Flow field-flow fractionation coupled with multidetector: A robust approach for the separation and characterization of resistant starch.

The unique properties of resistant starch (RS) have made it applicable in the formulation of a broad range of functional foods. The physicochemical properties of RS play a crucial role in its applications. Recently, flow field-flow fractionation (FlFFF) has attracted increasing interest in the separation and characterization of different categories of RS. In this review, an overview of the theory behind FlFFF is introduced, and the controllable factors, including FlFFF channel design, sample separation conditions, and the choice of detector, are discussed in detail. Furthermore, the applications of FlFFF for the separation and characterization of RS at both the granule and molecule levels are critically reviewed. The aim of this review is to equip readers with a fundamental understanding of the theoretical principle of FlFFF and to highlight the potential for expanding the application of RS through the valuable insights gained from FlFFF coupled with multidetector analysis.

[Separation and characterization of Gastrodia elata polysaccharides based on asymmetrical flow field-flow fractionation: steric transition phenomenon].

Asymmetrical flow field-flow fractionation (AF4), a gentle tool for the separation and characterization of particles and macromolecules, has attracted increased interest in recent years owing to its broad dynamic size range and utilization of "open channel" voids in the packing or stationary phase. A steric transition phenomenon in which the sample elution mode change from the normal mode to the steric/hyperlayer mode occurs. Accurate characterization by AF4 requires the absence of steric transition, particularly when the sample has a broad size distribution, because the effect of the combination of different modes is difficult to interpret. In this study, the relative molecular mass (M), radius of gyration (Rg), and conformation of Gastrodia elata polysaccharides (GEPs) were characterized using AF4 coupled with online multi-angle light scattering (MALS) and differential refractive index (dRI) detection (AF4-MALS-dRI). Steric transition was observed during GEP separation by AF4 owing to the broad size distribution of the molecules. This phenomenon would result in the inaccurate characterization of the GEPs in terms of M and Rg because two GEP groups of different sizes may elute together. In this study, the effects of constant and exponentially decaying cross-flow rates, sample mass concentration, and spacer thickness on steric transition were systematically investigated. The results indicated that a high GEP mass concentration (i. e., 0.75 mg/mL) can lead to steric transition. The spacer thickness affected the resolution and retention time of the GEPs and changed the steric transition point (di). An exponentially decaying cross-flow rate not only adjusted the di of the polydisperse GEP samples but also improved the GEP resolution and shortened the analysis time. The influence of steric transition was solved under the following operating conditions: injected GEP mass concentration=0.5 mg/mL; injection volume=50 µL; spacer thickness=350 µm; detector flow rate=1.0 mL/min; and cross-flow rate exponentially decayed from 0.2 to 0.05 mL/min with a half-life of 2 min. Moreover, the influence of GEP origins and ultrasound treatment time on the M and Rg distributions and conformation of GEPs were investigated under the optimized operating conditions. The results showed that the M and Rg distributions of Yunnan and Sichuan GEPs decreased with increasing ultrasound time. When the ultrasound treatment time was 15 min, the Yunnan GEPs had a loosely hyperbranched chain conformation, whereas the Sichuan GEPs had a spherical conformation. When the ultrasound treatment time was increased to 30 or 60 min, the GEPs from both Yunnan and Sichuan had a hyperbranched chain conformation, indicating that ultrasound treatment resulted in GEP degradation. Under the same extraction conditions, GEPs from Yunnan had larger M and Rg values than those from Sichuan. AF4-MALS-dRI showed good repeatability for the characterization of GEPs under the optimized operating conditions. The relative standard deviations of Rg and M were 0.5% and 1.7%, respectively. The data presented in this study can be used as a starting point for in-depth studies on the structural bioactivity of GEPs.

Assessment of the Effects of Structural Modification of Gastrodia elata Polysaccharide on Anti-Breast Cancer Activity Using Asymmetrical Flow Field-Flow Fractionation.

