Bioconcentration potential of ionic liquids: New data on membrane partitioning and its comparison with predictions obtained by COSMOmic.

Ionic liquids (ILs) have recently gained significant attention in both the scientific community and industry, but there is a limited understanding of the potential risks they might pose to the environment and human health, including their potential to accumulate in organisms. While membrane and storage lipids have been considered as primary sorption phases driving bioaccumulation, in this study we used an in vitro tool known as solid-supported lipid membranes (SSLMs) to investigate the affinity of ILs to membrane lipid - phosphatidylcholine and compare the results with an existing in silico model. Our findings indicate that ILs may have a strong affinity for the lipids that form cell membranes, with the key factor being the length of the cation's side chain. For quaternary ammonium cations, increase in membrane affinity (logMA) was observed from 3.45 ± 0.06 at 10 carbon atoms in chain to 4.79 ± 0.06 at 14 carbon atoms. We also found that the anion can significantly affect the membrane partitioning of the cation, even though the anions themselves tend to have weaker interactions with phospholipids than the cations of ILs. For 1-methyl-3-octylimidazolium cation the presence of tricyanomethanide anion caused increase in logMA to 4.23 ± 0.06. Although some of our data proved to be consistent with predictions made by the COSMOmic model, there are also significant discrepancies. These results suggest that further research is needed to improve our understanding of the mechanisms and structure-activity relationships involved in ILs bioconcentration and to develop more accurate predictive models.

Insights into the interaction of human serum albumin with ionic liquids - Thermodynamic, spectroscopic and molecular modelling studies.

Human serum albumin (HSA) effectively binds different types of low-molecular-weight compounds and thus enables their distribution in living organisms. Recently, it has been reported that the protein-ligand interactions play a crucial role in bioaccumulation processes and provide an important sorption phase, especially for ionogenic compounds. Therefore, the binding interactions of such compounds with proteins are the subject of an ongoing interest in environmental and life sciences. In this paper, the influence of some counter-ions, namely [B(CN)4]- and [C(CN)3]- on the affinity of the [IM1-12]+ towards HSA has been investigated and discussed based on experimental methods (isothermal titration calorimetry and steady-state fluorescence spectroscopy) and molecular dynamics-based computational approaches. Furthermore, the thermal stability of the resulting HSA/ligand complexes was assessed using DSC and CD spectroscopy. As an outcome of the work, it has been ascertained that the protein is able to bind simultaneously the ligands under study but in different regions of HSA. Thus, the presence in the system of [IM1-12]+ does not disturb the binding of [C(CN)3]- and [B(CN)4]-. The presented results provide important information on the presence of globular proteins and some ionogenic compounds in the distribution and bioaccumulation of ILs in the environment and living organisms.

Bioconcentration of imidazolium ionic liquids: In vivo evaluation in marine mussels Mytilus trossulus.

Although imidazolium ionic liquids (ILs) are beginning to be used more widely in many industrial fields e.g., as reaction media, electrolytes, stationary phases in gas chromatography), there is still little information about their potential environmental fate. Among the uncertainties regarding the risks associated with these compounds, bioconcentration is one of the key issues, about which many doubts have been raised in recent years. While in vitro data suggest that permanently charged compounds can also bioconcentrate, conclusive evidence in the form of studies on organisms, at least for selected compounds, is needed. Therefore, the main objective of this work was to determine whether imidazolium cations of ILs, namely 1-methyl-3-octylimidazolium ([IM18]+) and 1-methyl-3-dodecylimidazolium ([IM1-12]+), can bioconcentrate in marine invertebrates tissues. During 21-day experiments, Mytilus trossulus mussels were exposed to these cations individually, at a concentration of 10 µg/L. In our study, it has been demonstrated for the first time during in vivo study, that long-chain imidazolium ionic liquids can bioconcentrate. The determined BCF value for [IM1-12]+ of 21,901 ± 3400 L/kg makes this compound to be considered highly bioaccumulative according to commonly accepted criteria. However, the obtained BCF for [IM18]+ (with the value below 100) suggests that this cation has little potential for bioconcentration. On the other hand, no salinity or anion influence on the bioconcentration of the tested cations was observed. Our tests also confirm that imidazolium ILs exhibit acute toxicity only at relatively high concentration levels, as LC50 reached 0.68 mg/L for [IM1-12][Br], and 11.66 mg/L for [IM18][C(CN)3]. This further confirms that the risks associated with the potential presence of these compounds in the environment should be attributed to their high persistence and potential bioconcentration, rather than acute toxicity.

