Mixture Effects of 3 Mechanistically Different Steroidogenic Disruptors (Prochloraz, Genistein, and Ketoconazole) in the H295R Cell Assay.

Mixture effects of 3 model endocrine disruptors, prochloraz, ketoconazole, and genistein, on steroidogenesis were tested in the adrenocortical H295R cell line. Seven key steroid hormones (pregnenolone, progesterone, dehydroepiandrosterone, androstenedione, testosterone, estrone, and 17ß-estradiol) were analyzed using gas chromatography and tandem mass spectrometry (GC-MS/MS) to investigate the effects throughout the steroidogenic pathway. Current modeling approaches often rely on models assuming compounds acting independently and that the individual effects in some way can be summarized to predict a mixture effect. In H295R cells with an intact steroidogenic pathway, such assumptions may not be feasible. The purpose of this study was therefore to evaluate whether effects of a mixture with differing modes of action followed or deviated from additivity (concentration addition) and whether the H295R cell line was suitable for evaluating mixture toxicity of endocrine disruptors with different modes of action. The compounds were chosen because they interfere with steroidogenesis in different ways. They all individually decrease the concentrations of the main sex steroids downstream but exert different effects upstream in the steroidogenic pathway. Throughout the study, we observed lowest observed effect concentrations of mixtures at levels 2 to 10 times higher than the predicted EC(50), strongly indicating antagonistic effects. The results demonstrate that chemical analysis combined with the H295R cell assay is a useful tool also for studying how mixtures of endocrine disruptors with differing modes of action interfere with the steroidogenic pathway and that existing models like concentration addition are insufficient in such cases. Furthermore, for end points where compounds exert opposite effects, no relevant models are available.

Determination of steroid hormones in blood by GC-MS/MS.

This paper presents the development, optimization and validation of a methodology to determine nine key steroid hormones (viz. pregnenolone, progesterone, dehydroepiandrosterone, androstenedione, testosterone, dihydrotestosterone, estrone, 17α-estradiol and 17ß-estradiol) expressed in the steroidogenesis in biological fluids. The analytical method allows for the determination of steroid hormones in blood plasma and serum down to 0.08-0.16 ng/mL for estrogens, 0.20-0.36 ng/mL for androgens and 0.36-0.43 ng/mL for progestagens. These limits of detection were obtainable using a two-step solid-phase clean-up for fractionation and elimination of interfering lipids (fatty acids, phospholipids, glycerides and sterols) from the steroid hormones. The accuracy of the method was 50-112% in the range 0.10 to 2.00 ng/mL.

Multiresidue method for the determination of 32 human and veterinary pharmaceuticals in soil and sediment by pressurized-liquid extraction and LC-MS/MS.

An analytical chemical method has been developed for the simultaneous determination of 32 different pharmaceuticals in soils and sediments. The pharmaceuticals cover a varity of different compound groups. Soil samples were extracted with different solvents with the help of pressurized-liquid extraction (PLE) followed by clean-up using a solid-phase extraction (SPE) procedure. The purified extracts were analyzed by LC-MS/MS. The extraction method was evaluated by testing the following variables: extraction solvents, solvent pH, and temperature. Applying 20 g of soil/sediment and extracting with a mixture of methanol with aqueous ammonia solution (0.1 mol L(-1)) at 80 degrees C for 5 min in five cycles provided satisfactory recoveries between 66 and 114% with SD of between 1 and 14%. For preconcentration and purification tandem MAX-HLB cartridges were used. The volume and composition was optimized and the highest recoveries were obtained with a combination of methanol-aqueous ammonia solution. The limits of quantification (LOQs) were between 0.2 and 2 ng g(-1) and linearity higher than 0.98 for the majority of the selected pharmaceuticals. The method was successfully applied to soil samples collected from the Jerez de la Frontera agricultural region, irrigated with treated wastewater, and to sediment samples from the River Guadalete. The detection of nine pharmaceuticals including stimulants, antirheumatics, analgesics, anti-inflammatories, tranquilizers, and veterinary medicines at ng g(-1) concentration levels was achieved.

