Interference with xenobiotic metabolic activity by the commonly used vehicle solvents dimethylsulfoxide and methanol in zebrafish (Danio rerio) larvae but not Daphnia magna

Organic solvents, such as dimethylsulfoxide (DMSO) and methanol are widely used as vehicles to solubilise lipophilic test compounds in toxicity testing. However, the effects of such solvents upon innate detoxification processes in aquatic organisms are poorly understood. This study assessed the effect of solvent exposure upon cytochrome P450 (CYP)-mediated xenobiotic metabolism in Daphnia magna and zebrafish larvae (4 d post fertilisation). Adult D. magna were demonstrated to have a low, but detectable, metabolism of ethoxyresorufin in vivo and this activity was not modulated by pre-exposure to DMSO or methanol (24 h, up to 0.1% and 0.05% v/v, respectively). In contrast, the metabolism of ethoxyresorufin in zebrafish larvae was significantly reduced by both solvents (0.1% and 0.05% v/v, respectively) after 24 h of exposure. In zebrafish, these observed decreases in activity towards ethoxyresorufin were accompanied by decreased expression of a variety of genes coding for drug metabolising enzymes (corresponding to CYP1, CYP2, CYP3 and UDP-glucuronyl transferase [UGT] family enzymes), measured by quantitative PCR. Reduction of gene expression and CYP1 enzyme activities by methanol (0.05% v/v) in zebrafish larvae was partially reversed by co-exposure with Aroclor 1254 (100 μg L⁻¹). Overall this study suggests that relatively low concentrations of organic solvents can impact upon the biotransformation of certain xenobiotics in zebrafish larvae, and that this warrants consideration when assessing compounds for metabolism and toxicity in this species.     

Photochemical behavior of fenpropathrin and λ-cyhalothrin in solution

The photodegradation processes of fenpropathrin and λ-cyhalothrin were studied in hexane, methanol/water (1:1, v/v), and acetone in both ultraviolet light and simulated sunlight. Intermediates in the photodegradation process were identified using gas chromatography/mass spectrometry (GC/MS), and the analysis of intermediates was used to speculate on possible photodegradation pathways. The photodegradation processes of fenpropathrin and λ-cyhalothrin followed pseudo first-order kinetics. The photodegradation rates varied according to the solvent in decreasing order: hexane?>?methanol/water (1:1, v/v)?>?acetone. The effects of substances coexisting in the environment on the photodegradation of pyrethroids were also investigated in the research. Acetone, humic acid, and riboflavin increased photodegradation rates while l-ascorbic acid slowed the process. This study provides a theoretical basis for the removal of pyrethroid pollution from the natural environment.     

Evaluation of an extraction method for a mixture of endocrine disrupters in sediment using chemical and in vitro biological analyses

Aquatic sediments are contaminated by a wide diversity of organic pollutants such as endocrine-disrupting chemicals (EDCs) which encompass a broad range of chemical classes having natural and anthropogenic origins. The use of in vitro bioassays is now widely accepted as an alternative method for their detection in complex samples. However, based on the diversity of EDC chemical properties, their common extraction is difficult and comprehensive validation of extraction methods for a bioanalysis purpose is still weakly documented. In this study, we compared the performance of several organic solvents, i.e., acetone, methanol, dichloromethane, heptane, dichloromethane/acetone (50:50, v/v), dichloromethane/methanol (50:50, v/v), heptane/acetone (50:50, v/v), and heptane/methanol (50:50, v/v), to extract a diversity of active chemicals from a spiked sediment matrix using pressurized liquid extraction. For this purpose, we defined a mixture of 12 EDCs with a wide range of polarity (2?<?log Kow?<?8) (i.e., estrone, 17β-estradiol, bisphenol A, o,p′DDT, 4-tert-octylphenol, fenofibrate, triphenyl phosphate, clotrimazole, PCB-126, 2,3,7,8 TCDD, benzo[k]fluoranthene, and dibenzo[a,h]anthracene). Working concentrations of each individual compound in the mixture were determined as equipotent concentrations on the basis of the concentration-addition (CA) model applied to in vitro estrogenic, dioxin-like, and pregnane X receptor (PXR)-like activities. Extraction efficiencies based on both chemical and biological analyses were assessed in triplicate in artificial blank sediment spiked with this mixture and in natural sediment contaminated by native EDCs. In both spiked and natural sediment, MeOH/DCM yields the best recovery while heptane was the least efficient solvent. Our study provided the validation of a sediment extraction methodology for EDC bioanalysis purposes, which can be used for comprehensive environmental contamination characterization.     

