Biphasic Liquid System Research Articles

Optimisation of parameters for the extraction of β-amylase from sweet potato via liquid biphasic floatation

This study explores the application of liquid biphasic flotation (LBF) for the extraction of β-amylase from sweet potato (Ipomoea batatas), aiming to provide a greener and more sustainable alternative to traditional extraction methods. The LBF method combines solvent sublation with liquid biphasic systems, offering potential advantages in terms of energy-efficient extraction, reduced environmental impact, and improved resource efficiency. Experimental investigations were conducted to assess the effects of salt concentration, solvent amount, and flotation time on β-amylase extraction efficiency and separation performance. Results indicate that optimal conditions for β-amylase extraction were achieved at a salt concentration of 0.3 g/mL, a solvent amount of 60 mL, and a flotation time of 15 minutes with a salt/solvent combination of trisodium citrate and tert-butanol. These optimal conditions resulted in an amylase yield of 18.29%, with a separation efficiency of 0.06% and a purification factor of 291.90. Comparative analysis with liquid biphasic systems (LBS) revealed that the LBF method provided higher efficiency and yield, contributing to more sustainable practices in enzyme extraction. The LBF method shows promise due to its improved performance and potential for reducing the environmental footprint of industrial processes, supporting both innovation and environmental sustainability. This research also studied the effect of various parameters on phase formation and separation efficiency, offering a more thorough understanding of the LBF system that can be applied to future studies with broader sustainability goals.

Innovations in Nanoemulsion Technology: Enhancing Drug Delivery for Oral, Parenteral, and Ophthalmic Applications.

Nanoemulsions (NEs) are submicron-sized heterogeneous biphasic liquid systems stabilized by surfactants. They are physically transparent or translucent, optically isotropic, and kinetically stable, with droplet sizes ranging from 20 to 500 nm. Their unique properties, such as high surface area, small droplet size, enhanced bioavailability, excellent physical stability, and rapid digestibility, make them ideal for encapsulating various active substances. This review focuses on recent advancements, future prospects, and challenges in the field of NEs, particularly in oral, parenteral, and ophthalmic delivery. It also discusses recent clinical trials and patents. Different types of in vitro and in vivo NE characterization techniques are summarized. High-energy and low-energy preparation methods are briefly described with diagrams. Formulation considerations and commonly used excipients for oral, ocular, and ophthalmic drug delivery are presented. The review emphasizes the need for new functional excipients to improve the permeation of large molecular weight unstable proteins, oligonucleotides, and hydrophilic drugs to advance drug delivery rapidly.

Expression of concern: Liquid biphasic electric partitioning system as a novel integration process for betacyanins extraction from red-purple pitaya and antioxidant properties assessment.

Expression of concern: Liquid biphasic electric partitioning system as a novel integration process for betacyanins extraction from red-purple pitaya and antioxidant properties assessment.

Assessment of organic solvents for 2,5-furandicarboxylic acid (FDCA) and distribution in water/cyclohexanone biphasic system

2,5-furandicarboxylic acid (FDCA) is one of the most promising sugar-derived building blocks aimed to produce greener polymers, such as the 100 % recyclable bioplastic polyethylene furanoate (PEF). One incipient field of research is the development of liquid–liquid biphasic systems for FDCA production from 5-hydroxymethylfurfural (HMF) to increase selectivity and minimize undesirable by-product formation. In this work, we first performed an assessment of potential organic green solvents to form biphasic systems considering operability, along with safety, health, and environmental implications. This analysis guided the selection of eight organic solvents in which the solubility of FDCA was measured: cyclohexanone, diethyl ether, isobutyl acetate, methyl isobutyl ketone, methyl ethyl ketone, methoxycyclopentane, tert-butylmethyl ether, and octan-1-ol. The results revealed the superior performance of cyclohexanone as a solvent for the organic phase/water FDCA distribution owing to its higher FDCA solubility (1.364 g/L at 293.15 K). Thus, the FDCA distribution coefficient (KFDCA) between water and cyclohexanone was examined at several temperatures (293.15–313.15 K) and various initial aqueous concentrations to gain deeper insight into the thermodynamics of the phase transfer process and the influence of pH on FDCA distribution between water and cyclohexanone. The resulting enthalpy and entropy of transfer were −15.3 ± 1.0 kJ mol−1 and −44.9 ± 4.6 J K−1 mol−1, thus the highest value of KFDCA (4.83) was obtained at 293.15 K, together with a very high separation factor (81.5) which shows the great potential of cyclohexanone to extract FDCA from aqueous solutions.

