Aqueous Two-phase Extraction Process Research Articles

Ultrasonic-assisted aqueous two-phase extraction of polysaccharides from the fruits of Ilex cornuta (Lindl. Et Paxt.): Process optimization, structural characterization and antioxidant activity study

The aqueous two-phase extraction process (APTE) was utilized for the extraction of polysaccharides from the fruits of Ilex cornuta (Lindl. et Paxt.) (PFIC). After screening and selection of isopropanol/ammonium sulfate as the aqueous two-phase system, various parameters of crude extract amount, ultrasonic time, extraction temperature and extraction time on the extraction process was investigated in detail and optimized by response surface methodology (RSM). Under the optimal conditions in which crude extract amount was 2.23 g, ultrasonic time was 17.5 min, extraction temperature was 69 °C and extraction time was 1.55 h, the yield of polysaccharides reached 50.87% which was close to the predicted value of 50.78%. Structural features of PFIC were investigated by GPC, GC–MS and FT-IR analysis. And the results indicated that PFIC with a molecular weight of 3.89 kDa was composed of monosaccharides including rhamnose (1.83%), arabinose (11.70%), xylose (3.30%), mannose (3.55%), glucose (61.88%) and galactose (17.74%). The bioactivity experiments showed that PFIC had strong scavenging ability on DPPH radical, fine scavenging ability on hydroxyl radical and certain scavenging ability on superoxide anion radical. Overall, these results suggested that PFIC exhibiting fine to good antioxidant capacities could be potentially developed as a natural antioxidant.

Amoxicillin removal from medical wastewater using an eco-friendly aqueous two-phase extraction system

ABSTRACT Antibiotics are a common medical waste that can cause antibiotic resistance in humans and harm aquatic life. One of the most effective approaches for treating medical wastewater is the aqueous two-phase extraction process (ATPS). ATPS is considered an ecologically benign technology. Nine ATPS chemical systems were developed to extract Amoxicillin from simulated hospital wastewater using various salts and organic solvents. The criteria that determine the most efficient system were explored in order to maximize the separation process. After assessing the performance of the considered systems, amoxicillin extraction was carried out utilizing an ATPS comprised of PEG6000 as an extractant and K2HPO4 as a separating agent. Complete amoxicillin removal was attained at room temperature and a mixing speed of 500 rpm for 540 mins. The extraction efficiency was found to increase with temperature, salt concentration, and the volume ratio of the solvent (PEG6000). Based on the experimental design methodology, the optimum amoxicillin removal efficiency of 96.4% was attained at a temperature of 44.3°C, 42.6 wt.% of salt, and a 0.5 volume fraction ratio of PEG6000. The stagewise application of the process was also performed within five separation stages. This approach outperforms traditional extraction methods in terms of efficacy and duration.

Study on Aqueous Two-phase Extraction Process of Volatile Oil from Cinnamon and Its Flavor Composition

Study on Aqueous Two-phase Extraction Process of Volatile Oil from Cinnamon and Its Flavor Composition

On-chip aqueous two-phase system (ATPS) formation, consequential self-mixing, and their influence on drop-to-drop aqueous two-phase extraction kinetics

Aqueous two-phase systems (ATPSs) allow an advantageous aqueous two-phase extraction process (ATPE), a special type of liquid–liquid extraction. Compared with conventional liquid–liquid extraction using aqueous/organic extraction media, ATPE is known to provide relatively easy mass transfer and a gentle environment for biological separation applications. Considering the recent interest in microscale ATPE, we aimed to study (i) the potential of preparing ATPS droplets on a digital microfluidic device, and (ii) the influence of the fluidic dynamics created during the formation of ATPS, with the goal of enhancing on-chip ATPE process. On-chip ATPS formation was evaluated by preparing a series of ATPSs on electrowetting on dielectric digital microfluidic chips and comparing their characteristics with the same ATPSs prepared at macroscale using conventional procedures. An enhanced on-chip drop-to-drop ATPE process was achieved by incorporating a self-mixing condition created during ATPSformation. Results indicate a successful on-chip ATPS preparation as well as enhanced extraction performance by self-mixing in the absence of forced mixing. Findings of this research suggest an alternative, simple, yet adequate technique to provide mixing for on-chip applications, such as sample preparation in portable microfluidics, for which it is unfavorable to implement complicated mixing sequences or complex device geometries.