Photolytic degradation of decabromodiphenyl ethane (DBDPE)

The photolytic degradation of decabromodiphenyl ethane (DBDPE), an alternative flame retardant to decabromodiphenyl ether, was investigated in a variety of matrixes (n-hexane, tetrahydrofuran, methanol/water, humic acid/water, and silica gel) by irradiation under ultraviolet light and in n-hexane under natural light. Photolytic degradation of DBDPE occurs in all the matrixes investigated within the irradiation period (<320 min). The degradation experiments showed varied reaction rates, dependent on the matrixes, with increasing half-lives (t_1/2) in the order of tetrahydrofuran (t_1/2 = 6.0 min) > n-hexane (t_1/2 = 16.6 min) > humic acid/water (30 < t_1/2 < 60) > silica gel (t_1/2 = 75.9 min) > methanol/water (t_1/2 > 240 min). The reaction in tetrahydrofuran, n-hexane, and silica gel matrixes can be described by the pseudo first order kinetics. Nevertheless, the matrixes have little effect on the degradation product distributions of DBDPE. A numbers of debrominated intermediates were identified. The degradation involves the initial formation of nona-BDPEs and the subsequent decomposition of these congeners to lower brominated congeners (octa- and hepta-BDPEs) within the irradiation time. To our knowledge, the present work is the first attempt to investigate the photolytic degradation kinetics and the identification of intermediates, as well as the degradation mechanism, during the degradation of DBDPE. Further research is needed to understand the photolytic degradation pattern of DBDPE in the natural environment.     

1H NMR-based metabolomics of time-dependent responses of Eisenia fetida to sub-lethal phenanthrene exposure

¹H NMR-based metabolomics was used to examine the response of the earthworm Eisenia fetida after exposure to sub-lethal concentrations of phenanthrene over time. Earthworms were exposed to 0.025 mg/cm² of phenanthrene (1/64th of the LC₅₀) via contact tests over four days. Earthworm tissues were extracted using a mixture of chloroform, methanol and water, resulting in polar and non-polar fractions that were analyzed by ¹H NMR after one, two, three and four days. NMR-based metabolomic analyses revealed heightened E. fetida responses with longer phenanthrene exposure times. Amino acids alanine and glutamate, the sugar maltose, the lipids cholesterol and phosphatidylcholine emerged as potential indicators of phenanthrene exposure. The conversion of succinate to fumarate in the Krebs cycle was also interrupted by phenanthrene. Therefore, this study shows that NMR-based metabolomics is a powerful tool for elucidating time-dependent relationships in addition to the mode of toxicity of phenanthrene in earthworm exposure studies.     

Visible-light driven oxidation of gaseous aliphatic alcohols to the corresponding carbonyls via TiO2 sensitized by a perylene derivative

Sensitized P25 TiO₂ was prepared by wet impregnation with a home-prepared perylene dye, i.e., N,N′-bis(2-(1-piperazino)ethyl)-3,4,9,10-perylene-tetracarboxylic acid diimide dichloride (PZPER). Energy levels of PZPER were found to be compatible with those of TiO₂ allowing fast electron transfer. The obtained catalyst has been characterized and used in the gas-phase partial oxidation of aliphatic primary and secondary alcohols, i.e., methanol, ethanol, and 2-propanol. The reaction was carried out under cut-off (λ?>?400 nm) simulated solar radiation in O₂ atmosphere. The perylene derivative allowed a good absorbance of visible radiation thanks to its low optical energy gap (2.6 eV) which was evaluated by cyclic voltammetry. The optimal organic sensitizing amount was found to be 5.6 % w/w in terms of yield in carbonyl derivatives. Moreover, no change in reactivity/selectivity was observed after 10-h irradiation thus confirming the catalyst stability. Yields into formaldehyde, acetaldehyde, and acetone were 67, 70, and 96 %, respectively. No significant amounts of organic byproducts were detected but for methanol oxidation, whereas a minor amount of the substrate degraded to CO₂.     