Deep dewatering of sludge and resource recovery of hydroxyapatite: A recyclable approach via ionic liquid biphasic system and hydrogen bonds reformation

Deep dewatering of Waste Activated Sludge (WAS) through mechanical processes remains inefficient, primarily due to the formation of a stable hydrogen bonding network between the biopolymers and water, which consequently leads to significant water trapped by Extracellular Polymeric Substances (EPS). In this study, a novel and recyclable treatment for WAS based on Ionic Liquids (ILs) was established, named IL-biphasic aqueous system (IL-ABS) treatment. Specifically, the IL-ABS formed in WAS facilitated rapid and efficient in-situ deep dewatering while concurrently recovering hydroxyapatite. The water content decreased from an initial 98.27 % to 65.35 % with IL-ABS, formed by 1-Butyl-3-methylimidazolium bromide (BmimBr) and K3PO4 synthesized from waste H3PO4. Moreover, the recycled BmimBr maintaining the water content of the dewatered sludge consistently between 65.61 % and 67.25 % across five cycles, exhibited remarkable reproducibility. Through three-dimensional excitation-emission matrix, lactate dehydrogenase analyses and confocal laser scanning microscopy, the high concentration of BmimBr in the upper phase effectively disrupted the cells and EPS, which exposed protein and polysaccharide on the EPS surface. Subsequently, the K3PO4 in the lower phase led to an enhanced salting-out effect in WAS. Furthermore, FT-IR analysis revealed that K3PO4 disrupted the original hydrogen bonds between EPS and water. Then, BmimBr formed numerous hydrogen bonds with the sludge flocs, leading to deep dewatering and agglomeration of the sludge flocs during the unique phase separation process of IL-ABS. Notably, sludge-derived hydroxyapatite product exhibited remarkable adsorption capacity for prevalent heavy metal contaminants such as Pb2+, Cd2+ and Cu2+, with efficiencies comparable to those of commercial hydroxyapatite, thereby achieving the resource utilization of waste H3PO4. Moreover, economic calculations demonstrated the suitability of this novel treatment. This innovative treatment exhibits potential for practical applications in the non-mechanical deep dewatering of WAS.

Optimization of Chlorophyll Extraction from Ulva Sp. with Ultrasound-Assisted Liquid Biphasic Systems Method

Ulva sp. is a type of green algae that is easily found in the shallow seas of Indonesia and contains various bioactive compounds. Chlorophyll is a bioactive compound that functions as a natural dye, free radical scavenger, and antioxidant in the body. Chlorophyll extraction with conventional methods requires a relatively long time and large amount of solvent. In this research, chlorophyll extraction from Ulva sp. with the Ultrasound-Assisted Liquid Biphasic System (UALBS) method. Ulva sp. as much as 2.5 grams mixed in acetone solvent, added K2HPO4 solution, and carried out the sonification process in a dark room. The extraction process performs at a parameter range of 5-15 minutes, ratio 0.05-0.1 g/mL, and 60-100 mesh particle size. The extracted filtrate was added with petroleum ether and distilled water to form a biphasic condition. The extracted chlorophyll was analyzed with chromatography and spectrophotometry method. Antioxidant activity was analyzed using the DPPH method. The optimum result of Ulva sp. chlorophyll extraction at an extraction time of 10 minutes, the ratio of solids to solvent was 0.1, the particle size of 60 mesh produced a yield of 1.88% with chlorophyll a 20.13664%, chlorophyll b 21.58672% and total chlorophyll 41.71012%, and the percentage inhibition 45.32%.