Transport and fate of estrogenic hormones in slurry-treated soil monoliths.

The naturally occurring hormones, such as 17-beta-estradiol, 17-alpha-estradiol, and estrone, present in livestock manure may have detrimental environmental effects if released into surface waters. In areas where manure application is intensive, estrogens have been found in surface waters in concentrations known to affect the endocrine system of fish and amphibians. How the estrogens reach the surface waters is unclear. To investigate whether leaching through the soil profile plays a significant role, we conducted leaching experiments on intact soil cores. Lysimeter soil monoliths (60 cm in diameter and 100 cm long) were excavated from two sites in Denmark (one loamy and one sandy soil). The soil monoliths were treated with pig slurry containing estrogenic hormones and amended with an estrogen tracer (17-alpha-ethinylestradiol) and a conservative tracer (bromide). 17-alpha-ethinylestradiol is a synthetic analog of 17-beta-estradiol with sorption characteristics and molecular structure similar to those of the naturally occurring estrogens in slurry. The monoliths were exposed to a short-term irrigation event (12 h) followed by a long-term semi-field experiment (16 wk), during which leaching of natural estrogens and tracers was followed. Estrogens from slurry were transported to a depth of 1 m in loamy soil and sandy soil. The estrogen concentrations in the leachate were at a level known to affect the endocrine system of aquatic organisms.

Solvent diversity in polymorph screening.

Selecting a diverse set of solvents to be included in polymorph screening assignments can be a challenging task. As an aid to decision making, a database of 218 organic solvents with 24 property descriptors was explored and visualized using multivariate tools. The descriptors included, among others, log P, vapor pressure, hydrogen bond formation capabilities, polarity, number of pi-bonds and descriptors derived from molecular interaction field calculations (e.g., size/shape parameters and hydrophilic/hydrophobic regions). The data matrix was initially analyzed using principal component analysis (PCA). Results from the PCA showed 57% cumulative variance being explained in the first two principal components (PCs), although relevant information was also found in the third, fourth and fifth component, revealing distinct clusters of solvents. Since five dimensions were not suitable for visual presentation, a nonlinear method, self-organizing maps (SOMs), was applied to the dataset. The constructed SOM displayed features of clusters observed in the first three PCs, however in a more compelling way. Thus, the SOM was chosen as the visually most convenient way to display the diversity of the 218 solvents. In addition, it was demonstrated how safety aspects can be considered by labeling a large fraction of the solvents in the SOM with toxicological information.

Risk assessment of topically applied products.

The human risk of harmful substances in semisolid topical dosage forms applied topically to normal skin and broken skin, respectively, was assessed. Bisphenol A diglycidyl ether (BADGE) and three derivatives of BADGE previously quantified in aqueous cream and the UV filters 3-BC and 4-MBC were used as model compounds. Tolerable daily intake (TDI) values have been established for BADGE and derivatives. Endocrine disruption was chosen as endpoint for 3-BC and 4-MBC. Skin permeation of the model compounds was investigated in vitro using pig skin membranes. Tape stripping was applied to simulate broken skin associated with various skin disorders. BADGE and derivatives had a tendency to permeate pig skin membranes in vitro with higher fluxes in the tape stripped membranes compared to the non-treated membranes. Data from the in vitro skin permeation study and from the literature were used as input parameters for estimating the risk. The immediate human risk of BADGE and derivatives in topical dosage forms was found to be low. However, local treatment of broken skin may lead to higher exposure of BADGE and derivatives compared to application to normal skin. 3-BC permeated skin at higher flux than 4-MBC. Both UV filters are endocrine disrupting compounds with 3-BC being the more potent. UV filters in sunscreen are often present in high concentrations, which potentially may lead to high systemic exposure dosages. Thus, the risk associated with use of 3-BC and 4-MBC containing sunscreen with regards to endocrine disrupting effects was found to be high and more data is urgently needed in order to fully assess the human risk of 3-BC and 4-MBC in commercial sunscreen.

