Flow Field-flow Fractionation Technique Research Articles

Delicate Analysis of Interacting Proteins and Their Assemblies by Flow Field-Flow Fractionation Techniques.

We study the efficiency of several asymmetrical flow field-flow fractionation (AF4) techniques to investigate self-associating wheat gluten proteins. We compare the use of a denaturing buffer including sodium dodecyl sulfate (SDS) and a mild chaotropic solvent, water/ethanol, as the eluent, on a model gluten sample. Through a thorough analysis of the data obtained from coupled light scattering detectors and with the identification of molecular composition of the eluted protein, we evidence coelution events in several conditions. We show that the focus step used in conventional AF4 with the SDS buffer leads to the formation of aggregates that coelute with monomeric proteins. By contrast, a frit-inlet device enables the fractionation of individual wheat proteins in the SDS buffer. Interestingly conventional AF4, using water/ethanol as eluent, is an effective method for fractionating gluten proteins and their complex dynamic assemblies, which involve weak forces and are composed of both monomeric and polymeric proteins.

Electrochemical treatment for leachate membrane retentate: Performance comparison of electrochemical oxidation and electro-coagulation technology.

With the widespread use of membrane in advanced treatment of leachate, China produces a large amount of leachate membrane retentate (LMR) (≈23.4 million tons) annually, which is usually treated by incineration or recirculation in engineering, but these technologies have many drawbacks. LMR is suitable for electrochemical treatment due to its high electrical conductivity. This study compared the performances of electrochemical oxidation (EO) and electro-coagulation (EC) technology on LMR treatment under different experimental conditions, including anode material, current density, initial pH and reaction time. We found that EO optimal conditions achieved 70.1%, 83.1%, 78.7%, 98.7%, and 69.7% removal of total organic carbon (TOC), UV absorption (at 254nm), chromaticity, ammonia nitrogen (NH3-N), and total nitrogen (TN), respectively. Compared with EO, EC exhibited a similar removal ability for orgainics and better removals of chroma, but much less performance for removing nitrogen pollutants in the same reaction time, that is, removals of NH3-N and TN were only 31.5% and 36.2%, respectively. Meanwhile, EC showed much higher instantaneous current efficiency of COD than EO under its optimal reaction time (120min). In addition, the UV-Vis spectra and 3D fluorescence spectra indicated that EO exhibited relatively outstanding performance in decomposing dissolved organic matter (DOM) with rather complicated structures than EC. Also, the flow field-flow fractionation technique demonstrated that EO preferentially destroy humic-like, large molecular weight DOM, and converting them to smaller molecules, which resulted in more volatile organic compounds in EO samples than EC samples. While EC had little selectivity in the removal of organics, except humic-like DOM with relative small molecular. These findings can provide a theoretical basis for the electrochemical treatment of LMR.

Limitations of Nanoparticles Size Characterization by Asymmetric Flow Field\u2011Fractionation Coupled with Online Dynamic Light Scattering

The development of reliable protocols suitable for the characterisation of the physical properties of nanoparticles in suspension is becoming crucial to assess the potential biological as well as toxicological impact of nanoparticles. Amongst sizing techniques, asymmetric flow field flow fractionation (AF4) coupled to online size detectors represents one of the most robust and flexible options to quantify the particle size distribution in suspension. However, size measurement uncertainties have been reported for on-line dynamic light scattering (DLS) detectors when coupled to AF4 systems. In this work we investigated the influence of the initial concentration of nanoparticles in suspension on the sizing capability of the asymmetric flow field-flow fractionation technique coupled with an on-line dynamic light scattering detector and a UV–Visible spectrophotometer (UV) detector. Experiments were performed with suspensions of gold nanoparticles with a nominal diameter of 40 nm and 60 nm at a range of particle concentrations. The results obtained demonstrate that at low concentration of nanoparticles, the AF4-DLS combined technique fails to evaluate the real size of nanoparticles in suspension, detecting an apparent and progressive size increase as a function of the elution time and of the concentration of nanoparticles in suspension.

Characterisation of the Interaction among Oil-In-Water Nanocapsules and Mucin.

Mucins are glycoproteins present in all mucosal surfaces and in secretions such as saliva. Mucins are involved in the mucoadhesion of nanodevices carrying bioactive molecules to their target sites in vivo. Oil-in-water nanocapsules (NCs) have been synthesised for carrying N,N′-(di-m-methylphenyl)urea (DMTU), a quorum-sensing inhibitor, to the oral cavity. DMTU-loaded NCs constitute an alternative for the treatment of plaque (bacterial biofilm). In this work, the stability of the NCs after their interaction with mucin is analysed. Mucin type III from Sigma-Aldrich has been used as the mucin model. Mucin and NCs were characterised by the multi-detection asymmetrical flow field-flow fractionation technique (AF4). Dynamic light scattering (DLS) and ζ-potential analyses were carried out to characterise the interaction between mucin and NCs. According to the results, loading DMTU changes the conformation of the NC. It was also found that the synergistic interaction between mucin and NCs was favoured within a specific range of the mucin:NC ratio within the first 24 h. Studies on the release of DMTU in vitro and the microbial activity of such NCs are ongoing in our lab.