Application of high shear-assisted liquid biphasic system for protein extraction from Chlorella sp

Microalgae is a sustainable alternative source to traditional proteins. Existing pretreatment methods for protein extraction from microalgae still lack scalability, are uneconomical and inefficient. Herein, high shear mixing (HSM) was applied to disrupt the rigid cell walls and was found to assist in protein release from microalgae. This study integrates HSM in liquid biphasic system with seven parameters being investigated on extraction efficiency (EE) and protein yield (Y). The highest EE and Y obtained are 96.83 ± 0.47 % and 40.98 ± 1.27 %, respectively, using 30% w/v K3PO4 salt, 60 % v/v alcohol, volume ratio of 1:1 and 0.5 % w/v biomass loading under shearing rate of 16,000 rpm for 1 min.

Influence of ionic liquid on polar organic compounds solubility in dense CO2 phase

Accurate measurement and prediction of the phase behaviour of mixtures involved in a chemical process are crucial for its optimisation. Given the importance of CO2 conversion technologies and considering possible benefits of CO2-ionic liquid biphasic systems, i.e., facilitating a product separation, we investigated the high-pressure behaviour of components of interest in a recently developed process of cyclic carbonate synthesis directly from CO2 and potentially bio-based alcohols. The solubility of 1,2-butanediol and 1,2-butylene carbonate in a dense carbon dioxide phase was determined experimentally at the temperature of 313.2 K and pressures between 6 and 18 MPa. The influence of 1-hexyl-3-methylimidazolium tris(pentafluoroethyl)trifluorophosphate ionic liquid, [hmim][FAP], as a solvent, on the solubility of these compounds in CO2-rich phase, in ternary (CO2 + 1,2-butanediol + [hmim][FAP]), CO2 + butylene carbonate + [hmim][FAP]) and quaternary (CO2 + 1,2-butanediol + butylene carbonate + [hmim][FAP]) mixtures was investigated. The experimental results of the two binary systems were correlated using the density-based Chrastil equation. The knowledge of phase equilibria behaviours reported in this work will be useful for designing chemical conversions of carbon dioxide using [hmim][FAP] ionic liquid as reaction solvents.

Extraction of phycocyanin from Spirulina using Deep Eutectic Solvent Liquid Biphasic System

BackgroundPhycocyanin (PC) is an intracellular protein produced in Spirulina sp. and is widely used as a natural dye in the cosmetic, food, and supplements industries. But the extraction of phycocyanin was limit by properties of proteins and technologies. Here, deep eutectic solvents were found to be an effective method of extraction, circumventing any usage of harmful organic solvents. MethodsIn particular, we explored the use of DES/K2HPO4 liquid biphasic system (LBS) as the extraction medium, where the extracted PC from the cell in bottom phosphate aqueous phase is selectively transferred to the top DES phase. FindingsAmong the different DES, the ChCl-Urea and ChCl-Glycerol showed the highest extraction efficiencies and yields, but the Glycerol synthesized DES has bad mass diffusion problems. Through continuous single-factor optimization, we finally achieved the best extraction efficiency and yield of 94.2% and 92.0%, respectively, using ChCl-Urea/K2HPO4. This proposed solution has the potential to be produced on a large scale and is environmentally friendly for the extraction of phycocyanin.

