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In the hierarchical model of soil aggregates, small soil microaggregates (small SMA; 40 μm up to 1700 μm in maximum. Our study on aggregation in wet-dry cycles revealed that the colloidal content has a controlling effect on the size distribution of resulting aggregates by acting as a binding agent and provides hereby new insights into the evolvement of aggregate hierarchy in soils.

Extracellular vesicles are small membrane-bound structures that are released by cells and play important roles in intercellular communication garnering significant attention in scientific society recently due to their potential as diagnostic and therapeutic tools. However, separating EVs from large-volume samples remains a challenge due to their small size and low concentration. In this manuscript, we presented a novel method for separating polystyrene beads as control and extracellular vesicles from large sample volumes using bifurcated asymmetric field flow fractionation in PDMS-free microfluidic devices. Separation characteristics were evaluated using the control system of polystyrene bead mix, which offers up to 3.7X enrichment of EV-sized beads. Furthermore, in the EV-sample from bioreactor culture media, we observed a notable population distribution shift of extracellular vesicles. Herein presented novel PDMS-free microfluidic device fabrication protocol resulted in devices with reduced EV-loss compared to size-exclusion columns. This method represented an improvement over the current state of the art in terms of EV separation from large sample volumes through the use of novel field flow fractionation design. [ABSTRACT FROM AUTHOR]

While the exact health risks associated with nanoplastics are currently the focus of intense research, there is no doubt that humans are exposed to nanoplastics and that food could be a major source of exposure. Nanoplastics are released from plastic materials and articles used during food production, processing, storage, preparation, and serving. They are also likely to enter the food chain via contaminated water, air, and soil. However, very limited exposure data for risk assessment exists so far due to the lack of suitable analytical methods. Nanoplastic detection in food poses a great analytical challenge due to the complexity of plastics and food matrices as well as the small size and expectedly low concentration of the plastic particles. Multidetector field flow fractionation has emerged as a valuable analytical technique for nanoparticle separation over the last decades, and the first studies using the technique for analyzing nanoplastics in complex matrices are emerging. In combination with online detectors and offline analysis, multidetector field flow fractionation is a powerful platform for advanced characterization of nanoplastics in food by reducing sample complexity, which otherwise hampers the full potential of most analytical techniques. The focus of this article is to present the current state of the art of multidetector field flow fractionation for nanoplastic analysis and to discuss future trends and needs aiming at the analysis of nanoplastics in food. [ABSTRACT FROM AUTHOR]

Soil contamination by multiple metals is a significant concern due to the interlinked mobilization processes. The challenges in comprehending this issue arise from the poorly characterized interaction among different metals and the complexities introduced by spatial and temporal heterogeneity in soil systems. We delved into these complexities by incubating size-fractionated paddy soils under both anaerobic and aerobic conditions, utilizing a combination of techniques for aqueous and colloidal analysis. The contaminated paddy soil predominantly consisted of particles measuring <53, 250-53, and 2000-250 μm, with the <53 μm fractions exhibiting the highest concentrations of multiple metals. Interestingly, despite their higher overall content, the <53 μm fractions released less dissolved metal. Furthermore, glucose enhanced the release of arsenic while simultaneously promoting the sequestration of other metals, such as Pb, Zn, and Cu. Utilizing asymmetric flow field-flow fractionation, we unveiled the presence of both fine (0.3-130 kDa) and large (130-450 nm) colloidal pools, each carrying various metals with different affinities for iron minerals and organic matter. Our results highlighted the pivotal role of the <53 μm fraction as a significant reservoir for multiple metal contaminants in paddy soils, in which the colloidal metals were mainly associated with organic matter. These findings illuminated the size-resolved dynamics of soil metal cycling and provided insights for developing remediation strategies for metal-contaminated soil ecosystems.

The combination of field-flow fractionation with powerful leading-edge detectors can be applied to the measurement of nanomaterial physicochemical properties, and the creation of harmonized robust measurement protocols. The Multiscale Metrology Suite (MMS) at the University of Strathclyde is a unique internationally leading facility combining multiple leading-edge field flow fractionation modalities(electric, asymmetric and centrifugal) with in-line Raman, inductively-coupled plasma (ICP) mass spectrometry and multimodal detector capability. Using exemplar case studies, we demonstrate the application of various FFF hyphenations for the analysis of a diverse materials portfolio. One of the goals for the MMS is to raise the standards of traditional academic analytical support underpinning pioneering academic engineering, physical and life sciences research exploring novel materials as diagnostics and therapeutics.

