Bioconcentration and Aquatic Toxicity of Superhydrophobic Chemicals: A Modeling Case Study of Cyclic Volatile Methyl Siloxanes.

Many chemicals in commerce are classified as "superhydrophobic", having log octanol-water partition coefficients (log KOW) approaching or exceeding 7. Examples include long-chain alkanes, halogenated aromatics, and cyclic volatile methylsiloxanes (cVMS). We show that superhydrophobic chemicals present unique assessment challenges because of their sparing solubility in water and difficulties in empirical determinations of bioconcentration factors (BCFs) and aquatic toxicity. Using cVMS as an example, BCFs are considerably lower than expected due to biotransformation. Reviewed aquatic toxicity test data for cVMS in a range of aquatic organisms show little or no toxic effects up to solubility limits in water and sediment. Explanations for this apparent lack of toxicity of cVMS, and by extension to other superhydrophobic chemicals, are explored using a conventional one-compartment uptake model to simulate bioconcentration and toxicity tests using an assumed baseline narcotic critical body residue (CBR) and a range of organism sizes. Because of the low aqueous concentrations, equilibration times are very long and BCFs are sensitive to even very slow rates of biotransformation. Most organisms fail to achieve the assumed CBR during feasible test durations even at the solubility limit. Regulatory evaluation of superhydrophobic substances requires specially designed test protocols addressing biotransformation and dietary uptake.

Determination of the dietary absorption efficiency of hexachlorobenzene with the channel catfish (Ictalurus punctatus).

This study was designed to experimentally measure the assimilation efficiency of hexachlorobenzene (HCB) in a warm-water, benthic-feeding fish species, the channel catfish (Ictalurus punctatus). Catfish were exposed to (14)C-radiolabeled HCB in catfish food over a 28-day exposure period, followed by a 14-day clearance period. Over the experimental period, the total (14)C residues were measured in fish tissue and a simple two-box kinetic model was applied to the data to simulate uptake and clearance dynamics. No detectable metabolism of HCB by catfish was found. A two-box kinetic model effectively modeled the uptake and clearance of (14)C-HCB in catfish, with a calculated assimilation efficiency of the chemical into the whole catfish of 67% (growth corrected). The growth-corrected pseudo first-order elimination half-life of (14)C-HCB from whole catfish was determined to be 29 days (k(2)=0.024 day(-1)).

Benthic invertebrate exposure and chronic toxicity risk analysis for cyclic volatile methylsiloxanes: Comparison of hazard quotient and probabilistic risk assessment approaches.

This study utilized probabilistic risk assessment techniques to compare field sediment concentrations of the cyclic volatile methylsiloxane (cVMS) materials octamethylcyclotetrasiloxane (D4, CAS # 556-67-2), decamethylcyclopentasiloxane (D5, CAS # 541-02-6), and dodecamethylcyclohexasiloxane (D6, CAS # 540-97-6) to effect levels for these compounds determined in laboratory chronic toxicity tests with benthic organisms. The concentration data for D4/D5/D6 in sediment were individually sorted and the 95th centile concentrations determined in sediment on an organic carbon (OC) fugacity basis. These concentrations were then compared to interpolated 5th centile benthic sediment no-observed effect concentration (NOEC) fugacity levels, calculated from a distribution of chronic D4/D5/D6 toxicologic assays per OECD guidelines using a variety of standard benthic species. The benthic invertebrate fugacity biota NOEC values were then compared to field-measured invertebrate biota fugacity levels to see if risk assessment evaluations were similar on a field sediment and field biota basis. No overlap was noted for D4 and D5 95th centile sediment and biota fugacity levels and their respective 5th centile benthic organism NOEC values. For D6, there was a small level of overlap at the exposure 95th centile sediment fugacity and the 5th centile benthic organism NOEC fugacity value; the sediment fugacities indicate that a negligible risk (1%) exists for benthic species exposed to D6. In contrast, there was no indication of risk when the field invertebrate exposure 95th centile biota fugacity and the 5th centile benthic organism NOEC fugacity values were compared.

Metabolism of 14C-octamethylcyclotetrasiloxane ([14C]D4) or 14C-decamethylcyclopentasiloxane ([14C]D5) orally gavaged in rainbow trout (Oncorhynchus mykiss).