Gastrodia elata ("Tian Ma" in Chinese) is used as a food and medical ingredient in traditional Chinese medicine. In this study, to enhance the anti-breast cancer activity of Gastrodia elata polysaccharide (GEP), GEPs were modified via sulfidation (SGEP) and acetylation (AcGEP). The physicochemical properties (such as solubility and substitution degree) and structural information (such as molecular weight Mw and radius of gyration Rg) of GEP derivatives were determined by Fourier transformed infrared (FTIR) spectroscopy and asymmetrical flow field-flow fractionation (AF4) coupled online with multiangle light scattering (MALS) and differential refractive index (dRI) detectors (AF4-MALS-dRI). The effects of the structural modification of GEP on the proliferation, apoptosis, and cell cycle of MCF-7 cell were studied systematically. The ability of MCF-7 cell for the uptake of GEP was studied by laser scanning confocal microscopy (LSCM). The results suggested that the solubility and anti-breast cancer activity of GEP were enhanced and the average Rg and Mw of GEP decreased after chemical modification. The AF4-MALS-dRI results showed that the chemical modification process simultaneously caused the degradation and aggregation of GEPs. The LSCM results revealed that more SGEP can enter the MCF-7 cell interior compared with AcGEP. The results indicated that the structure of AcGEP could play a dominating role in antitumor activity. The data obtained in this work can be used as a starting point for investigating the structure-bioactivity of GEPs.

Study on effects of preparation method on the structure and antioxidant activity of protein-Tremella fuciformis polysaccharide complexes by asymmetrical flow field-flow fractionation.

Oxidation is an essential biological process for human life. In this study, low density lipoprotein-Tremella fuciformis polysaccharide (LDL-TFP) complexes were prepared by electrostatic and covalent methods. The effects of preparation method on the structure and antioxidant activity of LDL-TFP complexes were investigated by asymmetrical flow field-flow fractionation (AF4) coupled with ultraviolet-visible (UV/Vis), multiangle light scattering (MALS), and differential refractive index (dRI) detectors. The results showed that the electrostatic LDL-TFP complexes had a spherical structure, while the covalent LDL-TFP complexes had a rod-like structure as indicated by the ratio of Rg (radius of gyration) to Rh (hydrodynamic radius). Moreover, the results revealed that the antioxidant activity of the LDL-TFP complexes on the HepG2 could be related to the structure of LDL-TFP complexes. The antioxidant activity of LDL-TFP complexes formed by LDL modified with phospholipase A2 was further enhanced. This study would help expand the application of TFP.

[Development of an asymmetrical flow field-flow fractionation system for the size characterization of starch granules].

Starch occurs naturally in the form of semicrystalline granules, and is composed of two types of carbohydrate molecules, amylose (AM) and amylopectin (AP). Starch granules and starch molecules have sizes in the range of 1-100 µm and 20-250 nm, respectively; these size ranges are among the key factors affecting the functional properties of starch. Asymmetrical flow field-flow fractionation (AF4) is a size-based separation technique. The major difference between AF4 and dynamic light scattering or microscopy techniques is that AF4 enables the separation of particles based on their size; consequently, the elution profile can be converted to the size distribution of the samples. In the last two decades, AF4 systems, when coupled online with multiangle light scattering (MALS) and differential refractive index (dRI) detectors (AF4-MALS-dRI), have demonstrated to be applicable for the size characterization of starch at the molecular level. Unlike size exclusion chromatography (SEC), AF4 systems use an open channel that does not require a stationary phase or packing materials. Thus, the shear scission of AP molecules during AF4 separation is minimized. The size detection range of a commercial AF4 system ranges from 1 nm to 10 µm, which is smaller than the size range of starch granules. In this study, a home-made AF4 system was developed, and its capability for the size characterization of starch granules extracted from sweet potato, lotus seed, and rice was investigated. The performance of the developed AF4 system was evaluated by running a mixture of polystyrene (PS) with diameter of 2, 6, 12, and 20 µm, respectively. Baseline separation of four PS samples was achieved, and the resolution for 6 µm PS and 12 µm PS was 1.40. The detection limit of the developed AF4 system was higher than that of commercial AF4 systems. Thus, the developed AF4 system is promising for the separation and characterization of starch granules. The effect of the composition of the carrier liquid on the AF4 separation of starch granules was also studied. Moreover, the accuracy of AF4 in terms of size characterization of the starch granules was evaluated by optical microscopy (OM). The results revealed that the type of dispersant and viscosity of the carrier liquid affect the accuracy of size characterization of the starch granules. The size distribution of rice starch granules obtained using a carrier liquid containing 0.01% (w/v) sodium dodecyl sulfate (used as a dispersant), 0.02% (w/v) NaN3 (used as a bactericide), and 0.001% (w/v) hydroxypropylmethylcellulose (used to adjust the viscosity of the carrier liquid) was in agreement with that obtained from OM. Furthermore, a commercial AF4 system coupled with MALS and dRI detectors was employed for the separation and characterization of starch molecules. A molecularly dispersed solution is necessary for the reliable molecular characterization of starch. The effect of the starch dissolution temperature on the AF4 characterization of starch was also investigated. The optimal dissolution temperature for lotus seed and rice starch granules was 75 ℃, while that for sweet potato starch granules was 78 ℃; this difference is mainly attributed to the different botanical origins of the granules. The results showed that the ratio of the radius of gyration (Rg) to the hydrodynamic radius (Rh) of rice starch and sweet potato starch is in the range of 0.9-1.1 over the molar mass range of 10 6-108 g/mol. For rice starch, the Rg/Rh ratio is between 1.2 and 1.4. Rice starch has the highest apparent density among the three starches, indicating that rice starch molecules have a dense structure. The results demonstrated that the AF4 system developed in this study is rapid and accurate for the size characterization of starch granules. The developed AF4 system, when combined with commercial AF4 systems coupled online with MALS and dRI detectors, can provide technical support to study the relationship between the size from the nanoscale to the microscale and functional properties of starch.