[Display omitted] • A novel characterization method for liposomes and lipid nanoparticles using CF3. • Particle size and drug-loading amounts alter the elution profiles of nanoparticles in CF3. • Density is one of the crucial properties of nanoparticles. • First evaluation method for lipid nanoparticles utilizing CF3. Nanomedicines comprise multiple components, and particle density is considered an important property that regulates the biodistribution of administered nanomedicines. The density of nanoparticles is characterized by centrifugal methods, such as analytical ultracentrifugation. Particle size and distribution are key physicochemical and quality attributes of nanomedicines. In this study, we developed a novel profiling method applicable to liposomes and lipid nanoparticles (LNPs), based on particle size and density, using centrifugal field-flow fractionation (CF3). We evaluated the elution profiles of PEGylated liposomes of different sizes with various doxorubicin (DOX)-loading amounts using CF3. This method was applied to evaluate the drug release of DOX-loaded liposomes, intra- and inter-batch variability, reconstitution reproducibility of AmBisome®, and elution characteristics of LNPs in COVID-19 vaccines (Comirnaty® and SpikevaxTM). The data obtained in the present study underscore the significance of the proposed methodology and highlight the importance of profiling and characterizing liposomes and LNPs using CF3 fractograms and a multi-angle light-scattering detector. [ABSTRACT FROM AUTHOR]

The study of the properties and composition of natural nanoparticles is an essential problem in analytical chemistry. Currently, there are no reference samples of natural nanoparticles, which complicates the study of the role of nanoparticles in specific processes, including geochemical ones, and makes it almost impossible to compare the results obtained by different researchers. In this work, nanoparticles of kaolinite, montmorillonite, and muscovite are studied as potential reference samples of natural nanoparticles. A complex of analytical methods has been proposed for the isolation and characterization of mineral nanoparticles, including the field flow fractionation of particles in a rotating coiled column, laser diffraction, scanning electron microscopy, and inductively coupled plasma atomic emission spectrometry. According to laser diffraction data, 93–98% of the isolated particles are from 40 to 300 nm in size; 2–7% of particles have sizes of up to 830 nm. The sizes of the isolated particles were confirmed by scanning electron microscopy. Based on the aluminum content, the concentrations of mineral nanoparticles in the isolated suspensions were estimated. The aggregation stability of nanoparticles in 5 mM phosphate buffer solutions of pH 6, 7, and 8 was studied. Muscovite nanoparticles are stable at pH 7 and 8 for at least 4 weeks, while montmorillonite nanoparticles have similar stability only at pH 8. For kaolinite nanoparticles, significant aggregation was observed at all studied pH values of buffer solutions. The obtained experimental results made it possible to clarify the problems and assess the prospects for developing reference samples of natural nanoparticles. [ABSTRACT FROM AUTHOR]

Nanoparticles (NPs) adsorb proteins when in the biological matrix, and the resulted protein corona could affect NP-cell interactions. The corona has a dynamic nature with the adsorbed proteins constantly exchanging with the free proteins in the matrix at various rates. The rapidly exchanging proteins compose the soft corona, which responds more dynamically to environment changes than the hard corona established by the ones with slow exchange rates. In the present study, the corona formed on the superparamagnetic iron oxide NPs (SPIONs) in human serum was studied by flow field-flow fractionation and ultracentrifugation, which rapidly differentiated the corona proteins based on their exchange rates. By varying the surface hydrophobicity of the SPIONs with a core size around 10 nm, we found out that, the more hydrophobic surface ligand attracted proteins with higher surface hydrophobicity and formed a more dynamic corona with a larger portion of the involved proteins with fast exchange rates. Increasing the core diameter of the SPIONs but keeping the surface ligand the same could also result in a more dynamic corona. A brief investigation of the effect on the cellular uptake of SPIONs using one selected corona protein, transferrin, was conducted. The result showed that, only the stably bound transferrin could significantly enhance cellular uptake, while transferrin bound in a dynamic nature had negligible impact. Our study has led to a better understanding of the relationship between the particle properties and the dynamic nature of the corona, which can help with design of nanomaterials with higher biocompatibility and higher efficacy in biosystems for biomedical applications.