Critical factors (uptake, distribution, metabolism and elimination) for understanding the bioaccumulation/biomagnification potential of Octamethylcyclotetrasiloxane (D4) and Decamethylcyclopentasiloxane (D5) siloxanes in fish were investigated to address whether these chemicals meet the "B" criteria of the Persistent, Bioaccumulative, and Toxic (PBT) classification. A metabolism study was conducted in rainbow trout whereby a 15mg [14C]D4/kg bw or [14C]D5/kg bw as a single bolus oral dose was administered via gavage. Of the administered dose, 79% (D4) and 78% (D5) was recovered by the end of the study (96-h). Eighty-two percent and 25% of the recovered dose was absorbed based on the percentage of recovered dose in carcass (69% and 17%), tissues, bile and blood (12% and 8%) and urine (1%) for D4 and D5, respectively. A significant portion of the recovered dose (i.e. 18% for D4 and 75% for D5) was eliminated in feces. Maximum blood concentrations were 1.6 and 1.4µg D4 or D5/g blood at 24h post-dosing, with elimination half-lives of 39h (D4) and 70h (D5). Modeling of parent and metabolite blood concentrations resulted in estimated metabolism rate constants (km(blood)) of 0.15 (D4) and 0.17day-1(D5). Metabolites in tissues, bile, blood, and urine totaled a minimum of 2% (D4) and 14% (D5) of the absorbed dose. The highest concentration of 14C-activity in the fish following D4 administration was in mesenteric fat followed by bile, but the opposite was true for D5. Metabolites were not detected in fat, only parent chemical. In bile, 94% (D4) and 99% (D5) of the 14C-activity was due to metabolites. Metabolites were also detected in the digestive tract, liver and gonads. Approximately 40% of the 14C-activity detected in the liver was due to the presence of metabolites. Urinary elimination represented a minor pathway, but all the 14C-activity in the urine was associated with metabolites. Clearance may occur via enterohepatic circulation of metabolic products in bile with excretion via the digestive tract and urinary clearance of polar metabolites.

Plasma sampling and freezing procedures influence vitellogenin measurements by enzyme-linked immunoassay in the fathead minnow (Pimephales promelas).

The present study compared three different methods for measuring plasma vitellogenin (VTG) in fathead minnow (FHM; Pimephales promelas): A procedure using liquid chromatography with electrospray ionization combined with tandem mass spectrometry (LC/ESI-MS/MS), and two commercial enzyme-linked immunoassay (ELISA) kits using either anti-carp or anti-FHM antibodies. The influence on plasma VTG measurements of using the protease-inhibitor aprotinin during blood sampling and of submitting the plasma samples to a freeze-thaw cycle before analysis also was evaluated. The addition of aprotinin to the blood during sampling significantly reduced the plasma VTG concentrations measured by ELISA, whereas the VTG values measured after plasma samples were submitted to a freeze-thaw cycle were significantly higher than those measured before freezing. This inflating effect of freezing on VTG measurements made by ELISA could be prevented if plasma samples were frozen diluted in citrate buffer containing 16 mg/ml of polyethylene glycol (PEG). In contrast, measurements of VTG made by LC/ESI-MS/MS were unaffected by freezing and, conceptually, are independent from enzymatic degradation. Although the use of aprotinin and PEG effectively reduced the influence of enzymatic and physical degradation caused by freezing and thawing on VTG measurements made by ELISA, it did not improve agreement between the three analytical techniques evaluated. More information is needed regarding the molecular structure and the existence of possible multiple forms of VTG before this protein can be measured adequately in FHM.

Evaluating the roles of biotransformation, spatial concentration differences, organism home range, and field sampling design on trophic magnification factors.

Trophic magnification factors (TMFs) are field-based measurements of the bioaccumulation behavior of chemicals in food-webs. TMFs can provide valuable insights into the bioaccumulation behavior of chemicals. However, bioaccumulation metrics such as TMF may be subject to considerable uncertainty as a consequence of systematic bias and the influence of confounding variables. This study seeks to investigate the role of systematic bias resulting from spatially-variable concentrations in water and sediments and biotransformation rates on the determination of TMF. For this purpose, a multibox food-web bioaccumulation model was developed to account for spatial concentration differences and movement of organisms on chemical concentrations in aquatic biota and TMFs. Model calculated and reported field TMFs showed good agreement for persistent polychlorinated biphenyl (PCB) congeners and biotransformable phthalate esters (PEs) in a marine aquatic food-web. Model testing showed no systematic bias and good precision in the estimation of the TMF for PCB congeners but an apparent underestimation of model calculated TMFs, relative to reported field TMFs, for PEs. A model sensitivity analysis showed that sampling designs that ignore the presence of concentration gradients may cause systematically biased and misleading TMF values. The model demonstrates that field TMFs are most sensitive to concentration gradients and species migration patterns for substances that are subject to a low degree of biomagnification or trophic dilution. The model is useful in anticipating the effect of spatial concentration gradients on the determination of the TMF; guiding species collection strategies in TMF studies; and interpretation of the results of field bioaccumulation studies in study locations where spatial differences in chemical concentration exist.