Analysis of imidazolium ionic liquids in biological matrices: A novel procedure for the determination of trace amounts in marine mussels.

Imidazolium ionic liquids (ILs) are chemical compounds beginning to be used on a mass scale. Although their presence in the environment is usually treated as only potential threat, there are already first evidences that this has become a real case. Taking into account their increasing use it might be expected that this problem will also increase in the nearest future. Given that some of the imidazolium cations exhibit high potential for bioconcentration, it is likely that they will accumulate in the tissues of wild organisms. Thus, there is no doubt that monitoring the presence of these compounds in organisms from potentially contaminated waters will be needed. Therefore, the aim of our study was to develop and fully validate a novel and reliable analytical procedure for the determination of the mixture of imidazolium ILs in Mytilus trossulus mussels. For this purpose, different extraction techniques were tested such as: microwave-assisted extraction (MAE), accelerated-solvent extraction (ASE) and bullet-blender homogenization (HOMO). Finally, the proposed procedure is based on the application of MAE technique for the extraction of imidazolium cations and SPE technique using Oasis HLB cartridges for the purification of the obtained extracts and LC-MS/MS technique with QqQ analyzer for their final determination. Absolute recoveries of the proposed analytical procedure reached 71-90%. The developed procedure is characterized with low limits of quantification, at 50-100 ng g-1 dry tissue and allows for reliable determination of trace amounts of the tested compounds in complex biological matrix. Matrix effects obtained for the optimized procedure ranged from 7.8 to 37.5%. As a result, this is the first study presenting the analytical procedure for the analysis of imidazolium ILs in aquatic animal tissues.

Ionic liquids as potentially hazardous pollutants: Evidences of their presence in the environment and recent analytical developments.

Ionic liquids (ILs) are considered to be very promising group of chemicals and the number of their potential applications is growing rapidly. However, while these compounds were originally proposed as a green alternative to classical solvents, there are certain doubts as to whether this classification is correct. Although in recent years there have been first reports published proving the presence of some ILs in the environment and even in human blood, at this point the scale of this possible problem is not yet fully understood. However, there is no doubt that as the number of ILs applications increases, analytical capabilities for rapid detection of possible environmental contamination should be also considered. Therefore, in this review paper, recent evidences for the ILs environmental contamination as well as analytical achievements related to the extraction of ILs from various environmental matrices have been summarized and important gaps and future perspectives have been pointed out. Based on the presented data it might be concluded that there is the urgent need for further development towards risk assessment of these potential environmental contaminants.

Transformation products of pharmaceuticals in the environment: Their fate, (eco)toxicity and bioaccumulation potential.

Nowadays, a huge scientific attention is being paid to the chemicals of emerging concern, which may pose a significant risk to the human and whole ecosystems. Among them, residues of pharmaceuticals are a widely investigated group of chemicals. In recent years it has been repeatedly demonstrated that pharmaceuticals are present in the environment and that some of them can be toxic to organisms as well as accumulate in their tissues. However, even though the knowledge of the presence, fate and possible threats posed by the parent forms of pharmaceuticals is quite extensive, their transformation products (TPs) have been disregarded for long time. Since last few years, this aspect has gained more scientific attention and recently published papers proved their common presence in the environment. Also the interest in terms of their toxicity, bioconcentration and stability in the environment has increased. Therefore, the aim of our paper was to revise and assess the current state of knowledge on the fate and effects resulting from the presence of the pharmaceuticals' transformation drugs in the environment. This review discusses the metabolites of compounds belonging to six major pharmaceutical groups: SSRIs, anticancer drugs, antibiotics, antihistamines, NSAIDs and opioids, additionally discussing other individual compounds for which literature data exist. The data presented in this paper prove that some TPs may be as harmful as their native forms, however for many groups of drugs this data is still insufficient to assess the risk posed by their presence in the environment.