Sorption of polycyclic aromatic hydrocarbons (PAHs) on glass surfaces

Sorption of polycyclic aromatic hydrocarbons (PAHs) to glass commonly used in laboratories was studied. Sorption coefficients (K_d) of five selected PAHs to borosilicate glass surfaces were measured using column chromatography. A linear relationship between log K_d and the corresponding water solubility of the subcooled liquid (log S_w) of the investigated PAHs was observed. Based on the determined sorption coefficients our data revealed that mass loss caused by sorption on glass walls strongly depends on the ratio of solution volume to contacted surface area (V/S). The influence of solution chemistry such as ionic strength, solution pH, presence of cosolvent, and the influence of temperature on the sorption process were investigated. In the presence of ionic strength, sorption coefficients concurrently increased but less than a factor of 2 up to 0.005 M calcium chloride concentration. However, further increasing ionic strength had no influence on K_d. The cosolvent reduced sorption at a concentration of methanol in water above 0.5% (v/v); however, for benzo[a]pyrene even with 10% (v/v) methanol the mass loss would be still higher than 10% (with a V/S ratio less than 0.25). Significant effects of the solution pH and temperature were not observed. These results suggest that van der Waal’s forces dominate the sorption process. In the analysis of highly hydrophobic PAHs in aqueous samples, mass loss due to sorption on glass walls should be accounted for in the final result if untreated glass is used. The presented relationship between log K_d and log S_w may help to decide if such a correction is necessary. Furthermore, the frequently used silanization of glass surfaces may not be sufficient to suppress sorption for large PAHs.     

Fast determination of sulfonamides and their acetylated metabolites from environmental water based on magnetic molecularly imprinted polymers

Group-selective magnetic molecularly imprinted polymers (MMIPs) that can extract four widely used sulfonamide antibiotics and their acetylated metabolites from environmental water were synthesized in this study. The MMIPs with saturation magnetization value of 16.7 emu g^-1 could be separated from the environmental water samples easily by the application of an adscititious magnetic field, reducing the time consumption of pretreatment. The extraction conditions were evaluated, and optimal extraction conditions were as follows: extraction time, 25 min; amount of polymers, 90 mg; washing solvent, 30 % methanol aqueous solution; and elution solvent, methanol–acetic acid (95:5, v/v). The target analytes were detected by liquid chromatography–tandem mass spectrometry, and the detection limits of the method are in the range of 0.38–1.32 ng L^-1. The relative standard deviations of intra- and inter-day are in the range of 1.3–6.8 % and 1.7–9.1 %, respectively. The proposed method is suitable for the analysis of environmental water samples.     

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.     

Effect of the temperature and the exclusion of UVB radiation on the phenolics and iridoids in Menyanthes trifoliata L. leaves in the subarctic

The long-term effects of UVB exclusion and temperature on the methanol extractable (ME) phenolics (flavonoids, phenolic acids) and iridoids of Menyanthes trifoliata L. (Mt) leaves were studied in northern Finland (68°N) using wooden frames covered with filters for UVB exclusion (polyester filter), control (cellulose acetate filter) and ambient (no filter) conditions. Analysis of ambient plots showed no effect of the daily mean temperature (2σ = 1.58 °C) on the leaf ME compound content and composition, but minimum temperatures decreased the flavonol content. UVB exclusion did not affect the total ME compound content but significantly decreased the proportion of flavonols concomitantly with an increase in iridoids. Due to its high iridoid content, Mt appears as an interesting model plant for studying the iridoid biosynthesis and its regulation under stress conditions.     

Reaktion von Pendimethalin in Lösung bei UV-bestrahlung

Pendimethalin (I) decomposed in hexane, methanol and aqueous methanol upon irradiation with UV-light (λ > 290 nm) to form “polar” products. The main products formed are the dinitroaniline (II) and the isomers mono-nitroanilines (III). Kinetic studies indicate that the photodegradation of pendimethalin is strongly dependent of solvent and concentration.     

Discrepancies in the acute versus chronic toxicity of compounds with a designated narcotic mechanism

In this study, it was illustrated that even for certain simple organic compounds with a designated mode of action (MOA) (i.e. narcotic toxicity) unexpected differences in acute and chronic toxicity can be observed. In a first part of the study, species sensitivity distributions (SSDs) based on either acute or chronic toxicity data of three narcotic test compounds (methanol, ethanol and 2-propanol) were constructed. The results of the acute SSDs were as expected for narcotic compounds: rather similar sensitivity and small differences in toxicity were observed among different species. On the contrary, the chronic SSDs of methanol and ethanol indicated larger interspecies variation in sensitivity. Furthermore, the chronic toxicity trend (ethanol > methanol > 2-propanol) was unexpectedly different from the acute toxicity trend (2-propanol > ethanol > methanol) and acute versus chronic extrapolation could not be successfully described for methanol and ethanol using an ACR of 10 (as suggested for narcotic compounds). In contrast to the interspecies approach in the first part of this study, the second part of the study was focused on the assessment of acute and chronic toxicity of the three test compounds in Daphnia magna, which was identified as one of the most sensitive organisms to methanol and ethanol. Here, the differences in acute and chronic toxicity trend were in accordance to the results of the SSDs. The enhancement of membrane penetration due to the small molecular size of methanol and ethanol, in combination with the higher toxicity of their respective biotransformation products were suggested as potential causes of the increased chronic toxicity. Furthermore, it was stressed that larger awareness of these irregularities in acute to chronic extrapolations of narcotic compounds is required and should receive additional attention in further environmental risk assessment procedure.     