Potentiation of the vitellogenic response to 17alpha-ethinylestradiol by cortisol in the fathead minnow Pimephales promelas.

The effects of elevated plasma cortisol levels on vitellogenin (VTG) induction were examined in the fathead minnow (Pimephales promelas) in an attempt to evaluate the potential influence of stress on this commonly used biomarker of estrogenicity. Two separate experiments were conducted in which fish plasma cortisol was elevated to various levels for 14 d by noninvasive additions of cortisol to the aquaria water. Fathead minnows were exposed to either cortisol alone, 17alpha-ethinylestradiol (EE2) alone, or a combination of the two hormones, and plasma levels of VTG as well as liver expression of VTG mRNA were measured. Both experiments gave similar results, with an exposure to 4 ng/L of EE2 resulting in significantly greater levels of plasma VTG in the presence of, compared to that in absence of, cortisol, whereas exposure to cortisol alone at concentrations between 144 and 800 microg/L had no effect on plasma VTG levels. This potentiation of the EE2-induced vitellogenesis by cortisol was dose-dependent, with plasma VTG reaching 125, 167, and 295% of the values obtained with EE2 alone when 144, 360, and 800 microg/L of cortisol, respectively, were added to the water. Liver mRNA results were consistent with plasma VTG, although they generally were more variable. The present study demonstrates that cortisol does not independently induce vitellogenesis but can potentiate estrogen-induced VTG synthesis in fathead minnow. The implications of these findings for the use of VTG as a biomarker of estrogenicity are discussed.

Bioaccumulation of decamethylpentacyclosiloxane (D5): A review.

Decamethylpentacyclosiloxane (D5) is a widely used, high-production volume personal care product with an octanol-water partition coefficient (log K(OW)) of 8.09. Because of D5's high K(OW) and widespread use, it is subject to bioaccumulation assessments in many countries. The present study provides a compilation and an in-depth, independent review of bioaccumulation studies involving D5. The findings indicate that D5 exhibits depuration rates in fish and mammals that exceed those of extremely hydrophobic, nonbiotransformable substances; that D5 is subject to biotransformation in mammals and fish; that observed bioconcentration factors in fish range between 1040 L/kg and 4920 L/kg wet weight in laboratory studies using non-radiolabeled D5 and between 5900 L/kg and 13 700 L/kg wet weight in an experiment using C(14) radiolabeled D5; and that D5 was not observed to biomagnify in most laboratory experiments and field studies. Review of the available studies shows a high degree of internal consistency among findings from different studies and supports a broad comprehensive approach in bioaccumulation assessments that includes information from studies with a variety of designs and incorporates multiple bioaccumulation measures in addition to the K(OW) and bioconcentration factor.

Decamethylcyclopentasiloxane (D5) environmental sources, fate, transport, and routes of exposure.

The environmental sources, fate, transport, and routes of exposure of decamethylcyclopentasiloxane (D5; CAS no. 541-02-6) are reviewed in the present study, with the objective of contributing to effective risk evaluation and assessment of this and related substances. The present review, which is part of a series of studies discussing aspects of an effective risk evaluation and assessment, was prompted in part by the findings of a Board of Review undertaken to comment on a decision by Environment Canada made in 2008 to subject D5 to regulation as a toxic substance. The present review focuses on the early stages of the assessment process and how information on D5's physical-chemical properties, uses, and fate in the environment can be integrated to give a quantitative description of fate and exposure that is consistent with available monitoring data. Emphasis is placed on long-range atmospheric transport and fate in water bodies receiving effluents from wastewater treatment plants (along with associated sediments) and soils receiving biosolids. The resulting exposure estimates form the basis for assessments of the resulting risk presented in other studies in this series. Recommendations are made for developing an improved process by which D5 and related substances can be evaluated effectively for risk to humans and the environment.

Fugacity and activity analysis of the bioaccumulation and environmental risks of decamethylcyclopentasiloxane (D5).