Experiment and simulation of an aqueous two-phase extraction process for the purification of a monoclonal antibody

This work presents the continuous, multistage aqueous two-phase extraction of an Immunoglobulin G1 from a Chinese Hamster Ovary cell supernatant. An aqueous two-phase extraction process combining a sodium chloride-rich extraction step, followed by a sodium chloride-poor back extraction and washing step is investigated experimentally. A polyethylene glycol 2000–phosphate salt aqueous two-phase system was applied for extraction. Different process configurations determine the influences of flow rates, stage number and cell supernatant loading. Immunoglobulin G1 yields of up to 88% and purities of up to 95% are obtained and indicate the potential of continuous, multistage aqueous two-phase extraction. To reduce the raw material consumption recycle batch experiments are conducted to analyze the influence of phase recycling on the extraction performance. Phase recycling is also carried out at multistage scale. An equilibrium stage model is developed and successfully validated within this work. The model is able to predict the continuous, multistage aqueous two-phase extraction on the basis of single stage equilibrium data representing a useful tool to decrease experimental process development effort.

Aqueous two-phase extraction and spectrofluorimetric determination of nitrite with o-phenylenediamine enhanced by hydroxypropyl-β-cyclodextrin in urine samples

An aqueous two-phase extraction process composed of tetrahydrofuran/ammonium sulfate to extract nitrite from urine samples is investigated. The method is based on the reaction of nitrite with o-phenylenediamine in acid media to form benzotriazole and the aqueous two-phase extraction of benzotriazole in the presence of salt as an inducing agent in phase separation, followed by spectrofluorimetric detection. The system of fluorescence intensity was enhanced by complexation of hydroxypropyl-β-cyclodextrin. The optimal reaction and extraction conditions were studied. The linear calibration range for nitrite was 5–1000 ng mL−1 with a detection limit of 1.5 ng mL−1 (S/N = 3). The relative standard deviations for six replicate measurements varied between 0.3% and 1.7%. The interference effects of some anions and cations were also tested. The proposed method was successfully applied for the determination of nitrite in the urine samples with recoveries of 93.3% to 105.0%.

Aqueous two-phase extraction for bovine serum albumin (BSA) with co-laminar flow in a simple coaxial capillary microfluidic device

Microfluidic extraction based on a co-laminar flow of aqueous two-phase system is used to separate bovine serum albumin (BSA). Mass transfer between the continuous two-phase flows is demonstrated by the extraction of BSA in a microfluidic device. The protein concentrations of the BSA samples were determined using the Bradford method. Polyethylene glycol 4000 and ammonium sulfate ((NH4)2SO4) served as model aqueous two-phase solutions. The appropriate flow rates of the aqueous two phases were thus determined. We can flexibly control the mass transfer area and time by simply adjusting the flow rate. It takes only 3.6 s for three extraction cycles in a coaxial microfluidic device to achieve a BSA recovery yield of 71.1 %, which is superior to the traditional beaker aqueous two-phase extraction process. In this study, co-laminar flow-based continuous microextraction is demonstrated and its mass transfer is analyzed by solving the diffusion model, based on a large specific interfacial area and surface renewal.

Application of Water-Miscible Alcohol-Based Aqueous Two-Phase Systems for Extraction of Dyes

The water-miscible alcohols (ethanol, 1-propanpol, 2-propanol) were used to form aqueous two-phase systems with K2HPO4 and Na2HPO4. Meanwhile, the organic citrates were also investigated as the phase-separation salts. The effects of the types and concentrations of phase-forming substances as well as temperature on the formation of aqueous two-phase systems were discussed. The alcohol (ethanol, 1-propanpol, 2-propanol)–salt (K2HPO4, K3C6H5O7, (NH4)3C6H5O7) aqueous two-phase systems were used to extract Rhodamine B and methyl orange in aqueous solution. It proved that the water-miscible alcohol–salt (inorganic salt, organic salt) aqueous two-phase systems can be both used to extract hydrophilic targets directly without adding any reaction reagents. The aqueous two-phase extraction process was also discussed.