Hydrolytic stability of anticancer drugs and one metabolite in the aquatic environment.

Due to the genotoxic, carcinogenic and teratogenic mechanism of action, anticancer drugs are highly hazardous compounds. Their occurrence, fate, and effects in the environment have not been systematically studied as compared to other medicaments. Therefore, reliable data, including their stability and persistency, is required in order to assess it. Taking into account, that hydrolysis is one of the most important factors regarding stability of chemicals in water, the aim of our study was to investigate the hydrolytic stability of five commonly used anticancer drugs (ifosfamide, cyclophosphamide, 5-fluorouracil, imatinib, and methotrexate) and one metabolite (7-hydroxymethotrexate), as the systematized and coherent data available is limited. The hydrolysis studies have been prepared according to the OECD 111 procedure to obtain standardized and comparable results. The preliminary tests at pH 4, 7, and 9 and 50 °C show that only cyclophosphamide and ifosfamide are unstable, whereas the estimated t1/2 at 25 °C is >1 year for other investigated compounds. Moreover, much more detailed experiments were performed and indicate that at environmentally relevant temperatures, cyclophosphamide, and ifosfamide would be quite persistent in the terms of hydrolytic stability. Moreover, the preliminary investigation on the hydrolysis products was performed.

Insight into the Sorption of 5-Fluorouracil and Methotrexate onto Soil-pH, Ionic Strength, and Co-Contaminant Influence.

Nowadays anticancer drugs (ADs), like other pharmaceuticals, are recognized as new emerging pollutants, meaning that they are not commonly monitored in the environment; however, they have great potential to enter the environment and cause adverse effects there. The current scientific literature highlights the problem of their presence in the aquatic environment by publishing more and more results on their analytics and ecotoxicological evaluation. In order to properly assess the risk associated with the presence of ADs in the environment, it is also necessary to investigate the processes that are important in understanding the environmental fate of these compounds. However, the state of knowledge on mobility of ADs in the environment is still very limited. Therefore, the main aim of our study was to investigate the sorption potential of two anticancer drugs, 5-fluorouracil (5-FU) and methotrexate (MTX), onto different soils. Special attention was paid to the determination of the influence of pH and ionic strength as well as presence of co-contaminants (cadmium (Cd2+) and another pharmaceutical-metoprolol (MET)) on the sorption of 5-FU and MTX onto soil. The obtained distribution coefficient values (Kd) ranged from 2.52 to 6.36 L·kg-1 and from 6.79 to 12.94 L·kg-1 for 5-FU and MTX, respectively. Investigated compounds may be classified as slightly or low mobile in the soil matrix (depending on soil). 5-FU may be recognized as more mobile in comparison to MET. It was proved that presence of other soil contaminants may strongly influence their mobility in soil structures. The investigated co-contaminant (MET) caused around 25-fold increased sorption of 5-FU, whereas diminished sorption of MTX. Moreover, the influence of environmental conditions such as pH and ionic strength on their sorption has been clearly demonstrated.

Interaction of pharmaceutical metabolites with blood proteins and membrane lipids in the view of bioconcentration: A preliminary study based on in vitro assessment.

Pharmaceuticals pose a real threat to the environment, which has been proven in many studies to date. However, still little is known about the transformation products (TPs) of these compounds, which can also interact with organisms, causing adverse effects like noticeable toxicity or bioconcentration. Many recent works confirm that metabolites of pharmaceuticals are present in the environment, and preliminary studies suggest that they may be equally dangerous to or even more so than their parent compounds. Additionally, it has been proven that some of them have high hydrolytic stability, thus they may be persistent in the environment. This property also increases the likelihood that these compounds will be uptaken and accumulated in the tissues of organisms. Therefore, the aim of the present study was to preliminarily estimate the affinity of the transformation products of selected drugs for blood proteins and cell membrane-forming lipids, considered as important sorption phases during distribution in a living organism. In this study, it was shown that although the examined metabolites do not have a strong affinity for membrane lipids, they exhibit relatively strong binding to proteins, which may considerably affect the distribution of TPs in an organism and may indicate a non-classical process of bioconcentration. The results obtained confirm that the TPs of pharmaceuticals should be given much more attention and their potential for bioconcentration should be further determined.