This study is about linking preparative processes of nanoparticles with the morphology of the nanoparticles and with their efficiency in delivering payloads intracellularly. The nanoparticles are composed of hyaluronic acid (HA) and chitosan; the former can address a nanoparticle to cell surface receptors such as CD44, the second allows both for entrapment of nucleic acids and for an endosomolytic activity that facilitates their liberation in the cytoplasm. Here, we have systematically compared nanoparticles prepared either A) through a two-step process based on intermediate (template) particles produced via ionotropic gelation of chitosan with triphosphate (TPP), which are then incubated with HA, or B) through direct polyelectrolyte complexation of chitosan and HA. Here we demonstrate that HA is capable to quantitatively replace TPP in the template process and significant aggregation takes place during the TPP–HA exchange. The templated chitosan/HA nanoparticles therefore have a mildly larger size (measured by dynamic light scattering alone or by field flow fractionation coupled to static or dynamic light scattering), and above all a higher aspect ratio (Rg/RH) and a lower fractal dimension. We then compared the kinetics of uptake and the (antiluciferase) siRNA delivery performance in murine RAW 264.7 macrophages and in human HCT-116 colorectal tumor cells. The preparative method (and therefore the internal particle morphology) had little effect on the uptake kinetics and no statistically relevant influence on silencing (templated particles often showing a lower silencing). Cell-specific factors, on the contrary, overwhelmingly determined the efficacy of the carriers, with, e.g., those containing low-MW chitosan performing better in macrophages and those with high-MW chitosan in HCT-116.

Broad diversity and heterogeneity are inherently showcased by both natural and synthetic macromolecular structures. The high application potential for such structures and their combinations calls for novel analytical approaches that allow for comprehensive characterization and a full understanding of their complex composition. This review gives an overview of recent advances in designing and fabricating bioconjugated and self‐assembled polymer structures, and introduces adequate characterization protocols for sufficient elucidation of their specific molecular properties. Possible pitfalls in their analysis are demonstrated, and potential alternatives are discussed. The primary focus is on addressing the highlights, and future prospects of applying field‐flow fractionation coupled and/or hyphenated to different detection methods as a powerful separation and analytical technique for bioconjugate and self‐assembled nanostructures. [ABSTRACT FROM AUTHOR]

Resveratrol (RSV) is a naturally occurring polyphenolic phytoalexin with beneficial effects on human health, including the treatment and prevention of ulcerative colitis and cancer. This work proposes the formulation and characterization of new nanoparticles (NPs) for the specific colon delivery of RSV. RSV was encapsulated in zein (Z) NPs, coated by a hydrophilic pectin (P) shell to significantly increase the dissolution rate of RSV in aqueous environments. Since the dissolved drug was strongly degraded in a simulated gastrointestinal tract, the ZP-NPs were coated with Eudragit S 100. The coated particles, even if aggregated, had a spherical morphology with diameters in the 100–200 nm range. Release studies of RSV from the ZP coated NPs evidenced their potential ability to induce a specific colon delivery of this drug so that this formulation can be proposed to enrich natural food products to enhance their preventive and therapeutic effects in the lower GI tract. [ABSTRACT FROM AUTHOR]

Liposomal formulations for the treatment of cancer and other diseases are the most common form of nanotechnology enabled pharmaceuticals (NEPs) submitted for market approval and in clinical application today. The accurate characterization of their physical-chemical properties is a key requirement; in particular, size, size distribution, shape, and physical-chemical stability are key among properties that regulators identify as critical quality attributes. Here we develop and validate an optimized method, based on multi-detector asymmetrical-flow field flow fractionation (MD-AF4) to accurately and reproducibly separate liposomal drug formulations into their component populations and to characterize their associated size and size distribution, whether monomodal or polymodal in nature. In addition, the results show that the method is suitable to measure liposomes in the presence of serum proteins and can yield information on the shape and physical stability of the structures. The optimized MD-AF4 based method has been validated across different instrument platforms, three laboratories, and multiple drug formulations following a comprehensive analysis of factors that influence the fractionation process and subsequent physical characterization. Interlaboratory reproducibility and intra-laboratory precision were evaluated, identifying sources of bias and establishing criteria for the acceptance of results. This method meets a documented high priority need in regulatory science as applied to NEPs such as Doxil and can be adapted to the measurement of other NEP forms (such as lipid nanoparticle therapeutics) with some modifications. Overall, this method will help speed up development of NEPS, and facilitate their regulatory review, ultimately leading to faster translation of innovative concepts from the bench to the clinic. Additionally, the approach used in this work (based on international collaboration between leading non-regulatory institutions) can be replicated to address other identified gaps in the analytical characterization of various classes of NEPs. Finally, a plan exists to pursue more extended interlaboratory validation studies to advance this method to a consensus international standard. Unlabelled Image [ABSTRACT FROM AUTHOR]

