Continuous Countercurrent Extractive Biocatalysis in Aqueous Surfactant Two‐Phase Systems

To evaluate the possibility of performing extractive biocatalysis in continuous mode, the hydrolysis of penicillin G in a micellar solution containing the nonionic surfactant Tergitol NP‐7 was chosen as model system. While the product phenylacetic acid distributes into the micellar phase, 6‐aminopenicillanic acid moves preferably into the aqueous phase. The yield in the continuous multi‐step process was higher in comparison to equivalent batch systems. Overall, the results demonstrate the suitability of aqueous surfactant two‐phase systems for continuous biocatalytic reactive extraction processes.     

Multi‐Stage Aqueous Two‐Phase Extraction for the Purification of Monoclonal Antibodies

Using monoclonal antibodies in a cell culture harvest as an example for complex biomolecules, a mini‐plant‐scale aqueous two‐phase purification process was studied. The results of this study indicate that antibodies can be concentrated and purified in a single extraction step employing a small phase ratio. Following the extraction step, a multi‐stage wash‐extraction for further purification was investigated. Starting at a test tube‐scale cross‐current wash, the process was advanced towards a continuous counter‐current mixer‐settler and column wash process. It was shown that a test tube cross‐current extraction operation can predict the multi‐stage purity reasonably well. The hydrodynamic process performance for the multi‐stage wash column was evaluated and related to the separation performance.     

Integration of Aqueous Two‐Phase Extraction into Downstream Processing

Aqueous two‐phase extraction (ATPE) is increasingly considered to be a feasible unit operation, e.g., for the capture of monoclonal antibodies or recombinant proteins. So far, knowledge on the applicability of ATPE in antibody processes has been collected mostly in lab‐scale. In contrast, approaches for the integration of ATPE into a downstream process are investigated. A complete process sequence including extraction, washing, ultrafiltration, and ion‐exchange chromatography is discussed and suggested for antibody purification. Excellent antibody purities can be achieved. Additionally, a model is applied that allows early‐on prediction of a multistage ATPE with high prediction accuracy. Finally, an economic evaluation between ATPE and Protein A chromatography is performed, reaching up to five‐fold cost‐saving factors.     

Application of Aqueous Two‐Phase Systems for the Potential Extractive Fermentation of Cyanobacterial Products

The potential use of aqueous two‐phase systems (ATPS) to establish a viable protocol for the in situ recovery of cyanobacterial products was evaluated. The evaluation of system parameters such as poly (ethylene glycol) (PEG) molecular mass, concentration of PEG and salt was carried out to determine the conditions under which Synechocystis sp. PCC 6803 cell and cyanobacterial products, i.e., β‐carotene and lutein, become concentrated in opposite phases. PEG‐phosphate ATPS proved to be unsuitable for the recovery of cyanobacterial products due to the negative effect of the salt upon the cell growth. The use of ATPS PEG‐dextran (6.6 % w/w PEG 3350, 8.4 % w/w dextran 66900, TLL 17.3 % w/w, V_R 1.0, pH 7) and (4.22 % w/w PEG 8000, 9.77 % w/w dextran 66900, TLL 18 % w/w, V_R 1.0, pH 7) resulted in the growth of cyanobacteria (Synechocystis sp. PCC 6803) and the concentration of lutein in opposite phases. However, β‐carotene was seen to concentrate in the top phase together with the biomass. The results reported here demonstrate the potential application of ATPS to establish the conditions for an extractive fermentation prototype process for the recovery of cyanobacterial products.     

A Hydration Shell‐Based Thermodynamic Model for Aqueous Two‐Phase Systems

In this work a Flory‐Huggins model modified to account for some unique features of Aqueous Two‐Phase Systems is presented. The model takes into account observed solvation between water and polymer molecules through the incorporation of a hydration shell to express the number of water molecules bonded to each polymer molecule. The parameters of the modified equation were determined using experimental data of ATPS containing poly (ethylene glycol) and dextran. The results revealed remarkable improvement in the correlation ability of the model. A general expression that defines the number of water molecules in the hydration shell was also obtained.     