Ionic liquids as environmental hazards - Crucial data in view of future PBT and PMT assessment.

Ionic liquids (ILs) constitute a large group of chemical compounds. They have gained much attention among scientists and industry due to their unique properties. Due to the fact that ILs are purely ionic compounds, there is the possibility to design an enormous number of cation and anion combinations, making them designer solvents. Thus it also creates the possibility of producing more environmentally benign solvents. However, significant drawbacks related mainly to their toxicity and persistence have already been noticed. Furthermore the interest in these compounds is constantly growing and their impact on the environment should be defined. More and more ILs are produced or imported in the amount higher than 10 tonnes per year and the group of ILs registered in REACH is still expanding. Thus for an increasing number of compounds, it will be necessary to perform a PBT and PMT assessment using the criteria described in REACH. Therefore the data collected in this work thoroughly sort out the information on the toxicity, bioconcentration/bioaccumulation, biodegradation and mobility of ILs in the context of PBT and PMT assessment.

In vitro methods for predicting the bioconcentration of xenobiotics in aquatic organisms.

The accumulation of anthropogenic chemical substances in aquatic organisms is an immensely important issue from the point of view of environmental protection. In the context of the increasing number and variety of compounds that may potentially enter the environment, there is a need for efficient and reliable solutions to assess the risks. However, the classic approach of testing with fish or other animals is not sufficient. Due to very high costs, significant time and labour intensity, as well as ethical concerns, in vivo methods need to be replaced by new laboratory-based tools. So far, many models have been developed to estimate the bioconcentration potential of chemicals. However, most of them are not sufficiently reliable and their predictions are based on limited input data, often obtained with doubtful quality. The octanol-water partition coefficient is still often used as the main laboratory tool for estimating bioconcentration. However, according to current knowledge, this method can lead to very unreliable results, both for neutral species and, above all, for ionic compounds. It is therefore essential to start using new, more advanced and credible solutions on a large scale. Over the last years, many in vitro methods have been newly developed or improved, allowing for a much more adequate estimation of the bioconcentration potential. Therefore, the aim of this work was to review the most recent laboratory methods for assessing the bioconcentration potential and to evaluate their applicability in further research.

Hydrolytic stability of selected pharmaceuticals and their transformation products.

The fact that pharmaceuticals are present in the environment has been proven in numerous publications. Nevertheless, their transformation products (mainly metabolites) are detected significantly less often, mainly because they are not included in the detecting methods, even though many of them are excreted from organisms at high rates and may be biologically active or have other properties that make them a potential threat to the environment. One of the most common processes that occur in the aqueous environment is hydrolysis, which may be one of the most important factors influencing the persistency of pharmaceuticals. Therefore four pharmaceuticals (carbamazepine, ibuprofen, tramadol and naproxen) as well as ten metabolites (10,11-dihydro-10-hydroxy carbamazepine, 10,11-dihydro-2-hydroxy carbamazepine, carbamazepine epoxide, 2-hydroxy ibuprofen, ibuprofen carboxylic acid, O-desmethyltramadol, hydroxy metronidazole, N-acetylsulfamethoxazole, 4'-hydroxy diclofenac, and O-desmethylnaproxen) were selected for the hydrolytic stability tests in accordance to OECD 111 Guideline. The preliminary test showed that only carbamazepine epoxide at pH 4, hydroxy metronidazole at pH 9 and 4'-hydroxy diclofenac at pH 4 and 9 were unstable and were included in the extended tests, which resulted in calculation of rate constants and half-lives at the temperature of 20, 50 and 70 °C as well as the activation energy at the pH values in which these compounds were unstable. The obtained results show that carbamazepine epoxide is quite unstable and the half-life at 20 °C was a little more than 8 days. Nevertheless, hydroxy-metronidazole and 4'-hydroxy diclofenac did not degrade at 20 °C for 30 days.