Insights into the correlations between the size of starch at nano- to microscale and its functional properties based on asymmetrical flow field-flow fractionation.

In this study, the starches were isolated from three botanical sources (i.e., rice, sweet potato, and lotus seed). The size distributions of starch granules and molecules were determined by asymmetrical flow field-flow fractionation (AF4), and compared with those measured from optical microscopy (OM) and dynamic light scattering (DLS). Furthermore, the starches were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). AF4 coupled online with UV-visible, multiangle light scattering (MALS), and differential refractive index (dRI) detectors (AF4-UV-MALS-dRI) was employed for the investigation of the digestion and retrogradation properties of starches. Meanwhile, the relationships between the size of starch at nano- to microscale and its functional properties (i.e., digestibility, retrogradation, and thermal properties) were studied by Pearson correlation analysis. AF4-UV-MALS-dRI was proved to be a rapid and gentle method for the separation and size characterization of starches at both micro- and nano-molecule levels. Moreover, it was demonstrated that AF4-UV-MALS-dRI is a useful tool for the monitoring of the digestion and retrogradation properties of starches. The results suggested that the sizes of starch granules and molecules were to some extent correlated with their thermal properties and digestibility, but not with retrogradation property.

Towards a better understanding of the relationships between the structure and antitumor activity of Gastrodia elata polysaccharides by asymmetrical flow field-flow fractionation.

To better understand the structure-function relationship of Gastrodia elata polysaccharides (PGEs), PGEs were extracted by ultrasound-assisted extraction method and the effects of extraction time on the structure and conformation of PGEs were evaluated by asymmetrical flow field-flow fractionation (AF4) coupled online with multiangle light scattering (MALS) and differential refractive index (dRI) detectors (AF4-MALS-dRI). Besides separation, AF4-MALS-dRI can provide more information about PGEs, such as size and molecular weight (Mw) distributions, apparent density, and conformation. The effects of PGEs on the proliferation, apoptosis, and cell cycle of MCF-7 cells were investigated. The cell activity assay indicated that the PGEs can inhibit the growth of MCF-7 cells by inducing late apoptosis. The results indicated that PGEs with a spherical conformation and compact structure seem to be beneficial to inducing MCF-7 cells late apoptosis. Moreover, results demonstrated that the information obtained by AF4-MALS-dRI is valuable for better understanding of the relationship of structure-activity of PGEs.

Insights into the structure and conformation of potato resistant starch (type 2) using asymmetrical flow field-flow fractionation coupled with multiple detectors.

Insight into the structure and conformation characteristics of starch that influence its enzyme susceptibility is import for its potential application. In this study, the capacity of asymmetrical flow field-flow fractionation (AF4) coupled online with multi-angle light scatting (MALS) and differential refractive index (dRI) detectors (AF4-MALS-dRI) for monitoring of change in structure and conformation of potato starch during enzymatic hydrolysis was evaluated. The dissolution behavior of potato resistant starch (type 2) (PRS) was investigated. The effect of incubation time and amyloglucosidase concentration on the structure and conformation of potato starch was studied. The apparent density and the ratio of Rg (radius of gyration) to Rh (hydrodynamic radius) obtained from AF4-MALS-dRI were proven to be important parameters as they offer an insight into conformation of PRS at molecular level. Results suggested that gelatinization process made potato amylose molecules have a loose and random coil conformation which could contribute to an acceleration of enzymatic hydrolysis of potato starch. Furthermore, an intermediate with an elongated branched conformation was found between amylose and amylopectin populations, which may play a role in digestion property of potato starch. The results demonstrated that AF4-MALS-dRI is a powerful tool for better understanding of conformation of PRS.