Cloud Point Extraction of Glycyrrhizic Acid from Licorice Root

Abstract

An attempt has been made to extract glycyrrhizic acid (GA) from licorice root by surfactant mediated cloud point extraction (CPE) using non‐ionic surfactant (Triton X‐100). Almost all of the GA molecules were concentrated in the surfactant‐rich phase (also called coacervate phase) after phase separation. The pH is the most critical factor regulating the distribution of GA in the micelle which related to the ionization form. The other effects of the concentration of GA and the surfactant, the temperature, and the salt concentration on the extraction efficiency of GA in the coacervate phase and aqueous phase have been studied. The mechanism of CPE of GA was explored with transmitting electron microscopy. It was deduced that aggregate GA molecules were adsorbed on micelles' outer poler mantle and inner cross‐linked micelles at high GA concentrations in coacervate phase.     

Cloud Point Extraction (CPE) of Parabens using Nonionic Surfactant Phase Separation

A cloud point extraction (CPE) method was developed using silicone-ethylene oxide surfactant, DC193C, to study the equilibrium phase separation of parabens in environmental water samples. The volume of the surfactant rich phase (V_s), aqueous phase (V_w), preconcentration factor (C_F), distribution coefficient (K_d), and percentage of recovery (%R) were evaluated to obtain the optimum condition for phase separation of parabens. Satisfactory results were obtained whereby the preconcentration factor for methylparaben, ethylparaben, propylparaben, and benzylparaben were close to 65, 60, 80, and 45, respectively, at 0.5% weight of surfactant. The presence of DC193C gave a low phase volume ratio but insignificant result was observed with the increased of surfactant concentration. The parabens were well separated on a reverse-phase Chromolith C₁₈ column (100 mm × 4.6 mm) with a gradient elution (acetonitrile: water; 30:70 v/v) and detected at 254 nm. The proposed method was successfully applied to environmental samples such as river water, treated wastewater, sea water, and tap water with satisfactory results. The method detection limit was 0.1–0.2 ppb and the recoveries were 71.2–97.7% with the relative standard deviations of 0.3–2.1%.     

Development of Two‐point Dynamic Method for Evaluating Extraction Columns

The steady‐state method by measuring the concentration profile along the column height is an effective way, but it is a time and material consumption method for large extraction columns. In order to investigate the axial‐mixing and mass transfer performances in a large pulsed‐sieve‐plate extraction column with the diameter of 150mm, a two‐point dynamic method with mass transfer based on the diffusion model has been developed. The results proved that the two‐point dynamic method has the advantages of good accuracy, simple boundary equations and flexible sampling position over the traditional single‐point dynamic method. It is a reliable tool for studying the axial‐mixing and the mass transfer performances.     

Cloud Point Extraction of Four Triphenylmethane Dyes by Triton X-114 as Nonionic Surfactant

A method for removing four triphenylmethane dyes from wastewater by cloud point extraction with the nonionic surfactant Triton X-114 (TX-114) was developed. The triphenylmethane dyes were crystal violet, ethyl violet, malachite green and brilliant green. The cloud point of TX-114 generally increased in the presence of any of the four dyes. In the cloud point system, these dyes were solubilized into a coacervate phase that left a color-free dilute phase. The extraction efficiency of the dyes increased with the temperature, TX-114 concentration, and salt (NaCl and CaCl₂) concentration. More than 97% TX-114 in the dilute phase was recovered by adjusting the volume ratio of dichloromethane to the dilute phase. The Langmuir-type adsorption isotherm was used to describe the dye solubilization. The Langmuir constants m and n were calculated as functions of temperature. The results showed that the solubilization of the triphenylmethane dyes in the cloud point system was related to the partition coefficient and their molecular structures.     