As part of an initiative to evaluate commercial chemicals for their effects on human and environmental health, Canada recently evaluated decamethylcyclopentasiloxane (D5; CAS no. 541-02-06), a high-volume production chemical used in many personal care products. The evaluation illustrated the challenges encountered in environmental risk assessments and the need for the development of better tools to increase the weight of evidence in environmental risk assessments. The present study presents a new risk analysis method that applies thermodynamic principles of fugacity and activity to express the results of field monitoring and laboratory bioaccumulation and toxicity studies in a comprehensive risk analysis that can support risk assessments. Fugacity and activity ratios of D5 derived from bioaccumulation measures indicate that D5 does not biomagnify in food webs, likely because of biotransformation. The fugacity and activity analysis further demonstrates that reported no-observed-effect concentrations of D5 normally cannot occur in the environment. Observed fugacities and activities in the environment are, without exception, far below those corresponding with no observed effects, in many cases by several orders of magnitude. This analysis supports the conclusion of the Canadian Board of Review and the Minister of the Environment that D5 does not pose a danger to the environment. The present study further illustrates some of the limitations of a persistence-bioaccumulation-toxicity-type criteria-based risk assessment approach and discusses the merits of the fugacity and activity approach to increase the weight of evidence and consistency in environmental risk assessments of commercial chemicals.

Characterization of ecological risks from environmental releases of decamethylcyclopentasiloxane (D5).

Decamethylcyclopentasiloxane (D5) is used in personal care products and industrial applications. The authors summarize the risks to the environment from D5 based on multiple lines of evidence and conclude that it presents negligible risk. Laboratory and field studies show that D5 is not toxic to aquatic organisms or benthic invertebrates up to its solubility limit in water or porewater or its sorptive capacity in sediment. Comparison of lipid-normalized internal concentrations with measured concentrations in benthos indicates that field-collected organisms do not achieve toxic levels of D5 in their tissues, suggesting negligible risk. Exposure to D5 resulted in a slight reduction of root biomass in barley at test concentrations 2 orders of magnitude greater than measured D5 levels in biosolids-amended soils and more than twice as high as the maximum calculated sorptive capacity of the soil. No effects were observed in soil invertebrates exposed to similar concentrations, indicating that D5 poses a de minimis risk to the terrestrial environment. High rates of metabolism and elimination of D5 compared with uptake rates from food results in biodilution in the food web rather than biomagnification, culminating in de minimis risk to higher trophic level organisms via the food chain. A fugacity approach substantiates all conclusions that were made on a concentration basis.

Quantitative measurement of fathead minnow vitellogenin by liquid chromatography combined with tandem mass spectrometry using a signature peptide of vitellogenin.

Vitellogenin (VTG) has been proposed as a sensitive biomarker of exposure to environmental estrogenic contaminants that induce VTG production in oviparous species. Enzyme-linked immunosorbent assay (ELISA) methods are currently widely used to measure the VTG levels. In this paper, a new liquid chromatography combined with tandem mass spectrometry (LC/ESI-MS/MS) method for the quantitative analysis of VTG in the plasma of fathead minnows exposed to 17alpha-ethinylestradiol (EE2) has been developed. This method includes, first, the selection of the signature peptide, which involves sodium dodecyl sulfate-polyarylamide gel electrophoresis separation, in-gel digestion, LC/ESI-MS/MS analysis with an ion trap mass spectrometer, and sequence determination with the TurboSEQUEST MS/MS database application; second, optimization of the selected signature peptide in multireaction monitor (MRM) mode with a triple quadrupole mass spectrometer; and third, trypsin digestion of plasma and VTG quantitation via MRM-mode LC/ESI-MS/MS. A series of plasma samples from fathead minnows following exposure to EE2 was assayed. A good correlation was found when EE2-induced plasma samples from fathead minnows were analyzed with ELISA and the described new method. Although used here with fathead minnow, the new LC/ESI-MS/MS method could be easily applied to the analysis of VTG expressed in any other fish species. Quantitation of VTG by this method was found to be highly specific and linear. The absence of potential artifactual measurements of VTG at low exposure levels could also be critical in future studies that evaluate weakly estrogenic compounds in aquatic species.

The bioconcentration and metabolism of chlorpyrifos by the eastern oyster, Crassostrea virginica.