Regular algal blooms are occurring in Taihu lake, which may be triggered by resuspension of sediments containing relevant amounts of phosphorus. Therefore, our study aims at quantification of phosphorus concentrations bound to suspended particulate matter in Taihu water samples to investigate this hypothesis. A field flow fractionation (FFF) method online with ICP-MS detection was developed to achieve an overview on particulate fractions of phosphorus and related elements including Fe, Al and C from the low nanometer to the low micrometer size range. Mass balance of dissolved and particulate elemental contents was established for quality control purpose and indicated low recovery of Fe, Al and P. Complementary determination of volume based particle size distribution by dynamic imaging analysis showed a majority of particle volume and thus mass in particles with size >5 μm. In order to address this challenge, the upper particle size limit of FFF online with ICP-MS was for the first time investigated in detail using well characterised monodisperse latex particles as model for organic matter in the low micrometer size range including microalgae. The effect of pre-filtration of the sample as well as the contribution of sample introduction via three different interfaces including micromist nebuliser/spray chamber, direct injection nebulisation and APEX with heated spray chamber and solvent removal by condensation on the particulate carbon recovery was studied by ICP-MS detection. The same instrumental setup was also applied for the characterisation of particulate elemental contents in the Taihu water samples as far as possible. Significant improvement of the detected particulate fraction in Taihu water samples was achieved by increasing the membrane pore size for pre-filtration and by using the APEX for introduction of the eluate from FFF into ICP-MS. Image 1 • First study on systematic investigation of the upper particle size limit of flow-FFF online with ICP-MS. • Full mass balance for water samples including particulate and dissolved elemental species both with and without filtration. • Study of the permeability of 5 μm and 10 μm filters for latex particles using carbon detection by ICP-MS and for Taihu water. • First application of direct injection nebulisation and APEX for online hyphenation of flow-FFF to ICP-MS. [ABSTRACT FROM AUTHOR]

• In Ap horizons of arable Luvisols, colloids control the aggregation of small soil microaggregates (<20 μm) in wet-dry cycles. • The presence of <450 nm colloids favored the formation of 1–40 μm soil microaggregates. • In absence of <1 μm colloids, >40 μm soil micro- and macroaggregates were preferentially formed. • The particle size is decisive for the aggregate formation under the current experimental condition. In the hierarchical model of soil aggregates, small soil microaggregates (small SMA; <20 μm) are often considered to be fundamental building units at the micron scale. Below which, soil colloids (<1 µm) have recently been proposed as binding agents of (micro)aggregates. However, the way in which soil colloids contribute to the formation and stability of soil micro- and macroaggregates remains largely unknown. For clarification, we evaluated potential impacts of the colloidal content, particularly the <450 nm colloids, on the aggregation of small SMA. Free water stable small SMA and <450 nm colloids were isolated from Ap-horizons of Stagnic Luvisols under different management (cropped and bare fallow). The size-resolved elemental composition of the <450 nm colloids was analyzed by asymmetric flow field-flow fractionation in combination with an inductively coupled plasma mass spectrometer and an organic carbon detector. To vary the colloidal content in small SMA, (1) suspensions containing different amounts of <450 nm colloids were added in small SMA, or (2) <1 µm colloids were removed from small SMA by centrifugation. In the maximum colloidal addition treatment, the mass ratios of added colloids to small SMA were 3.0 and 5.1 wt% for the cropped and bare fallow soil samples, respectively. Aggregation of small SMA with different colloidal amounts was performed in three successive wet-dry cycles. Afterwards, the size distribution of the resulting aggregates was measured by laser diffraction. Our results indicated that, in wet-dry cycles, colloids were important binding agents for the formation of SMA. Their presence, especially those <450 nm, was likely to support the formation of solid bridges during drying at particle contacts of 1–10 µm small SMA, favoring hereby SMA build-up in a relatively small size range of 1–40 µm. In contrast, the absence of <1 μm colloids in small SMA led to a preferential generation of relatively large aggregates in wet-dry cycles, i.e., typically with sizes >40 μm up to 1700 μm in maximum. Our study on aggregation in wet-dry cycles revealed that the colloidal content has a controlling effect on the size distribution of resulting aggregates by acting as a binding agent and provides hereby new insights into the evolvement of aggregate hierarchy in soils. [ABSTRACT FROM AUTHOR]