Preparation of plasmid DNA polyplexes from alkaline lysates by a two-step aqueous two-phase extraction process

This work studied the possibility of using polyethyleimine (PEI) as an affinity ligand for the purification of plasmid DNA (pDNA) from alkaline lysates using aqueous two-phase systems (ATPSs). The goal was to find conditions under which this cationic polymer could steer the partition of pDNA to the phase where less impurities accumulate. In poly(ethylene glycol) (PEG)/ammonium sulphate systems, neither free nor PEGylated PEI (pPEI) were able to change the partition of pDNA. This is probably due to the high salt concentration present in these systems that impair the interaction between pDNA and PEI. In PEG 3350/dextran 110 systems, the desired effect could be observed but 0.2–0.5 M ammonium sulphate had to be added to prevent the co-partition of RNA to the same phase. These results were used to develop a methodology to obtain polyplexes from alkaline lysates in a two-step ATPSs extraction process. In the first step, a PEG 600/ammonium sulphate system is used to remove most impurities to the top phase. The pDNA-containing bottom phase is then isolated and contacted with a second PEG 3350/dextran 110 system supplemented with a small amount of pPEI (0.2%). Plasmid yield was 100% and the final preparation had no RNA and only small amounts of contaminant protein. Additionally, pDNA was obtained in the form of 53 nm-sized polyplexes which are likely to suit specific gene delivery applications.

Recent Advances in Study on Thermodynamic Models for Real Systems Including Electrolytes

A comprehensive review of recent advances in study on thermodynamic models for real electrolyte solutions is presented. The differences between primitive and non-primitive electrolyte models are demonstrated. Some new thermodynamic models for electrolyte solutions based on the mean spherical approximation and perturbation theory are introduced. An extended scaled-particle theory and modified Clegg-Pitzer equation are presented for physical and chemical absorption processes with mixed solvents, respectively. A pseudo one-component two-Yukawa equation of state is used for the aqueous two-phase extraction process in charged colloidal systems.

Aqueous two-phase extraction as an effective tool for isolation of geniposide from gardenia fruit.

Natural products are normally obtained by organic solvent extraction and many subsequent chromatographic separations. Compounds of interest are often isolated with very low yield and limited purity. An aqueous two-phase extraction process combined with a simple ethanol treatment, for removing excess inorganic salt, has been developed for preparation of geniposide from gardenia. The system was comprised of PE62, a random copolymer composed of 20% ethylene oxide and 80% propylene oxide, KH2PO4 and ethanol. To find optimal conditions, the partition behavior of geniposide under an aqueous two-phase system was investigated. Various factors were considered, including the concentration of salt, the concentration of polymer, the sample loading, and the addition of ethanol. The experimental results demonstrated that increasing salt concentration or decreasing PE62 concentration results in enhancement of the geniposide partition in the salt-rich phase. The addition of ethanol and higher sample loading also promoted the partition efficiency of geniposide. Based on this study, an optimized system containing 5% PE62, 7.5% KH2PO4, and 10% ethanol was tested on a large-scale extraction. A 39.0-g aliquot of final product (in powder form) with 77% purity of geniposide can be effectively extracted from 500 g of gardenia fruit. This process is proved to be useful for industrial application of geniposide preparation.

Recovery of endo-polygalacturonase using polyethylene glycol-salt aqueous two-phase extraction with polymer recycling.

The partitioning behaviour of endo-polygalacturonase (endo-PG) and total protein from a clarified Kluyveromyces marxianus fermentation broth in polyethylene glycol (PEG)-ammonium sulfate and PEG-potassium phosphate (pH=7) aqueous two-phase systems was experimentally investigated. Both the enzyme and total protein partitioned in the bottom phase for these two kinds of systems. The enzyme partitioning coefficient can be lower than 0.01 in PEG8000-(NH4)2SO4 ATPS with a large phase volume ratio and a moderate tie-line length, which implies the possibility of concentration operation using aqueous two phase partitioning. An ion-exchange separation of high purification efficiency was applied to analyze the clarified and dialyzed fermentation broth. A total purification factor of only 2.3 was obtained, which indicated the high enzyme protein content in the total protein of the fermentation broth. Consequently, the main purpose for separating endo-PG is concentration rather than purification. A separation scheme using an aqueous two-phase extraction process with polymer recycling and a dialysis was proposed to recover endo-PG from the fermentation supernatant of K. marxianus for commercial purpose. A high enzyme recovery up to 95% and a concentration factor of 5 to 8 with a purification factor of about 1.25 were obtained using the single aqueous two-phase extraction process. More than 95% polymer recycled will not affect the enzyme recovery and purification factor. Dialysis was used mainly to remove salts in the bottom phase. The dialysis step has no enzyme loss and can further remove small bulk proteins. The total purification factor for the scheme is about 1.7.