Investigation of zinc oxide particles in cosmetic products by means of centrifugal and asymmetrical flow field-flow fractionation

The dimensional characterization of insoluble, inorganic particles, such as zinc oxide ZnO, dispersed in cosmetic or pharmaceutical formulations, is of great interest considering the current need of declaring the possible presence of nanomaterials on the label of commercial products.This work compares the separation abilities of Centrifugal- and Asymmetrical Flow Field-Flow Fractionation techniques (CF3 and AF4, respectively), equipped with UV–vis, MALS and DLS detectors, in size sorting ZnO particles, both as pristine powders and after their extraction from cosmetic matrices.ZnO particles, bare and superficially modified with triethoxycaprylyl silane, were used as test materials. To identify the most suitable procedure necessary to isolate the ZnO particles from the cosmetic matrix, two O/W and two W/O emulsions were formulated on purpose. The suspensions, containing the extracted particles ZnO, were separated by both Field-Flow Fractionation (FFF) techniques to establish a common analysis protocol, applicable for the analysis of ZnO particles extracted from three commercial products, sold in Europe for the baby skin care.Key aspects of this study were the selection of an appropriate dispersing agent enabling the particles to stay in stable suspensions (>24h)and the use of multiple detectors (UV–vis, MALS and DLS) coupled on-line with the FFF channels, to determine the particle dimensions without using the retention parameters. Between the two FFF techniques, CF3 revealed to be the most robust one, able to sort all suspensions created in this work.

Frit inlet field-flow fractionation techniques for the characterization of polyion complex self-assemblies

Polymer self-assemblies joining oppositely charged chains, known as polyion complexes (PICs), have been formed using poly(ethyleneoxide – b – acrylic acid)/poly(l-lysine), poly(ethyleneoxide-b-acrylic acid)/dendrigraft poly(l-lysine) and poly[(3-acrylamidopropyl) trimethylammonium chloride – b – N – isopropyl acrylamide]/poly(acrylic acid). The self-assemblies have been first characterized in batch by Dynamic Light Scattering. In a second step, their analysis by Flow Field-Flow Fractionation techniques (FlFFF) was examined. They were shown to be very sensitive to shearing, especially during the focus step of the fractionation, and this led to an incompatibility with asymmetrical FlFFF. On the other hand, Frit Inlet FlFFF proved to be very efficient to observe them, either in its symmetrical (FI-FlFFF) or asymmetrical version (FI-AsFlFFF). Conditions of elution were found to optimize the sample recovery in pure water. Spherical self-assemblies were detected, with a size range between 70–400nm depending on the polymers. Compared to batch DLS, FI-AsFlFFF clearly showed the presence of several populations in some cases. The influence of salt on poly(ethyleneoxide-b-acrylic acid) (PEO-PAA) 6000-3000/dendrigraft poly(l-lysine) (DGL 3) was also assessed in parallel in batch DLS and FI-AsFlFFF. Batch DLS revealed a first process of swelling of the self-assembly for low concentrations up to 0.8M followed by the dissociation. FI-AsFlFFF furthermore indicated a possible ejection of DGL3 from the PIC assembly for concentrations as low as 0.2M, which could not be observed in batch DLS.

A critical evaluation of an asymmetrical flow field-flow fractionation system for colloidal size characterization of natural organic matter

Colloidal retention characteristics, recovery and size distribution of model macromolecules and natural dissolved organic matter (DOM) were systematically examined using an asymmetrical flow field-flow fractionation (AFlFFF) system under various membrane size cutoffs and carrier solutions. Polystyrene sulfonate (PSS) standards with known molecular weights (MW) were used to determine their permeation and recovery rates by membranes with different nominal MW cutoffs (NMWCO) within the AFlFFF system. Based on a ≥90% recovery rate for PSS standards by the AFlFFF system, the actual NMWCOs were determined to be 1.9kDa for the 0.3kDa membrane, 2.7kDa for the 1kDa membrane, and 33kDa for the 10kDa membrane, respectively. After membrane calibration, natural DOM samples were analyzed with the AFlFFF system to determine their colloidal size distribution and the influence from membrane NMWCOs and carrier solutions. Size partitioning of DOM samples showed a predominant colloidal size fraction in the <5nm or <10kDa size range, consistent with the size characteristics of humic substances as the main terrestrial DOM component. Recovery of DOM by the AFlFFF system, as determined by UV-absorbance at 254nm, decreased significantly with increasing membrane NMWCO, from 45% by the 0.3kDa membrane to 2–3% by the 10kDa membrane. Since natural DOM is mostly composed of lower MW substances (<10kDa) and the actual membrane cutoffs are normally larger than their manufacturer ratings, a 0.3kDa membrane (with an actual NMWCO of 1.9kDa) is highly recommended for colloidal size characterization of natural DOM. Among the three carrier solutions, borate buffer seemed to provide the highest recovery and optimal separation of DOM. Rigorous calibration with macromolecular standards and optimization of system conditions are a prerequisite for quantifying colloidal size distribution using the flow field-flow fractionation technique. In addition, the coupling of AFlFFF with fluorescence EEMs could provide new insights into DOM heterogeneity in different colloidal size fractions.