Characterization of natural nanomaterial (NNM) physicochemical properties – such as size, size distribution, elemental composition and elemental ratios - is often hindered by lack of methods to disperse NNMs from environmental samples. This study evaluates the effect of extractant composition, pH, and ionic strength on soil NNM extraction in term of recovery and release of primary particles/small aggregate sizes (i.e. , <200 nm). The extracted NNMs were characterized for hydrodynamic diameter and zeta potential by dynamic light scattering and laser Doppler electrophoresis, natural organic matter desorption by UV–Vis spectroscopy, element composition by inductively coupled plasma-mass spectroscopy (ICP-MS), size based elemental distribution by field flow fractionation coupled to ICP-MS, and morphology by transmission electron microscopy. The extracted NNM concentrations increased following the order of NaOH ≤ Na 2 CO 3 < Na 2 C 2 O 4 < Na 4 P 2 O 7. Na 4 P 2 O 7 was the most efficient extractant and results in 2–12 folds higher NNM extraction than other extractants. The Na 4 P 2 O 7 extracted NNMs exhibited narrower size distribution with smaller modal size relative to NaOH, Na 2 CO 3 , Na 2 C 2 O 4 extracted NNMs. Thus, Na 4 P 2 O 7 enhances the extraction of primary NNMs and/or smaller NNM aggregates (i.e. , size <200 nm). Na 4 P 2 O 7 promote soil microaggregates breakup and release of NNMs by reducing free multivalent cation concentration in soil pore water by forming metal-phosphate complexes and by enhancing NNM surface charge via phosphate sorption on NNM surfaces. Additionally, the extracted NNM concentrations increased with the increase in extractant concentration and pH, except at 100 mM where the high ionic strength might have induced NNM aggregation. The improved NNM-extraction will improve the overall understanding of the physicochemical properties of NNMs in environmental systems. This study presents the key properties of NNMs that can be used as background information to differentiate engineered nanomaterials (ENMs) from NNMs in complex environmental media. Unlabelled Image • Efficient extraction of natural nanomaterials from complex environmental matrices • Multimethod approach for characterization of natural nanomaterials from soils • Sodium pyrophosphate to efficiently disperse nanomaterials from soil heteroaggregates • Key nanomaterial properties to discriminate engineered from natural nanomaterials in soils [ABSTRACT FROM AUTHOR]

Magnetic particles (MPs) and magnetic nanoparticles (MNPs) are appealing candidates for biomedical and analytical applications due to their unique physical and chemical properties. Given that magnetic fields can be readily used to control the motion and properties of M(N)Ps, their integration in analytical methods opens new avenues for sensing and quantitative analysis. There is a large body of literature related to their synthesis, with a relatively small number of methods reporting the analysis of M(N)Ps using separation methods, which provide information on their purity and monodispersity. This review discusses analytical separation methods of M(N)Ps published between 2013 and June 2018. The analytical separation methods evaluated in this work include (i) field flow fractionation, (ii) capillary electrophoresis, (iii) macroscale magnetophoresis and (iv) microchip magnetophoresis. Among the trends in analytical separations of M(N)Ps an inclination towards miniaturization is moving from conventional benchtop methods to rapid and low-cost methods based on microfluidic devices. • The importance of analytical separations of magnetic (nano)particles is highlighted. • Analytical separations of magnetic (nano)particles during the period between 2013 and 2018 include field flow fractionation, macroscale magnetophoresis, capillary electrophoresis and microchip magnetophoresis. • Challenges and successes of different approaches on magnetic (nano)particle analytical separations are discussed and the future trends are explored. [ABSTRACT FROM AUTHOR]

Highlights • Polymer adsorption on GPC columns is observed for partially hydrolyzed PVA. • No adsorption for fully hydrolyzed PVA on these GPC columns is found. • Slow desorption of adsorbed PVA from porous packing particles is non-exponential. • Desorption of adsorbed PVA follows stretched exponential kinetics with index 0.36. • AF4 can be used for the characterization of PVA, regardless of the degree of hydrolysis. Abstract Both fully (98%) and partially (88%) hydrolyzed poly(vinyl alcohol) (PVA) are characterized by aqueous gel permeation chromatography (GPC) coupled with multi-angle laser light scattering. Polymer adsorption on hydrophilic GPC columns is observed for partially hydrolyzed PVA but not for fully hydrolyzed PVA, which is driven by the hydrophobic interactions between the residual acetate groups on PVA and the stationary phase. Desorption of these partially hydrolyzed PVA chains from the packing materials is monitored by decay of the light scattering signal. It is found that the desorption of these polymer chains from porous particles is non-exponential and follows stretched exponential kinetics with index 0.36. It is also shown that asymmetric flow field flow fractionation can be successfully used for the molar mass characterization of PVA, irrespective of the degree of hydrolysis. [ABSTRACT FROM AUTHOR]

