Ethinylestradiol (EE2) is a synthetic hormone that has been recognized as one of the most prominent endocrine disruptors found in the aqueous environment. Nevertheless, the low content of EE2 in wastewater makes its identification/quantification unfeasible - a major drawback for the evaluation of its persistence and environmental impact. In this context, a novel extraction/concentration method for EE2 from wastewater is proposed here based on aqueous biphasic systems composed of ionic liquids (ILs). Aqueous biphasic systems formed by several hydrophilic ILs and KNaCHO were initially screened and optimized, with extraction efficiencies of EE2 for the IL-rich phase ranging between 92 and 100%. Remarkable results were obtained with systems that allow the complete extraction of EE2 in a single-step, and without loss of EE2 or the saturation of the extractive phase. Further, the concentration factors of EE2 attainable with these systems were investigated by a suitable manipulation of the composition of the phase-forming components and the corresponding volumes of the coexisting phases. An outstanding concentration of EE2 up to 1000-fold (from ng L to μg L) in a single extraction and concentration step was achieved for the first time with IL-based aqueous biphasic systems. These systems are straightforwardly envisaged for the monitoring of wastewater as one-step extraction and concentration routes for a wide array of endocrine disrupting chemicals while allowing an adequate evaluation of their environmental impact.

Aqueous biphasic systems (ABS) composed of ionic liquids (ILs) and conventional salts have been largely investigated and successfully used in separation processes, for which the determination of the corresponding ternary phase diagrams is a prerequisite. However, due the large number of ILs that can be prepared and their high structural versatility, it is impossible to experimentally cover and characterize all possible combinations of ILs and salts that may form ABS. The development of tools for the prediction and design of IL-based ABS is thus a crucial requirement. Based on a large compilation of experimental data, a correlation describing the formation of IL-based ABS is shown here, based on the hydrogen-bonding interaction energies of ILs (EHB) obtained by the COnductor-like Screening MOdel for Real Solvents (COSMO-RS) and the molar entropy of hydration of the salt ions. The ability of the proposed model to predict the formation of novel IL-based ABS is further ascertained.

Human activities are responsible for the release of innumerous substances into the aquatic environment. Some of these substances can be used as pollution tracers to identify contamination sources and to prioritize monitoring and remediation actions. Thus, their identification and quantification are of high priority. However, due to their presence in complex matrices and at significantly low concentrations, a pre-treatment/concentration step is always required. As an alternative to the currently used pre-treatment methods, mainly based on solid-phase extractions, aqueous biphasic systems (ABS) composed of ionic liquids (ILs) and KCHO are here proposed for the simultaneous extraction and concentration of mixtures of two important pollution tracers, caffeine (CAF) and carbamazepine (CBZ). An initial screening of the IL chemical structure was carried out, with extraction efficiencies of both tracers to the IL-rich phase ranging between 95 and 100%, obtained in a single-step. These systems were then optimized in order to simultaneously concentrate CAF and CBZ from water samples followed by HPLC-UV analysis, for which no interferences of the ABS phase-forming components and other interferents present in a wastewater effluent sample have been found. Based on the saturation solubility data of both pollution tracers in the IL-rich phase, the maximum estimated concentration factors of CAF and CBZ are 28595- and 8259-fold. IL-based ABS can be thus envisioned as effective pre-treatment techniques of environmentally-related aqueous samples for a more accurate monitoring of mixtures of pollution tracers.

Deoxyribonucleic acid (DNA) carries the genetic information essential for the growth and functioning of living organisms, playing a significant role in life sciences research. However, the long-term storage and preservation of DNA, while ensuring its bioactivity, are still current challenges to overcome. In this work, aqueous solutions of ionic liquids (ILs) were investigated as potential preservation media for double stranded (dsDNA). A screening of several ILs, by combining the cholinium, tetrabutylammonium, tetrabutylphosphonium, and 1-ethyl-3-methylimidazolium, cations with the anions bromide, chloride, dihydrogen phosphate, acetate, and glycolate, was carried out in order to gather fundamental knowledge on the molecular features of ILs that improve the dsDNA stability. Different IL concentrations and the pH effect were also addressed. Circular dichroism (CD) spectroscopy was used to evaluate the conformational structure and stability of dsDNA. IL-DNA interactions were appraised by UV-Vis absorption spectrophotometry and 31 P nuclear magnetic resonance (NMR) spectroscopy. The results obtained demonstrate that pH has a significant effect towards the dsDNA stability. Amongst the ILs investigated, cholinium-based ILs are the most promising class of ILs to preserve the dsDNA structure, in which electrostatic interactions between the cholinium cation and the DNA phosphate groups play a significant role as demonstrated by the 31 P NMR data, being more relevant at higher IL concentrations. On the other hand, the denaturation of dsDNA mainly occurs with ILs composed of more hydrophobic cations and able to establish dispersive interactions with the nucleobases environment. Furthermore, the IL anion has a weaker impact when compared to the IL cation effect to interact with DNA molecules. The experimental data of this work provide relevant fundamental knowledge for the application of ILs in the preservation of nucleic acids, being of high relevance in the biotechnology field.

Aiming at the development of self-buffering and benign extraction/separation processes, this work reports a novel class of aqueous biphasic systems (ABS) composed of ionic liquids (ILs) and organic biological buffers (Good's buffers, GBs). A large array of ILs and GBs was investigated, revealing than only the more hydrophobic and fluorinated ILs are able to form ABS. For these systems, the phase diagrams, tie-lines, tie-line lengths, and critical points were determined at 25 °C. The ABS were then evaluated as alternative liquid-liquid extraction strategies for two amino acids (L-phenylalanine and L-tryptophan). The single-step extraction efficiencies for the GB-rich phase range between 22.4 and 100.0 % (complete extraction). Contrarily to the most conventional IL-salt ABS, in most of the systems investigated, the amino acids preferentially migrate for the most biocompatible and hydrophilic GB-rich phase. Remarkably, in two of the studied ABS, L-phenylalanine completely partitions to the GB-rich phase while L-tryptophan shows a preferential affinity for the opposite phase. These results show that the extraction efficiencies of similar amino acids can be tailored by the design of the chemical structures of the phase-forming components, creating thus new possibilities for the use of IL-based ABS in biotechnological separations.