Leaching behavior of pharmaceuticals and their metabolites in the soil environment.

Pharmaceuticals constitute a significant group of emerging pollutants (EPs). The use of pharmaceuticals in animal breeding causes them to reach the soil environment in excrement and fertilizers. Depending on their chemical properties, pharmaceuticals can be sorbed to the soil or be washed out with rainfall and eventually be entered into groundwater. This paper evaluates the mobility of tramadol (TRA) and carbamazepine (CBZ), and two transformation products, O-desmethyltramadol (O-DMTRA) and 10,11-dihydro-10-hydroxycarbamazepine (10-OH-CBZ) in soils. Both pharmaceuticals are applied in human and animal treatment, which makes them enter the environment in native and metabolized form in high doses. Experiments were carried out in accordance with the OECD 106 procedure (batch tests) and DIN 19528:2009-01 procedure (percolation column test). The adsorption coefficients (Kd) for TRA, CBZ, O-DMTRA and 10-OH-CBZ were, respectively, 1.41 ±â€¯0.10, 1.87 ±â€¯0.06, 0.90 ±â€¯0.03 and 0.37 ±â€¯0.07 for sandy soil RS04, and 18.09 ±â€¯0.78, 2.56 ±â€¯0.05, 10.89 ±â€¯0.17 and 0.56 ±â€¯0.38  L kg-1 for loamy soil RS06. The percolation column test was carried out for sandy soil RS04. The results obtained for TRA and O-DMTRA under static conditions indicated a high mobility of these compounds in soil, whereas the column leaching experiment showed that these compounds bind strongly to soil particles. A correlation between static and dynamic tests was observed in the case of CBZ and 10-OH-CBZ. These compounds will probably be characterized by a high or moderate mobility in soil.

Preliminary evaluation of the application of carbon nanotubes as potential adsorbents for the elimination of selected anticancer drugs from water matrices.

The presence of anticancer drugs in the environment raises a major concern due to their potentially negative impact on living organisms, as they cause, inter alia, teratogenic, mutagenic, carcinogenic effects on growing cells. The main source of these pharmaceuticals in the environment is the ineffectively treated wastewaters, hence the efficient methods for their removal are required. In this study, we have evaluated for the first time the applicability of different Multi-Walled Carbon Nanotubes (MWCNTs) as alternative adsorbents for the removal of three popular anticancer drugs from water matrices, which are detected in waste/hospital waters at the concentrations even up to µg L-1: cyclophosphamide (CP), ifosfamide (IF) and 5-fluorouracil (5-FU). It was observed that adsorption equilibrium time was reached after 20 min for each compound and CNTs. Moreover, CP has the highest sorption potential (Kd up to 1597 L kg-1) for all CNTs and CNTs with the highest surface area have the greatest adsorption capacity. Isotherms were fitted best to the Freundlich model. Significant influence of pH in the range 4-9 and ionic strength was not observed what may indicate that adsorption is not dependent on fluctuation of basic environmental conditions.

A new approach for the extraction of tetracyclines from soil matrices: application of the microwave-extraction technique.

The widespread use of tetracyclines (TCs) in animal husbandry is associated with their constant penetration into the environment and the threat they might pose to living organisms. While the literature data on the analysis of these substances in such matrices as food, tissues, or wastewater are quite extensive, there are still only a few methods presented for the determination of these compounds in soil samples. Moreover, among the literature methods for the extraction of TCs from soil samples, microwave-assisted solvent extraction (MAE) was used only once and in combination with liquid chromatography with spectrophotometric detection (LC-UV). However, according to the EU Commission Decision 2002/657/EC, the use of LC-UV for the determination of compounds, including pharmaceuticals, in environmental samples is not sufficient. Therefore, the development and application of a sensitive and selective method using the MAE-SPE-LC-MS/MS(MRM) technique for the isolation and identification of a mixture of oxytetracycline (OTC), tetracycline (TC), and chlortetracycline (CTC) in soils is presented in our study. The credibility of this method has been confirmed with good parameters of validation. The optimal extraction conditions of three TCs using MAE techniques were to conduct double extraction for 10 min each, at 60 °C, using a mixture of ACN:McIlvaine buffer:0.1 M EDTA (2:1:1, v/v/v) and an additional cleaning of the extracts by SPE. The effectiveness of the extraction of these compounds was assessed based on two different ways (absolute recovery from 46 to 65.1% and relative recovery from 101.1 to 109.5%). Finally, the validated MAE-SPE-LC-MS/MS(MRM) method was used for the analysis of six samples from agricultural areas of northern Poland. OTC and TC, at concentrations of 11.7 and 14.5 µg kg-1 were determined in two different samples. An assessment of risk quotients was also performed. The presented method was proven to be a useful tool in the analysis of residues of selected TCs in the soil ecosystem. Obtained data on the presence of these drugs in Polish soils is the first report for this country.