Applications of asymmetrical flow field-flow fractionation for separation and characterization of polysaccharides: A review.

Polysaccharides are the most abundant natural biopolymers on the earth and are widely used in food, medicine, materials, cosmetics, and other fields. The physicochemical properties of polysaccharides such as particle size and molecular weight often affect their practical applications. In recent years, asymmetrical flow field-flow fractionation (AF4) has been widely used in the separation and characterization of polysaccharides because it has no stationary phases or packing materials, which reduces the risk of shear degradation of polysaccharides. In this review, the principle of AF4 was introduced briefly. The operation conditions of AF4 for the analysis of polysaccharides were discussed. The applications of AF4 for the separation and characterization of polysaccharides from different sources (plants, animals, and microorganisms) over the last decade were critically reviewed.

Adoptive Immunotherapy of iNKT Cells in Glucose-6-Phosphate Isomerase (G6PI)-Induced RA Mice.

Rheumatoid arthritis (RA) is a complex chronic inflammatory autoimmune disease. The pathogenesis of the disease is related to invariant natural killer T (iNKT) cells. Patients with active RA present fewer iNKT cells, defective cell function, and excessive polarization of Th1. In this study, an RA animal model was established using a mixture of hGPI325-339 and hGPI469-483 peptides. The iNKT cells were obtained by in vivo induction and in vitro purification, followed by infusion into RA mice for adoptive immunotherapy. The in vivo imaging system (IVIS) tracking revealed that iNKT cells were mainly distributed in the spleen and liver. On day 12 after cell therapy, the disease progression slowed down significantly, the clinical symptoms were alleviated, the abundance of iNKT cells in the thymus increased, the proportion of iNKT1 in the thymus decreased, and the levels of TNF-α, IFN-γ, and IL-6 in the serum decreased. Adoptive immunotherapy of iNKT cells restored the balance of immune cells and corrected the excessive inflammation of the body.

[Separation and characterization of millet starch based on asymmetrical flow field-flow fractionation].

A method based on asymmetrical flow field-flow fractionation (AF4) coupled with a multi-angle light scattering detector and a differential refractive detector was developed for the separation and characterization of millet starch. In this study, the effects of sample loading, cross-flow rate, half-life, and ionic strength and pH value of the carrier liquid on the AF4 analysis of millet starch were investigated. In addition, the molecular structure of millet starch was determined under the optimum conditions. The optimized operation conditions for the AF4 analysis of millet starch were as follows:injection mass concentration, 0.50 g/L; injection volume, 50 µL; cross-flow rate, 1.2 mL/min; half-life, 3 min; and the carrier liquid, demonized water containing 10 mmol/L pH 7.00 NaNO3 (add 3 mmol/L NaN3). The method developed in this study showed good reproducibility. The relative standard deviations for the radius of gyration (Rg) and molar mass (Mw) were 3.4% and 7.0%, respectively.

Study on structure-function of starch by asymmetrical flow field-flow fractionation coupled with multiple detectors: A review.

Starch is one of the most abundant, renewable, non-toxic, and biodegradable polysaccharides, mainly comprising amylose and amylopectin. The physicochemical properties and structure of starch affect the quality of starch-based products. Therefore, the accurate characterization of the physicochemical properties and structure of starch is important to obtain products that meet specific functions. Recently, asymmetrical flow field-flow fractionation coupled with multi-angle light scattering (MALS) and differential refraction detectors (dRI) (AF4-MALS-dRI) has attracted great attention in the separation and characterization of starch. In this review, the structure and functions of starch characterized by AF4-MALS-dRI are summarized. The principle and important characterization parameters of AF4 are introduced. The starch structure-function relationship is deeply investigated owing to utilization of AF4-MALS-dRI. Besides separation of starch, AF4-MALS-dRI can provide more information about starch sample, such as radius of gyration (Rg), hydrodynamic radius (Rh), molar mass (Mw), apparent density (ρ), and conformation (Rg/Rh) of starch molecules. These parameters can be further used to investigate the structure-function relationship of starch.

Study of the adoptive immunotherapy on rheumatoid arthritis with Thymus-derived invariant natural killer T cells.