Susceptibility of bacteria isolated from pigs to tiamulin and enrofloxacin metabolites.

Susceptibilities to metabolites of tiamulin (TIA) and enrofloxacin (ENR) were tested using selected bacteria with previously defined minimal inhibitory concentrations (MIC). The TIA metabolites tested were: N-deethyl-tiamulin (DTIA), 2beta-hydroxy-tiamulin (2beta-HTIA) and 8alpha-hydroxy-tiamulin (8alpha-HTIA), and the ENR metabolites were: ciprofloxacin (CIP) and enrofloxacin N-oxide (ENR-N). Bacteria, all of porcine origin, were selected as representatives of bacterial infections (Staphylococcus hyicus and Actinobacillus pleuropneumoniae), zoonotic bacteria (Campylobacter coli) and indicator bacteria (Escherichia coli and enterococci). Furthermore the effects of these compounds were tested on the microbial community of active sludge to test any negative effect on colony forming units (CFU). DTIA had a potency of 12.5-50% of the potency of TIA. 2beta-HTIA and 8alpha-HTIA had potencies less than 1% of the potency of TIA. ENR-N had a potency of 0.75-1.5% of the potency of ENR, while CIP and ENR had similar potencies. Results obtained here indicate that CIP and DTIA could contribute to the selective pressure for upholding antimicrobial resistant bacteria in animals under ENR or TIA treatment. The most potent metabolites CIP and DTIA showed considerable potencies against activated sludge bacteria compared to the parent compounds. EC(50) (microg/ml) for ENR, CIP, TIA and DTIA were 0.018 [95% CI: 0.028-0.149], 0.064 [95% CI: 0.007-0.046], 6.0 [95% CI: 3.6-9.8], and 9.7 [95% CI: 5.8-16.3], respectively. This indicates that the compounds can change the bacterial population in the sludge, and hereby alter the properties of the sludge.

The six most widely used selective serotonin reuptake inhibitors decrease androgens and increase estrogens in the H295R cell line.

Selective serotonin reuptake inhibitors (SSRIs) used as first line of treatment in major depressive disorder (MDD) are known to exert negative effects on the endocrine system and fertility. The aim of the present study was to investigate the possible endocrine disrupting effect of six SSRIs, fluoxetine, paroxetine, citalopram and its active enantiomer escitalopram, sertraline and fluvoxamine using the OECD standardized and validated human in vitro adrenocortical H295R cell assay. All the major steroids, including progestagens, corticosteroids, androgens and estrogens were analysed using a fully validated LC-MS/MS method. All 6 SSRIs were found to exert endocrine disrupting effects on steroid hormone synthesis at concentrations just around Cmax. Although the mechanisms of disruption were all different, they all resulted in decreased testosterone levels, some due to effects on CYP17, some earlier in the pathway. Furthermore, all SSRIs relatively increased the estrogen/androgen ratio, indicating stimulating effects on the aromatase. Our study demonstrates the potential of SSRIs to interfere with steroid production in the H295R cells around Cmax levels and indicates that these drugs should be investigated further to determine any hazards for the users.

Distribution of the UV filter 3-benzylidene camphor in rat following topical application.