相分离辅助方法对有机硅表面活性剂浊点萃取PAHs的影响

相分离性能和萃取率是评价浊点萃取(CPE)性能的两个重要因素.对于采用非离子表面活性剂的CPE,当溶液加热至浊点以上,仅依靠热运动来促使其发生相分离是十分缓慢的,因此通常需要一定的辅助方法来加速其相分离的进程.今利用两种聚醚型有机硅表面活性剂DC-193和DC-190,对蒽、菲、芘和苊四种多环芳烃(PAHs)进行浊点萃取研究,分别使用加热、加盐、离心和搅拌等辅助方法改变相分离过程的速度.通过测定表面活性剂富集相的体积百分率Vs/(Vs Vw)随时间的变化和最终的萃取率,研究上述四种不同辅助方法对相分离行为和萃取性能的影响.结果表明,相分离速度:加盐＜加热＜离心＜搅拌;Vs/(Vs Vw):搅拌＜离心＜加盐＜加热;萃取率:加热＜加盐≈离心＜搅拌.可见,离心与搅拌辅助CPE具有较大的优势,可同时提供更高的相分离性能和萃取率.与离心相比,搅拌的引入所需设备更简单,而且对于容器的体积没有限制,易于实现连续操作,更有望应用于实际的大规模水处理之中.     

Multistage Processing of Tunable Aqueous Polymer Phase Impregnated Resins (TAPPIR®)

A promising concept of aqueous two‐phase extraction (ATPE) is the Tunable Aqueous Polymer Phase Impregnated Resins (TAPPIR®) technology. Thereby, one phase of an aqueous two‐phase system (ATPS) is immobilized inside porous solids, which are then dispersed in the second aqueous phase. Beside extraction performance, the reusability of the impregnated solids for several cycles decides upon the competitiveness of TAPPIR®. Multistage extraction experiments using a polyethylene glycol 4000/citrate ATPS to partition lysozyme were investigated for TAPPIR® and ATPE. A stable impregnation was demonstrated for multiple cycles. Moreover, it was found that multistage extraction processes should not be designed on basis of the first extraction as the yield changes in the following stages.     

多元渗透维里方程通式的表达：双水相系统的液液平衡计算

提出了基于三元系修正Ｐｉｔｚｅｒ模型的多元渗透维里方程通式，并应用于双水相系统液液平衡性质的关联和预测，用汽液平衡数据关联模型参数，预测了聚乙二醇／硫酸镁双水相系统的液液平衡；用汽液平衡数据和尽可能少的液液平衡数据关联模型参数。同时预测了焦点或不含电解质的ＰＥＧ／葡萄糖双水相系统的液液平衡和ＫＣｌ电解质在ＰＥＧ／ＤＥＸ系统中的分配系数。计算结果表明，模型有较好的预测功能。     

Tuning of Cloud Point by the Nature of Surfactant Headgroup: Influence of Counterion and Additives

Work was performed to distinguish the role of sulfonate (–SO₃ ^?) and sulfate (–OSO₃ ^?) with respect to the micellization and clouding phenomenon in ionic surfactant solutions. The clouding phenomenon is a recent addition to the conventional one observed with nonionic surfactants. Three ionic surfactants [sodium dodecylsulfate (SDS), sodium dodecylbenzenesulfonate (SDBS), and sodium dodecylsulfonate (SDSo)] are chosen and the effects of added tetra-n-pentylammonium bromide (TPeAB) and benzyl tributylammonium bromide (BTAC) have been studied on micellization and clouding behaviors in aqueous solution. Based on critical micelle concentration (CMC) and cloud point (CP) measurements, the following order has been observed: SDBS < SDS < SDSo. Though both SDBS and SDSo contain sulfonate groups, they are found at the two ends of the ordering. Therefore, the role of the phenyl ring is also having importance in clouding phenomena. For a typical surfactant, TPeAB was found to be more effective than BTAC. Based on the CP studies, two compositions of SDSo + TPeAB/BTAC were chosen and the effects of different additives (carbohydrate, amino acid, and l-ascorbic acid) on the CP were investigated. Additive may either decrease or increase CP, depending on the structure of the counterion or additive. The present work shows a few novelties: (1) headgroup/counterion dependence of CP and (2) hydrophobicity of counterion/surfactant has an important bearing on the phenomenon. The data can be utilised in improving cloud point extraction methodologies (CPEMs).     

