Brazil. This region has been modified by the increase of industrial activities during the last 30 years. Among species sampled the brown algae Padina gymnospora contained the greatest amount of Zn. The Bioconcentration Factor for Zn in P. gymnospora was found to be 10(4) under laboratory conditions. The absorption of (65)Zn depended on exposure time and increased with Zn concentrations in the medium. An insignificant amount of Zn desorbed from algae suggested a very high affinity with cellular binding sites. Zinc uptake by P. gymnospora is discussed by considering field and laboratory conditions.     

Estrogenicity profile and estrogenic compounds determined in river sediments by chemical analysis, ELISA and yeast assays

An effects-directed strategy was applied to bed sediments of a polluted tributary in order to isolate and identify the major estrogenic chemicals it discharges into the River Po, the principal Italian watercourse. Sediment extract was concentrated by solid phase extraction and then fractioned into 10 fractions by reversed phase high performance liquid chromatography (RP-HPLC). Estrogenic activity of whole extract and fractions were determined using a recombinant yeast assay containing the human estrogen receptor (YES). The 10 fractions and whole extract were analysed for target compounds, e.g. estrone (E1), 17beta-estradiol (E2), estriol (E3), 4-nonylphenol (NP), 4-tert-octylphenol (t-OP), bisphenol A (BPA), using both liquid chromatography-tandem mass spectrometry (LC-MS/MS) and non-competitive enzyme-linked immunosorbent assays (ELISA). The YES assay determined high estrogenic activity in whole sediment (15.6 ng/g EE2 equivalents), and positive results for fractions nr 1, 2, 6, 7 and 8. E1, E3 and NP were the main estrogenic chemicals, however, other unidentified compounds contributed to sediment estrogenicity, particularly for polar fractions nr 1 and 2. A GC-MS screening performed in scan mode identified other potential contributors such as phthalates (DBP, BBP), and OP isomers. A next sampling campaign extended to other tributaries and receiving stretches of the River Po confirmed E1, E3 and NP as major estrogenic chemicals potentially threatening other sites of the main river. In general, target compound ELISAs have been shown to be suitable tools for a rapid screening of wide areas or large numbers of environmental samples for estrogenic risk. The potential for interferences suggests however to use cautiously the concentration values obtained from some of the immunoassays.     

Juvenile sea bass biotransformation, genotoxic and endocrine responses to beta-naphthoflavone, 4-nonylphenol and 17 beta-estradiol individual and combined exposures

Juvenile sea bass, Dicentrarchus labrax L., were exposed during 2, 4, 8, and 24 h to 0.9 microM beta-naphthoflavone (BNF), 131 nM 17 beta-estradiol (E(2)), 4.05 microM 4-nonylphenol (NP), as well as to BNF combined either to E(2) or NP (maintaining the previous concentrations). Liver cytochrome P450 content (P450), ethoxyresorufin-O-deethylase (EROD), and glutathione S-transferase (GST) activities were measured in order to evaluate biotransformation responses. Genotoxicity was assessed as erythrocytic nuclear abnormalities (ENA) frequency. The effects on endocrine function were evaluated as plasma cortisol and glucose. Cortisol was not affected by xeno/estrogens tested, either in single exposure or mixed with BNF. Nevertheless, the intermediary metabolism was affected since glucose concentration increased after 4 h exposure to E(2), and after all BNF+NP exposure lengths. Moreover, a synergism between BNF and NP was thoroughly demonstrated, whereas a sporadic antagonistic interaction was found at 4 h BNF + E(2) exposure. Liver EROD and GST activities were not significantly altered by single E(2) or NP exposure. However, both compounds were able to induce EROD activity in the presence of BNF. NP single exposure was able to significantly increase liver P450 content, while its mixture with BNF displayed an antagonistic interference. Considering the xeno/estrogens single exposures, only NP induced an ENA increase; however, both mixtures (BNF + E(2) and BNF + NP) displayed genotoxic effects. Fish responses to mixtures of xenobiotics are complex and the type of interaction (synergism/potentiation or antagonism) in a particular mixture can vary with the evaluated biological response.