A straightforward analytical method for determination of 3-benzylidene camphor (3-BC) in rat adipose tissue, brain, liver, muscle, plasma and testis following topical application was developed and validated. Three exposure levels (60, 180 and 540 mg kg(-1) day(-1)) were tested for 65 days in male Sprague-Dawley rats (24 days postnatal). Sample preparation involving homogenization and n-heptane or methanol extraction of the tissue was applied before injection into the LC-ESI-MS-MS system. The response was linear from 2 to 100 microg l(-1) for the qualifier and the quantifier MRM transitions (R(2) (quantifier) > 0.994). Detection limit of the method corresponded to 0.005 microg g(-1) tissue and 12.5 microg l(-1) plasma, respectively. Recovery was determined for all tissues (adipose tissue: 40%; all other tissues: 80-100%) at three individual levels. 3-(4-Methyl benzylidene camphor) (4-MBC) was used throughout the study as internal standard. 3-Benzylidene camphor was detected in all tissues at all exposure levels at concentrations between 0.05 microg g(-1) (liver) and 36 microg g(-1) (adipose tissue) and in plasma at 16-89 microg l(-1). The method allowed for the quantification of 3-benzylidene camphor in all tested tissues following topical application. Furthermore, it was shown that 3-benzylidene camphor can be found in various tissues in the rat following topical application. These findings may suggest that following use of 3-benzylidene camphor containing sunscreen, similar disposition and distribution may occur in humans.

Effect of tetracycline residues in pig manure slurry on tetracycline-resistant bacteria and resistance gene tet(M) in soil microcosms.

Effects of tetracycline residues from pig manure slurry on the prevalence of tetracycline-resistant bacteria and the tetracycline resistance gene, tet(M), were studied in soil microcosms. Four types of soil microcosms were established for a period of 152 days, supplemented with combinations of pig manure slurry and a tetracycline-resistant Enterococcus faecalis, CG110, containing the tetracycline resistance gene tet(M) (on the conjugative transposon, Tn916). The prevalence of both tetracycline-resistant aerobic bacteria and tetracycline-resistant enterococci declined rapidly until day 45 where no significant differences in the levels of tetracycline-resistant bacteria in any of the four types of microcosms could be detected. tet(M) could be detected in microcosms supplemented with either pig manure slurry and/or E. faecalis CG110 (tet(M)) for the whole period (152 days). tet(M) could be detected longer than tetracycline-resistant enterococci could be isolated (limit of detection 100 CFU/g soil) probably due to viable but not culturable (VBNC) bacteria with tet(M), horizontal gene transfer of tet(M) to indigenous soil bacteria or presence of "free" DNA. The concentration of chlortetracycline and oxytetracycline were almost stable through out the experimental period, but the tetracycline concentrations had no effect on prevalence of tetracycline-resistant bacteria. The presented microcosm approach simulated natural farmland conditions well and supported results from previous field studies.

Determination of bisphenol diglycidyl ethers in topical dosage forms.

A method involving extraction and LC-ESI-MS-MS detection of BADGE, BFDGE, BADGE*H2O, BADGE*2H2O, BADGE*HCl, BADGE*H2O*HCl, BADGE.2HCl and BFDGE*2HCl in aqueous cream was developed and validated. Initially, empty internally lacquered aluminum container closure systems were extracted with isopropanol as an attempt to estimate the upper limit of extractable bisphenol diglycidyl ethers present in lacquer. Six of the eight potential bisphenol diglycidyl ethers were quantified. In an accelerated experiment, on aqueous cream stored in lacquered aluminum tubes at 70 degrees C, all derivatives except BADGE*2HCl and BFDGE*2HCl were extracted from cream samples and quantified as an attempt to estimate the upper limit of compounds leaching to the cream. Detection limits were from 0.3+/-0.2 to 3.4+/-0.7 microgl(-1). Recoveries were determined for all compounds at three concentration levels (mean 63+/-6%). Mean inter-day and mean intra-day precision was 7+/-2 and 13+/-6%, respectively. Three commercially available creams were obtained from a local community pharmacy and analysed for bisphenol diglycidyl ethers. BADGE, BADGE*H2O, BADGE*2H2O and BADGE*H2O*HCl were detected and quantified. In conclusion, the developed method allows for the extraction and detection of bisphenol diglycidyl ethers originating from the epoxy phenol lacquer used in aluminum tubes. This study does not indicate that they leach into aqueous cream in significant amounts under normal storage conditions.

Isolation and structural elucidation of tiamulin metabolites formed in liver microsomes of pigs.

