380 research outputs found
Aqueous two-phase systems: Towards novel and more disruptive applications
Aqueous two-phase systems (ATPS) have been mainly proposed as powerful platforms for the separation
and purification of high-value biomolecules. However, after more than seven decades of research, ATPS
are still a major academic curiosity, without their wide acceptance and implementation by industry,
leading to the question whether ATPS should be mainly considered in downstream bioprocessing.
Recently, due to their versatility and expansion of the Biotechnology and Material Science fields, these
systems have been investigated in novel applications, such as in cellular micropatterning and bioprinting,
microencapsulation, to mimic cells conditions, among others. This perspective aims to be a reflection on
the current status of ATPS as separation platforms, while overviewing their applications, strengths and
limitations. Novel applications, advantages and bottlenecks of ATPS are further discussed, indicating
some directions on their use to create innovative industrial processes and commercial products.publishe
Insights on the DNA stability in aqueous solutions of ionic liquids
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 31P 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 31P 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.publishe
The role of ionic liquids in the pharmaceutical field: an overview of relevant applications
Solubility, bioavailability, permeation, polymorphism, and stability concerns associated to solid-state pharmaceuticals demand for effective solutions. To overcome some of these drawbacks, ionic liquids (ILs) have been investigated as solvents, reagents, and anti-solvents in the synthesis and crystallization of active pharmaceutical ingredients (APIs), as solvents, co-solvents and emulsifiers in drug formulations, as pharmaceuticals (API-ILs) aiming liquid therapeutics, and in the development and/or improvement of drug-delivery-based systems. The present review focuses on the use of ILs in the pharmaceutical field, covering their multiple applications from pharmaceutical synthesis to drug delivery. The most relevant research conducted up to date is presented and discussed, together with a critical analysis of the most significant IL-based strategies in order to improve the performance of therapeutics and drug delivery systems.publishe
Distinct roles of salt cations and anions upon the salting-out of electro-positive albumin
Precipitation experiments of electro-positive albumin by the action of a wide number of salts, and at differentconcentrations, were performed at a constant temperature (25 °C). The pH range studied covered extreme acidicconditions up to hydronium concentrations where the dissociation of the protein carboxyl groups becomes no-ticeable. The time required for the clouding phenomenon to occur and the quantity of salted-out protein werealso ascertained. The results here reported show that the salt anion is the main salting-out species for the posi-tively charged protein, where their efficacy in salting-out albumin from aqueous solution increases in theorder: F−bCl−bBr−bNO3−bI−bSCN−~ ClO4−bSO42−. Although at extreme pH conditions the salt cationhas no significant influence on the protein salting-out, experiments performed at higher pH values, where thecarboxyl groups starts to dissociate, revealed a non-monotonic effect of the salt upon protein precipitation. Weinterpret this observation as a result of the presence of different protein forms, with which the salt cation partic-ipates in chemical equilibrium. Overall, the proteins salting-out phenomenon induced by salt can be rationalizedby a general mechanism driven by electrostatic interactions and chemical equilibrium concepts.publishe
Progress in the Development of Aqueous Two-Phase Systems Comprising Ionic Liquids for the Downstream Processing of Protein- Based Biopharmaceuticals
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Extraction of vanillin using ionic-liquid-based aqueous two-phase systems
To explore proper extractive solvents and to design an optimized separation process it is highly imperative to understand the molecular-based phenomena governing the solutes partitioning. Moreover, the development of new techniques for the biomolecules separation and purification, while maintaining their functional characteristics unchanged, is still ongoing. Therefore, in this work, the partition coefficients of vanillin, a compound with well-known organoleptic properties, were determined using improved ionic-liquid-based aqueous two-phase systems (ATPS). Three main parameters were evaluated through the vanillin partitioning process: the ionic liquid (IL) cation and anion structure, the temperature of equilibrium and the available concentration of vanillin in the global system. In all systems and conditions tested, vanillin preferentially migrates for the IL-rich phase. In addition, the three studied parameters largely influence the vanillin partitioning. In an attempt to elucidate the thermodynamics of the partitioning process, the standard molar thermodynamic functions of transfer of vanillin were also determined based on the temperature dependence data. These data indicate that the partition of vanillin results from an interplay between enthalpic and entropic contributions where both the IL anion and more complex cations play an essential role. Moreover, viscosities and densities of both aqueous phases were experimentally measured at the mass fraction compositions for which the partition coefficients were determined. The results gathered in this work indicate that IL-based ATPS can be further employed in the extraction and purification of vanillin from different matrices, as confirmed by the large partition coefficients obtained and improved low viscosity systems. (C) 2010 Elsevier B.V. All rights reserved.BIIPP projec - QREN 11551FCT - SFRH/BPD/41781/200