Surface chemistry of dihydromyrcenol (2,6-dimethyl-7-octen-2-ol) with ozone on silanized glass,glass, and vinyl flooring tiles

The surface-phase reaction products of dihydromyrcenol (2,6-dimethyl-7-octen-2-ol) with ozone (O₃), air, or nitrogen (N₂) on silanized glass, glass and vinyl flooring tile were investigated using the recently published FACS (FLEC (Field and Laboratory Emission Cell) Automation and Control System). The FACS was used to deliver ozone (100 ppb), air, or N₂ to the surface at a specified flow rate (300 mL min^?1) and relative humidity (50%) after application of a 2.0% dihydromyrcenol solution in methanol. Oxidation products were detected using the derivatization agents: O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine hydrochloride (PFBHA) and N,O-bis(trimethysilyl)trifluoroacetamide (BSTFA). The positively identified reaction products were glycolaldehyde, 2,6-dimethyl-5-heptenal, and glyoxal. The proposed oxidation products based on previously published VOC/O₃ reaction mechanisms were: 2,6-dimethyl-4-heptenal, 6-methyl-7-octen-2-one and the surface-specific reaction products: 6-methyl-6-hepten-2-one, 6-methyl-5-hepten-2-one, and 6-hydroxy-6-methylheptan-2-one. Though similar products were observed in gas-phase dihydromyrcenol/O₃ reactions, the ratio, based on peak area, of the reaction products was different suggesting stabilization of larger molecular weight species by the surface. Emission profiles of these oxidation products over 72 h are also reported.     

BVOC emissions, photosynthetic characteristics and changes in chloroplast ultrastructure of Platanus orientalis L. exposed to elevated CO2 and high temperature

To investigate the interactive effects of increasing [CO₂] and heat wave occurrence on isoprene (IE) and methanol (ME) emissions, Platanus orientalis was grown for one month in ambient (380 μmol mol⁻¹) or elevated (800 μmol mol⁻¹) [CO₂] and exposed to high temperature (HT) (38 °C/4 h). In pre-existing leaves, IE emissions were always higher but ME emissions lower as compared to newly-emerged leaves. They were both stimulated by HT. Elevated [CO₂] significantly reduced IE in both leaf types, whereas it increased ME in newly-emerged leaves only. In newly-emerged leaves, elevated [CO₂] decreased photosynthesis and altered the chloroplast ultrastructure and membrane integrity. These harmful effects were amplified by HT. HT did not cause any unfavorable effects in pre-existing leaves, which were characterized by inherently higher IE rates. We conclude that: (1) these results further prove the isoprene's putative thermo-protective role of membranes; (2) HT may likely outweigh the inhibitory effects of elevated [CO₂] on IE in the future.     

Characterization of an efficient catalytic and organic solvent-tolerant azoreductase toward methyl red from Shewanella oneidensis MR-1

The acyl carrier protein (ACP) phosphodiesterase gene (SO 4396) of Shewanella oneidensis MR-1 which was analyzed to have azoreductase activity was heterologously expressed in Escherichia coli. The ACP phosphodiesterase was found to reach maximum enzyme velocity 220.59 U/mg, named azoreductase in this study. The azoreductase had highest specific activity (153.16 U/mg) at pH 6.5, which showed a preference for nicotinamide adenine dinucleotide (NADH) as electron donor. The phylogenetic tree analysis indicated that the azoreductase had preference for NADH and dependence for flavin mononucleotide (FMN). However, the azoreductase from S. oneidensis MR-1 still had high enzyme activity in the absence of FMN. The Mg²⁺ had a positive influence on the enzyme activity with 25 mM concentration, whereas Cr³⁺, Cd²⁺ usually had significantly negative effect on enzyme activity. The purified azoreductase retained nearly 100 % activity after incubating in 30 % dimethyl sulfoxide (DMSO), 30 % acetone, 30 % methanol, 20 % ethanol, 20 % isopropanol, and 10 % propanol.     

Photochemical behavior of chlorsulfuronR in water and in adsorbed phase

The photochemistry of the herbicide chlorsulfuron^R (2-chloro-N-[[4-methoxy-6-methyl-1.3.5-triazinyl-2-amino]carbonyl]benzene sulfonamide) was investigated in methanol, distilled water, natural creek-water, on silica gel and on montmorillonit surfaces. The rate of degradation was determined in outdoor and indoor experiments simulating tropospheric conditions (λ>290 nm). Two major photoproducts were identified in all cases.     