Although the antimicrobial tiamulin is extensively metabolized in pigs, the metabolism is not well investigated. In this work the NADPH dependent metabolism of tiamulin in liver microsomes from pigs has been studied. The tiamulin metabolites formed in the incubations were analysed using LC-MS, and three major metabolites were isolated using solid phase extraction and preparative HPLC. The final structure elucidations were performed by tandem mass spectrometry and (1)H and (13)C NMR. The structures of the metabolites were found to be 2beta-hydroxy-tiamulin, 8alpha-hydroxy-tiamulin and N-deethyl-tiamulin. In addition, the LC-MS chromatograms revealed two other minor metabolites. From their chromatography and from MS(2) analysis the structures were estimated to be 2beta-hydroxy-N-deethyl-tiamulin and 8alpha-hydroxy-N-deethyl-tiamulin, but the structures were not confirmed by NMR. In these studies approximately 20% of tiamulin was deethylated, 10% was hydroxylated in the 2beta-position and 7% was hydroxylated in the 8alpha-position. About 40% of tiamulin was metabolized during the incubation conditions used. The protein precipitation in the incubations was performed using perchloric acid, and the preparative purification was performed under alkaline conditions. Therefore, the stability of the metabolites under these conditions was studied. The metabolites were found to be stable in the acid solution, but under alkaline conditions, particularly at room temperature, the stability of especially 8alpha-hydroxy-tiamulin was considerably reduced (40% loss after 1 week).

Assessment of the importance of sorption for steroid estrogens removal during activated sludge treatment.

Distribution coefficients (K(d)) between water and activated sludge particles (f(oc)=27.7+/-0.1%) were measured for the steroid estrogens (SE), estrone (E1), 17beta-estradiol (E2) and 17alpha-ethinylestradiol (EE2) in batch experiments. Experimental concentration levels ranged from environmentally realistic low ng/l to the high microg/l. In this range K(d)s were independent of their water concentration. The experimentally obtained K(d)s (with 95% confidence intervals) were 402+/-126 l/kg, 476+/-192 l/kg and 584+/-136 l/kg for E1, E2 and EE2, respectively. K(d)s were used to estimate the fraction of the total SE concentration that is expected to be sorbed in the activated sludge treatment tanks of a typical STP assuming equilibrium conditions. Assuming a suspended solids concentration of 4 g/l dissolved solids (ds), it was estimated that 61+/-9%, 66+/-13% and 70+/-6% of the total concentration of E1, E2 and EE2, respectively, would be sorbed during activated sludge treatment. The fraction of the SEs that was expected to be sorbed to suspended sludge particles in the effluents from a typical Danish STP was estimated to be only 0.20+/-0.06%, 0.24+/-0.10% and 0.29+/-0.07% of the total concentration of E1, E2 and EE2, respectively, at a suspended solids concentration of 5 mg/lds. For a typical STP the removal of steroid estrogens with excess sludge was estimated to be only 1.5-1.8% of the total loading if equilibrium conditions exists. Sorption is therefore not important for the fate of SEs in STPs compared to biodegradation.

Dissipation of oxytetracycline, chlortetracycline, tetracycline and doxycycline using HPLC-UV and LC/MS/MS under aquatic semi-field microcosm conditions.

A mixture of four tetracyclines; oxytetracycline (OTC), chlortetracycline (CTC), tetracycline (TC), and doxycycline (DC) was applied in fifteen 12000l outdoor microcosms at four treatment levels plus controls each with three replicates (n = 3). The dissipation times of parent compounds were monitored and half-lives (DT50) of 1-4 days, depending on treatment level were recorded. This is in accordance with half-lives from previous findings in bench-top experiments. Parent compound DT50, were determined using HPLC-UV. Furthermore, the samples were analyzed for ten different tetracycline products using LC/MS/MS. Two products were found for chlortetracycline; 4-epi-anh-chlortetracyline and the iso-chlortetracycline. Iso-forms were only found for CTC and only at the highest treatment (300 microg l(-1)). The half-lives, trajectories, and relative amounts of the products were analogous for all four tetracyclines. DT50 for products were less than 1.2 days. Formation of 4-epi-anh-tetracyclines, occurred at neutral to weak alkaline conditions.

