Leonard Böhm

Sorption of Highly Hydrophobic Organic Chemicals to Organic Matter Relevant for Fish Bioconcentration Studies

With regard to a potential underestimation of bioconcentration factors (BCF) in flow-through fish tests, sorption of 11 highly hydrophobic organic chemicals (HOCs) (log KOW 5.5-7.8) from different substance classes was systematically investigated for the first time in the presence of fish feed (FF) and filter residues (FR), the organic matter (OM) most relevant for fish bioconcentration studies. Sorption was investigated in batch-equilibrium experiments by solid-phase microextraction (SPME) resulting in partitioning coefficients of solid-water (Kd), total organic carbon-water (KTOC), and dissolved organic carbon-water (KDOC). Results prove a high affinity of HOCs for FF and FR supporting a significant impact on BCF studies and differing from sorption to Aldrich-humic acid (AHA) utilized as reference sorbent. Sorption is influenced by interactions between HOCs and OM characteristics. For FF, KDOC values were higher than KTOC values. Results help to assess the relevance of interaction of HOCs from different substance classes with OM relevant for BCF studies.

Passive Dosing in Chronic Toxicity Tests with the Nematode Caenorhabditis elegans

In chronic toxicity tests with Caenorhabditis elegans, it is necessary to feed the nematode with bacteria, which reduces the freely dissolved concentration (Cfree) of hydrophobic organic chemicals (HOCs), leading to poorly defined exposure with conventional dosing procedures. We examined the efficacy of passive dosing of polycyclic aromatic hydrocarbons (PAHs) using silicone O-rings to control exposure during C. elegans toxicity testing and compared the results to those obtained with solvent spiking. Solid-phase microextraction and liquid-liquid extraction were used to measure Cfree and the chemicals taken up via ingestion. During toxicity testing, Cfree decreased by up to 89% after solvent spiking but remained constant with passive dosing. This led to a higher apparent toxicity on C. elegans exposed by passive dosing than by solvent spiking. With increasing bacterial cell densities, Cfree of solvent-spiked PAHs decreased while being maintained constant with passive dosing. This resulted in lower apparent toxicity under solvent spiking but an increased apparent toxicity with passive dosing, probably as a result of the higher chemical uptake rate via food (CUfood). Our results demonstrate the utility of passive dosing to control Cfree in routine chronic toxicity testing of HOCs. Moreover, both chemical uptake from water or via food ingestion can be controlled, thus enabling the discrimination of different uptake routes in chronic toxicity studies.

Solid-phase microextraction for bioconcentration studies according to OECD TG 305

An important aim of the European Community Regulation on chemicals and their safe use is the identification of (very) persistent, (very) bioaccumulative, and toxic substances. In other regulatory chemical safety assessments (pharmaceuticals, biocides, pesticides), the identification of such (very) persistent, (very) bioaccumulative, and toxic substances is of increasing importance. Solid-phase microextraction is especially capable of extracting total water concentrations as well as the freely dissolved fraction of analytes in the water phase, which is available for bioconcentration in fish. However, although already well established in environmental analyses to determine and quantify analytes mainly in aqueous matrices, solid-phase microextraction is still a rather unusual method in regulatory ecotoxicological research. Here, the potential benefits and drawbacks of solid-phase microextraction are discussed as an analytical routine approach for aquatic bioconcentration studies according to OECD TG 305, with a special focus on the testing of hydrophobic organic compounds characterized by log KOW > 5.

Fish bioconcentration studies with column‐generated analyte concentrations of highly hydrophobic organic chemicals

The performance of aqueous exposure bioconcentration fish tests according to Organisation for Economic Co-operation and Development (OECD) guideline 305 requires the possibility of preparing stable aqueous concentrations of the test substances. For highly hydrophobic organic chemicals (HOCs; octanol-water partition coefficient [log KOW ] > 5), testing via aqueous exposure may become increasingly difficult. A solid-phase desorption dosing system was developed to generate stable concentrations of HOCs without using solubilizing agents. The system was tested with hexachlorobenzene (HCB), o-terphenyl (oTP), polychlorinated biphenyl (PCB) 153, and dibenz[a,h]anthracene (DBA) (log KOW 5.5-7.8) in 2 flow-through fish tests with rainbow trout (Oncorhynchus mykiss). The analysis of the test media applied during the bioconcentration factor (BCF) studies showed that stable analyte concentrations of the 4 HOCs were maintained in the test system over an uptake period of 8 wk. Bioconcentration factors (L kg-1 wet wt) were estimated for HCB (BCF 35 589), oTP (BCF 12 040), and PCB 153 (BCF 18 539) based on total water concentrations. No bioconcentration could be determined for DBA, probably because of the rapid metabolism of the test item. The solid-phase desorption dosing system is suitable to provide stable aqueous concentrations of HOCs required to determine the bioconcentration in fish and represents a viable alternative to the use of solubilizing agents for the preparation of test solutions. Environ Toxicol Chem 2017;36:906-916.

Can solid‐phase microextraction replace solvent extraction for water analysis in fish bioconcentration studies with highly hydrophobic organic chemicals?

With the aim to refine water analysis in fish bioconcentration studies, automated solid-phase microextraction (SPME) was used as an alternative approach to conventional solvent extraction (liquid-liquid extraction [LLE]) for the extraction of 3 hydrophobic organic chemicals (HOCs; log KOW 5.5-7.8) from flow-through studies with rainbow trout (Oncorhynchus mykiss). The results showed that total concentrations extracted by SPME combined with internal standards and LLE are equal. The results further verify the possibility of simultaneous extraction of total and freely dissolved HOC concentrations by SPME. Freely dissolved concentrations allow the assessment of sorption and bioavailability of HOCs in bioconcentration studies and their potential impact on resulting bioconcentration factors (BCFs). Reduction in freely dissolved water concentrations can result in an underestimation of BCFs if they are calculated based on total water concentrations. For polychlorinated biphenyl (PCB) 153, a significant increase in BCF value was observed when freely dissolved concentrations were taken into account. However, log BCF values calculated based on freely dissolved concentrations did not correlate linearly with log KOW values above 5 to 6. This pointed to further influences besides a reduction in freely dissolved water concentrations by sorption to organic matter. The results can aid in assessment of the factors that influence bioconcentration systems and also give important information regarding the possible replacement of LLE by SPME for water analysis of highly HOCs in fish bioconcentration studies. Environ Toxicol Chem 2017;36:2887-2894.