BACKGROUND: The therapeutic effect of adoptive infusion of specific thymus-derived invariant natural killer T (iNKT) cells in a mouse model of rheumatoid arthritis (RA) was observed, and the mechanism of cellular immunotherapy was preliminarily explored. METHODS: Thymus-derived iNKT cells were infused to RA model mice, with α-GalCer as a positive control. Then, ankle swelling was examined, as well as inflammatory cell infiltration to the joint tissue (hematoxylin-eosin [H&E] staining). Flow cytometry (FCM) was used to assess iNKT cell and helper T lymphocyte (Th) subsets. Serum cytokine levels were determined with cytometric bead array (CBA), with protein expression levels of related transcription factors assessed by Western blot. RESULTS: The joint swelling in RA model animals were significantly improved in the cell therapy and α-GalCer positive control groups (P < 0.05). In addition, iNKT frequencies in peripheral blood, the thymus and spleen were increased significantly (P < 0.05). Meanwhile, iNKT1 subset frequencies in the thymus and spleen were decreased, as well as splenic Th1 and Th17 cell subset rates, and serum TNF-α, IFN-γ and IL-6 levels. The rates of iNKT2 and Th2 subsets as well as IL-4 and IL-10 levels were increased (P < 0.05). Thymus GATA-3 and splenic PLZF protein levels were increased (P < 0.05). CONCLUSIONS: Adoptive infusion of thymus-derived iNKT cells exerts therapeutic effects in RA mice by increasing iNKT frequency, altering the proportions of iNKT cell subsets, correcting Th cell subset imbalance and reducing the amounts of inflammatory cytokines.

Study on the retrogradation behavior of starch by asymmetrical flow field-flow fractionation coupled with multiple detectors.

In this study, the capacity of asymmetrical flow field-flow fractionation (AF4) coupled with multiangle light scattering (MALS) and differential refractive index (dRI) detectors, to monitor starch retrogradation behavior in situ was evaluated. Meanwhile, the starch samples were characterized by differential scanning calorimetry (DSC) and fourier transform infrared spectroscopy (FTIR). The effect of the botanical origin, amylose/amylopectin ratio, storage conditions (i.e., temperature and time), and salt on starch retrogradation was systematically investigated. The starch aggregation and disassociation is a kinetic equilibrium process that is dependent on the storage conditions and the content of amylose. Moreover, it was found that nitrate ions retarded starch retrogradation behavior by inhibiting the formation of hydrogen bonds between amylose molecules. The results suggest that the formation of the small amylose aggregates plays an important role in starch retrogradation and maize amylopectin degradation. The information obtained by AF4-MALS-dRI is valuable for understanding the starch retrogradation mechanism.

[Application of asymmetrical flow field-flow fractionation for size characterization of low density lipoprotein in egg yolk plasma].

Home-made asymmetrical flow field-flow fractionation (AF4) system, online coupled with ultraviolet/visible (UV/Vis) detector was employed for the separation and size characterization of low density lipoprotein (LDL) in egg yolk plasma. At close to natural condition of egg yolk, the effects of cross flow rate, sample loading, and type of membrane on the size distribution of LDL were investigated. Under the optimal operation conditions, AF4-UV/Vis provides the size distribution of LDL. Moreover, the precision of AF4-UV/Vis method proposed in this work for the analysis of LDL in egg yolk plasma was evaluated. The intra-day precisions were 1.3% and 1.9% (n=7) and the inter-day precisions were 2.4% and 2.3% (n=7) for the elution peak height and elution peak area of LDL, respectively. Results reveal that AF4-UV/Vis is a useful tool for the separation and size characterization of LDL in egg yolk plasma.

Study on external factors affecting egg yolk plasma by asymmetrical flow field-flow fractionation.

In this work, the ability of asymmetrical flow field-flow fractionation (AF4) coupled with UV-vis, multiangle light scattering (MALS), and quasi-elastic light scattering (QELS) (AF4-UV-MALS-QELS) for monitoring aggregation of components of egg yolk plasma was evaluated. The effect of external factors (i.e., pH, storage conditions, freezing and heat treatments) on the egg yolk plasma was studied. The results reveal that the aggregation mechanism of components of egg yolk plasma during heat and freezing treatment is different. The results suggest that the low density lipoproteins (LDLs) in egg yolk plasma undergo a 'clusters-fusion-gel' process under heat treatment at pH10. The alkaline conditions promote the formation of LDL aggregates. Also, storage conditions play a role in the formation of LDL aggregates. It was found that the hen eggs stored for 7days at room temperature contain less aggregate than those stored at 4°C for the same period. The combination of AF4 with online MALS-QELS provided conformational information in terms of the shape and size distribution of LDL aggregates. AF4-UV-MALS-QELS was proved to be a rapid and gentle method for the separation and characterization of egg yolk plasma aggregates.