Cascade utilization of water chestnut: recovery of phenolics, phosphorus, and sugars

Overgrowth of aquatic plants, such as water chestnut, has been reported as a regional problem in various areas. We proposed cascade utilization of water chestnut through the recovery of phenolics, phosphorus, and sugars. Phenolics were extracted using 50 g (wet weight) of biomass with 300 mL of acetone, methanol, or hot water, and the yields of total phenolics were 80.2, 56.2, and 49.7 mg?g^?1 dry weight of native biomass, respectively. The rate of eluted phosphorus in the phenolic extraction step was 8.6, 14.8, and 45.3 % of that in the native biomass, respectively, indicating that the use of polar organic solvents suppressed phosphorus elution at the phenolic extraction step. Extraction of phosphorus following the phenolic extraction was combined with alkaline pretreatment (1 % NaOH solution) of biomass for saccharification; 64.1 and 51.0 % of phosphorus in the native biomass were extracted using acetone and methanol for the phenolic extraction, respectively. Saccharification following the alkaline pretreatment showed that the glucose recovery rates were significantly increased (p?<?0.05) with the phenolic extraction step compared to alkaline pretreatment alone. This finding indicates that extraction of phenolics not only provides another useful material but also facilitates enzymatic saccharification.     

Removal of acutely hazardous pharmaceuticals from water using multi-template imprinted polymer adsorbent

Molecularly imprinted polymer adsorbent has been prepared to remove a group of recalcitrant and acutely hazardous (p-type) chemicals from water and wastewaters. The polymer adsorbent exhibited twofold higher adsorption capacity than the commercially used polystyrene divinylbenzene resin (XAD) and powdered activated carbon adsorbents. Higher adsorption capacity of the polymer adsorbent was explained on the basis of high specific surface area formed during molecular imprinting process. Freundlich isotherms drawn showed that the adsorption of p-type chemicals onto polymer adsorbent was kinetically faster than the other reference adsorbents. Matrix effect on adsorption of p-type chemicals was minimal, and also polymer adsorbent was amenable to regeneration by washing with water/methanol (3:1, v/v) solution. The polymer adsorbent was unaltered in its adsorption capacity up to 10 cycles of adsorption and desorption, which will be more desirable in cost reduction of treatment compared with single-time-use activated carbon.     

A flow-through passive dosing system for continuously supplying aqueous solutions of hydrophobic chemicals to bioconcentration and aquatic toxicity tests

A continuous supply of water with defined stable concentrations of hydrophobic chemicals is a requirement in a range of laboratory tests such as the OECD 305 protocol for determining the bioconcentration factor in fish. Satisfying this requirement continues to be a challenge, particularly for hydrophobic chemicals. Here we present a novel solution based on equilibrium passive dosing. It employs a commercially available unit consisting of ∼16 000 polydimethylsiloxane (PDMS) tubes connected to two manifolds. The chemicals are loaded into the unit by repeatedly perfusing it with a methanol solution of the substances that is progressively diluted with water. Thereafter the unit is perfused with water and the chemicals partition from the unit into the water. The system was tested with nine chemicals with log K_OW ranging from 4.1 to 6.3. The aqueous concentrations generated were shown to be largely independent of the water flow rate, and the unit to unit reproducibility was within a factor of ∼2. In continuous flow experiments the aqueous concentrations of most of the study chemicals remained constant over 8 d. A model was assembled that allows prediction of the operating characteristics of the system from the log K_OW or PDMS/water partition coefficient of the chemical. The system is a simple, safe, predictable and flexible tool that generates stable aqueous concentrations of hydrophobic chemicals.     

Methanol measurements in the lower troposphere near Innsbruck (047°16′N; 011°24′E), Austria

Diurnal variations on a time scale of minutes of the mixing ratios of methanol were measured using proton transfer reaction mass spectrometry (PTR-MS) technique during 67 days throughout the time span from November 1996 to July 1998 together with benzene and other volatile organic compounds at the western outskirts of Innsbruck, Austria. Comparison with the course of the mixing ratio of benzene, which served as marker for traffic emissions, as well as the observation of a seasonal variation allowed to distinguish between different sources for methanol release into the troposphere. Strong evidence for methanol removal via deposition on dew-wetted surfaces is obtained from the comparison of meteorological data with methanol mixing ratios. The mean volume mixing ratio of total methanol was 7.5 nmol mol⁻¹. Mixing ratios ranged from 0.03 up to 45 nmol mol⁻¹.