Abstract An acoustic field flow fractionation (FFF) device was developed to fractionate a micro-particle mixture on the basis of the particle diameter using an acoustic force field in a carrier liquid flow. In the acoustic FFF channel used in the device, ultrasound waves generated from piezoelectric transducers driven by a sinusoidal signal of 2.02 Mhz propagated into the carrier liquid flow and built up a quarter-wavelength ultrasound standing wave field across the channel height. It was experimentally demonstrated that the acoustic field with a pressure node plane at the bottom surface of the channel reduced the thickness of the particle diffusion layer in a stagnant liquid proportional to the applied voltage driving the piezoelectric transducer. In the size-dependent particle separation, the particle mixture flowing through the acoustic FFF channel experienced an acoustic radiation force in the gravitational direction. As a result, suppressing the diffusion of small particles, particles were transported along the bottom surface of the channel with the local velocity of the carrier liquid at the particle center. The developed acoustic FFF device successfully fractionated a fluorescent micro-particle mixture (1, 3, 5, and 10 μm diameter), whereas the 3 and 5 μm particles were not fractionated in the FFF device using only the gravitational force field due to the diffusion of 3 μm particles. Graphical abstract Image 1 Highlights • Develops a field flow fractionation technique using an acoustic radiation force. • Analyzes the induced acoustic force field across the height of the field flow fractionation channel. • Demonstrates the effect of the induced acoustic force field on the resolution of field flow fractionation. • Predicts the resolution and the relaxation time of an acoustic field flow fractionation. [ABSTRACT FROM AUTHOR]

Abstract For best photosensitizer activity phthalocyanine dyes used in photodynamic therapy should be molecularly dispersed. Polyethylene glycol- block -polylactide derivatives presenting benzyl side-groups were synthesized to encapsulate a highly lipophilic phthalocyanine dye (AlClPc) and evaluate the effect of π-π interactions on the nanocarrier colloidal stability and dye dispersion. Copolymers with 0, 1, 2 and 6 mol% of benzyl glycidyl ether (BGE) were obtained via polyethylene glycol initiated ring-opening copolymerization of D, l -lactide with BGE. The block copolymers formed stable, monodisperse nanospheres with low in vitro cytotoxicity. AlClPc loading increased the nanosphere size and affected their colloidal stability. The photo-physical properties of the encapsulated dye, studied in batch and after separation by field flow fractionation, demonstrated the superiority of plain PEG-PLA over BGE-containing copolymers in maintaining the dye in its monomeric (non-aggregated) form in aqueous suspension. High dye encapsulation and sustained dye release suggest that these nanocarriers are good candidates for photodynamic therapy. Graphical abstract Unlabelled Image Highlights • Polyethylene glycol- block -poly(lactide- co -benzyl glycidyl ether) were obtained. • The copolymers form stable non-cytotoxic nanospheres below 100 nm. • Less benzylated copolymer meant better chloroaluminum phthalocyanine dispersion. • Controlled release, stable and bright phthalocyanine nanocarriers were achieved. [ABSTRACT FROM AUTHOR]

Applications of asymmetrical flow field-flow fractionation (AF4) continue to expand rapidly in the fields of nanotechnology and biotechnology. In particular, AF4 has proven valuable for the separation and analysis of particles, biomolecular species (e.g., proteins, bacteria) and polymers (natural and synthetic), ranging in size from a few nanometers to several micrometers. The separation of non-spheroidal structures (e.g., rods, tubes, etc.) with primary dimensions in the nanometer regime, is a particularly challenging application deserving of greater study and consideration. The goal of the present study was to advance current understanding of the mechanism of separation of rod-like nano-objects in the AF4 channel. To achieve this, we have systematically investigated a series of commercially available cetyltrimethylammonium bromide stabilized gold nanorods (AuNRs), with aspect ratios from 1.7 to 10. Results show clearly that the retention time is principally dependent on the translational diffusion coefficient of the AuNRs. Equations used to calculate translational and rotational diffusion coefficients (cylinder and prolate ellipsoid models) yield similarly good fits to experimental data. Well characterized gold nanorods (length and diameter by transmission electron microscopy) can be used as calibrants for AF4 measurements allowing one to determine the aspect ratio of nanorod samples based on their retention times.ᅟ [ABSTRACT FROM AUTHOR]