Asymmetrical flow field-flow fractionation coupled with multiple detections: A complementary approach in the characterization of egg yolk plasma.

The capability of asymmetrical flow field-flow fractionation (AF4) coupled with UV/VIS, multiangle light scattering (MALS) and quasi-elastic light scattering (QELS) (AF4-UV-MALS-QELS) for separation and characterization of egg yolk plasma was evaluated. The accuracy of hydrodynamic radius (Rh) obtained from QELS and AF4 theory (using both simplified and full expression of AF4 retention equations) was discussed. The conformation of low density lipoprotein (LDL) and its aggregates in egg yolk plasma was discussed based on the ratio of radius of gyration (Rg) to Rh together with the results from bio-transmission electron microscopy (Bio-TEM). The results indicate that the full retention equation is more relevant than simplified version for the Rh determination at high cross flow rate. The Rh from online QELS is reliable only at a specific range of sample concentration. The effect of programmed cross flow rate (linear and exponential decay) on the analysis of egg yolk plasma was also investigated. It was found that the use of an exponentially decaying cross flow rate not only reduces the AF4 analysis time of the egg yolk plasma, but also provides better resolution than the use of either a constant or linearly decaying cross flow rate. A combination of an exponentially decaying cross flow AF4-UV-MALS-QELS and the utilization of full retention equation was proved to be a useful method for the separation and characterization of egg yolk plasma.

Study on aggregation behavior of low density lipoprotein in hen egg yolk plasma by asymmetrical flow field-flow fractionation coupled with multiple detectors.

In this study, asymmetrical flow field-flow fractionation (AF4) coupled online with UV, multiangle light scattering (MALS), and fluorescence (FS) detectors (AF4-UV-MALS-FS) was employed for separation and characterization of egg yolk plasma. AF4 provided separation of three major components of the egg yolk plasma i.e. soluble proteins, low density lipoproteins (LDL) and their aggregates, based on their respective hydrodynamic sizes. Identification of LDL was confirmed by staining the sample with a fluorescent dye, Nile Red. The effect of carrier liquids on aggregation of LDL was investigated. Collected fractions of soluble proteins were characterized using sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). Moreover, the effect of heat and enzymatic treatment on egg yolk plasma was investigated. The results suggest that enzymatic treatment with phospholipase A2 (PLA2) significantly enhances the heat stability of LDL. The results show that AF4-UV-MALS-FS is a powerful tool for the fractionation and characterization of egg yolk plasma components.

Sedimentation field-flow fractionation for characterization of citric acid-modified Hß zeolite particles: Effect of particle dispersion and carrier composition.

In this study, sedimentation field-flow fractionation (SdFFF) was, for the first time, applied for determination of size distribution of Hß zeolite particles modified by citric acid (CA-Hß). Effects of the particle dispersion and the carrier liquid composition (type of dispersing reagent (surfactant) and salt added in the carrier liquid, ionic strength, and pH) on SdFFF elution behavior of CA-Hß zeolite particles were systematically investigated. Also the SdFFF separation efficiency of the particles was discussed in terms of the forces such as van der Waals, hydrophobic, and induced-dipole interactions. Results reveal that the type of salt and pH of the carrier liquid significantly affect the SdFFF separation efficiency of the zeolite particles. It was found that addition of a salt (NaN3) into the carrier liquid affects the characteristic of the SdFFF channel surface. It was found that the use of an acidic medium (pH 3.2) leads to a particle-channel interaction, while the use of a basic medium (pH 10.6) promotes an inter-particle hydrophobic interaction. Result from SdFFF was compared with those from scanning electron microscopy (SEM) and dynamic light scattering (DLS). It seems that, once the experimental conditions are optimized, SdFFF becomes a valuable tool for size characterization of the zeolite particles.

Factors affecting measurement of channel thickness in asymmetrical flow field-flow fractionation.

Asymmetrical flow field-flow fractionation (AF4) has been considered to be a useful tool for simultaneous separation and characterization of polydisperse macromolecules or colloidal nanoparticles. AF4 analysis requires the knowledge of the channel thickness (w), which is usually measured by injecting a standard with known diffusion coefficient (D) or hydrodynamic diameter (dh). An accurate w determination is a challenge due to its uncertainties arising from the membrane's compressibility, which may vary with experimental condition. In the present study, influence of factors including the size and type of the standard on the measurement of w was systematically investigated. The results revealed that steric effect and the particles-membrane interaction by van der Waals or electrostatic force may result in an error in w measurement.