Eastern oysters (Crassostrea virginica) were exposed to [14C]chlorpyrifos (O,O-diethyl-O-[3,5,6-trichloro-2-pyridyl] phosphorothioate) at an average measured seawater concentration of 0.6 microg/L under flow-through conditions for 28 d. The compound O,O-diethyl-O-(3,5-dichloro-6-methylthio-2-pyridyl)phosphorothioate (DMP) was extracted and identified as the single metabolite observed, and this metabolite constituted the majority of the total [14C] activity in the oyster at all sampling times. Once oysters were exposed to clean water, both chlorpyrifos and DMP residues cleared rapidly from whole oysters, with elimination half-lives of <3 d. A simple two-compartment uptake/elimination model was adequate to describe total [14C] activity in whole oysters, edible tissue, and oyster liquor. The average bioconcentration factors (BCFs) for total [14C] activity in whole oysters, edible tissue, and oyster liquor were 565, 1,400, and 35 ml/g, respectively. The parent [14C]chlorpyrifos accumulated to a peak residue concentration of 135 microg/kg in whole oyster tissue, representing an empirical [14C]chlorpyrifos BCF value in the oyster of approximately 225 ml/ g; the BCF value for [14C]chlorpyrifos was lower than the BCF for total [14C] activity in whole oysters and edible tissue because of extensive metabolism to DMP and oyster elimination processes.

Comparing laboratory- and field-measured biota-sediment accumulation factors.

Standardized laboratory protocols for measuring the accumulation of chemicals from sediments are used in assessing new and existing chemicals, evaluating navigational dredging materials, and establishing site-specific biota-sediment accumulation factors (BSAFs) for contaminated sediment sites. The BSAFs resulting from the testing protocols provide insight into the behavior and risks associated with individual chemicals. In addition to laboratory measurement, BSAFs can also be calculated from field data, including samples from studies using in situ exposure chambers and caging studies. The objective of this report is to compare and evaluate paired laboratory and field measurement of BSAFs and to evaluate the extent of their agreement. The peer-reviewed literature was searched for studies that conducted laboratory and field measurements of chemical bioaccumulation using the same or taxonomically related organisms. In addition, numerous Superfund and contaminated sediment site study reports were examined for relevant data. A limited number of studies were identified with paired laboratory and field measurements of BSAFs. BSAF comparisons were made between field-collected oligochaetes and the laboratory test organism Lumbriculus variegatus and field-collected bivalves and the laboratory test organisms Macoma nasuta and Corbicula fluminea. Our analysis suggests that laboratory BSAFs for the oligochaete L. variegatus are typically within a factor of 2 of the BSAFs for field-collected oligochaetes. Bivalve study results also suggest that laboratory BSAFs can provide reasonable estimates of field BSAF values if certain precautions are taken, such as ensuring that steady-state values are compared and that extrapolation among bivalve species is conducted with caution.

Comparing laboratory and field measured bioaccumulation endpoints.

An approach for comparing laboratory and field measures of bioaccumulation is presented to facilitate the interpretation of different sources of bioaccumulation data. Differences in numerical scales and units are eliminated by converting the data to dimensionless fugacity (or concentration-normalized) ratios. The approach expresses bioaccumulation metrics in terms of the equilibrium status of the chemical, with respect to a reference phase. When the fugacity ratios of the bioaccumulation metrics are plotted, the degree of variability within and across metrics is easily visualized for a given chemical because their numerical scales are the same for all endpoints. Fugacity ratios greater than 1 indicate an increase in chemical thermodynamic activity in organisms with respect to a reference phase (e.g., biomagnification). Fugacity ratios less than 1 indicate a decrease in chemical thermodynamic activity in organisms with respect to a reference phase (e.g., biodilution). This method provides a holistic, weight-of-evidence approach for assessing the biomagnification potential of individual chemicals because bioconcentration factors, bioaccumulation factors, biota-sediment accumulation factors, biomagnification factors, biota-suspended solids accumulation factors, and trophic magnification factors can be included in the evaluation. The approach is illustrated using a total 2393 measured data points from 171 reports, for 15 nonionic organic chemicals that were selected based on data availability, a range of physicochemical partitioning properties, and biotransformation rates. Laboratory and field fugacity ratios derived from the various bioaccumulation metrics were generally consistent in categorizing substances with respect to either an increased or decreased thermodynamic status in biota, i.e., biomagnification or biodilution, respectively. The proposed comparative bioaccumulation endpoint assessment method could therefore be considered for decision making in a chemicals management context.

Evaluation of bioaccumulation using in vivo laboratory and field studies.