Membrane fouling indicator of effluent organic matter with nanofiltration for wastewater reclamation, as obtained from flow field-flow fractionation

Membrane filtration processes, due to their great performance, have been recognized as one of the effective technologies for the reclamation of wastewater, especially for the removals of inorganic/organic micropollutants and wastewater effluent organic matter (EfOM). Although the attraction of membrane filtration is remarkable, membrane fouling is a major obstacle to its usage; thus, the degree of fouling can have a significant impact on the cost, design and operation of full-scale facilities. In this study, the propensity for membrane fouling by EfOM was investigated using a flow field-flow fractionation (FlFFF) technique by comparing two different nanofiltration (NF) membranes: meta-phenylene diamine (MPD) versus piperazine based polyamides. From the characterizations of EfOM with FlFFF, the mutual interaction between the membrane surface and solutes was elucidated from the shape of elution peak and the hydrodynamic sizes with respect to the different membrane properties. Therefore, the propensities for fouling of the two NF membranes by EfOM were evaluated in relation to the membrane characteristics. Finally, several indicators of the potential for membrane fouling have been suggested in this study. This FlFFF technique can not only characterize the physicochemical properties of EfOM based on certain membrane properties, but can also provide a quick and easy evaluation of the potential for membrane fouling in terms of the suggested indicators. Furthermore, this technique is expected to contribute to the optimal design of membrane systems through the optimum selection of membranes for water and wastewater treatment.

Micro-thermal focusing field-flow fractionation

Focusing mechanism was effectively exploited to separate large (micrometer-size) particles by using new micro-thermal field-flow fractionation (micro-TFFF). It has been shown that the retention order of micrometer-size particles at high field strength can be explained by the mechanism of steric exclusion only at lowest flow rates of the carrier liquid. A simplistic, purely mechanical model of steric exclusion is not accurate to describe the retention at higher flow rates where the focusing phenomenon appears. Despite the fact that the thickness of the channel for micro-FFF cannot be reduced without taking into account a possible deterioration of the separation due to the contribution of “steric exclusion” mechanism, this paper demonstrates, in agreement with our previous results, that if the operational conditions were conveniently chosen, namely a low flow rate, a reasonable fit of the experimental retention data with the theory of steric exclusion mechanism in FFF was found and the separation of micron-size particles can be accomplished. However, high selectivity and resolution and high-speed separation were achieved if the focusing effect has clearly dominated the FFF mechanism. As a result, it seems that the micro-TFFF is the most universal technique which can be applied for the separation of the synthetic and natural macromolecules within an extended range of molar masses up to ultra-high molar masses and for the particles of various chemical nature and origin in a nano-size range as well as for large (micrometers) particles. Until nowadays, only sedimentation and flow field-flow fractionation techniques in so called “steric” modes were applied for the separations of large size particles. This application of micro-TFFF in focusing mode for the separation of large size particles is the first one described in the literature.

Separation and characterization of cationic, anionic, and nonionic water-soluble polymers by flow FFF: Sample recovery, overloading, and ionic strength effects

The flow field-flow fractionation (FFF) separation, characterization, and molecular weight distribution of a wide variety of synthetic water-soluble polymers is reported. Cationic polyvinylpyridine (PVP) anionic polystyrene sulfonate (PSS), and neutral polyacrylamide (PAAm) polymers of molecular weight from a few thousands to several millions were separated by flow FFF and their molecular weight distribution evaluated by software. The good correspondence of the molecular weight distribution curves extrapolated from two different FFF techniques, flow FFF and thermal FFF, either for narrow PVP samples with molecular weight 28,000, 110,000, and 240,000 or for the broad-disperse component of nominal molecular weight above one million demonstrates consistency with theory. Some critical issues in the separation and characterization of polyelectrolytes by flow FFF, such as the recovery of the polymer sample, and the effect of the injected load and of ionic strength on the retention parameters, are also discussed here. Study of the recovery of sample shows that under the experimental conditions used, the absolute recovery of the PVP polymers of molecular weight 240,000 is close to 100%, and the proportionate recovery of either polystyrene sulfonate or polyacryalmide samples is always obtained. The change in retention time (and the parameters derived from it) with injected mass for polyelectrolytes is related to intermolecular electrostatic interactions. The changes are found to be suppressed with an increase of ionic strength. The polymer equivalent Stokes diameter measured by flow FFF for polyelectrolytes at ionic strength from 10−7 to 0.15 M shows a dependence expected for charged polymers. © 1997 John Wiley & Sons, Inc. J Micro Sep9: 479–495, 1997