Dissipation and effects of chlortetracycline and tylosin in two agricultural soils: a field-scale study in southern Denmark.

Presently, there is a basic lack of information concerning the accumulation of antibacterial agent residues in agricultural soils. In this field study, performed in southern Denmark, we assess the dissipation of chlortetracycline (CTC), and tylosin A (TYL A) as a function of time. Field soils were classified as a sandy loam soil (field A) and a sandy soil (field B) and each field was sampled on six occasions during the 155-d experimental period from May to October 2000 for chemical analysis and counts of colony-forming units (CFU) detecting the level of aerobic bacteria surviving antibiotic exposure. Colony-forming units and TYL A were detected throughout the entire sampling period, with respective starting soil concentrations of 30 and 50 microg kg(-1) soil declining to 1 and 5 microg kg(-1) soil, on day 155. Compound half-lives (95% confidence limits in parentheses) were estimated for both fields and T1/2 for CTC was 25 d (20-34) and 34 d (28-42) in fields A and B, respectively, and T1/2 for TYL A was 67 d (54-86) and 49 d (40-64) in fields A and B, respectively. No significant difference was determined between compound half-lives on the two fields. The level of aerobic antibiotic-resistant bacteria in the soil over time and soil fauna community was assessed in relation to application of manure containing antibacterial agents to the agricultural fields. The level of both CTC- and TYL-resistant bacteria was affected in the soil by amendment of manure, but declined during the study to the same level as observed at the beginning.

Effects of selective serotonin reuptake inhibitors on three sex steroids in two versions of the aromatase enzyme inhibition assay and in the H295R cell assay.

Selective serotonin reuptake inhibitors are known to have a range of disorders that are often linked to the endocrine system e.g. hormonal imbalances, breast enlargement, sexual dysfunction, and menstrual cycle disorders. The mechanisms behind most of these disorders are not known in details. In this study we investigated whether the endocrine effect due to SSRI exposure could be detected in well adopted in vitro steroidogenesis assays, two versions of the aromatase enzyme inhibition assay and the H295R cell assay. The five drugs citalopram, fluoxetine, fluvoxamine, paroxetine and sertraline, were shown to inhibit the aromatase enzyme in both types of aromatase assays. The IC50 values ranged from 3 to 600 µM. All five SSRIs, were further investigated in the H295R cell line. All compounds altered the steroid secretion from the cells, the lowest observed effect levels were 0.9 µM and 3.1 µM for sertraline and fluvoxamine, respectively. In general the H295R cell assay was more sensitive to SSRI exposure than the two aromatase assays, up to 20 times more sensitive. This indicates that the H295R cell line is a better tool for screening endocrine disrupting effects. Our findings show that the endocrine effects of SSRIs may, at least in part, be due to interference with the steroidogenesis.

Simvastatin decreases steroid production in the H295R cell line and decreases steroids and FSH in female rats.

Endocrine modulating effects of Simvastatin (SV) and its metabolite, Simvastatin ß-hydroxy acid (SVA), were investigated in H295R cells and in female Sprague-Dawley (SPRD) rats. H295R cells were exposed to SV and SVA concentrations from 0 to 10µM for 48h. Four groups of SPRD rats received 0 (CT), 1.3 (L), 5.0 (M), and 20.0 (H)mg SV/kg bw/day for 14 days. 10 Steroids were investigated in H295R growth media, and in tissues and plasma from rats using GC-MS/MS. Plasma LH and FSH were quantified by ELISA. In the H295R assay, SV and SVA particularly decreased progestagens with IC50-values from 0.10-0.13µM for SV and from 0.019-0.055µM for SVA. In rats, SV decreased progestagens in ovaries, brain and plasma, and plasma FSH in the M (72.4% decrease) and H group (76.6% decrease). Because progestagens and gonadotropins are major players in fertility, administration of SV might exert negative effects on female reproduction.