Evaluation of the sorption mechanism of ionic liquids onto multi-walled carbon nanotubes.

The knowledge of the sorption mechanism of different chemicals onto third generation carbon sorbents such as carbon nanotubes (CNTs) is needed in order to project systems for the effective removal of pollutants from the environment. This paper reports evaluation of the sorption mechanism of selected ionic liquids (ILs), being considered as potential pollutant in environment, onto various CNTs. CNTs characterized by the smallest diameter and the biggest surface area showed the highest sorption capacity to isolate ILs from an aqueous solution. CNTs with a bigger diameter, a functionalized surface and particularly a helical shape showed a lower sorption capacity. The sorption mechanism has been defined as complex, including van der Waals, π-π and electrostatic interactions with dominating π-π interactions. Due to the relatively high sorption coefficient (355.98 ± 20.69-6397.10 ± 355.42 L kg-1 depending on the IL) the study showed that multi-walled carbon nanotubes can potentially be used to effectively isolate ILs from an aqueous solution. Moreover, proved in this study, the fast sorption kinetic, and uncomplicated regeneration process, leading to an even higher sorption capacity, means that CNTs are promising material which could find potential applications in the treatment of water contaminated by ILs.

Assessment of soils contamination with veterinary antibiotic residues in Northern Poland using developed MAE-SPE-LC/MS/MS methods.

Among the wide range of compounds reaching the soil are the veterinary antimicrobials. Since no regulations regarding acceptable levels of drug concentrations in the environment exist, monitoring tests, particularly concerning soils, are carried out very rarely. This study presents a preliminary assessment of the contamination of agricultural soils in Northern Poland with seven antimicrobial veterinary medicines which has never been carried out before. Veterinary drugs were detected in 54% of the examined soil samples; the most commonly detected drugs were sulfonamides and trimethoprim. The highest indicated concentrations refer to enrofloxacin (57.0 µg kg-1) and trimethoprim (47.8 µg kg-1). The presence of these target drugs in the soil environment confirms the need for further monitoring studies. The analytical methods developed in this study are an excellent tool to achieve this goal and allow an estimation of the risk connected with the presence of veterinary antimicrobials in soils.

Membrane partitioning of ionic liquid cations, anions and ion pairs - Estimating the bioconcentration potential of organic ions.

Recent efforts have been directed towards better understanding the persistency and toxicity of ionic liquids (ILs) in the context of the "benign-by-design" approach, but the assessment of their bioaccumulation potential remains neglected. This paper reports the experimental membrane partitioning of IL cations (imidazolium, pyridinium, pyrrolidinium, phosphonium), anions ([C(CN)3]-, [B(CN)4]-, [FSO2)2N]-, [(C2F5)3PF3]-, [(CF3SO2)2N]-) and their combinations as a measure for estimating the bioconcentration factor (BCF). Both cations and anions can have a strong affinity for phosphatidylcholine bilayers, which is mainly driven by the hydrophobicity of the ions. This affinity is often reflected in the ecotoxicological impact. Our data revealed that the bioconcentration potential of IL cations and anions is much higher than expected from octanol-water-partitioning based estimations that have recently been presented. For some ILs, the membrane-water partition coefficient reached levels corresponding to BCFs that might become relevant in terms of the "B" (bioaccumulation potential) classification under REACH. However, this preliminary estimation need to be confirmed by in vivo bioconcentration studies.