Nonlinear liquid-liquid chromatography: Modeling a binary mixture separation

In liquid-liquid chromatography (LLC), mixture components are separated due to their different distribution between the phases of a biphasic liquid system composed of three or four solvents. LLC separations are typically modeled assuming that only the solutes distribute between the two liquid phases and their distribution can be described with a concentration-independent distribution constant. With increasing solute concentration, the physicochemical properties of the biphasic system change, and the distribution of the solutes becomes a function of their concentration. However, the experimental determination of liquid-liquid equilibria in multicomponent systems is time-intensive, and its prediction using thermodynamic models is often not sufficiently accurate for process design purposes. Thus, in this work, we propose a simple approach to model and simulate LLC separations in the nonlinear (concentration-dependent) range of the solutes’ distribution equilibria, namely cannabidiol (CBD) and cannabigerol (CBG). Using the inverse method, the distribution equilibrium equation parameters were estimated from pulse injection experiments of single solutes at concentrations ranging from 1 to 100 mg/mL and 1–50 mg/mL for CBD and CBG, respectively. The obtained parameters were then successfully used to predict the elution profiles of binary mixtures of different compositions at 40 mg/mL total cannabinoid concentration. The approach was demonstrated and validated for CBD and CBG as model compounds and n-hexane/methanol/water 10/7.5/2.5 (v/v/v) as the biphasic solvent system. It should be noted that the applicability of the proposed approach is system-dependent, and hence, it should be evaluated for each separation task individually.

Prediction of potential energy profiles of molecular dynamic simulation by graph convolutional networks

A graph convolutional networks (GCN)-based machine learning (ML) model is constructed to predict physical properties of metallic materials from graph representation of atomic configuration of molecular dynamics (MD) simulation. The developed ML model is employed for the prediction of time variation of the potential energy of a solid–liquid biphasic system of nickel. The learned ML model gives a good prediction on the property of training data. Moreover, it is confirmed that the ML model has generalization performance sufficient to make adequate predictions on unknown graph structures despite the lack of information on interatomic distances in the graph representation. It is significant in this study to show that the graph representation can be a good notation for the prediction of various properties from MD simulations since there is no established notation for atomic configuration of MD simulation especially for large-scale system of metallic materials.

Biphasic systems of mutual amine solution with tunable phase separation for CO2 capture: Optimal compositions and volume of rich phase

Research and development of biphasic systems with significant absorption performance and low energy penalties is the key to achieving CO2 capture. In this paper, the effect of diethylenetriamine (DETA) and diethylethanolamine (DEEA) as amine solution dissolved in N-Methyl-2-pyrrolidone (NMP) and water with various molar ratios on the CO2 absorption was investigated. The formation of saturated solution including CO2-rich and lean phases was studied with scrutiny via 13C nuclear magnetic resonance (13C NMR). Under the optimal conditions, the CO2 loading of fresh absorbents could reach 0.97 and 0.95 mol CO2/mol amine solution with the final solid–liquid and liquid–liquid systems, respectively. Also, overcoming the tradeoff between the volume of rich-phase and CO2 loading as one of the biggest challenges in the biphasic liquid system occurred. The addition specific amount of NMP to the solution (when the molar ratio of NMP to solution is 1) decreased the rich phase volume to 50%, while the CO2 loading is 0.93 mol.mol−1.

Lipid Recovery from Microalgae Biomass Using Sugaring-Out Extraction in Liquid Biphasic Flotation System

The increase in global temperature calls for ambitious action to reduce the release of greenhouse gases into the atmosphere. The transportation sector contributes up to 25% of the total emissions released, mainly from the burning of vehicle fuel. Therefore, scientists from all around the world are focusing on finding a sustainable alternative to conventional vehicle fuel. Biofuel has attracted much attention, as it shows great potential for the replacement of traditional fossil fuels. However, the main bottlenecks of biofuel are the ongoing controversial conflict between food security with biofuel production. Therefore, this study focuses on a sustainable extraction of lipids from microalgae for the production of biofuel using a liquid biphasic flotation system coupled with sugaring-out method. This is the first study to combine the methods of liquid biphasic flotation system with the sugaring-out technique. It represents a holistic study of optimum and effective conditions needed to extract lipids from the system and to understand the reliability of sugar solution as the agent of cell disruption. At the 15-min flotation time, 150 g/L of fructose solution with a 1:2 mass separating agent-acetonitrile ratio successfully extracted up to 74% of lipid from Chlorella sorokiniana CY-1. Two types of fatty acid methyl esters were recovered from the study, with C5:0 being the main component extracted.