Abstract The interaction of polymer nanocapsules (NC) prepared from four biodegradable polyesters with variable polymer hydrophobicity (PCL, PLA, PLGA and PLA-PEG) was investigated in the non-phagocytic Vero, Caco-2 and HepG2 cell lines. The NC, labeled with the highly lipophilic fluorescent indocarbocyanine dye DIL, had very similar sizes (approx. 140 nm) and negative zeta-potentials. Asymmetric flow field-flow fractionation evidenced NC colloidal stability and negligible transfer of the dye to serum proteins in the incubation medium. The cytotoxicity of the NC was evaluated via MTT assay over a large polymer concentration range (1–1000 μg/mL) and time of exposure (2, 24 and 48 h). The NC were safe in vitro up to a concentration of approx. 100 μg/mL or higher, depending on the cell line and nature of the polymer. Vero cells were more sensitive to the NC, in particular NC of the more hydrophobic polymer. The cells were exposed to endocytosis inhibitors, incubated with NC, and the cell-associated fluorescence was quantified by spectrofluorometry. HepG2 cells presented a 1.5–2-fold higher endocytic capacity than Caco-2 and Vero cells. The main mechanism of NC uptake was caveolin-mediated endocytosis in HepG2 and Vero cells, and macropinocytosis in Caco-2 cells. Polymer hydrophobicity had an effect on the level of NC associated to HepG2 cells and to a lesser extent on the endocytosis mechanisms in Vero and Caco-2 cells. The NC uptake levels and endocytosis mechanisms differed significantly between cell lines tested. Graphical abstract Unlabelled Image [ABSTRACT FROM AUTHOR]

Basic and applied virus research requires specimens that are purified to high homogeneity. Thus, there is much interest in the efficient production and purification of viruses and their subassemblies. Advances in the production steps have shifted the bottle neck of the process to the purification. Nonetheless, the development of purification techniques for different viruses is challenging due to the complex biological nature of the infected cell cultures as well as the biophysical and -chemical differences in the virus particles. We used bacteriophage ϕ6 as a model virus in our attempts to provide a new purification method for enveloped viruses. We compared asymmetrical flow field-flow fractionation (AF4)-based virus purification method to the well-established ultracentrifugation-based purification of ϕ6. In addition, binding of ϕ6 virions to monolithic anion exchange columns was tested to evaluate their applicability in concentrating the AF4 purified specimens. Our results show that AF4 enables one-hour purification of infectious enveloped viruses with specific infectivity of ~1 × 10 13  PFU/mg of protein and ~65–95% yields. Obtained purity was comparable with that obtained using ultracentrifugation, but the yields from AF4 purification were 2–3-fold higher. Importantly, high quality virus preparations could be obtained directly from crude cell lysates. Furthermore, when used in combination with in-line light scattering detectors, AF4 purification could be coupled to simultaneous quality control of obtained virus specimen. [ABSTRACT FROM AUTHOR]

The use of nanotechnology in medical products has been demonstrated at laboratory scale, and many resulting nanomedicines are in the translational phase toward clinical applications, with global market trends indicating strong growth of the sector in the coming years. The translation of nanomedicines toward the clinic and subsequent commercialization may require the development of new or adaptation of existing standards to ensure the quality, safety and efficacy of such products. This work addresses some identified needs, and illustrates the shortcomings of currently used standardized methods when applied to medical-nanoparticles to assess particle size, drug loading, drug release and in vitro safety. Alternative physicochemical, and in vitro toxicology methods, with the potential to qualify as future standards supporting the evaluation of nanomedicine are provided. [ABSTRACT FROM AUTHOR]

Silver nanoparticles (Ag-NP) are used in various consumer products and are one of the most prevalent metallic nanoparticle in commodities and are released into the environment. Transport behavior of Ag-NP in groundwater is one important aspect for the assessment of environmental impact and protection of drinking water resources in particular. Ag-NP transport processes in saturated single-fractured sandstones using triaxial flow cell experiments with different kind of sandstones is investigated. Ag-NP concentration and size are analyzed using flow field-flow fractionation and coupled SEM-EDX analysis. Results indicate that Ag-NP are more mobile and show generally lower attachment on rock surface compared to experiments in undisturbed sandstone matrix and partially fractured sandstones. Ag-NP transport is controlled by the characteristics of matrix porosity, time depending blocking of attachment sites and solute chemistry. Where Ag-NP attachment occur, it is heterogeneously distributed on the fracture surface. [ABSTRACT FROM AUTHOR]