A primary consideration in the evaluation of chemicals is the potential for substances to be absorbed and retained in an organism's tissues (i.e., bioaccumulated) at concentrations sufficient to pose health concerns. Substances that exhibit properties that enable biomagnification in the food chain (i.e., amplification of tissue concentrations at successive trophic levels) are of particular concern due to the elevated long-term exposures these substances pose to higher trophic organisms, including humans. Historically, biomarkers of in vivo chemical exposure (e.g., eggshell thinning, bill deformities) retrospectively led to the identification of such compounds, which were later categorized as persistent organic pollutants. Today, multiple bioaccumulation metrics are available to quantitatively assess the bioaccumulation potential of new and existing chemicals and identify substances that, upon or before environmental release, may be characterized as persistent organic pollutants. This paper reviews the various in vivo measurement approaches that can be used to assess the bioaccumulation of chemicals in aquatic or terrestrial species using laboratory-exposed, field-deployed, or collected organisms. Important issues associated with laboratory measurements of bioaccumulation include appropriate test species selection, test chemical dosing methods, exposure duration, and chemical and statistical analyses. Measuring bioaccumulation at a particular field site requires consideration of which test species to use and whether to examine natural populations or to use field-deployed populations. Both laboratory and field methods also require reliable determination of chemical concentrations in exposure media of interest (i.e., water, sediment, food or prey, etc.), accumulated body residues, or both. The advantages and disadvantages of various laboratory and field bioaccumulation metrics for assessing biomagnification potential in aquatic or terrestrial food chains are discussed. Guidance is provided on how to consider the uncertainty in these metrics and develop a weight-of-evidence evaluation that supports technically sound and consistent persistent organic pollutant and persistent, bioaccumulative, and toxic chemical identification. Based on the bioaccumulation information shared in 8 draft risk profiles submitted for review under the United Nations Stockholm Convention, recommendations are given for the information that is most critical to aid transparency and consistency in decision making.

An ecological risk assessment for chlorpyrifos in an agriculturally dominated tributary of the San Joaquin River.

Single-species toxicity testing of ambient water samples and national-scale probabilistic risk assessment have implicated the organophosphorous (OP) insecticide chlorpyrifos (O, O-diethyl O-(3,5,6-trichloro-2-pyridyl)-phosphorothioate) as a potential chemical stressor of aquatic organisms residing in the lower San Joaquin River basin. This site-specific aquatic ecological risk assessment was conducted to determine the probability of adverse effects occurring from exposure to chlorpyrifos in an agriculturally dominated tributary of the San Joaquin River and to assess the ecological significance of such effects. Assessment endpoints were fish population persistence and invertebrate community productivity. Daily chemical measurements collected over a period of one year were analyzed temporally for frequency, duration, and spacing between events for acute and chronic exposure episodes. Effects thresholds for fish and freshwater lotic invertebrates were determined from single-species laboratory toxicity tests. Potential risk was characterized by the degree of overlap of distributions of exposure events and effects, with consideration given to additive toxicity of other OP insecticides, recovery periods, and duration of chronic exposure (> or = 21 d). Ecological significance was determined by analysis of fish assemblage dietary and reproductive habits in relation to the surrogate invertebrate taxa judged at risk. Results of analysis indicated no direct effects on fish, and indirect effects on fish through elimination of invertebrate food items were considered unlikely. Biological survey information will be necessary to address uncertainty in this risk conclusion, especially as it relates to the benthic invertebrate community. Results of this site-specific risk analysis suggest that fish population persistence and invertebrate community productivity were not adversely affected by measured chlorpyrifos residues during a year-long monitoring period.

Quantitation of 17alpha-ethinylestradiol in aquatic samples using liquid-liquid phase extraction, dansyl derivatization, and liquid chromatography/positive electrospray tandem mass spectrometry.

A rapid and sensitive method for the analysis of 17alpha-ethinylestradiol (EE2) in environmental aqueous samples has been developed. Aquatic samples were extracted using liquid-liquid extraction, and organic phase extracts were concentrated and derivatized with dansyl chloride. Analysis was performed using high-performance liquid chromatography with positive electrospray ionization and tandem mass spectrometry (HPLC/ESI-MS/MS). Deuterated 17alpha-ethinylestradiol was used as internal standard and was added to samples before extraction. A limit of quantitation of 1 ng/L was obtained using a 25 mL aqueous sample. The average recovery of EE2 spiked into a 25 mL tapwater sample was 100%. This highly sensitive quantitation method is useful for measuring low levels of EE2 in aqueous environmental samples.