Simultaneous extraction of tetracycline, macrolide and sulfonamide antibiotics from agricultural soils using pressurised liquid extraction, followed by solid-phase extraction and liquid chromatography-tandem mass spectrometry.

The veterinary antibacterial agents chlortetracycline (CTC), oxytetracycline (OTC), sulfadiazine (SDZ), erythromycin (ERY) and tylosin (TYL A, B, C and D) were extracted from soil using pressurized liquid extraction (PLE). Citric acid (pH 4.7) and methanol was used as extraction buffer, followed by tandem-solid-phase extraction (SPE) clean-up (SAX + HLB) for all compounds. For quantification two slightly different methods were employed using LC-MS-MS with MRM detection. The soil extraction method was validated using a loamy sand soil and a sandy soil, representing two typical Danish agricultural soils. Recoveries were 50-80% for the tetracyclines (CTC and OTC) and sulfadiazine (SDZ) and 60-100% for the macrolides (TYL and ERY). Limits of detection for the soil extraction method (LOD(soil)) were 0.6-5.6 microg kg(-1) soil for CTC and OTC, 0.9-2.9 microg kg(-1) soil for SDZ and 2.4-5.5 microg kg(-1) soil for TYL A and ERY. Furthermore, the method was applied to field samples taken from two agricultural fields fertilised with liquid manure containing CTC and TYL A. These results showed a decline in the content of antibacterial agents throughout the sampling period of 155 days from 10 to 15 microg CTC kg(-1) soil and 20-55 microg TYL A kg(-1) soil to below or near the LOD(soil) listed above. Finally, the method was applied to barley grains harvested from the fields. None of the antibacterial agents were measured in grain samples, but recoveries for spiked grain samples were similar to soil recoveries.

Liquid chromatography-mass spectrometry method to determine alcohol ethoxylates and alkylamine ethoxylates in soil interstitial water, ground water and surface water samples.

Alcohol ethoxylates (AEs) and alkylamine ethoxylates (AMEs) are used as adjuvants in pesticide formulations. Analytical procedures for these compounds in environmental aqueous samples using LC-MS are presented. Sample preparation uses solid-phase extraction with Porapak Rdx cartridges. Detection limits and recoveries in ground water and surface water are, respectively, AEs: 16-60 ng/l, 35-93% and AMEs: 0.3-6 microg/l, 28-96%. The lower recoveries are obtained for the apolar surfactants. The procedure was employed on samples of ground water and soil interstitial water collected from farming areas. The individual AEs were detected at concentration levels ranging from 33 to 189 ng/l water.

Fast and robust simultaneous determination of three veterinary antibiotics in groundwater and surface water using a tandem solid-phase extraction with high-performance liquid chromatography-UV detection.

A simple and robust analytical method is presented in which the three veterinary antibiotics oxytetracycline (OTC), sulfachloropyridazine (SCP) and tylosin (TYL) were simultaneously determined in surface water and groundwater. The three compounds were simultaneously extracted from the water samples using a mixture of methanol, EDTA and McIlvaine buffer (citric acid and sodium orthophosphate) and then cleaned-up and pre-concentrated by solid-phase extraction using sacrificial Isolute strong anion-exchange cartridges, to remove interfering organic material, and Waters Oasis hydrophilic-liphophilic balance polymer cartridges, to retain the compounds, in tandem. Analysis was performed using liquid chromatography with ultraviolet detection. Recoveries for river water samples spiked at 10 and 1 microgl(-1) were respectively 99.6+/-4.6 and 99.4+/-8.4% for OTC; 99.9+/-2.2 and 105.0+/-5.7% for SCP; and 94.9+/-2.4 and 71.6+/-8.2% for TYL. Overall limits of detection based on pre-concentrating 400 ml of sample were 0.35 microgl(-1) for OTC and TYL and 0.25 microgl(-1) for SCP.