This paper describes the development of an integrated system using a dry film resistant (DFR) microfluidic channel consisting of pulsed field dielectrophoretic field-flow-fractionation (DEP-FFF) separation and optical detection. The prototype chip employs the pulse DEP-FFF concept to separate the cells (Escherichia coli and Saccharomyces cerevisiae) from a continuous flow, and the rate of release of the cells was measured. The separation experiments were conducted by changing the pulsing time over a pulsing time range of 2–24 s and a flow rate range of 1.2–9.6 μ L min − 1 . The frequency and voltage were set to a constant value of 1 M Hz and 14 V pk-pk, respectively. After cell sorting, the particles pass the optical fibre, and the incident light is scattered (or absorbed), thus, reducing the intensity of the transmitted light. The change in light level is measured by a spectrophotometer and recorded as an absorbance spectrum. The results revealed that, generally, the flow rate and pulsing time influenced the separation of E. coli and S. cerevisiae. It was found that E. coli had the highest rate of release, followed by S. cerevisiae. In this investigation, the developed integrated chip-in-a lab has enabled two microorganisms of different cell dielectric properties and particle size to be separated and subsequently detected using unique optical properties. Optimum separation between these two microorganisms could be obtained using a longer pulsing time of 12 s and a faster flow rate of 9.6 μ L min − 1 at a constant frequency, voltage, and a low conductivity.

In the last 10 years, asymmetrical flow field flow fractionation (AF4) has been one of the most promising approaches to characterize colloidal particles. Nevertheless, despite its potentialities, it is still considered a complex technique to set up, and the theory is difficult to apply for the characterization of complex samples containing submicron particles and nanoparticles. In the present work, we developed and propose a simple analytical strategy to rapidly determine the presence of several submicron populations in an unknown sample with one programmed AF4 method. To illustrate this method, we analyzed polystyrene particles and fullerene aggregates of size covering the whole colloidal size distribution. A global and fast AF4 method (method O) allowed us to screen the presence of particles with size ranging from 1 to 800 nm. By examination of the fractionating power F , as proposed in the literature, convenient fractionation resolution was obtained for size ranging from 10 to 400 nm. The global F values, as well as the steric inversion diameter, for the whole colloidal size distribution correspond to the predicted values obtained by model studies. On the basis of this method and without the channel components or mobile phase composition being changed, four isocratic subfraction methods were performed to achieve further high-resolution separation as a function of different size classes: 10-100 nm, 100-200 nm, 200-450 nm, and 450-800 nm in diameter. Finally, all the methods developed were applied in characterization of nanoplastics, which has received great attention in recent years. [ABSTRACT FROM AUTHOR]

The effect of polymeric nanocapsule dose on plasmatic and liver concentrations 20 min after intravenous administration in mice was evaluated. Nanocapsules were prepared with different polymers, namely, poly(D,L-lactide) (PLA), polyethylene glycol- block -poly(D,L-lactide) (PLA-PEG), and PLA with chitosan (PLA-Cs) and compared with a nanoemulsion. These formulations were labelled with a phthalocyanine dye for fluorescent detection. The nanostructures had narrow size distributions upon separation by asymmetric flow field flow fractionation with static and dynamic light scattering detection, with average hydrodynamic diameters in the 130–300 nm range, negative zeta potentials, except PLA-Cs nanocapsules, which had a positive zeta potential. Flow cytometry revealed uptake mostly by monocytes and neutrophils in mice and human blood. PLA nanocapsules and the nanoemulsion showed dose-dependent plasma concentrations, where the percentage of plasmatic fluorescence increased with increasing administered dose. In contrast, PLA-PEG nanocapsules led to a dose-independent plasmatic profile. PLA-Cs nanocapsules showed the lowest plasmatic and liver levels of fluorescence at all administered doses and significant intravenous toxicity in mice. This work demonstrates the importance of considering the nanocarrier dose when evaluating pharmacokinetic and biodistribution data and emphasizes the role of surface features in determining the plasmatic and liver concentrations of a poorly soluble lipophilic encapsulated compound. [ABSTRACT FROM AUTHOR]

Field flow fractionation (FFF) techniques are used to successfully characterize several nanomaterials by sizing nano-entities and producing information about the aggregation/agglomeration state of nanoparticles. By coupling FFF techniques to specific detectors, researchers can determine particle-size distributions (PSDs), expressed as mass-based or number-based PSDs. This review considers FFF applications in the food, biomedical, and environmental sectors, mostly drawn from the past 4 y. It thus underlines the prominent role of asymmetrical flow FFF within the FFF family. By concisely comparing FFF techniques with other techniques suitable for sizing nano-objects, the advantages and the disadvantages of these instruments become clear. A consideration of select recent publications illustrates the state of the art of some lesser-known FFF techniques and innovative instrumental set-ups. [ABSTRACT FROM AUTHOR]