Enhanced conversion of xylan into furfural using acidic deep eutectic solvents with dual solvent and catalyst behavior
An efficient process for the production of furfural from xylan by using acidic deep eutectic solvents (DESs), which act both as solvents and catalysts, is developed. DESs composed of cholinium chloride ([Ch]Cl) and malic acid or glycolic acid at different molar ratios, and the effects of water and γ-valerolactone (GVL) contents, solid/liquid (S/L) ratio, and microwave heating are investigated. The best furfural yields are obtained with the DES [Ch]Cl:malic acid (1:3 molar ratio)+5 wt % water, under microwave heating for 2.5 min at 150 °C, a S/L ratio of 0.050, and GVL at a weight ratio of 2:1. Under these conditions, a remarkable furfural yield (75 %) is obtained. Direct distillation of furfural from the DES/GVL solvent and distillation from 2-methyltetrahydrofuran (2-MeTHF) after a back-extraction step enable 89 % furfural recovery from 2-MeTHF. This strategy allows recycling of the DES/GVL for at least three times with only small losses in furfural yield (>69 %). This is the fastest and highest-yielding process reported for furfural production using bio-based DESs as solvents and catalysts, paving the way for scale-up of the process.publishe
Supported ionic liquids used as chromatographic matrices in bioseparation: an overview
Liquid chromatography plays a central role in biomanufacturing, and, apart from its use as
a preparative purification strategy, either in biopharmaceuticals or in fine chemicals industries, it is
also very useful as an analytical tool for monitoring, assessing, and characterizing diverse samples.
The present review gives an overview of the progress of the chromatographic supports that have been
used in the purification of high-value products (e.g., small molecules, organic compounds, proteins,
and nucleic acids). Despite the diversity of currently available chromatographic matrices, the interest
in innovative biomolecules emphasizes the need for novel, robust, and more efficient supports
and ligands with improved selectivity. Accordingly, ionic liquids (ILs) have been investigated as
novel ligands in chromatographic matrices. Given herein is an extensive review regarding the
different immobilization strategies of ILs in several types of supports, namely in silica, Sepharose,
and polymers. In addition to depicting their synthesis, the main application examples of these
supports are also presented. The multiple interactions promoted by ILs are critically discussed
concerning the improved selectivity towards target molecules. Overall, the versatility of supported
ILs is here considered a critical point to their exploitation as alternatives to the more conventional
liquid chromatographic matrices used in bioseparation processes.publishe
Extraction of high value triterpenic acids from eucalyptus globulus biomass using hydrophobic deep eutectic solvents
Triterpenic acids (TTAs), known for their promising biological properties, can be found in different biomass sources and related by-products, such as Eucalyptus globulus bark, and have been extracted using organic volatile solvents such as dichloromethane. Recently, deep eutectic solvents (DES) have been identified as promising alternatives for the extraction of value-added compounds from biomass. In the present work, several hydrophobic DES were tested for the extraction of TTAs from E. globulus bark. Initial solubility studies revealed that DES based on menthol and thymol as the most promising solvents for these compounds given the highest solubilities obtained for ursolic acid (UA) at temperatures ranging from room temperature up to 90 °C. Accordingly, an eutectic mixture of menthol:thymol (1:2) was confirmed as the best candidate for the TTAs extraction from E. globulus outer bark, leading to extraction yields (weight of TTA per weight of biomass) at room temperature of 1.8 wt% for ursolic acid, 0.84 wt% for oleanolic acid and 0.30 wt% for betulinic acid. These values are significantly higher than those obtained with conventional organic solvents under similar conditions. The results obtained using these DES are promising for the recovery of TTAs for nutraceutical and pharmacological applications, while reinforcing the potential of DES as promising solvents to be applied in biorefinery processes.publishe
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