Partitioning behavior of rutin in novel liquid–liquid biphasic systems formed by choline chloride/ maltose natural deep eutectic solvents and n-propanol

The development of promising environmentally friendly extraction tools has been one of the hottest topics in green chemistry. Among these, natural deep eutectic solvents (NADESs) as green extraction media have received widespread recognition. In this paper, with the aim of exploring the potential applications of NADESs in the extraction field, water-soluble n-propanol was used to form liquid–liquid biphasic systems with choline chloride/ maltose NADESs. The phase behavior of those systems was designed by adjusting the molar proportion of the hydrogen bond acceptors (HBA): hydrogen bond donors (HBD) and temperature. This allows for the systems tailored to the specific requirements of the target molecule, as shown by their application in the extraction of rutin. Investigation of the distribution behavior of rutin suggested that the partition coefficient and the extraction efficiency of rutin in the bottom phase increase significantly with decreasing temperature and increasing HBA: HBD molar ratio. And with the temperature of 288.15 K, the highest TLL of NADES (ChCl: Mal = 1:2) + NPA + water system achieved higher E% of 94.27 %. The interaction force between NADES and rutin was studied using the Hansen solubility parameter, it turned out that Δδi,j of the 2:1 and 1:2 M ratio of NADES was only 8.91, 5.12, which meaning that there may be strong interactions between NADES and rutin. In addition, the hydrogen bond (ΔδH) of the NADES (HBA: HBD = 1:2) was 25.47, which is greater than other interaction force, revealing that the hydrogen bond is the maximum force between rutin and the NADES (HBA: HBD = 1:2). The energy (ΔGT), enthalpy (ΔHT) and entropy (ΔST) calculated from the partition data of rutin was consistent with the free energy (ΔGC), enthalpy (ΔHC) and entropy (ΔSC) changes of system, which confirmed that the driving force of the phase separation of the system drives the transfer of rutin to the lower phase. In addition, we used eco-friendly water to regenerate NADES. The recovery efficiency of rutin was explored while NADES was reused in three cycles. This result showed there was no change observed in the extraction capability for rutin in those system. Finally, a larger aggregate of size 1315 nm was observed by dynamic light scattering, which furthermore demonstrated that the hydrogen bonding interaction between NADES (HBA: HBD = 1:2) and rutin is the main driving force tool for enhancing rutin partitioning. This method presents a newly system constructed by NADES and NPA for the green extraction, revealing the feasibility of the system for rutin extraction and facilitating industrial applications.

A synergistic ‘push and pull’ ionic liquid biphasic system for enhanced extraction separation of cholic acid and deoxycholic acid

A ‘push and pull’ strategy is applied to enhance the liquid–liquid extraction separation efficiency of cholic acid (CA) and deoxycholic acid (DCA) based on ionic liquid (IL).

A Review on Progressive Trends in Pharmaceutical Nano Emulsions and their Assessment

This review aimed to deliver the progressive trends and need for research activities in the area of liquid bi-phasic systems i.e., Nano emulsion (NE). These categories of drug delivery system (DDS) are progressive modes for providing and increasing the bioavailability of non-aqueous drugs and the drug which have increased the first-pass metabolism. The NE's can be framed by either high or low energy techniques. High-pressure homogenization, micro fluidization, and ultra-sonication are involved in High energy techniques whereas the phase inversion emulsion forming method and the self-NE method are involved in low energy techniques. High energy techniques are having lower usage when compared to low energy techniques due to their high consumption of energy, hence low energy techniques are more operative and do not need any sophisticated devices. Even though high energy techniques are more suitable for food-grade emulsion as they need a reduced amount of surfactant than low energy techniques. Methods for formulation of NE DDS are overlying in nature, exclusively in the process of low energy techniques. This review gives the eminence of NE'S by comparing previous research carried over it.