Aqueous two‐phase systems for the recovery of a recombinant viral coat protein from Escherichia coli

In this study the use of an aqueous two‐phase system (ATPS) following the direct chemical extraction of a recombinant viral coat protein, from the cytoplasm of Escherichia coli, is evaluated. The driving force is the need to establish an economically‐viable process for the manufacture of a vaccine against human papilloma infection. The partition behaviour of recombinant L1 protein, the major structural protein of the virus, and DNA was investigated in a polyethylene glycol (PEG)–phosphate system. An evaluation of system parameters including PEG molecular mass and the concentrations of PEG and phosphate was conducted, to estimate conditions under which the L1 protein and DNA partition to opposite phases. ATPS extraction comprising a volume ratio of 1.00, PEG 1000 (18.0%(w/w)) and phosphate (15.0%(w/w)) provided the conditions for accumulation of DNA into the bottom phase and concentration of L1 protein into the opposite phase (ie partition coefficient of DNA; ln K_DNA < 0.0 and partition coefficient of L1; ln K_L1 > 2.5). The findings reported here demonstrate the potential of ATPS to recover recombinant protein released from E coli by direct chemical extraction. © 2002 Society of Chemical Industry     

Cloud point and solvent extraction study of uranium(VI) by 8‐hydroxyquinoline

BACKGROUND: Cloud point extraction (CPE) is an attractive alternative to solvent extraction. However, comparisons between both techniques are lacking. In this paper, the extraction of uranium(VI) using 8‐hydroxyquinoline (HQ) as chelating agent was studied by CPE using Triton X‐114 as non‐ionic surfactant and by solvent extraction using CHCl₃ as diluent. RESULTS: Using CPE, a quantitative extraction was observed for pH higher than 4.5 with a HQ/U ratio of 10. Using solvent extraction an increase in the HQ/U ratio up to 50 is necessary to obtain a near quantitative extraction. Both extraction systems were then compared with respect to the nature of extracted species, and the extraction constants determined using log‐log analysis of the extraction data. In the solvent extraction system, the extracted species were identified as UO₂Q₂ and the corresponding extraction constant was found to be log k_ex = ? 3.6 ± 0.2 on the molar scale. Considering that UO₂Q₂ is also the extracted species in CPE, a slightly higher extraction constant, i.e. log k_ex = ? 2.5 ± 0.3, was found. CONCLUSION: Such a small difference in favour of the CPE system may arise from the combination of various phenomena, including effects of temperature and effects of ‘extractant environment’. However, a change in the nature of the extracted species, namely from UO₂Q₂ in the solvent extraction system to the formation of adducts, i.e. UO₂Q₂(HQ) and UO₂Q₂(HQ)₂ in the CPE system, due to higher HQ concentration in the surfactant‐rich phase compared with chloroform, cannot be precluded, but requires confirmation. Copyright © 2012 Society of Chemical Industry     

Partitioning equilibria in multicomponent surfactant systems for design of surfactant-based extraction processes

Aqueous biphasic systems based on nonionic surfactants have perspective applications in extraction processes, in particular, cloud point extraction of hazardous compounds or high valued products, especially biomolecules. Additives (e.g., ionic surfactants, salts) and variations in pH can significantly affect the surfactant-based separation processes, representing an additional degree of freedom for their optimization. However, there are few systematic studies of phase and partition behavior for these multicomponent surfactant systems.In this study we examined the clouding, phase compositions and partitioning equilibria for aqueous mixed surfactant systems of a nonionic surfactant (Triton X-114), ionic surfactants (cetyltrimethylammonium bromide or sodium dodecyl sulfate) and NaCl, in order to improve the extraction efficiency. Vanillin was used as a model substance at three different pH values, specifically in (partly) dissociated or non-dissociated states. The partition coefficients obtained in the batch experiments were compared to the predictions by the thermodynamic model COSMO-RS. Based on this knowledge a continuous multistep extraction process was carried out.To the best of our knowledge this is the first demonstration of using a mixed surfactant system for continuous countercurrent cloud point extraction.     

Improved Void Fraction Correlation for Two‐Phase Flow in Large‐Diameter Annuli

A flow pattern‐independent void fraction correlation for gas‐liquid two‐phase flow in vertical large‐diameter annuli is established. Two equations are proposed for the parameters of a drift flux model‐based correlation: the distribution parameter and the drift flux velocity. These equations are expressed as a function of two‐phase flow variables including void fraction, fluid properties, pipe geometry, and phase flow rates. Experiments were performed to study the void fraction of vertical air‐water two‐phase flow in large‐diameter annuli. The obtained experimental data along with the literature data of Caetano are used to verify the performance of the proposed void fraction correlation. The accuracy of this correlation is compared with nineteen frequently used correlations in literature. The proposed correlation was found to predict the void fraction consistently with a better accuracy.     

Void Fraction Measurement for Two‐Phase Flow Using Electrical Resistance Tomography

Measurement of void fraction of two‐phase flows remains a challenging area. In this paper the application of an electrical resistance tomography (ERT) system for this purpose has been studied. A new approach through the direct use of the voltage data measured by the ERT system is presented. The measured voltage data are first compressed through a feature extraction, and a polynomial regression procedure is followed to obtain the relationship between the void fraction and the feature extracted. Both simulation and experiment are carried out to verify the approach. The methodology of the new approach, simulation and experimental results are presented in the paper.     

Improvement of Impedance Imaging for Two‐Phase Systems with Boundary Estimation Approach in Electrical Impedance Tomography

Numerical and experimental works are conducted to develop a visualization technique for the phase distribution in a two‐phase system by electrical impedance tomography technique. For the two‐phase flow system, the impedance of each phase can be known but instead the phase boundary depending on the distribution of dispersed phase is of interest. Here, the image reconstruction problem is derived as a boundary estimation one and a novel algorithm is developed for the estimation of phase boundary. Based on the synthetic and real experimental data, the images of the simulated bubble distributions are reconstructed successfully.     

Cloud Point Extraction and Preconcentration for the Determination of Cu and Ni in Natural Water by Flame Atomic Absorption Spectrometry

Abstract

In this work, a new cloud point extraction (CPE) method was developed for the separation and preconcentration of copper and nickel. The analyte was complexed with 3‐[(8‐{[(E)‐2‐hydroxyimino‐1‐methylpropylidene] amino}‐1‐naphthyl) imino]‐2‐butanone oxime (H₂mdo) in the initial aqueous solution and octylphenoxy polyethoxy ethanol (Triton X‐114) was added as a surfactant. After phase separation, based on the cloud point in the mixture, and dilution of the surfactant‐rich phase with methanol containing 0.1 mol l^?1 HNO₃, the enriched analytes were determined by flame atomic absorption spectrometry. After optimization of the complexation and extraction conditions (i.e. pH=8.5, H₂mdo=3×10^?4 mol l^?1, Triton X‐114=0.1% (w/v)) the enhancement factors of 65 and 59 and detection limits of 0.14 and 0.2 ng ml^?1 were obtained for copper and nickel respectively. The proposed method was applied to the determination of copper and nickel in several natural water samples with satisfactory results.