Wormlike micelles of CTAB with phenols and with the corresponding phenolate derivatives - When hydrophobicity and charge drive the coacervation

HypothesisHydrophobicity and the presence or absence of charge in phenol derivatives are relevant on the rheology and phase behavior when they are assembled with a cationic surfactant, forming wormlike micelles. The incorporation of phenols with a greater number of rings into the micellar palisade is entropically favored, but a solubilization limit or coacervation are two paths followed by the solutions, depending on the electrical nature of the aromatic co-solutes. ExperimentsThe investigations were carried out with systems formed by a fixed concentration of hexadecyltrimethylammonium bromide (CTAB) and increasing concentrations of neutral phenols (1-naphthol, 2-naphthol, 2,3-dihydroxynaphthalene and R and S-binol) and with their corresponding phenolate derivatives. The monophasic limits of the systems were established, as well as their linear and non-linear rheology. The structural investigation of the coacervates formed with the phenolates were done using SAXS and Cryo-TEM. FindingsThe zero-shear viscosity of the solutions reaches maxima values close to the solubility limit of the aromatics, which depends on the numbers of rings and hydroxyl groups (position and number). However, when the correspondent ionized phenols were investigated, beyond the maxima values for the zero-shear viscosity, liquid–liquid biphasic systems are formed, in which the upper phase contains a coacervate, associated with branched wormlike micelles. However, when the ratio between phenolate and CTAB is around 3:1 the coacervate evolves to a lamellar structure.

Liquid Biphasic System

Many researchers have been interested in a substitute bioseparation technology for different biomolecules, namely the liquid biphasic system (L.B.S.). The analysis starts with a detailed discussion of the basic concept of L.B.S. and follows by a debate on the further creation of the different components for phase-forming of L.B.S. Also, various advance L.B.S. technologies will be introduced. It was addressed that is beneficial to L.B.S.'s e-function for bimolecular extraction, separation, and purification. The L.B.S. key parameters for link were obtainable and assessed. Furthermore, a guideline for future growth was described as future opportunities and challenges of L.B.S. The measures outlined in this review gives an insight into potential studies of the techniques of liquid-liquid split.

Microfluidics approach for determination of the equilibrium phase composition in multicomponent biphasic liquid systems

Based on the fundamentals of microfluidics, a novel approach for the determination of liquid-liquid equilibria (LLE) of ternary systems was proposed. The system studied here consists of the compounds water, acetone, and toluene. This method was realized in a microfluidic set-up that consists of a microchannel, a camera for the determination of the position of the phase interface, and a micro density meter for measuring the densities of the conjugated phases at the outlet of the microchip. To determine the equilibrium phase compositions, an optimization problem was defined that minimized the difference between the experimentally determined and calculated ratio of the volumetric flow rates of the conjugated phases. The developed procedure uses information on the position of the phase interface in the microchannel and the phase densities, and requires previous knowledge of the binodal curve, which as shown in our previous work (Hübner and Minceva, 2019) can be also determined using the same set-up. The obtained equilibrium phase compositions were in good agreement with literature data. Eventually, once the procedure is automatized to reduce the required measuring time and achieve full user independency, the proposed approach poses a cheap and fast alternative to conventional methods for measuring LLE.

Asymmetric reduction of 4-trimethylsilyl-3-butyn-2-one to (R)-4-trimethylsilyl-3-butyn-2-ol catalyzed by a novel strain lyophilized Acetobacter sp. CCTCC M209061 in an aqueous/ionic liquid biphasic system

Asymmetric reduction of 4-trimethylsilyl-3-butyn-2-one to (R)-4-trimethylsilyl-3-butyn-2-ol catalyzed by a novel strain lyophilized Acetobacter sp. CCTCC M209061 in an aqueous/ionic liquid biphasic system